lifesciences-plus_01-16

01 I 2016 CHF 24,90 I E 23,80 www.chemieplus.ch

ZIKA-VIRUS: SMALL BITE – BIG IMPACT P. 49

BRAIN-PLASTICITY: TRAIN THE BRAIN P. 8

MEDICAL-TECHNOLOGY: SENSORS FOR NON-INVASIVE CANCER DETECTION P. 12 START UP: SCIENCE FROM OUTER SPACE P. 36

Innovation:

How to Foster Fruitful Dialogue P. 30

EDITORIAL

“Switzerland specifically offers good conditions for constructive collaboration between life sciences and industry.”

Brilliant Minds in Dialogue

“I

see deficits concerning the interaction between

In the past, doctors have often tried to identify which

sciences and private companies,” notes our inter-

illness the patient was suffering from based on the smell

view partner Peter Pichler, President and CEO of Brech-

of exhaled breath. In modern breath diagnostics, instead

bühler AG. He continues: “The challenge is to intensify the

of the human sense of smell, so-called “electronic noses”

dialogue between these two players.” How to master this

are used. Substances can thus be detected which are pres-

challenge in practice is the subject of the networking dis-

ent in the breath when someone has certain illnesses. In-

cussion in “Lifesciences plus” between Mr. Pichler, Su-

terpretation of the measurement results using gas chro-

sanne Lauber Fürst, Inartis Network Vice President, and

matography und mass spectrometry is nevertheless diffi-

Dr Helmut Traitler, former Head of Innovation Partner-

cult and expensive. At the Swiss Nanoscience Institute at

ships at Nestlé Group and now Innovation Connector at

the University of Basel, a research team is now using a

Swissnex in San Francisco. These experts also describe

new method based on exhaled breath analysis. Nanome-

how the participating partners’ all too different goals along

chanical membrane surface stress sensors (MSS) are used.

with organisational bottlenecks can take the wind out of

The penetration of the polymer-coating of the membranes

the sails of the dialogue process. But there is one thing all

by the volatile organic compounds (VOCs) contained in

of them agree on: knowledge transfer and networking are

breath bends these membranes in a specific way. The dif-

important incentives for innovation, and Switzerland spe-

fractions are measured piezoresistively. The voltage sig-

cifically offers good conditions for constructive collabora-

nals found give information on the VOC structure of the

tion between life sciences and industry (page 30).

exhaled air, thus allowing conclusions to be drawn on the course of head and neck cancer patients’ therapy. Sensory

Moreover, this edition of “Lifesciences plus” also offers an

VOC analysis could become an important non-invasive al-

in-depth look at important research projects underway at

ternative for the diagnosis of certain types of cancer and

Swiss universities which may someday significantly im-

other illnesses (page 12).

prove people’s quality of life. Two highlights in the field of oncology are mentioned as examples:

Besides the aforementioned examples, this edition of “Lifesciences plus” will give information on many other

Radio-immunotherapy is a promising new form of therapy,

exciting research projects. We do not limit ourselves in any

in particular for the treatment of patients with refractory

way to fundamental research at universities. In this con-

tumours which can often not be fought effectively with

text, we would like to draw your attention to the

chemotherapy. Contrary to radiation from outside which

“Lab & Process” and “Players & Products” columns where

damages healthy tissues along with the tumour, radio-im-

we present current developments in appliance technology

munotherapy very selectively acts on the tumour cells

and new applications.

“from inside”. In this method, a radionuclide is bound to a specific antibody by means of a chelator which transports the radio-isotope straight to the tumour. The radionuclide-labelled antibodies dock to specific antigens ex-

RALF MAYER

pressed on the surface of the tumour cells which are then destroyed by extremely localised radiation. Researchers at Swiss universities are testing the potential of radio-immunotherapy for the treatment of various types of cancer. The results are encouraging (page 22).

Editor in Chief

S U M M A RY

8

BRAIN PLASTICITY

Humans and many animals could not survive without a special ability of their brains: learning. This ability is possible because the brain is not a static organ but a malleable network of communicating nerve cells. The formation of new neurons in the brain and their interaction pattern can be influenced by a simple and very natural movement: Mice running in wheels seem to have better long-term memory than sitting animals. The corresponding studies conducted by Professor Josef Bischofberger and his team in Basel yielded some more exciting results.

16 SYSTEMS BIOLOGY

Tissue complexity has been largely ignored in infection research. Biologists from the University of Basel are now investigating how pathogen’s tolerance to antibiotic substances evolves in infected host tissues. Their focus is on Gram-negative bacteria, which actually start running out of control.

SWISS RESEARCH

04 News 12

Medical Technology: Nanomechanical Piezoresistive Membrane Sensors for Non-Invasive Cancer Detection

20

Oncology: Avicenna Oncology and the Big Challenge: The Fight against Aggressive and Resistant Cancers

22

30

PLAYERS & PRODUCTS

36

Microgravity and Drug Design: Science from Outer Space

40

Autoimmune Diseases: Allocyte – New Hope for Patients Suffering from Autoimmune Diseases

44

HPAPIS: Trends in Aseptic Manufacturing

47

Exhibition: Get ready for Chemspec Europe 2016 in Basel

Radioimmunotherapy: Not just for Lymphomas anymore

48 Tools

Switzerland definitely stands for excellence in research

LAB & PROCESS

FILTRATE

and innovation and is globally renowned to be a driving force. This is especially related to the area of chemistry, pharma and food. But is the drive as powerful as it might be? In our interview three experts from different regions and professions discuss their individual views on current trends, bottlenecks and on possible innovation catalysts: Susanne Lauber Fürst, INARTIS Network Vice President, Renens; Peter Pichler, President and CEO Brechbühler AG, Schlieren/Echallens; Dr Helmut Traitler, former Head of Innovation Partnerships at Nestlé Group and now Innovation Connector at Swissnex in San Francisco as well as a textbook author for all those who are engaged in the field of food, water, and nutrition.

26 Drug Discovery: Advances

Zika Virus: Small Bite – Big Impact

NETWORKING

Lifesciences plus 01 I 2016

in Microplate Reader High-Throughput Screening

28 Systems Biology: Innovative Research for Future Production of Biopharmaceuticals

3

49

54 News 57 Edition Notice

SWISS RESEARCH

NEWS

Medigene and University of Lausanne Collaborate to Explore New Technologies for the Characterisation T Cell Receptors

Medigene AG,

JE Viruses Can Be Passed Directly from Pig to Pig

Japanese

Encephalitis (JE) virus causes serious inflammation of the brain in people in Asia and fertility problems in pigs. Mosquitoes were previously the only known transmission route.

Now a team of researchers from the Institute of Virology and Immunology and the University of Bern at the Vetsuisse Faculty led by Dr Meret Ricklin and Prof Artur Summerfield have shown that JE viruses can also be passed directly from pig to pig.

As the researchers have been able to show, infected pigs discharge the virus in their saliva for several days, and the animals are also susceptible to infection through the mouth or nose with very low doses of the virus. In pigs – as in humans – the virus was found to spread through the brain and cause inflammation. The virus was, however, found to grow most in the tonsils, where it was detectable for several weeks or even months. The authors suggest that JE viruses could possibly circulate in pigs and survive for up to months. When the virus is secreted again, for example as a result of a different infection that weakens the immune system, a new infection cycle could then begin. However, the researchers say that further studies are needed to prove this link.

The study published in “Nature Communications” shows that even for viruses that are spread by insect bites, direct transmission through animal to animal contact cannot be ruled out.

“This means that the virus could circulate within the pig population without mosquitoes, and thus spread even to regions with a temperate climate”, says Artur Summerfield. This would theoretically also mean a higher risk to humans. A vaccine is, however, available for both people and animals. # www.unibe.ch

a clinical stage immune-oncology company focusing on the development of T cell immune-therapies for the treatment of cancer, recently announced a collaboration agreement with the University of Lausanne in order to establish a better and faster method for selecting tumourspecific T cells characterised by expression of suitable T cell receptors (TCRs) for Medigene's expanding TCR library. The agreement provides Medigene access to the state-of-the-art NTAmer technology that has been established by TCMetrix, a spin-off company of the Ludwig Institute for Cancer Research in Lausanne, and further developed in collaboration with leading scientists of the University of Lausanne. Financial terms of the agreement were not disclosed.

Prof Dr Dolores Schendel, CSO and designated CEO of Martinsried / Munich based Medigene, comments: “With this cooperation we are able to explore a cutting-edge technology in order to complement our comprehensive TCR platform. The NTAmer technology supports us to gain additional information about the structural avidity of TCRs by using direct assessment of living T cells. This will help us to select TCRs with optimal target antigen sensitivity for inclusion in our TCR library that is developed in order to treat patients with different MHC backgrounds and various types of cancer.” Prof Dr George Coukos, Professor at the University of Lausanne and Director of the Ludwig Institute for Cancer Research in Lausanne, added: “The development of personalised, cell-based immunotherapies is a major objective of Ludwig Lausanne, as such approaches hold great promise for the treatment of cancers. We are very pleased to be working with Medigene to help advance its innovative TCR technology and are hopeful that it will ultimately be of significant benefit to cancer patients, especially those with advanced and refractory disease.” # www.medigene.com

Diabetes: A Smart Shoe to Help Reduce Amputations

EPFL researchers

have developed a shoe sole with valves that electronically control the pressure applied to the arch of the foot. The goal is to relieve foot ulcers commonly caused by diabetes and help them heal to avoid dangerous secondary infections. Every year in Europe, 250,000 diabetics have a leg amputated, and the mortality rate is 30 % at 30 days and 50 % at one year. Foot ulcers are the root cause of most of the amputations. To avoid these eventualities, a team of researchers from EPFL is collaborating with the Geneva University Hospitals (HUG) on a smart shoe. The technology is based on actuators that electronically control the stiffness of the sole in order to reduce the onset of diabetic foot ulcers and prevent existing ones from worsening. An initial prototype has just been completed.

The challenge for doctors is threefold. They need to find a way to remove all pressure from the ulcers as soon as they appear, keep it off as long as necessary for them to fully heal, and quickly react to new ulcers, which can reappear at any time elsewhere on the arch of the foot. In response to this challenge, researchers from EPFL’s Integrated Actuators Laboratory (LAI) in Neuchâtel came up with an idea: embed the sole of a shoe with around 50 small electromagnetic valves filled with magnetorheological material. “We can control the viscosity of the material, which is made up of suspended iron microparticles,” said Yves Perriard, the director of LAI. “When we apply a magnetic field, the particles react immediately and align themselves with it, causing the material to change from liquid to solid state in a fraction of a second.”

This means that the rigidity in different parts of the shoe sole can be controlled separately depending on where the sensitive areas and wounds are.

The system should not only help the wounds heal quickly but also prevent the onset of new ulcers. # www.epfl.ch

4

Lifesciences plus 01 I 2016

The newly developed gene circuit can recognise two different molecules at the same time. (Visualisations: Science Translational Medicine / Chris Bickel.

Reprinted with permission from AAAS)

Implant Acts as a Countermeasure

ETH Professor

Martin Fussenegger calls them molecular prosthetics: cells with specially developed gene circuits that can be implanted into an organism, where they take over metabolic functions that the organism cannot perform itself. Fussenegger and his team at ETH Zurich’s Department of Biosystems Science and Engineering in Basel have now succeeded in developing a molecular prosthesis of this kind where the functions are far more complex than before. The prosthesis is tailored to the treatment of psoriasis, a complex and chronic inflammatory disease of the skin. Gene circuits created in the past typically monitored only whether a metabolic molecule A was present in their environment; if so, they produced a metabolic molecule X as a response. The new, more complex circuit can detect two molecules, A and B, simultaneously, and only if both are present does it produce the molecules X and Y. “We have used cellular components to build an AND logic gate, as is familiar in electronics and without which computers could not function,” says Fussenegger. When researchers implanted a circuit with an AND gate of this kind into mice, the circuit was able to successfully suppress phases of psoriasis in the mouse model.

The different cells of the immune system are involved in two ways during a psoriasis phase: first, they are responsible for triggering an inflammatory response by increasing the production of various messengers, including those referred to as TNF and IL-22. Second, at a later point, they produce a series of messengers that cause the inflammation to fade away again, among them IL-4 and IL-10. The circuit developed by the ETH researchers can detect the inflammatory molecules TNF and IL-22. If (and only if) these two messengers are present simultaneously, the circuit produces the anti-inflammatory molecules IL-4 and IL-10. “In this way, our molecular prosthesis helps the immune system to suppress the inflammatory response,” explains Fussenegger. # www.ethz.ch

SWISS RESEARCH I News

Prejudices Make the Brain Linger

New Substance Specifically Blocks Alzheimer’s Enzyme

For the first time,

UZH Researchers have shown an international team headed by scientists from the University of Zurich has found a way that it is possible to inhibit the Alzheimer’s disease to specifically inhibit an enzyme that is partly process specifically without responsible for Alzheimer’s disease. Protein affecting the physiological deposits in the brain are hallmarks of processes using induced Alzheimer’s disease and partly responsible pluripotent stem cell-derived for the chronically progressive necrosis of neurons. (Picture: UZH) the brain cells. The protein clumps mainly consist of the β amyloid peptide (Aβ), a protein fragment that forms when two enzymes, β and γ secretase, cleave the amyloid precursor protein (APP) into three parts, including Aβ, which is toxic. If β or γ secretase is blocked, this also inhibits the production of any more harmful β amyloid peptide. The problem is that the enzymes are also involved in other key cell processes.

A large number of substances have been developed, including some highly promising ones that reduced the amount of Aβ in mouse models effectively. The current β secretase inhibitors however don’t just block the enzyme function that drives the course of Alzheimer’s, but also physiologically important cell processes.

A soccer fan

needs more time to associate a positive word with an opposing club than with his own team. And supporters of a political party associate a favourable attribute faster with their party than with political rivals – even if they endeavour towards the opposite.

It is long since known that a positive association with one’s own group, an “in-group,” happens unconsciously faster than with an “outgroup.” These different reaction times become visible in the Implicit Association Test (IAT). But why the effort to address a friendly word to an outgroup takes more time was not clear up to now.

Now a team headed by Prof Daria Knoch from the Department of Social Psychology and Social Neuroscience at the Institute of Psychology, University of Bern, shows that an additional mental process is not responsible for this, as has often been postulated – but rather the brain lingers longer in certain processes. The researchers relied on a unique combination of methods for their study: they conducted an Implicit Association Test with 83 test subjects who are soccer fans or political supporters. While the test persons had to associate positive terms on the screen by means of a button click, either with their in-group or with an outgroup, the brain activity was recorded by means of an EEG. “We analysed these data with a microstate analysis. It enabled us to depict all processes in the brain for the first time – from the presentation of a word up to pressing the button – temporally and also spatially,” explains co-lead author Dr Lorena Gianotti. The analysis shows the following: the brain runs through seven processes, from the presentation of stimulus – i.e. a word – up to button click, in less than one second. The number and sequences of these processes remain exactly the same, regardless of whether the test subject had to associate positive words with the in-group or with an outgroup. The reaction time with the outgroup situation is therefore longer, because some of the seven processes take longer. # www.unibe.ch

To address this, the Zurich scientist studied how β secretase might be inhibited selectively. In a series of experiments, the scientists were able to demonstrate that the Alzheimer’s protein APP is cleft by β secretase in endosomes, special areas of the cells that are separated by membrane envelopes, while the other vital proteins are processed in other areas of the cell. The researchers exploited this spatial separation of the protein processing within the cell. “We managed to develop a substance that only inhibits β secretase in the endosomes where the β amyloid peptide forms. The specific efficacy of our inhibitor opens up a promising way to treat Alzheimer’s effectively Preparation of an EGG recording: in future, without causing the patients any serious side effects,” It is used for a microstate analysis says Professor Lawrence Rajendran from the Systems and Cell Biology which can depict processes in the of Neurodegeneration at the Institute of Regenerative Medicine of brain temporally and spatially. the University of Zurich. # www.uzh.ch (Picture: University of Bern / Adrian Moser)

SWISS RESEARCH

BRAIN PLASTICITY

Train the Brain: Physical Excercise Influences Memory The ability of learning and memory in animals and humans is possible because the brain is not a static organ but a malleable network of communicating nerve cells. The formation of new neurons in the brain and their interaction pattern can be influenced by a simple and very natural movement: Mice running in wheels seem to have better long-term memory than sitting animals.

The Patterns of Brain Plasticity Although neuroscience has unraveled a lot of secrets of the mammalian brain, there are still a lot of processes to discover and understand. It was only in the later 20th century, when the old dogma of the “static brain” was corrected. The previous scientific consensus was that the brain develops during a critical period in early childhood and then remains relatively unchangeable (static) afterward. However, research brought to light that the brain remains changeable or “plastic” even into adulthood.

Today, the term brain plasticity describes lasting changes to the brain throughout an animal’s or man’s life course.

Plasticity is most important for learning and memory. Without the constant formation of new circuits in the cerebral network of neurons, the brain of an adult would never be able to learn and

SONJA BICHSEL-KÄSER

H © istockphoto.com

recall new things. A demonstrative example for umans and many animals could not sur-

brain plasticity is the recovery of brain injured or

vive without a special ability of their

stroke patients. Movements or language can in-

brains: learning. This process of investi-

deed be re-learned in an adult phase and not only

gating and memorizing new objects or

during childhood or embryogenesis.

situations and being able to recall them is crucial

A human brain contains several billion neurons,

for evolution, since memories of past experience

each connected by synapses to several thousand

continuously influence behavior and thus interac-

other neurons. These neurons communicate via

tion with the environment.

their axons, which carry action potentials to distant

8

Lifesciences plus 01 I 2016

SWISS RESEARCH I Brain Plasticity

parts of the brain or body. Nerve cells

pocampal network. Furthermore, the

stem cell proliferation and adult neuro-

form different biological neural net-

dentate gyrus of the hippocampus is

genesis. The more cells are formed, the

works, series of interconnected neurons

special, as it is the only region in the

higher the possibility of integration and

whose activations define a recognizable

adult human brain, where new neurons

formation of new circuits. Indeed, ex-

pathway. Among these networks, there

can be formed even until 80 years of

periments with mice showed that hip-

are neural circuits, functional entities of

life. Adult neurogenesis, the formation

pocampal memory formation was im-

interconnected neurons, which are able

of new neurons in the adult hippocam-

proved when the animals could run in

to regulate their own activity using feed-

pus, further supports pattern separa-

wheels.

back loops.

tion during memory tasks. Especially

In 2015, the team of Professor Josef

Learning bases on the plasticity of

newly generated cells called hippocam-

Bischofberger, group leader at the De-

these circuits in the brain because syn-

pal granule cells influence pattern sep-

partment of Biomedicine at the Univer-

aptic communication can be dynami-

aration.

sity of Basel, published new findings on

cally adjusted. Dependent on experi-

But life of a newborn granule cell

the effect of physical exercise on neuro-

ence and brain activity, new synapses

isn’t easy: There are already exist-

genesis in the journal “Brain Plasticity”.

can be formed and pre-existing syn-

Bischofberger was not pleased with the

apses can be weakened or strength-

fact that “many behavioral tests assess-

ened to make lasting changes in the ef-

ing

ficiency of their synaptic transmission.

Simply

stores

put,

the

information

in

brain

electrical

the

flow

activity

separation

in

rodents

feedback (food rewards, electric

net-

shocks) and thus, involved

works of modified synapses controlling

pattern

worked with positive or negative

emotion.” Since emotion al-

of

so evokes extensive hip-

and

pocampal activity, these

thereby shaping infor-

tests do not show clear-

mation processing. Af-

ly whether the pattern

terwards,

separation

memories

observed

are retrieved by reac-

in

tivating

net-

curred due to emo-

works. An important

tion or learning tasks.

these

process called pattern

the

studies

oc-

Novel Object Recognition

separation is the process of making similar

In order to test effects of

patterns of neural activ-

physical exercise on pat-

ity more distinct to gen-

tern separation without the

erate a unique representa-

influence of reward or pun-

tion of each memory item.

ishment, hence without emo-

The process is highly relevant

tion, scientists of Bischofbergers’

since we have many objects/expe-

group used a method called novel ob-

riences that are similar to each other

ject recognition (NOR). This test is

(for example human faces) but none-

based on a natural behavior of rodents,

theless must be remembered as distinct. In a nutshell: Different memories

ing cell-networks in which they have to

known to preferentially explore novel

need different patterns.

become integrated, comparable to a

objects in their environment. They

new kid on the block in the school yard.

monitored the mice’s exploration of ob-

However, granule cells are most sensi-

jects either similar or very distinct to fa-

The hippocampus, a specific, sea-horse-

tive to integration into the hippocampal

miliar objects. During the tests, mice

shaped region of the brain, plays a cru-

circuitry in a period of 4 weeks after mi-

were either sitting (control group) or

cial role in the learning and consolida-

tosis. If they do not manage to connect

running in a wheel voluntarily. After

tion of information for short- and long-

with the adult cells, they die off within a

several weeks, the researchers tested

term memories. It is located in the me-

few weeks. In this context, it makes

learning capabilities of the different an-

dial temporal lobe underneath the

sense for the newbies to be very active

imals and placed two identical objects

cortical surface and contains two main

and thus being able to connect to their

(cones and pyramids) in front of an ani-

interlocking parts called Ammon’s horn

neighbors more easily than the older

mal for a few minutes. For the test phas-

and the dentate gyrus.

surrounding cells.

es 90 minutes and 24 hours later, they

The Active Survives

The latter is known to be important

Interestingly, physical exercise has

exchanged one of the objects by a new

for pattern separation within the hip-

been shown to increase hippocampal

one with different shape and different

Lifesciences plus 01 I 2016

9

SW SW I SISS S RE RSEESA ER AC RH CH I BI rUan i nt ePr lrausbtri ci ki t y

color (distinct) or of the same color but

ilar sample objects. In their experi-

In the control animals, the recognition

different shape (similar) and presented

ments, Bischofberger and his team did

memory

them again to the animals. Finally, they

not find a difference in sensory percep-

hours. This decay was prevented in run-

evaluated memory by comparison of the

tion of details in the test groups. After

ning mice.

time spent on exploring the novel and

90 minutes, details could be retrieved in

In the long-term retention test, sed-

the familiar objects.

both groups. “What we found interest-

entary animals were able to remember

Both, runners and control animals

ing, was a temporal gradient for the re-

distinct objects. In other words, they

were able to distinguish distinct or sim-

tention with similar objects,” he states.

recognized which of the objects was fa-

clearly

decayed

within

24

miliar and which was novel. However, when it came to similar novel objects, running mice remembered the familiar

Further Explanations

objects significantly better. Apparently, sedentary animals did not consider the

Membrane Potentials

similar novel object as novel. This im-

Voltage is a difference in positive and negative electric charges on opposite

plies that they had simply forgotten the

sides of a resistive barrier such as the cell membrane. A typical resting

exact shape.

membrane potential of a cell arises from the separation of potassium ions

Taken together, running did not af-

from intracellular, relatively immobile anions across the membrane of the cell.

fect NOR memory of highly distinct ob-

(The separation occurs because of the different permeability for the two ions

jects, sedentary mice could still remem-

and because of the work of ion pumps embedded in the membrane).

ber them, albeit less precisely. But only

However, excited neurons are able to communicate via an increase of voltage:

running mice were able to distinguish fa-

The resulting action potentials (spikes) are generated by voltage-gated ion

miliar objects from similar novel objects.

channels in the plasma membrane. These channels open upon stimuli (for example the release of neurotransmitter in the synaptic cleft or an electric

Take a Closer Look

impulse) and allow an influx of sodium ions. This in turn changes the

In order to investigate the effects of

membrane potential, causing more channels to open, and a further rise

physical exercise on brain areas on a

in membrane potential occurs. Thus, a signal is transmitted along the cell

cellular level, the researchers examined

membrane and the axon.

hippocampal samples of the test ani-

The influx of sodium ions causes the polarity of the plasma membrane to

mals in a later step. They labeled brain

reverse. The ion channels are then inactivated. In a refractory period,

slices with an antibody against Double-

potassium channels cause an outward current of potassium ions, returning

cortin, (a protein expressed selectively

the membrane potential to the resting state.

by neuronal precursor cells and young

Spike-timing-dependent plasticity (STDP)

neurons), and analyzed the samples by fluorescence microscopy. As previously

STDP refines the strength of connections between hippocampal neurons.

shown by other researchers, Bischof-

It is induced by tight temporal correlations between the spikes of pre- and

berger and his team could confirm, that

postsynaptic neurons. Repeated presynaptic spike arrival (a few milliseconds)

voluntary

before postsynaptic action potentials may lead to long-term potentiation

creased the number of young granule

(LTP) of the synapses.

cells in rodents due to increased hip-

running

significantly

in-

pocampal cell stem cell proliferation.

Long-Term Potentiation (LTP)

“In the dorsal hippocampus, the in-

LTP is a persistent strengthening of synapses based on recent patterns of

crease was about 2-fold”, states Bis-

activity. These are patterns of synaptic activity that produce a long-lasting

chofberger. Moreover, the researchers

increase in signal transmission between two neurons.

analyzed the cell growth by simply counting the dendrites formed in the in-

Pattern separation

ner and outer molecular layer of the

Pattern separation is a computational process (associated with the dentate

dentate gyrus. (Dendrites are the parts

gyrus) that occurs when the output firing patterns of a network are less similar

of a neuron which form connections to

to one another than the input firing patterns. In the hippocampus, it has been

other neurons in the surrounding area).

proposed to be an essential step in information processing to avoid interfer-

Whereas in sedentary mice the den-

ence between memories. In associative networks memory cues activate a set of

drites grew into the inner molecular

neurons that then activate those neurons that represent the stored memory

layer, dendrites of neurons of running

itself (pattern completion). Therefore, memories have to be stored “far apart”.

mice reached the outer molecular layer.

If two events are encoded too similarly, the stored memories may converge

Thus, the dendrites of young granule

into a single, inappropriate memory. Future recall would then be impossible.

cells were on average longer in mice using running wheels. By calculation of a

10

Lifesciences plus 01 I 2016

SWISS RESEARCH I Brain Plasticity

fluorescence ratio, the scientists found a 2-fold in-

ter) even before they had formed synapses with

crease of dendrites in the outer molecular layer in

other cells. “However, the exact mechanisms for

running mice. Added up, a 4-fold increase in num-

neurogenesis-dependent consolidation of memo-

ber of young granule cell dendrites occurs, leading

ry items are still not fully understood,” the pro-

to a 4-fold higher chance for the formation of new

fessor says. “Especially on the molecular level,

synapses with axon terminals of the pre-existing

many open questions remain.”

mature cells.

Although there are many indications that

These findings suggest that exercise-induced

running and physical exercise also improves

increase in neurogenesis improves memory: The

memory in humans, adult neurogenesis in hu-

more cells and dendrites, the higher the possibility

mans is much more difficult to investigate and

of synapses being formed, the higher the precision

many details are still unknown. But since physi-

of pattern separation and hence, the better long-

cal exercise has an important impact on human

time memory. With his studies, Bischofberger could

well-being, it would be a nice side-effect if it also

show that running improves hippocampal pattern

trained our brains at the same time.

separation needed for object-recognition during spontaneous behavior.

Bischofberger published earlier that young granule cells in the adult hippocampus differ from

REFERENCES

mature cells regarding active and passive membrane properties. In young neurons, Ca2+ channels

1

www.wikipedia.com

generate isolated spikes and boost fast Na -action

2

www.nature.com

potentials. Their excitability pattern is unique and

3

Bolz L, Heigele S, Bischofberger J: Running Improves Pattern Separation during Novel

+

supports enhanced synaptic plasticity which in turn enables easier formation and tuning of new

Object Recognition. Brain Plasticity. 2015 (1), 125–137 4

synapses. Other publications showed that new neurons are activated by GABA (a neurotransmit-

Schmidt-Hieber C, Jonas P, Bischofberger J: Enhanced synaptic plasticity in newly generated granule cells of the adult hippocampus. Nature. May 2004 (429), 184–187

4

Miller G: New Neurons Strive to Fit In. Science. February 2006 (311), 938–940

02

The microscope image shows section through a part of the dentate gyrus of a young mouse’s hippocampus. Neural stem cells are dyed in green. The red cells are the astrocytes surrounding them. Together the cells help in the storage of memory in mammals.

(Picture: Wikimedia)

01

Neuron and Synapse in detail. (Picture: Wikimedia)

03

Newborn neurons in the transgenic mouse hippocampus (dentate gyrus) labeled with green fluorescent marker. (© istockphoto.com)

Lifesciences plus 01 I 2016

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SWISS RESEARCH

MEDICAL TECHNOLOGY

Nanomechanical Piezoresistive Membrane Sensors for Non-Invasive Cancer Detection More than a century ago, medical practitioners asked the patient to exhale in order to figure out whether the breath contains specific smells possibly related to a disease. This old idea is adopted to investigate breath samples of cancer patients using a nanomechanical electronic nose device.

HANS PETER LANG

I

n many diseases of the respiratory tract system, chemical by-products are found in the patient’s exhaled breath.

Such specific chemical tracer substances are often analyzed using gas chromatography and mass spectrometry

methods, but interpretation of results is difficult and time-consuming. Here, an electronic nose technique is presented to characterize patients’ exhaled breath samples in a non-invasive way which allows a simpler analysis than the abovementioned classical standard analytical procedures.

Microfabrication and Nanomechanics In recent years, mechanics has experienced a revival, as microfabrication technologies and nanotechnology are applied to produce tiny structures. The development started with a novel imaging technique named atomic force microscopy1, which provides ultrahigh resolution of a surface on the atomic scale. This technology is based on raster-scanning a surface with a microfabricated cantilever beam that has a tiny tip at its free end. While keeping the distance constant between tip and surface by controlling their interaction force, a topography map of the surface is produced, revealing details on the atomic scale. Most frequently, a laser is used to determine the tiny deflection of the cantilever in the nanometer range by reflecting the laser beam at the apex of the cantilever and measuring the position with a lateral photodiode. Albeit the cantilever is very small, the readout still requires tabletop-sized equipment.

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S W I S S R E S E A R C H I M e d i c a l Te c h n o l o g y

A cantilever beam is an excellent force sensor

volatile organic compounds present in patients’

for ultrasmall forces in the nano Newton range.

breath diffuse into the polymer layers and deform

The high sensitivity can not only be used for atom-

the membranes by changes in surface stress. The

ic force microscopy, but also allows to apply the

bending of the membranes is measured piezoresis-

cantilever beam for measuring surface forces during molecule adsorption processes on the cantilever surface, thus enabling cantilevers as chemical sensors2.

Over the years, cantilever sensors turned out to be very useful for detecting DNA hybridization with single point mutation sensitivity3, protein and antibody recognition4 and even for assessing patient eligibility for cancer treatment5. The only drawback is that the equipment for optical cantilever deflection readout is still quite bulky. This disadvantage can be overcome by employing a different method for deflection detection, namely the use of piezoresistor elements to determine bending. The required readout electronics then fits in a box of 10 cm × 10 cm × 16 cm, including data acquisition and gas handling.

Membrane Surface Stress Sensors Our type of sensors consists of a circular diskshaped membrane held by 4 piezoresistive arms, which are electrically connected in a Wheatstone bridge circuit [Figure 1]. Here we use an array of such nanomechanical membrane surface stress sensors (MSS). Each membrane is coated by inkjet spotting with a different polymer layer of a thick-

01

Schematics of a membrane-type surface stress sensor (MSS). The actual diameter of the round membrane (shown in blue) is 500 µm and its thickness

ness of typically 1 micrometer. The applied coat-

is 2,5 µm. The membrane is suspended by four sensing beams with integrated

ings include polymers such as carboxymethylcel-

p-type piezoresistors (shown in red), representing a full Wheatstone bridge.

lulose, polyvinylpyridine or polyethylenimine. By

A solid supporting frame (green) holds the sensor. MSS are arranged in arrays

pumping

for detection of VOCs in a gas stream passing through the measurement

previously

collected

breath

from

a

breath-sample bag into a measurement chamber,

02

chamber. The numbers indicate the scale in millimeters.

Measured MSS bending response curves of the 8 membranes in an array.

4 cycles of breath sample injection and purging with dry nitrogen gas are shown, whereby the first is discarded.

The deflection values after 10, 15, 20 and 25 seconds after beginning of each injection were subtracted from the value at the beginning of the injection (0 seconds) to reduce the influence of possible drifts in the measurement. The resulting data set of deflection values of all 8 sensors (“fingerprint”) characterizes the breath sample.

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S W I S S R E S E A R C H I M e d i c a l Te c h n o l o g y

03

Data evaluation in a principal component analysis plot. Each injection/purging cycle in Figure 2 is represented here as a dot.

Clear cluster forming is observed, allowing the distinction of HNSSC patients before surgery (within red ellipse) and healthy control persons (green ellipse). Head and neck cancer patients after surgery (blue ellipse) are found to be completely cured and appear in the sample cluster as the healthy control persons. Principal component analysis (PCA) is a multivariate statistics method, in which multidimensional data is projected into the two-dimensional space in such a way that different samples are maximally spaced in the graph, whereas similar samples form narrow clusters.

tively and the signals are converted to voltages,

In a clinical study, we investigated breath samples from

which are then further amplified and recorded.

head and neck cancer patients and healthy donors (smokers)

MSS represents a non-invasive method that de-

as control persons in a double blind trial. The patient inclu-

tects volatile organic compounds (VOCs) in ex-

sion criteria were based on histologically confirmed carcino-

haled breath. Such VOCs are associated with the

ma at a comparable stage. The patients were selected from the

occurrence of certain cancer types, in particular

same age groups. Patients and donors were asked to breathe

head and neck cancer.

into a 1 liter Tedlar bag. The bags were then stored at 4 °C until analysis. Each breath sample has been measured 7 times,

Head and Neck Cancer

whereby the first injection-purge cycle has been discarded to

Cancer is a malicious disease where cells are grow-

avoid influence of previous measurements. Evaluation using

ing in an uncontrolled way forming a tumor, invad-

principal component analysis (PCA) allowed to clearly distin-

ing and destroying adjacent healthy tissues and or-

guish between the group of head and neck cancer patients

gans. Cancerous cells spread to other locations in

and the group of healthy control persons. As head and neck

the body via lymph or blood vessels to form metas-

cancer can be completely removed by surgery, the breath of

tases, the most common cause of cancer-related

cured patients was also investigated after surgery and the re-

death in patients with solid tumors.

sults were found to be similar to those of the healthy control

Head and neck squamous cell carcinoma (HN-

group indicating that surgery was successful [Figure 3].

SCC) is the fifth most important cancer type world-

Other evidence that VOC profiles in exhaled breath can be

wide and is closely related to smoking. HNSCC is

used to detect diseases has been shown by Phillips et al. for

highly curable if spotted early. Early detection of

lung and breast cancer6. A pilot study of analysis of air exhaled

HNSCC and identification of residual or recurrent

by HNSSC patients using an array of 5 gold nanoparticle sen-

disease in treated patients allow early therapeutic

sors and gas chromatography has shown promising results7.

intervention and may result in a survival advantage.

Diagnosis is normally performed by endoscopy and taking a biopsy of suspect lesions.

Medical applications favor the routine use of a compact, small-sized, portable and non-invasive device. A MSS prototype was used to examine pa-

Acknowledgements

tients’ exhaled breath samples in search for VOC patterns associated with head and neck cancer. Patients’ breath samples are transported using micropumps into the measurement chamber. The bend-

We thank

ing response of all 8 polymer-coated membrane

(SNI), the NanoTera Program, the Cleven Foundation

sensors to the VOCs in the breath sample is record-

and the Swiss National Science Foundation for financial

ed. Consecutive purging with dry nitrogen gas re-

support. We acknowledge the continuous technical

sets the response back to the baseline and the sen-

support from A. Tonin, R. Maffiolini (electronics work-

sors are fully regenerated. Thus a series of expo-

shop), H. Breitenstein, S. Martin and his crew (mechani-

sure/purge cycles is recorded [Figure 2]. Analyzing

cal workshop) in the Department of Physics, University

these deflection patterns allows characterization of

of Basel.

the Swiss Nanoscience Institute

the condition of the patient.

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S W I S S R E S E A R C H I M e d i c a l Te c h n o l o g y

Outlook Early detection of primary tumors and of recurrences after surgical removal of the primary tumor is crucial for patients with HNSSC. Invasive analyses, e. g. endoscopies, give clear indication on the treatment success, but are a hassle for the patient.

Detecting the VOCs in exhaled breath represents a non-invasive method to follow the success of a treatment/surgery. We have shown that MSS are capable to distinguish HNSSC patients before surgery from healthy control persons and HNSSC patients after surgery by monitoring VOCs in patients’ breath samples. The measurement device used is portable and powered by a laptop computer’s universal serial bus port.

Detecting VOCs associated with cancer growth will ultimately lead to a simple, easily performable and non-invasive screening technique that can be used in conjunction with, or as alternative to standard more invasive techniques. The technique could eventually be adapted to other pathologies affecting the respiratory tract.

AUTHOR Hans Peter Lang. Physicist and Research Group Leader at the Swiss Nanoscience Institute, Dept. Physics, Univ. Basel

REFERENCES 1

G. Binnig, C. F. Quate, Ch. Gerber. 1986. Atomic Force Microscope.

Phys. Rev. Lett 56: 930–933.

2

M. K. Baller, H. P. Lang, J. Fritz, Ch. Gerber, J. K. Gimzewski, U. Drechsler, H. Rothuizen, M. Despont, P. Vettiger, F. M. Battiston, J.-P. Ramseyer, P. Fornaro, E. Meyer, H.-J, Güntherodt. 2000. A cantilever array-based artificial nose. Ultramicroscopy 82: 1–9.

3

J. Fritz, M.K. Baller, H.P. Lang, H. Rothuizen, P. Vettiger, E. Meyer, H.-J. Güntherodt, Ch. Gerber, J.-K. Gimzewski. 2000. Translating Biomolecular Recognition into Nanomechanics. Science 288: 316–318.

4

N. Backmann, C. Zahnd, F. Huber, A. Bietsch, A. Plückthun, H.P. Lang, H.-J. Güntherodt, M. Hegner, Ch. Gerber. 2005. A label-free immunosensor array using single-chain antibody fragments. Proc. Natl. Soc. Sci. (USA) 102: 14587–14592.

5

F. Huber, H.P. Lang, N. Backmann, D. Rimoldi, Ch. Gerber. 2013.

Direct detection of a BRAF mutation in total RNA from melanoma cells using cantilever arrays. Nat. Nanotechnol. 8: 125–129.

6

M. Phillips, N. Altorki, J.H. Austin, R.B. Cameron, R.N. Cataneo, J. Greenberg, R. Kloss, R.A. Maxfield, M.I. Munawar, H.I. Pass, A. Rashid, W.N. Rom, P. Schmitt.

2007. Prediction of lung cancer using volatile biomarkers in breath.

Cancer Biomark. 3: 95–109.

7

M. Hakim, S. Billan, U. Tisch, G. Peng, I. Dvrokind, O. Marom, R. AbdahBortnyak, A. Kuten, H. Haic. 2011. Diagnosis of head-and-neck cancer from exhaled breath. Brit. J. Cancer 104, 164–165.

Lifesciences plus 01 I 2016

SWISS RESEARCH

Illustration of flagellated Escherichia coli cells (© istockphoto.com)

SYSTEMS BIOLOGY

Bacterial Resistance – Getting to know the Enemy Tissue complexity has been largely ignored in infection research. Biologists from the University of Basel are now investigating how pathogen’s tolerance to antibiotic substances evolves in infected host tissues. Their focus is on Gram-negative bacteria, which actually start running out of control.

BEATE PEISELER

reus (MRSA), a classical hospital pathogen, could

A

be reduced following severe hospital hygiene rules. ntibiotic drugs cause a selection pres-

But multidrug-resistant tuberculosis (MDR-TB) is

sure favoring the growth of bacteria,

on the rise: 480,000 new cases have been reported

which can escape the drug’s poisonous

in 2013. Especially infections with multi-resistant

effect. Both Gram-positive and -nega-

Gram-negative bacteria like gonorrhoea and uri-

tive bacteria possess efflux pumps: transport pro-

nary tract infections actually start running out of

teins, which are involved in the expulsion of toxic

control (see box Eskape pathogens on the rise).

substrates like antibiotics from within cells into the external environment. They also produce en-

Persistent Pathogens

zymes, which are able to transform antibiotic sub-

“The situation is grave and new strategies for a suc-

stances into more tolerable molecules. Genes en-

cessful control of infectious diseases are not in

coding for efflux pumps, drug-modifying enzymes

sight,” states infection biologist Dirk Bumann. In

and other resistance mechanisms are spread from

2006, Bumann, who is a Professor at the Biozentrum

one species to another by horizontal gene transfer.

of the University of Basel since 2007, had published

Several resistance genes may be taken up at one

a network analysis on Salmonella enterica in Na-

go. As a result, the routine use of antibiotics in the

ture. 64 Salmonella enzymes had been identified as

healthcare sector and animal husbandry has led to

essential and are conserved in other important hu-

considerable problems with multi-resistant patho-

man pathogens. Unfortunately, all these potential

gens all over the world. The number of new infec-

targets for new drugs belong to metabolic pathways,

tions with methicillin-resistant Staphylococcus au-

which are already inhibited by current antibiotics or

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SWISS RESEARCH I Systems Biology

have previously been considered for antimi-

Microenvironment Influences Bacterial Growth

crobial development. This is only one reason

Bumann’s Timerbac reports on single-cell growth

why there is so little progress in the field of

rates at various stages of disease. The experiments

identification and development of new po-

revealed extensive phenotypic variation in Salmo-

tent antibiotics with new modes of action for

nella single-cell growth rates. Rapidly growing

combating multi resistant pathogens. In fact,

subsets dominated overall Salmonella prolifera-

the problem is far more complex. Whereas

tion and disease progression. The researchers ar-

an entire bacterial population can be killed

gue that these Salmonella might have superior ac-

by one single dose of appropriate antibiotic

cess to nutrients, but don’t exclude other yet-un-

when grown in a test tube, antibiotic thera-

identified factors. Afterwards, infected mice were

pies have to be continued for many days. For

treated with a fluoroquinolone antibiotic. “Salmo-

instance scarlet fever: Symptoms subside af-

nella eradication was slow. Fast-growing Salmonel-

ter three days, and the responsible pathogen

la survived poorly, whereas non-dividing and

Streptococcus pyogenes does not show any

slow-growing Salmonella survived best but were

antimicrobial resistance yet. But therapy

rare; this limits their impact. Instead, Salmonella

with penicillin has to be strictly continued

with moderate growth rates of one to four divisions

for ten days, in order to minimize the risk for

per day and intermediate levels of tolerance domi-

cardiac and other long-term complications.

nated throughout the therapy. “More effective tar-

“Antibiotic treatment of patients often

geting of this large subset could substantially ac-

leaves a fraction of transiently refractory

celerate therapy,” Bumann concludes from the re-

bacteria unaffected. Such persisters are hy-

sults. There is emerging evidence that distinct host

pothesized to cause relapse and chronic in-

microenvironments are linked to disparate out-

fections and require long periods of admin-

comes of local host-pathogen interactions. Disease

Dirk Bumann has been

istration. Our research group is investigat-

progression can result from failure to control indi-

Professor at the Biozentrum

ing how pathogen’s tolerance to antibiotic

vidual infection foci, despite successful eradication

of the University of Basel

substances evolves in infected host tissues,”

of others. Bumann’s team uses proteomics, micro-

since 2007. His research

Bumann explains. The researchers from

focusses on the Systems

Basel work with in vitro and in vivo infec-

biology of Salmonella

tion models, which they obtain by infecting

and Shigella Infections.

cells (cell culture infection) or mice (animal model) with well-understood model pathogens like Gram-negative Salmonella or Shigella. Both cause diarrhea, while Salmonella infections additionally may lead to severe typhoid fever and non-typhoidal Salmonella bacteremia. In order to monitor Salmonella growth and survival during disease and treatment, Bumann’s team has re-

Microscopic picture of

fined an in-vivo labeling method published

Gram-stained Pseudomonas

by Alexey Terskikh in 2000. Terskikh’s

aeruginosa cells.

“Timer” is a variant of the red fluorescent

(Picture: Wikimedia)

protein and used as a reporter to estimate terms of promoter activity. This reporter-protein time-delayed emits light at two different wavelengths. It indicates activation as well as down-regulation of target promoters on the whole-organism scale.

After detergent-treatment of the samples, pathogen cells are sorted from infected host cells by flow cytometry, while at the same time executing a reporter gene expression read-out at the single-cell level.

Sorted pathogen subsets can be subjected to further analysis (proteomics, metabolomics, molecular genetics).

Lifesciences plus 01 I 2016

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SWISS RESEARCH I Systems Biology

➜

Eskape pathogens on the rise

Infections

“The situation is grave and new

with multi-resistant Gram-nega-

strategies for a successful control of

tive bacteria like gonorrhoea and urinary tract infections

infectious diseases are not in sight.”

annual cases of sexually transmitted gonorrhoea infections,

DIRK BUMANN. Professor at the Biozentrum of the University of Basel.

worldwide. N. gonorrhoeae strains display resistance to

(UTIs) actually start running out of control. 100 million new provoked by the cocci Neisseria gonorrhoeae, are reported several antibiotic classes including penicillins, sulphonamides, tetracyclines, quinolones, and macrolides. The situation is aggravated by a recent decrease of susceptibility

bial genetics, competitive infections, and computa-

to the last-line treatment with extended-spectrum

tional approaches in order to obtain a comprehen-

cephalosporins.

sive overview on Salmonella infections in animal models like the mouse typhoid fever model. They

Since the late 1990s, there is a rise in community acquired

found out that Salmonella access at least 31 differ-

urinary tract infections (UTIs) due to multidrug-resistant

ent host nutrients in infected tissues. In order to si-

Escherichia coli, a usually harmless bowel bacterium.

multaneously exploit these different host nutri-

Multidrug-resistant E. coli produce extended-spectrum

ents, Salmonella express versatile catabolic path-

β-lactamases (ESBLs) causing resistance to all penicillins, to

ways. There is evidence that such complex host/

cephalosporins, and to aztreonam. Cross-resistances to

pathogen nutritional interfaces are a common fea-

trimethoprim/sulfamethoxazole and quinolones are present.

ture in many infectious diseases.

Together with Enterococcus faecium, Staphylococcus

From time to time, Bumann’s basic research

aureus, Klebsiella pneumoniae, Acinetobacter baumannii,

discloses new Achilles’ heels in pathogens with po-

Pseudomonas aeruginosa and Enterobacter spp., ESBL-pro-

tential for application in drug discovery and devel-

ducing E. coli have become the “highly problematic

opment. Within the framework of the SystemsX.ch,

antibiotic-resistant Eskape pathogens”. The Review on

he coordinated the RTD project BattleX from 2010

Antimicrobial Resistance from December 2014, commis-

to 2013. BattleX was focusing on pathogen and host

sioned by UK Prime Minister David Cameron, estimates that

metabolism in Shigella infections. One finding was

antimicrobial resistance is killing about 50,000 people a

that host cells maintain normal fluxes through gly-

year in the US and Europe, and more than 700,000 people

colytic pathways during infection. But it is not the

worldwide. Economic modellers, who were asked to make a

host who gains benefit from glycolysis. Instead, the

projection, argue that, if action was not taken to master

entire output is captured by the pathogen in form

resistance, a continued rise in resistance

of pyruvate, which is degraded in three steps to ac-

by 2050 would lead to 10 million people dying every year.

etate. “This pathway is crucial for Shigella’s intra-

For comparison only: Cancers, one of the leading causes

cellular growth. Two of the three enzymes, which

of morbidity and mortality worldwide, were related to

catalyze the degradation of pyruvate, do not occur

8,2 million deaths in 2012. Approximately 14 million new

in the host, what makes them suitable targets for

cancer cases are reported every year.

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SWISS RESEARCH I Systems Biology

Superbug Pseudomonas aeruginosa

rial agents. Drugs which manage to penetrate the barrier are often expelled from the cell by means of numerous broadly acting efflux

In 2008,

pumps. Membrane’s impenetrability and efflux pumps’ efficiency the “European Federation of Pharmaceutical

represent an enormous problem for drug development. Bumann’s

Industries and Associations” (Efpia) and the EU have launched the

Basel-based research team is contributing in the Translocation program

“Innovative Medicines Initiative” (IMI) as a Public-Private-Partnership.

by examining the outer membrane of Gram-negative bacteria.

IMI has funded several large projects including the “New Drugs for Bad Bugs” (ND4BB) program. ND4BB was launched in 2012 and disposes of

“Pseudomonas and other Gram-negative bacteria readily adapt to

a 680 million Euro total budget funded by Efpia and the EU. Among

different conditions by comprehensively remodeling their cell envelope

other things, ND4BB aims to demonstrate effective new antibacterial

properties. We observe differential expression of one or more of the

strategies for the treatment and prevention of infections. Its subpro-

some 30 porins, induction of one or more of their approximately

gram “Translocation” is primarily focusing on Gram-negative pathogens

20 efflux pumps, or modifications of lipopolysaccharides,” says Bumann.

like Pseudomonas aeruginosa and Escherichia coli. 18 academic and

The researchers are determining in vivo envelope properties with a focus

industrial partners are cooperating on identifying new ways of getting

on porins and efflux systems. They work with Gram-negative Salmonella

antibiotics into Gram-negative cells and preventing them from

in a mouse typhoid fever model and examine Salmonella’s outer cell

discharging the drugs before they can exert their antibiotic effect.

membrane during disease progression as well as during treatment with antibiotics. Experiments on P. aeruginosa are programmed, too.

“In immune compromised humans, P. aeruginosa causes infections

“We plan to collect and investigate patient’s sputum before and during

of the respiratory and the urinary tract and of wounds. It is often resistant

therapy with antibiotics,” says Bumann. His team had previously

to multiple antibiotics and has an enormous capacity to engender

developed a powerful tool for pathogen purification from infected host

resistance. P. aeruginosa demonstrates decreased susceptibility to most

tissue, an approach that reveals comprehensive quantitative data on

antibiotics due to low outer membrane permeability coupled to adap-

pathogen properties in vivo when coupled to mass spectrometry-based

tive mechanisms and can readily achieve clinical resistance,” informs

proteomics. “The aim is to generate an envelope model that accurately

infection biologist Dirk Bumann. The envelope of Gram-negative bacteria

predicts antibiotic penetration under in vivo conditions and identifies

is comprised of two membranes with significantly different properties.

potential targets for perturbation and increased antibiotic efficacy,”

The asymmetric outer membrane consists of phospholipids and lipo-

resumes Dirk Bumann.

polysaccharides and provides an excellent physical barrier to antibacte-

antibiotic therapy”, resumes the former

zyme adenylate cyclase. Research re-

lence and found in many Gram-nega-

project leader. In 2014, Bumann et al.

sults from Bumann’s lab suggest that

tive bacteria like Shigella, E. coli and

disclosed that EIIAGlc, a metabolic pro-

EIIAGlc binds to Salmonella’s type

Pseudomonas aeruginosa. The unex-

tein from Salmonella, is essential for

three secretion system 2 (TTSS-2), thus

pected role of this protein in Salmonella

acute infection. EIIAGlc was known to

stabilizing its cytoplasmic part and acti-

virulence makes it a potential target for

participate in carbohydrate transport

vating

drug discovery.

and activation of the key regulatory en-

TTSS-2 is involved in systemic viru-

virulence

factor

secretion.

-

Diagrammatic drawing of a Gram-negative bacterial cell wall.

(Picture: Wikimedia)

Lifesciences plus 01 I 2016

19

SWISS RESEARCH

ONCOLOGY

Avicenna Oncology and the Big Challenge: The Fight against Aggressive and Resistant Cancers Antibodies can be linked with the toxins that are released in a tumour and gradually destroy it by apoptosis, the term used to refer to programmed cell death. That’s how antibody drug conjugates (ADCs) work in cancer therapy. ADCs are the focus of interest of Avicenna Oncology GmbH in Basel, which is developing a new generation of toxic payloads associated with antibodies or tumour-targeting agents especially to treat aggressive and resistant cancers.

chemotherapy and antibody-based therapy. While

ELSBETH HEINZELMANN

I

the use of antibodies alone is not sufficiently effect’s something every physician is afraid of:

tive, and cytotoxic agents produce burdensome ad-

many cancers develop resistance to chemo-

verse effects during cancer treatments, a new

therapy drugs, and the treatment fails. This is

strategy involves the conjugation of a monoclonal

the case for a variety of blood cancers and sol-

antibody to a small cytotoxic molecule. This ap-

id tumours encountered in breast or ovarian, lung

proach has led to the development of ADCs. The

or lower gastrointestinal tract cancers. Generally,

toxic payloads used in the latest Antibody Drug

tumours consist of mixed populations of malignant

Conjugate therapies are focused on a small range

cells; some may be drug-sensitive while others are

of anti-tumour mechanisms and substances such

drug-resistant. While chemotherapy eliminates

as DNA damaging agents, RNA polymerase inhibi-

drug-sensitive cells, it leaves behind a large num-

tion, microtubule-disrupting agents or topoisomer-

ber of drug-resistant cells. Should the tumour start

ase inhibitors. “Today the ADC clinical pipeline is

to grow again, the chemotherapy no longer works

composed of more than 40 ADCs and 80 % of these

as the tumour cells are now resistant to this kind of

ADCs comprise only two toxic payloads developed

therapy and to irradiation.

by two leaders, namely Seattle Genetics and ImmunoGen”, the CEO reports.

Breaking New Ground in Cancer Therapy

© istockphoto.com

Some years ago, Zaki Sellam, who began his career

Focusing on a Special Toxin Platform

in research in Functional Genomics and Bioinfor-

“My vision was to found a company that offers an

matics, discovered his interest in Antibody Drug

alternative to this monopoly position of two sup-

Conjugates (ADCs). ADCs are composed of an anti-

pliers in the market”, says Zaki Sellam, who

body, a linker and a toxic pay-load, in other words

founded Avicenna Oncology GmbH in 2014 and is

cytotoxic entities targeted against cancer cell lines.

now its CEO. “We want to develop a new genera-

Numerous companies and institutions are working

tion of toxic payloads associated with antibodies

and developing antibodies and linkers, but very

or tumour-targeting agents especially for the ther-

few involve a toxic payload. But Zaki Sellam, with

apy of aggressive and resistant cancers. We are not

many years of experience in the fields of oncology

interested in working on ’me-too’ products. Our fo-

and biotechnology, wanted to pioneer an entirely

cus is on new toxic payloads to overcome aggres-

new approach to cancer.

siveness and resistance in cancer.” His aim is to

For the last decade conventional anti-cancer

identify several toxin families, to validate these

therapy has been focused on two approaches:

products preclinically and to demonstrate their

20

Lifesciences plus 01 I 2016

SWISS RESEARCH I Oncology

➜ mechanisms of action on therapy-re-

“The key issue of the operating mechanism is the efficient internalization of the antigen.”

ZAKI SELLAM. CEO of Aviecenna Oncology GmbH.

sistant tumour cells.

For the future toxin platforms, the only accessible compounds today involve two main modes of action, limiting

mour cells once the substance is ab-

biology. An inspired move was to appoint

the scope of efficacy against tumours.

sorbed. The conjugation of the cytotoxic

Dr Ivan Csendes to the Advisory Board

The objective of Avicenna Oncology is

molecule to the antibody inactivates the

of Avicenna Oncology. The long-time

to develop a new generation of toxic

drug while it is in circulation.

Head of Laboratory in antibiotic chemis-

pay-loads with differentiated mecha-

We must bear in mind that – like a

try was President of the Swiss Pharma

nisms of action. The company is also de-

Trojan horse – the toxin enters the can-

Licensing Group from 2004 to 2008 and

veloping a comprehensive mapping of

cer cell unnoticed. At this point the tox-

has extensive experience in business de-

the technology compatible with Avicen-

in is still firmly connected to the anti-

velopment. In Switzerland, Avicenna

na’s toxin platforms.

body. “This connection must be broken

Oncology has started to identify key

so that the active substance is able to do

opinion leaders in the field of ADC, fo-

How Do ADCs Work?

its deadly work within the tumour,” ex-

cusing on the clinical domain. “We are

ADCs have the potential to become a

plains Zaki Sellam. The linker causes

still evaluating potential partners in

new approach to the targeted and selec-

the toxin to be cleaved and released

Switzerland and are open to all ideas,”

tive treatment of cancer, provided that

within the targeted cancer cell, causing

concludes Zaki Sellam. “Our approach is

each component is effective. The mono-

cell death. “The key issue of the operat-

market driven. We use science to create

clonal antibody guides the toxic payload

ing mechanism is the efficient internali-

value and our interest is to achieve the

selectively to the tumour cell. This

zation of the antigen.”

best value as fast as possible – and

means that the antibody has to bind to

therefore we need innovative and relia-

malignant cells at a relatively high den-

Committed to Strong Partnerships

sity and merge in a way that allows the

From the outset, Avicenna Oncology has

drug to be released from the linker in

adopted an international approach and

the appropriate intracellular compart-

taken account of the global context. Zaki

ment. The linker attaches the active

Sellam believes in the benefits of co-

agent to the antibody – so as to ensure

operation with strong partners and com-

stability in the circulation – and releas-

mon efforts to generate value. He has

es the drug once it is internalized. The

built a solid network of scientific and in-

cytotoxic drug needs to be nontoxic

dustrial experts specialized in pharma-

Antibody-drug conjugates (ADCs) are highly

when linked to the mAb in the circula-

cology, chemical synthesis, conjugation,

potent biopharmaceutical drugs designed as

tion, but must be capable of killing tu-

antibody development and ADV target

a targeted cancer therapy. ADCs are complex

ble partners.”

molecules composed of an antibody (whole or fragment such as a single-chain variable fragment) linked, via a stable, chemical linker with labile bonds, to a biological active cytotoxic (anticancer) payload or drug. The anticancer drug (e. g. a cell toxin or cytotoxin) is coupled to an antibody that specifically targets a certain tumour marker (a protein that is, ideally, only present on the surface of tumour cells). Antibodies attach themselves to cells bearing this antigen marker. The biochemical reaction between the antibody and the antigen triggers a signal in the tumor cell, which then internalizes the antibody together with the cytotoxin. After the ADC is internalized, the cytotoxic drug is released Zaki Sellam, CEO of Avicenna Oncology

and kills the cancer. Due to this targeting,

GmbH. (Picture: avicenna oncology)

ideally the drug has lower side effects.

Lifesciences plus 01 I 2016

21

SWISS RESEARCH

RADIOIMMUNOTHERAPY

Not just for Lymphomas anymore Chemotherapy is often ineffective against refractory tumors. Scientists are working to overcome the defenses of the cancer cells in these tumors using radioimmunotherapy. It has been approved for the treatment of B-cell non-Hodgkin lymphomas. Several radioimmunotherapy agents have been studied for the treatment of a variety of other solid malignancies.

plex is not stable enough, transferrin may compete

CLAUDIA BORCHARD-TUCH

I

for the radionuclide and distribute it to the wrong n Switzerland, each year 16,200 people die of

sites in the body via circulation.

cancer. Chemotherapy is often ineffective

If the complex is stable, the radionuclide-labe-

against refractory tumors. Some forms of radia-

led antibodies dock to specific antigens expressed

tion penetrate deep through the body tissue

on the surface of the tumor cells with high affinity.

and can be used to treat tumors. However, they of-

The radioactive element decays, releasing the tu-

ten also damage healthy cells at the same time. Ra-

mor-destroying radiation without causing signifi-

dioimmunotherapy is an effective method to chan-

cant damage to the adjoining healthy tissue. The

nel this destructive energy so that it can be directed

radiation is extremely localized.

more effectively against tumors. A radionuclide is attached to an antibody, which transports the radio-

Non-Hodgkin Lymphomas

isotope to its site of action in the tumor.

In Switzerland, radioimmunotherapy with yttrium-

Chelators bind the radionuclide to the antibody.

90-ibritumomab-tiuxetan is approved for second-

Together they form an antibody-chelate complex,

line treatment of indolent follicular non-Hodgkin

which is extremely stable. And the strength of the

lymphomas [Figure 1]. A group of researchers

combination has a direct impact on the potential

around Valentin Koechli, University of Bern, exam-

success of the drug product. The body is home to

ined the effectiveness of yttrium-90-ibritumomab-

many other natural substances which are capable

tiuxetan in patients with Burkitt lymphoma [Figure

of forming complexes with the radionuclide such

2]. The patients had received rituximab during

as the iron-binding protein transferrin. If the com-

first-line treatment. Rituximab is a chimeric mono-

22

Lifesciences plus 01 I 2016

clonal antibody targeting CD20, a protein found on the surfaces of normal and malignant B lymphocytes [Figure 3]. CD20 plays an important role in the activation process for cell cycle initiation and differentation. The CD20 antigen is expressed on nearly all B-cell non-Hodgkin lymphomas but not in other bone marrow cells. Rituximab is effective in the treatment of non-Hodgkin lymphomas1. The role of additional anti-CD20 directed radioimmunotherapy for consolidation of first remission was examined. The data suggest that survival rates were excellent.

A research team around Flavio Forrer, University of Basel, explored the clinical response to lutetium-177 labeled chimeric anti-CD20-antibody rituximab in the treatment of patients with indolent lymphomas. The antibody was modified with DOTA-chelators with retained immunoreactivity.

DOTA is an organic compound with the formula (CH2CH2NCH2CO2H)4 [Figure 4]. DOTA can be conjugated to monoclonal antibodies by attachment of one of the four carboxyl groups as an amide. The remaining three carboxylate anions are available for binding to the radionucleotide2.

The researchers observed clinical responses at all dose levels and for all lymphoma entities. Some of the responses were durable; the longest follow-up is currently over eight years. The results demonstrated the safety and feasibility of 177Lu-

© istockphoto.com

DOTA-rituximab treatment.

01

05

Model of human

Skeletal formula of the chelator tiuxetan, attached to ibritumomab. (Pictures: Wikipedia)

03

antigen.

Crystal structure of rituximab

04 Lifesciences plus 01 I 2016

in complex with CD20.

02

DOTA.

Symptom of Burkitt

23

carcinoembryonic

lymphoma.

SWISS RESEARCH I Radioimmunotherapy

Colorectal Cancer

Thorium-227, for example, emits very ener-

There is a rich clinical experience of early phase clinical trials reported

gy-rich radiation, albeit over a short distance. Only

for radioimmunotherapy. Several promising targets have been identified,

two to ten cell layers are penetrated by thorium ra-

including CEA (carcinoembryonal antigen), TAG-72 (tumor-associated

diation, so its action on the tumor is localized and

glycoprotein 72), Ep-CAM (epithelial cell adhesion molecule), and A (an-

the surrounding, healthy tissue is left unharmed.

tigen) 33 [Figure 5]. In the setting of minimal residual disease, some

Targeted alpha therapy with 227thorium-tras-

studies have demonstrated favorable progression-free survival (PF) and

tuzumab of human ovarian cancer cells growing

overall survival (OS)3.

intraperitoneally in nude mice was clearly superior

Two studies with anti-CEA antibody have been reported in patients

to unlabeled trastuzumab therapy. The results war-

with small-volume disease. In both studies, 16 % of patients exhibited an

rant further studies of 227Th-radioimmunothera-

objective response, with 42 % to 45 % showing a mixed or minor response

py used as adjuvant treatment and for metastatic

to radioimmunotherapy. The median disease-free survival (DFS) in these

cancer6.

studies was approximately 18 months, with 5-year OS in nearly half of the treated patients3.

Hepatocellular Carcinoma

A team around Franz Buchegger, University of Lausanne, showed po-

The membrane antigen HAb18G/CD147 has been

tential ways of combining radioimmunotherapy and radiotherapy in pa-

identified as a potential radioimmunotherapy tar-

tients with limited liver metastatic disease from colorectal cancer. The

get, using the monoclonal antibody F(ab’)2 frag-

patients were treated with 6,9 GBq (range 4,7 to 8,4 GBq) 131I-labeled

ment 131I-metuximab as a hepatic artery infusion.

anti-CEA MAb F(ab’)2 fragments combined with 20 Gy RT to the liver4.

A phase I/II study determined the maximum tolerated dose to be 0,75 mCi/kg body weight per cycle

Breast Cancer

and, of the 73 patients receiving 2 cycles of radio-

Several active targets have been identified, including CEA, TAG-72,

immunotherapy, 20 had objective responses and 43

MUC-1 (mucin), L1 and L6. Modest responses have been seen, with

exhibited stable disease in response to therapy.

greater responses in the setting of high-dose radioimmunotherapy fol-

Median overall survival was 19 months. In a sepa-

lowed by autologous stem cell rescue. Interferon alfa may have the abili-

rate report, the group demonstrated highly specific

ty to upregulate expression of key radioimmunotherapy targets in pa-

tumor-to-normal-tissue absorbed doses3.

tients with breast cancer. Murine antibodies result in a high expression of human antimurine antibodies, limiting dosing strategies3.

In a study of a research group around Dennis Lindenblatt, Paul-Scher-

The Way Forward Radioimmunotherapy

for

solid

malignancies

rer Institute in Switzerland, combination of radioimmunotherapy with

shows promise, typically with fewer adverse events

177Lutetium-labeled anti-L1 cell adhesion molecule and the taxane

than traditional cytotoxic systemic therapy. The

paclitaxel was an effective therapy resulting in a prolonged overall surviv-

greatest efficacy will likely be in the adjuvant set-

al of human ovarian carcinoma-bearing nude mice compared with either

ting of minimal residual disease. Newer radionu-

monotherapy5.

clides, particularly alpha-emitters, offer increased antitumor potency with less toxicity.

Prostate Cancer A number of targets have been described for prostate cancer, including TAG-72, L6, MUC-1, and PSMA (prostate-specific membrane antigen).

REFERENCES

By contrast to most solid tumor types, patients with prostate cancer have exhibited some reasonable to radioimmunotherapy, particularly in the

1

combined modality setting when delivered with taxane chemotherapy.

with yttrium-90-ibritumomab-tiuxetan in adult patients with Burkitt

Patients with skeletal metastases experienced significant pain relief, even with radioimmunotherapy alone. The most promising current agent

Koechli V, et al. Consolidation of first remission using radioimmunotherapy

lymphoma. Leuk Res. 2015;39(3):307–10.

2

is 177Lu-J591 (lutetium-177 with monoclonal antibody J591), which tar-

Forrer F, et al. Radioimmunotherapy with 177Lu-DOTA-rituximab: final results of a phase I/II Study in 31 patients with relapsing follicular, mantle cell,

gets PSMA3.

and other indolent B-cell lymphomas. J Nucl Med. 2013;54(7):1045–52.

3

Ovarian Cancer

Tomblyn MB, et al. The new golden era for radioimmunotherapy: not just for lymphomas anymore. Cancer Control. 2013;20(1):60–71.

Radioimmunotherapy appears to be a potentially promising option, par-

4

ticularly in patients who have been optimally debulked following surgery

liver metastases from colorectal cancer: a feasibility study. Anticancer Res.

and chemotherapy but who still are at high risk for microscopic intraperitoneal disease. Alpha-emitting radionuclides hold particular promise

Buchegger F, et al. Combined radioimmunotherapy and radiotherapy of

2000;20(3B):1889–96.

5

in nonbulky disease, given the higher linear energy transfer and shorter

Lindenblatt D, et al. Paclitaxel improved anti-L1CAM lutetium-177 radioimmunotherapy in an ovarian cancer xenograft model.

path length compared with beta particles.

EJNMMI Res. 2014;4(1):54.

6

Heyerdahl H, et al. Targeted alpha therapy with 227Th-trastuzumab of intraperitoneal ovarian cancer in nude mice. Curr Radiopharm.

2013;6(2):106–16.

24

Lifesciences plus 01 I 2016

P L AY E R S & P R O D U C T S

NETWORKING

Enhancing Innovation Yield by Fruitful Dialogue Switzerland definitely stands for excellence in research and innovation and is globally renowned to be a driving force. This is especially related to the area of chemistry, pharma and food. But is the drive as powerful as it might be? In our interview three experts from different regions and professions discuss their individual views on current trends, bottlenecks and on possible innovation catalysts: Susanne Lauber Fürst, Inartis Network Vice President, Renens; Peter Pichler, President and CEO Brechbühler AG, Schlieren/Echallens; Dr Helmut Traitler, former Head of Innovation Partnerships at Nestlé Group and now Innovation Connector at Swissnex in San Francisco as well as a textbook author for all those who are engaged in the field of food, water, and nutrition.

© istockphoto.com

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Lifesciences plus 01 I 2016

P L AY E R S & P R O D U C T S I N e t w o r k i n g

01 02 03

Susanne Lauber Fürst. Inartis Network

01

Vice President, Renens.

Peter Pichler. President and CEO Brechbühler AG, Schlieren/Echallens.

Dr Helmut Traitler. Former Head of Innovation Partnerships at Nestlé Group and now Innovation Connector at Swissnex in San Francisco.

02

03

Lifesciences plus 01 I 2016

27

P L AY E R S & P R O D U C T S I N e t w o r k i n g

➜

INTERVIEW: CHRISTIAN EHRENSBERGER

L

“Although our country is always

ifesciences plus: Let us start with you, Mrs Lauber Fürst, what is your analysis of the status quo in innova-

rated among the best, this must not

tion in Switzerland, and how are you and your organization Inartis Network trying to rocket the country to-

mislead us to rest on our laurels.”

wards its possible optimum?

SUSANNE LAUBER FÜRST.

Susanne Lauber Fürst: The name Inartis Network is composed of two pieces: IN stands for innovation und ARS for art – the art of innovation. We are CTI’s National Thematic Network for the Life Sciences and make companies and universities team up in order to maintain Switzerland at the top of the world. Although our country is always rated among the

Dr Traitler, you live in California and are focusing,

best, this must not mislead us to rest on our laurels. We have

amongst other interests, on the food sector. What are you

to be one step ahead day by day. At Inartis Network we wish to

actually working on and what is your view on Switzerland

make our contribution to this challenge by bringing industrial

from a remote site?

researchers and university scientists into spheres of a new di-

Dr Helmut Traitler: I am what can be described as senior

alogue.

innovation connector at Swissnex in San Francisco but live in the greater Los Angeles area, close to institutions such as

Mr Pichler, in how far is it difficult to foster a fruitful dialogue

Caltech, the Jet Propulsion Laboratory of NASA, USC or the

between industry and science?

Art Center College of Design. Obviously “senior” means I am

Peter Pichler: The internal scientific discourse seems to have

of a certain age, but it may also hint at my treasure trove of ex-

reached an advanced level. However, I see deficits concerning

perience. I am leveraging it to bring fitting and motivated

the interaction between science and private enterprises. The

partners together.

challenge is to intensify the dialogue between these two play-

Usually the question is: Does it make sense for a company

ers. The problem can be tracked back to the different goals:

to look for an external partner for a challenging problem? If

generating insight and knowledge or reacting to the demands

the answer is yes, my task is to find maybe a small and very

of the market. At first glance both parties do not have a great

specialized company or a scientist who then carries out a suit-

deal in common. On the other hand scientific insight may gen-

able joint research project.

erate new market demands in the long run, but this process

Naturally, next to the local institutions that I have just

may last years. Small and medium-sized companies often lack

mentioned, one of my focus areas is Silicon Valley, which has

the power of endurance to leverage the chances emerging

multiple interactions with Switzerland. Think of the company

from scientific research. But also the big players do not have

Logitec which is well-known for their computer mice and

the time to exploit it, because they have to follow the quarterly

other personal interface products! One of its founders, Daniel

rhythms of the stock exchanges. Nevertheless, I am convinced

Borel, came from Lausanne. Since its creation, the company

that an intensified collaboration of private enterprises and

also had a main pillar in Silicon Valley. The trends you see

science is harbouring an enormous innovation potential for

here, as a rule, give direction to what will be manifesting itself

Switzerland. To raise it is a precondition for maintaining the

in ten years all over the world, and this even for example in

country’s wealth.

the food sector.

In this area, start-ups have traditionally been of great interest. In 2013 a total of approximately $30 billion were invested in Silicon Valley in start-ups, of which 330 million in the food sector – the latter with an upward tendency. The megatrends in food comprise urban gardening, organic food, clean label, no preservatives, no additives. Unlike in other areas such as some European countries, very little significance is given to the attributes “bio,” and “GMO-free.” Reality is that one can hardly find any major crop used today which is not genetically modified. However, there are increasing movements,

28

Lifesciences plus 01 I 2016

P L AY E R S & P R O D U C T S I N e t w o r k i n g

especially in the state of Hawaii that

looks for something different. He or she

neers was difficult to establish. After the

support the creation – or rather continu-

wants less industrially-looking food, and

kick-off meeting, the research partners

ation – of a GMO-free agriculture.

this is one of the reasons why innovation

pushed the idea to immediately split the

is increasingly emerging far away from

project in a technical-oriented and an

the well-known big players in the sector.

agronomy-driven part. Although it was

On another note, basic genetic information of plants is increasingly used for

a cross-industry and transdisciplinary

“computer breeding” in the traditional

This opens the gate for many small

way to achieve new properties such as

and medium-sized companies provided

for instance cold-hardiness or drought

they can find the suitable partners

resistance. This new type of plant re-

amongst globally active companies and

search leads to substantial time savings.

universities. Scouting by Inartis Net-

Maybe those advanced techniques could

work, Swissnex and my own activities in

in the near future be used to grow high

this vast field, but also for instance at-

value-added crops in areas such as in

tending a local trade fair, can be recom-

the State of Hawaii. In Maui the last cane

mended as a good starting point for a

sugar facility of the entire country will

Swiss company active in this field.

innovation is increasingly

quently the question arises: “Which

Mrs Lauber Fürst, one should take it for

emerging far away from

plants should we now cultivate here?”

granted that two partners with scientific

The current merger and acquisitions

expertise and common sense who have

activities, for example the recently an-

decided to start a joint project will easily

nounced

succeed. What is your experience?

take-over

of

Syngenta

by

ChemChina, are certainly a fight for

Lauber Fürst: I have had the opportu-

leadership in this industry. The large

nity to facilitate such a project involving

seeds producers will however, although

two research institutes and three com-

they are presently showing an air of ar-

panies for two years, but a fruitful

rogance, soon realize that the consumer

dialogue between biologists and engi-

trially looking food, and this is one of the reasons why

the well-known big players in the sector.”

DR HELMUT TRAITLER.

➜

be closed by the end of 2016, and conse-

“He or she wants less indus-

P L AY E R S & P R O D U C T S I N e t w o r k i n g

project, each group desired to start within their

Lab Innovations Lausanne 2016

own expert-field and only come together again, once they had advanced in their respective research. I was very passionate about first defining what we might achieve together, as team, with our collective brain.

Inartis Network (see also interview) is actively supporting “Lab Innovations 2016,” which will take place on 13 and 14 April 2016 in Lausanne.

Approximately 90 exhibitors and universities representing over 300 brands are waiting visitors at the Expo Beaulieu Lausanne.

It was tough for everybody. And it was extremely rewarding to finally reach the magic moment when the participants felt the benefit from actively listening to their counterpart, and enter into a learn-and-teach process.

This kind of a magic dialogue in science and technology strengthens the value chain from university to industry and from basic to applied research. Switzer-

Exhibition fields • Laboratory equipment • Cleanroom technology • Laboratory services • Laboratory facilites • Consumables

Thursday 14 April 2016: • ScienceCenter 2 – The lectures will provide you the latest news in the field “Jobs in Life Sciences.” Adecco will hold a speech and Heiko Bruhn from Mediatum Schweiz will give Show highlights you some “Tips & Tricks for the Wednesday 13 April 2016: Job Interview” •O fficial opening by Patrick • ScienceCenter 1 – Lectures, Barbey, Managing Director of workshops and an exhibition Innovaud, together with Benoît about “Phages.” You have to Dubuis, Director of Campus pre-register for the workshops.

Biotech and President of BioAlps, The lectures and the workshops Eric Préat, Head of Product will be held in French. The Development Group at Artexis exhibition was conceived to Easyfairs, Joseph Maisano, bring you up to date on the Secretary General of the Inartis current research status and give Network and Christin Rudin, you an opportunity to discuss Head of Swiss Event Units at the latest therapy options with Easyfairs Switzerland at 09.15 experts.

• ScienceCenter 1 – Interesting • Between 10.00 and 13.00 lectures about Chromatography you are invited to enjoy a tasty – “Use of mass spectrometry brunch at the show. for doping analyses” by R. Raoul Nicoli from the Swiss Laboratory Show: Lab Innovations for Doping Analyses (LAD) Lausanne 2016 is one topic among others Venue: Expo Beaulieu Lausanne • ScienceCenter 2 – Varied – Hall 36 speeches and workshops Date: 13 and 14 April 2016 about Innovation Technologies Opening hours: both days – in Labtech. Dr med. Thierry 09.00 – 17.00 Weber from Vivactis will hold a workshop about “Branding Use registration code 4202 Scientific communication for a free entrance. and Market launches.” Erika Györvary from CSEM will speak www.labinnovations.ch about “Trends for Innovation in Lab Technologies” • At noon – Innovaud Pitch Session • Daniel Brélaz, National Councillor of the Canton of Vaud and Lord Mayor of Lausanne, will present the Lab Innovations Award in the categories of Industry and Research. The ceremony begins at 15.00.

land is predestined to realize additional innovation potential, especially in the broad field of the Life Sciences, encompassing disciplines such as pharma, medtech, biotech, biomaterials, agronomy, food or cosmetics. It may be difficult to quantify the benefits of speeding up the flow of information and ideas along the value chain by such an intensified dialogue, but the dividend will – no doubt – be payed in the form of vibrant “Denk- und Werkplatz Schweiz.”

Mr Pichler, you are engaged in applied chromatography, a mature technique within chemical and pharmaceutical analytics. To what extent does experience in your every day business underline the difficulty of a scientific dialogue between university and industry?

Pichler: I must endorse what Mrs Lauber says about the magical moment when the members of an inhomogeneous research group come to listen, to teach and to learn from each other. We need a new culture like that, with the main characteristic that a sharing economy becomes chic and trendy.

“It has to be admitted that sometimes companies do not

➜

take enough courage to really think out of the box and venture into unknown waters.”

PETER PICHLER. 30

P L AY E R S & P R O D U C T S I N e t w o r k i n g

Today a typical situation is: We have identified a specific

ratory, to accurately determine MOSH/MOAH in food packag-

demand in the market but basic researchers in the university

ings. Soon, we identified a rising demand in the market giving

think the problem to be irrelevant. Vice versa, a young scien-

us the chance to present ourselves as a virtual competence

tist may convince us with a sophisticated concept, but after

center with an outstanding know-how in the area of chroma-

several interviews with opinion leaders and potential custom-

tography and mass spectrometry.

ers we feel obliged to decide against this project – although our criterion is not solely the return on invested capital! But

Dr Traitler, your mother was German, you are Austrian by

it has to be admitted that sometimes companies do not take

birth, became also Swiss citizen, you have for many years been

enough courage to really think out of the box and venture into

working in Switzerland and now in America. The MOSH / MOAH

unknown waters.

example we have just been presented with is a Swiss affair.

In our chromatography business, for example, we see the

Which are the implications for the international standing of the

most promising innovation potentials beyond the classical an-

country? Which role may the upcoming local Lab Innovations

alytical device. They lie in new proprietary products targeting

Lausanne trade fair play?

at a accelerated sample preparation and featuring significant-

Traitler: A successful dialogue and collaboration between

ly less single steps and a higher grade of automation. The

companies and public authority, which, just to give a small ex-

benefit is obvious: a gain both in accuracy and in speed and a

ample, may result in a new analytical technique useful to fos-

higher throughput. In this area a collaboration with a scien-

ter food safety, can have huge impact. This may become per-

tific partner would be especially desirable.

ceived even beyond the borders of the country where the in-

As a prime example, I would like to highlight our develop-

novation has originally been developed.

ment of an LC/GC system to determine mineral oil saturated

In general, from an international perspective, pioneers

hydrocarbons and mineral oil aromatic hydrocarbons in food

such as Patrick Aebischer, president of the École polytech-

packagings. This so called MOSH/MOAH application was the

nique fédérale de Lausanne (EPFL) since 2000, have already

result of a joint project of our company with the Cantonal

managed to implement global technology transfer especially

Laboratory of Zurich. They were concerned about MOSH and

in the Arc lémanique and thereby attracted huge amounts of

MOAH, because if they leach out of the packaging and enter

new talents. Subsequently, the region, in my eyes, enjoys a

into the food they may act as a carcinogenic.

higher reputation than comparable ones in Germany or other

Together, we managed to develop an LC/GC method en-

European countries. There is a lot of fascinating research

abling the Cantonal Laboratory, actually as the very first labo-

emerging in the triangle Lausanne – Geneva – Fribourg. A chemistry and laboratory trade fair in this area will naturally be an event of high importance and should be a great success.

Mrs Lauber Fürst, in how far is a trade fair like this also a good starting point for an intensified dialogue between industrial and basic research?

Lauber Fürst: In their every day work our researchers are increasingly driven by administration, SAP sheets and performance reviews, which can really kill curiosity and innovation spirit. To foster innovation one has to go outside, to the marketplace. A trade fair like the Lab Innovations Lausanne is such a marketplace. It brings researchers and industry actors from different fields and geographic places together. Inartis Network is supporting Lab Innovations Lausanne with a comprehensive conference program including interesting lectures and workshops to stimulate among the participants a mind set creating the best preconditions to enter into a new aera of catalyzing innovation by dialogue.

Lifesciences plus 01 I 2016

31

P L AY E R S & P R O D U C T S

© istockphoto.com

MICROGRAVITY AND DRUG DESIGN

SWISS START-U Ps

Science from Outer Space

Swiss Start-up company SpacePharma is not at all spaced out. The company provides a service for experiments and trials under microgravity condition in miniaturized lab systems. Studies under microgravity give insight into molecular and cellular processes or can even help developing and improving or producing drugs.

CHRISTIAN EHRENSBERGER

F

rom time to time some critical citizen will ask politicians and scientists: What the heck is our business in space? This is a

good question, but the pick of the bunch is that you need not go to the Moon or to the Mars to find the answer, but simply to Delémont, Switzerland. Sited there, in the Swiss Jura, a company named SpacePharma is leveraging the powers of microgravity to propel research in various areas of chemistry, pharmaceutics

and

life

sciences. SpacePharma has developed miniaturized lab systems capable of working

in

different

microgravity

platforms, including ground simulators, parabolic flights, and nano-satellites. All experiments can be commanded and remotely controlled by the users from their own location using SpacePharma’s proprietary Scientist Front End Software.

In order to pursue their goal, the team has to combine advances in several areas of research which seem to be quite far away from each other at first glance. But the company’s founder Yossi Yamin points out: “Change depends on unreasonable people.”

This spirit has led the SpacePharma crew, for example, into a current pro-

P L AY E R S & P R O D U C T S I M i c r o g r a v i t y a n d D r u g D e s i g n

ject, the aim of which is to develop a lab-on-a-chip research

Big Goals

model that enables scientists to study live human cells in mi-

This example is delineating at least two of the key principles

crogravity. Here, the company is collaborating with Sanford

of SpacePharma’s way of research: a really big goal and col-

Burnham Prebys Medical Discovery Institute Spin-Out Com-

laborations with potent national or international partners.

pany micro-gRx, Florida.

From this point of view, it is interesting to dive into the world of microgravity and learn some more about the ideas behind

More detailed, both partners are striving for a fully auto-

current research projects:

mated, miniaturized cell-culturing laboratory in order to track expression levels of protein bio-markers in human stem

A very basic aspect relates to the fundamentals of chemi-

cell-derived cell types over time. Therefore the scientists will

cal reactions. In microgravity they are driven only by molecu-

have to customize microfluidics and human cell-based assays

lar forces, but not at all affected by relative densities. This may

technology for applications under the condition of a reduced

give scientists a deeper insight into the nature of the chemical

gravity environment. The great hope is to be able to provide

reactions. One should be able to directly observe the effects of

new possibilities of disease modeling.

the pure molecular forces without the necessity to calculate away the influence of other parameters by means of complex statistics which inevitably includes a risk of wrong modeling.

Promising Success in the Pharma Area From chemistry to pharmacy: Microgravity conditions tend to increase bacterial virulence and related stress mechanisms.

SpacePharma’s team measuring

The accelerated biological processes may give way to a faster

the electromagnetic prior to

development of, e. g., vaccines fighting a disease which is trig-

selecting the antenna’s location.

gered by known bacteria. Furthermore, development of new

(Pictures: SpacePharma)

Current collaboration with Florida-based partner micro-gRx: SpacePharma microgravity experiments on their way in space. In detail, the mission comprises a full 3U satellite (dimensions = 34,05 cm × 10 cm × 10 cm, max. weight = 4 kg) containing a complete liquid-based laboratory. That will be able to execute in full orbit experiments controlled by clients from their own location.

A company in the Swiss Jura: SpacePharma scientist Alex Pekin (left) and Lotan Horev, Head of SW engineering, performing research activities on earth.

Lifesciences plus 01 I 2016

33

P L AY E R S & P R O D U C T S I M i c r o g r a v i t y a n d D r u g D e s i g n

lizing proteins revealing their true 3-D structure.

This

is

very

effectively

achieved in microgravity experiments – and has already resulted in obvious success!

Experiments sponsored by Schering-Plough have led to the crystalliza-

based cancer therapies are currently

➜

drugs is often based on ideally crystal-

being the subject of various patents and patent pending applications.

Be willing to fail but aim for

Even agriculture may benefit from microgravity experiments helping to reveal the fundamentals of plant biology

revolutionary success.

including basic processes such as gra-

YOSSI YAMIN. Founder and CEO of SpacePharma

viresponses, phototropism, and juvenil-

tion of the human interferon alfa-2b

ity. This may support the development

and to the discovery of a new drug for

of improved crop strains with increased

Hepatitis C. This disease is affecting

yield, disease resistance and the ability

150,000,000 people worldwide and kill-

testing under microgravity conditions,

to grow in severe conditions. As a short-

ing 350,000 people every year. Compared

Amgen has found out that “treatment

term objective, farmers may be given

to prior art, the improved form of inter-

with Prolia resulted in greater bone

advice concerning the optimal amounts

feron is more effective in treating Hepa-

density, stronger bones, and reduced

of water and nutrients.

titis C and shows fewer side effects. In

risk for vertebral, hip and non-vertebral

addition, the success of microgravity ex-

fractures.”

Proprietary Microgravity Expertise and Strong Local Networks

periments includes a new effective in-

A pioneering pharmaceutical for-

hibitor against influenza surface protein

mulation technology may be the Micro-

Microgravity seems to be an all-rounder.

Neuraminidase, broad spectrum antibi-

encapsulation Electrostatic Processing

Are there any limits? As a matter of fact,

otics against bacterial NAD synthetase,

System-II (MEPS-II). Its clue is the

from the point of view of a terrestrial

or an inhibitor against Malaria.

combination of dissimilar fluids. Where-

space experiments under microgravity

Another condition in focus of micro-

as the gravity field on earth forces two

conditions will always be, in a sense, ide-

gravity experiments is osteoporosis. It is

non-miscible liquids in a test-tube to sit

alized techniques. So, the results have to

well-known that astronauts undergo ac-

on top of one another, in space surface

be checked for their potential use here

celerated bone loss during their mis-

tension forces drive the fluids to inter-

on earth – for example in the Swiss Jura.

sions in space. This may provide a mod-

face and form microcapsules. Injecting

Here, SpacePharma is thriving based on

el for this disease. – Is this overexcited?

suitable

were

their expertise in microgravity experi-

Reality gives the answer: Amgen, one of

produced under microgravity condi-

ments. Any company, university or other

the world’s leading biotech companies,

tions and contained anti-tumor drugs,

organization may contact Yossi Yamin

is marketing an injectable drug (“Pro-

into human prostate tumors showed a

and his team for consulting in this area.

lia”) for postmenopausal women at risk

significant inhibition of tumor growth

They also benefit from a strong local

for bone fractures because of osteopo-

according to preclinical investigations.

network including, e. g., Inartis, one of

rosis and other factors. By means of

Thus,

the eight National Thematic Networks

microcapsules,

several

MEPS-II

which

technology

supported by the Commission for Technology and Innovation (CTI), the Swiss agency for the promotion of technology and innovation.

SpacePharma SA – brief information about the company

Maybe, not every promising project will result in the success one has ex-

• founded on May 2012

pected in the beginning. But as Yossi

• currently having 15 employees – 2 in Switzerland, 12 in R&D center in Israel, 1 in USA

Yamin says: “Be willing to fail but aim

• Courgenay Ground Station: SpacePharma will install its ground station to communicate all

for revolutionary success.”

Satellites’ telemetry passageways above Courgenay. The site will include hardware and software as well as client’s front labs and meeting rooms. The operators will do their job in this site as well. That will increase employees in Switzerland.

• satellite launch: The first launch is expected to be coming in the second quarter of 2016 using SpaceX Falcon9 launch vehicle, a two-stage rocket for the reliable and safe transport of satellites into orbit.

• status of parabolic flights activities: The full parabolic flight project integration along with S-3 (“Swiss Space Systems”), a company founded 2012 in Payerne with the goal to develop, build, certify, and operate suborbital shuttles for the deployment of satellites up to 250 kg.

All devices have already been installed and the program is now ready for presale. The third quarter of 2016 is expected to be the timeline for flights.

34

Lifesciences plus 01 I 2016

People with autoimmune diseases suffer from a hyper-active immune system, which has spiralled out of control and attacks them by mistake. (© istockphoto.com)

SWISS START-U Ps AUTOIMMUNE DISEASES

Allocyte – New Hope for Patients Suffering from Autoimmune Diseases AlloCyte Pharmaceuticals AG is developing a new class of small molecule drugs for patients suffering from severe autoimmune diseases. AlloCyte’s orally available “integrin silencers” are designed to restore patients’ self-protective immune balance, translating into lasting disease control.

© istockphoto.com

36

Lifesciences plus 01 I 2016

P L AY E R S & P R O D U C T S I Au t o i m m u n e D i s e a s e s

In 2011, a small group of Swiss- and US-based

ELSBETH HEINZELMANN

W

pre-clinical and clinical researchers, intrigued by hether our body is attacked by bacte-

recent insights into the roles of integrins in aber-

ria, viruses, toxins or cancer cells, our

rant immune processes, decided to tackle the chal-

immune system’s army of white blood

lenge of developing novel integrin-targeting medi-

cells is always ready to protect us

cations. These medications are designed to restore

from these dangerous aggressors known as anti-

the patient’s protective immune balance and, in

gens. However, if we are suffering from an auto-

consequence, to provide lasting disease control.

immune disease, it is precisely this immune system

AlloCyte’s co-founders were well-positioned to

that spirals out of control and attacks us by mistake.

meet this challenge. They brought with them a detailed understanding of integrin structural biology,

Action Required for Novel Medications

an in-depth knowledge of the roles of integrins in

Almost any organ can fall victim to this kind of im-

health and disease and, importantly, considerable ex-

mune attack. As a result, over 80 different autoim-

pertise in pre-clinical and clinical drug development.

mune diseases are known. A growing number often chronic, debilitating and sometimes life-

Opportunities and Challenges of Integrins as Therapeutic Targets

threatening. The underlying cause of autoimmuni-

Integrins are a family of 24 heterodimeric cell

of patients suffer from these diseases, which are

ty is an overactive immune response of our body

membrane glycoproteins composed of α- and β-

against structures that occur naturally in the body

chain subunits. Different cell types express differ-

and do not normally elicit immune responses. Inte-

ent integrins – or combinations of integrins – on

grins (a class of trans-membrane receptors) are

their surfaces. Therefore, integrins are attractive

known to play decisive roles in triggering and sus-

therapeutic targets that offer the potential to mod-

taining these aberrant and excessive immune re-

ulate the function of distinct cell populations and

sponses.

thus to selectively target cellular processes driving autoimmune diseases.

Professor Marianne Hürzeler and Martin Gut (B.Sc. in Molecular Life Sciences) at the School of Life Sciences, The Basel Inkubator provides space, infrastructure and support

FHNW Basel, examine the

for young start-up companies such as AlloCyte Pharmaceuticals AG.

purity of newly synthesized

From left to right: Dr Albrecht Schmidt, PD Dr Gabriele Weitz-

LFA-1 inhibitors.

Schmidt (both AlloCyte) and Dr Peter Burckhardt (EVA and Basel Inkubator). (Pictures: AlloCyte)

Riccardo Mancuso (M.Sc. Biotechnology), PhD student at the University Hospital Basel, isolates and cultures white blood cells, which are critical for testing biological activities of novel small molecule LFA-1 inhibitors.

Lifesciences plus 01 I 2016

37

P L AY E R S & P R O D U C T S I Au t o i m m u n e D i s e a s e s

AlloCyte’s first integrin target is the leukocyte

Integrins are known to play

➜

receptor

lymphocyte

function-associated

anti-

gen-1 (LFA-1). LFA-1 controls leukocyte migration

decisive roles in triggering

and T cell activation during inflammatory and im-

and sustaining aberrant and

mune responses. Furthermore, LFA-1 plays an im-

excessive immune responses.

towards disease-driving (pro-inflammatory) or

portant role in directing the fate of young T cells protective (regulatory) populations.

Experts join Forces and roll up their Sleeves Integrins are difficult targets from a pharmacologi-

When AlloCyte started, the team already had a de-

cal perspective, however. This is because integrins

tailed understanding of the molecular site within

are normally expressed on cell surfaces in an inac-

the LFA-1 heterodimer that they wanted to target,

tive or “silent” state. Intracellular signals are re-

based on prior research by Timothy Springer, Har-

quired to activate integrins. This activation is asso-

vard Medical School Boston and Gabriele Weitz-

ciated with massive conformational changes of the

Schmidt, AlloCyte Basel. This very high level of un-

α/β heterodimers. Subsequent ligand binding to

derstanding allowed for a virtual screening ap-

the activated integrins triggers complex signalling

proach for identifying compounds that bind to this

cascades that fundamentally change the cells’ mor-

site and potentially inhibit (silence) LFA-1.

phology and behaviour.

AlloCyte’s virtual screening performed by Gis-

It is a central pharmacological challenge to in-

bert Schneider’s group at the ETH Zurich (ETHZ)

hibit integrins without triggering unwanted signal-

rapidly led to the identification of virtual hits. They

ling into the cells. Unwanted signalling is of major

became the starting points of a focused chemistry

clinical concern because it could aggravate, rather

programme conducted by Marianne Hürzeler’s

than treat the disease. This phenomenon is also

group at the School of Life Sciences of the Univer-

known as paradoxical agonism, a risk associated

sity of Applied Sciences and Arts Northwestern

with all modalities that interact with the li-

Switzerland (FHNW) Basel. The pre-clinical phar-

gand-binding site of integrins. Paradoxical ago-

macological profiling of the compounds was per-

nism has haunted integrin pharmacology for dec-

formed by the groups headed by Daniel Gygax at

ades. AlloCyte aims to avoid paradoxical agonism

FHNW Basel, and Stephan Krähenbühl at the Uni-

by pursuing an elegant pharmacological approach

versity Hospital Basel. This efficient collaboration

in which integrins are stabilized in their inactive

of leading research groups from different Swiss ac-

state, referred to as “integrin silencing”. By their

ademic institutions rapidly advanced the initially

small molecule nature, steerability, selectivity and

virtual hits to produce a new chemical class of po-

their avoidance of paradoxical effects, Allocyte’s

tent, selective and orally available LFA-1 inhibitors

integrin silencers are expected to support innova-

with the desired mode of action.

tive therapeutic concepts that could not be realized

Working hand in hand with the company’s

with earlier integrin-targeting pharmacological

pre-clinical research consortium, Albrecht Schmidt,

approaches.

AlloCyte Basel, an experienced internist and clinical drug developer, reached out to leading clinicians to develop and advance Allocyte’s innovative translational concepts, thus building the second pillar of

BASEL INKUBATOR – the high-tech incubator in Basel

therapeutic innovation – right from the company’s start.

The Basel Inkubator,

is a service provided for start-ups by the University of Basel and the

Partners sharing the Vision and accepting the Risks

University of Applied Sciences and Arts Northwestern Switzerland

As a start-up company, AlloCyte needed financial

(FHNW), Muttenz. Created on the initiative of the University of Basel,

partners who shared the company’s vision and were

the FHNW and EVA – the Basel life sciences start-up agency – as well

prepared to accept inherent risks. The young com-

as the AWA (Office of Economy and Labour) Basel, the incubator

pany was lucky to gain the almost immediate sup-

offers coaching and low-cost business accommodation. The start-up

port of NTN Swiss Biotech and the Swiss Commis-

companies benefit from subsidized employment and consulting

sion for Technology and Innovation (CTI). The Ba-

services in order to further develop their business idea.

sel Incubator/EVA (see box) also stepped in to pro-

founded in 2009,

vide AlloCyte with an attractive home and the

38

Lifesciences plus 01 I 2016

P L AY E R S & P R O D U C T S I Au t o i m m u n e D i s e a s e s

support of a Basel-based pharmaceutical

up companies. Moreover, i-net Innova-

company that joined the CTI consortium.

tions Network Switzerland provided the

“Initially, it proved impossible to at-

company with access to expertise not

tract venture capital, as we did not own

available elsewhere. In addition, private

any substantive intellectual property,”

help was provided at an early stage, sup-

states Gabriele Weitz-Schmidt. She ap-

plemented later on in the project by the

preciates the generous financial sup-

➜

stimulating environment of other start-

“AlloCyte brings together an international team of experienced researchers with the common goal of exploiting the therapeutic opportunities

Questions to Gabriele WeitzSchmidt, PhD and co-founder of AlloCyte

offered by integrins.“

Interview conducted by Elsbeth Heinzelmann

port of CTI all the more, since it enabled

DR GABRIELE WEITZ-SCHMIDT. CSO and co-founder of AlloCyte

rapid progress during AlloCyte’s startLifesciences plus: As a pharmacist and biochemist you led global interdisciplinary projects in cardiovascular disease, transplant rejection and autoimmunity.

What is your personal motivation as a Chief Scientific Officer at AlloCyte?

Gabriele Weitz-Schmidt: AlloCyte brings together an international team of experienced researchers with the common goal of exploiting the therapeutic opportunities offered by integrins. It is a major motivation to work within such an interdisciplinary team and to translate research into therapeutic benefits for patients.

AlloCyte is a young company with a very productive and open-minded atmosphere. It’s fun to be a part of it.

What role does the inclusion of Daniel Gygax, President of biotechnet, in the AlloCyte team play and – thanks to him – the access to innovative technology and methodology infrastructure at FHWN? AlloCyte could not have started without Daniel Gygax’s early support. Daniel recognized the potential of AlloCyte’s pharmacological concept and joined the initial team that went on to found the company. He also provided AlloCyte with critical infrastructure during the start-up phase and helped AlloCyte’s consortium to gain the support of NTN Swiss Biotech and CTI. AlloCyte is privileged to have Daniel Gygax on board as a senior advisor, development partner and shareholder.

Before founding AlloCyte, you spent a sabbatical in the laboratory of Timothy Springer, a further co-founder of AlloCyte. The Boston area has a worldwide reputation in identifying innovative companies and putting them on a sound footing. Why did you decide to found AlloCyte in Basel?

Indeed, the Boston area provides an attractive and stimulating environment for start-up companies. We decided to found AlloCyte in Basel for a number of reasons: Firstly, the Basel location provided us with immediate access to exceptional medicinal chemistry and biology infrastructure and expertise.

We could hardly have found this expertise anywhere else. Secondly, the concept of public-private partnership (which made great sense from the co-founders’ perspectives) could be realized in Switzerland thanks to the generous support of CTI and NTN Swiss Biotech. Last but not least, the Basel area is an exceptionally privileged location for innovative drug development. There are major international pharmaceutical companies in close proximity, as well as successful budding biotech companies. The momentum that can be derived from this constellation should not be underestimated.

up phase, resulting in the prompt discovery of a new chemical class of integrin inhibitors leading to AlloCyte’s first patent application. CTI’s support was also critical in initiating the kind of public-private partnership that worked so productively in AlloCyte’s case. “The FHNW Basel, as well as the ETHZ and the University of Basel, provided competencies that synergized with our own expertise, not to mention infrastructures that would not have otherwise been accessible to AlloCyte,” remembers Weitz-Schmidt, who contributes her expertise in pharmacology and project management. There was another important element shaping the environment within AlloCyte’s team: The involvement of highly motivated and practice-oriented Bachelor, Master and PhD students. “They helped to create the ’can do’ spirit that characterizes our young company,” adds Weitz-Schmidt.

Meanwhile, AlloCyte has initiated collaborations within Switzerland and internationally to further assess its LFA-1 silencers. These activities will be an important next step in translating AlloCyte’s integrin pharmacology into innovative therapeutic concepts, expected to provide patients suffering from severe autoimmune diseases with sustained therapeutic benefit and, ultimately, freedom from disease.

Lifesciences plus 01 I 2016

39

P L AY E R S & P R O D U C T S

04

Isolator technology is modularly expandable.

HPAPIS (HIGHLY POTENT ACTIVE PHARMACEUTICAL INGREDIENTS)

Trends in Aseptic Manufacturing HPAPIs/ADCs (antibody-drug conjugates) are a new generation of highly active/ highly toxic pharmaceutical products, which are used among other things for the targeted treatment of cancer. Some of these HPAPIs/ADCs require occupational safety measures that ensure compliance with OELs (occupational exposure limits) below 100 nanograms per cubic meter of air in the workplace.

RICHARD DENK

especially in the US, while PDE is a European

T

designation by EMA (European Medicines Agenhe production of HPAPIs/ADCs represents

cy). Since 2015, the PDE is mandatory in the pro-

a new challenge, especially in aseptic pro-

duction of medicines. This obligation can be found

duction. Isolators have been successfully

in section 5 of the EU GMP guidelines and the arti-

used in aseptic production for many years

cle “Guideline on setting health based exposure

for product protection. These isolators must now al-

limits for use in risk identification in the manufac-

so provide active employee protection. A contradic-

ture of different medicinal products in shared fa-

tion? At first glance, “Yes”. Because in aseptic man-

cilities.” Among other things, the guideline defines

ufacturing, isolators are operated under positive

“Data requirements for hazard identification.”

pressure in order to protect the products (according to GMP). On the other hand, efficient containment

The following formula applies:

for operator protection requires a negative pres-

Worker-ADE/PDE (in mg/day) =

sure in the isolator to prevent dangerous substanc-

NOEL (mg/day)/UFc × BA

es from escaping. In particular, it needs for example a specific design on the seals and high end filter

NOEL = No Observable Effect Level

technologies on the isolator that both ( product and

UFc = Cumulative uncertainty factor for the worker

personal protection) can be reached.

BA = Bioavailability by inhalation ADE = Acceptable daily exposure in mg/day

How Is the OEL Worked out?

PDE = Permitted daily exposure in mg/day

The OEL (occupational exposure limit) is calculated from the ADE/PDE (acceptable daily exposure/

The ADE (Acceptable Daily Exposure) or (accord-

permitted daily exposure). The term ADE is used

ing to the new EMA guideline for shared facilities)

40

Lifesciences plus 01 I 2016

02

Modular isolator technology.

L-flange is connected to the isolator.

Containment Classification

01

Occu

patio

nal E

xposu

re Lim

it (μg

/m3)

Containment pyramid.

μg ur e( po s

ur e(

μg /

da y)

/d ay )

00 xp os

- 5000

Ex

1000

OEL

00

10

mi

© Ri c D ha SK enk rd AN AG

OEB Occupational Exposure Band

000

00

‘0

0 -1

ily

low hazard

1

‘0

1

100 - 1

00

10

yE

moderately hazardous

00

ail

2

10 - 100

0

- 10

Da

hazardous

10

03

ted

3

1 -10

10

highly hazardous

0

1 -1

dD

4

0,1 - 1

tte

very highly hazardous

Ac ce p

5

<0,1 1

>

extremely hazardous

P AD DE E Per

6

Photo of the L-flange equipped with a Bausch & Stroebel vial-filling unit.

the PDE (Permitted Daily Exposure) can be calculated from

mand new modular systems for the production of pharmaceu-

the NOEL (No Observable Effect Level). The relationship be-

tical substances.

tween ADE/PDE and OEL is shown in the containment pyra-

A new innovative technology for modular isolators and

mid [Figure 1]. In addition to the derived air concentration

filling machines has developed here, particularly in fill & fin-

limit OEL, the limits for residues from cleaning and for the

ish for smaller production quantities. This modular technolo-

limits of product carryover from one substance to another in

gy is achieved with customizable PSI-L isolator technology.

shared facilities are also calculated from the ADE/PDE.

PSI stands for Process Safety Isolator and the L for an interchangeable L-flange.

Modular Isolator Technologies

This modularity is characterized by:

• • Individually expandable to accommodate different

Furthermore, these new HPAPIs demand a high level of flexi-

Fast product and format changeovers possible

bility in production. Smaller production batches, more fre-

capacities

quent format change as a result of innovative dosage forms, as

• Process requirements can be individually customized.

well as easy to clean solutions when changing products, de-

Lifesciences plus 01 I 2016

41

P L AY E R S & P R O D U C T S I H PA P I S

The idea behind modular PSI-L technology arose to meet the

Possible filter systems:

• Bag-in/bag-out filter • Filter cartridge (FiPa).

demand for smaller production batches. The aim is to put together processes in a modular way, without needing a large number of aseptic isolators. To achieve this, a new isolator technology with a mobile L-flange was developed. The

All these filter systems are suitable for preventing the spread

L-flange is part of the isolator and is connected to the isolator

of the highly active/toxic substance from the isolator. How-

by a specially designed inflatable seal [Figure 2]. This design

ever, some filter systems represent a GMP risk. For example,

permits the individual equipping of the L-flange with differ-

recontamination from the contaminated filter to the new filter

ent process steps [Figure 3].

can occur with bag-in/bag-out filters during filter replacement. This is critical because recontamination often goes undetected. The filter cartridge (FiPa) excludes this risk. FiPa fil-

L-flanges can be equipped with the following systems:

•

Preparation vessel for the closed transfer of HPAPIs under

ter technology was developed as a filter isolator. The filter car-

aseptic conditions

tridge designed as a closed housing is docked to the isolator,

• Manual filling of vials as a base unit • Semi-automatic to fully automatic vial filling • Aseptic filling of powders in vials • Lyophilisation on mobile L-flange • Vial capping • Vial exterior washing station • Sterility testing • Stopper transfer and portioning into smaller bags.

and the connection on the FiPa to the inside of the isolator is opened from the outside. The airborn product particles can thus enter the interior of the FiPa, and particle are deposited on the filter. When changing products, the FiPa is sealed from the outside again. The isolator can now be cleaned and, after cleaning, the FiPa is removed without any risk to the operator or for the next product to be manufactured.

Summary

L-flange technology provides an individual configuration of

HPAPIs/ADCs are a new generation of highly active and high-

the isolator. For process formulation, the L-flange can thus be

ly dangerous substances in the pharmaceutical industry, for

equipped with the formulation tank and the relevant weigh-

the production of which a new kind of containment is re-

ing technology. After the L-flange has been connected to the

quired. Even though production batches are fairly low in the

isolator, the automatic H2O2 decontamination cycle starts.

early development stages, there is still a major risk of coming

Then the highly active or toxic agent (API) is weighed and

into contact with the product without the appropriate protec-

safely introduced into the formulation tank under aseptic

tive measures. Modular isolator technologies can meet the

conditions. On completion of the process and when the interi-

need for flexibility during process control as well as the han-

or of the isolator has been cleaned, the L-flange is disconnect-

dling of highly active/toxic substances.

ed and the L-flange module for vial filling is docked and the lyophilisation L-Flange is docked to a second isolator. While

www.skan.ch

the formulation continues, the decontamination cycle for the fill & finish area can run in the meantime. After the sterile filtration of the product, the filling of the vials and subsequent

AUTHOR

freeze-drying can take place.

Once this process is complete, the module for vial filling

Richard Denk. SKAN AG, Head Sales Containment

can be disconnected and the sterility testing module docked.

and ISPE Chair of the CoP Containment Working Group,

L-flange technology enables a wide range of possibilities.

DACH affiliate, richard.denk@skan.ch

Filtering Technologies Are a Key Factor for Handling HPAPIs in Aseptic Production Filter technology on the isolator is an important factor in preventing the spread of the substance inside the isolator, as well as to the outside. Inside the isolator, the distribution of the substance must be contained within the smallest possible space. The aim is to avoid the highly active/toxic substance entering the circulating air channels and thus leading to product carryover into uncontrollable areas. This is critical, especially for substances in the low OEL area because it can lead to possible cross-contamination during product changes.

42

Lifesciences plus 01 I 2016

P L AY E R S & P R O D U C T S

EXHIBITION

Get Ready for Chemspec Europe 2016 in Basel From 1 – 2 June 2016, the 31st edition of Chemspec Europe, International Exhibition for Fine and Speciality Chemicals, will open its gates at Basel Messe, Switzerland.

T

Diverse Selection of Fine and Speciality Chemicals

shops, presenting the latest findings

ous event in Cologne illustrated once more the high demand

Compared to the previous event, Chem-

includes the symposium of the Royal

for bespoke chemicals and in-

spec Europe 2016 shows an increase in

Society of Chemistry, the REACHReady

novative products. With 380 exhibitors

net exhibition space by 10 per cent. Half

Regulatory Services Conference, the

and 5,768 visitors from 56 countries, the

a year before the start of the exhibition, a

Agrochemical

event was the largest Chemspec of all

total of 320 exhibitors from 22 countries

ence and the Pharma Outsourcing Best

times. “In a time driven by innovation

had already booked their stand space at

Practices Panel, moderated by Dr Magid

and shorter product life cycles, the

this year’s event. Visitors of Chemspec

Abou-Gharbia from the Moulder Center

choice of the right suppliers and the ex-

Europe include production managers,

for Drug Discovery. The “Chemspec

change within international industry

process experts, R&D specialists, prod-

Careers Clinic,” organised by Chemical

networks play a major role. Due to its

uct designers, procurement managers

Search International, will offer the op-

specialised exhibition profile, Chem-

as well as chemists and consultants from

tion to discover new career opportuni-

spec Europe is a key event for buyers,

numerous industry sectors, like agricul-

ties. The programme for the symposium

traders and agents in search of innova-

ture / pesticides / fertilisers, biotechnolo-

of the Royal Chemical Society can be

tive substances. Here, leading interna-

gy, cleaning products, cosmetics, food in-

found on the exhibition website.

tional manufacturers and distributors

dustry, adhesives and sealants, dyes / Detailed information on the other con-

present a wide range of fine and speci-

dyestuffs, high-tech polymers, pharma-

ferences and workshops will be available

ality chemicals for diverse industrial

ceuticals / drug

paints,

shortly on the exhibition website. For

sectors,” explains Nicola Hamann, Man-

coatings, petrochemicals, pulp and pa-

visitors, participation in the conferences

aging Director of the organiser, Mack

per, textiles and water treatment.

is included in the entrance fee.

portant manufacturing location for the

High-Quality Conferences and Workshops

www.chemspeceurope.com

fine and speciality chemical industry,

In addition to stands from exhibitors

the organiser chose a central market-

from all over the world, visitors at

place amidst the tripoint of Germany,

Chemspec Europe 2016 will find a range

Switzerland, and France.

of high-quality conferences and work-

he great success of the previ-

development,

from R&D. The conference programme

Intermediates

Brooks Exhibitions. With Basel, an im-

The previous event in Cologne was the largest Chemspec of all times.

Confer-

P L AY E R S & P R O D U C T S

TOOLS ➜

Measure Cells Fuel Dependency and Flexibility in a Single Assay

New Suite of dsDNA Quantification Assays Improving Sensitivity and Dynamic Range

Seahorse Bioscience introduces the Seahorse XF Mito Fuel Flex Test Kits for XFp and Seahorse XFe Analyzers.

Taking less than two hours the test enables a unique measure of the dependency, capacity and flexibility of cells to oxidize three critical mitochondrial fuels: glucose, glutamine, or fatty acids. This can only be accomplished in the context of a living cell capable of switching from one mitochondrial fuel to the next in real time. Cells oxidize glucose, glutamine and fatty acids at different rates in the mitochondria to control a wide variety of biological processes, including energy production, proliferation, activation and differentiation. A single injection reveals dependency, a measure of the cells reliance on a particular fuel pathway for respiration. Injection of a combination of compounds measures cells flexibility in meeting metabolic demand. Flexibility indicates that the cells have the ability to compensate for the inhibited pathway(s) by using an alternative fuel pathway for energy production. With this information researchers can now: • identify fuel dependencies to uncover cancer cell vulnerabilities • explore how fuel preferences lead to cell fate decisions for differentiation and immune cell activation • determine whether/how cells can adjust fuel oxidation to match nutrient availability while meeting energy demand. The new XF Mito Fuel Flex Test provides a rapid, label-free solution for studying substrate metabolism. Now, researchers can measure both, fuel pathway preferences and compensation, by modulating all three pathways in one single assay.

Bucher Biotec AG | www.bucher.com

DeNovix Inc. launches three new dsDNA fluorescence quantification kits. The new kits are able to accurately measure sample concentrations covering a range of 0,5 pg/μL to 4000 ng/μL, providing a 20 × increase in sensitivity and greatly enhanced dynamic range over existing technology.

As Genomics is moving rapidly towards small volume and low concentration samples, Denovix recently launched a step-change in performance for fluorescence with the DS-11 FX Series instruments.

And it was clear that researchers needed the same improvement from their assays. This led Denovix to develop assays that address the needs of researchers working with single and rare cells. The combination of DeNovix FX Series instruments and assay technology provides an analytical package that removes existing barriers to the limit of detection of fluorescence quantification. A robust 2-point standard curve and simple mix-and-measure assay protocol delivers rapid, error-free set up. Full integration into the DeNovix EasyApps software, pre-installed on all DS-11 FX instruments means sample measurement, data analysis and exporting takes only seconds per sample. DeNovix dsDNA Fluorescence Quantification kits are available in 1000 assay or 50 assay evaluation size and include the following products: • DeNovix dsDNA Broad Range (0,1 to 4000 ng/μL) • DeNovix dsDNA High Sensitivity (5 pg/μL to 250 ng/μL) • DeNovix Ultra High Sensitivity (0,5 to 300 pg/μL).

Bucher Biotec AG | www.bucher.com

LLG uniTEXER – Available from stock at Faust One head –multiple uses: Vortexer with single carrier head, accommodating the most popular consumables and tubes: for 1 × assay plate, 2 × 50 ml conical tubes, 2 × 15 ml conical tubes, 2 × 5 ml conical tubes, 4 × 1,5 / 2 ml microcentrifuge tubes, 6 × 0,5 ml and 24 × 0,2 ml microcentrifuge tubes.

• Touch function for short-term operation • Carrier head is very easy to clean • Precise speed control via a variable rotary switch • Elastomeric feet ensure excellent stability on the bench and quieter operation • Modern design • Robust, ABS housing • 3 years warranty.

Specifications: • Type of movement: orbital/vortex • Orbital diameter: 3,7mm • Speed: 1000, 2000 und 3000rpm • Dimensions (W × D × H): 173 × 198 × 198mm • Weight: 3,8kg • Protection class: IP 20 • Supply requirements: 220 V, 50/60 Hz • Guarantee: 3 years. Faust Laborbedarf AG | www.faust.ch 44

➜ Lifesciences plus 01 I 2016

LA B C&EP LAB & P RO SR S OI C UE nSt eSr r u b r i k

DRUG DISCOVERY

Advances in Microplate Reader High-Throughput Screening

01

The new gold standard HTS

T

microplate reader Pherastar FSX.

High-Throughput Screening (HTS) in microplate readers is especially used in drug discovery and relevant to the pharmaceutical industry as well as academic core facilities. HTS allows researchers to quickly conduct millions of chemical, biochemical, or pharmacological tests and establish a basis for drug design. In order to maximize efficiency, screening facilities must constantly strive to increase throughput and minimize sample expenditure, while controlling costs. This leads to the key factors in HTS which are highest speed and flexibility without compromising in sensitivity.

o meet these requirements

next generation laser for TRF/TR-

tions, photobleaching, decaying kinetic

BMG Labtech developed the

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signals, or fluctuating conditions like

new

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reference

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creases throughput and precision of

multaneous Dual Emission detection

FSX for HTS applications. This reader

measurement and allows for a full

can be used in any assay that measures

was specifically conceived for the fast-

1536-well plate to be measured in only

two emission wavelengths or polariza-

est read times, the best sensitivity and

36 seconds [Figure 2].

tion vectors, including FP, FRET, HTRF

unmatched flexibility in all plate for-

and AlphaPlex.

mats up to 3456 wells [Figure 1].

Simultaneous Dual Emission Detection for Alpha Technology

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forms applications like protein-protein

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cation. Moreover, the microplate reader

high energy xenon flash lamp, a special

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Lifesciences plus 01â&#x20AC;&#x2030;Iâ&#x20AC;&#x2030;2016

detect

45

two

Best Performance in All HTS Assays

L A B & P R O C E S S I D r u g D i s c o v e r yL A B & P R O C E S S I U n t e r r u b r i k

Pherastar FSX especially provides excellent performance in all HTS applications, including NanoBRET, Alpha Technology and HTRF assays [Figure 3].

Automation in Robotic systems Automation is a key factor in HTS hence microplate readers have to be easily integrated in automated processes of modern laboratories. For HTS automation purposes, the Pherastar FSX offers improved robotic integration capabilities, multiuser control and MARS data analysis software with included digital signature and FDA 21 CFR part 11 compliance, and can be equipped with BMG Labtech’s Stacker. Moreover, data can be exported as Excel or ASCII files.

German Engineering at its Best BMG Labtech is a leading global developer and manufacturer of innovative, high-quality, and reliable microplate reader instrumentation. The company has been committed to producing microplate readers for more than twenty five years. By focusing on the needs of the scientific community, the company’s microplate readers have earned the company the reputation of being a technology leader in the field. All microplate readers are made in Germany and are conceived, developed, assembled, and tested entirely at BMG Labtech’s headquarters in Germany. www.bmglabtech.com

03

NanoBRET competitive binding experiments of propranolol-BY630 with increasing concentrations of known unlabeled ß2AR ligands.

02

Transcreener ADP2 TR-FRET Performance in 384w format using the advanced TRF laser of the PHERAstar FSX.

46

Lifesciences plus 01 I 2016

LA B C&EP LAB & P RO SR S OI C UE nSt eSr r u b r i k

SYSTEMS BIOLOGY

Innovative Research for Future Production of Biopharmaceuticals Global multi-technology company 3M develops innovative filtration solutions for various application fields, including biopharmaceutical production. Since 2014 a collaboration with the Department of Bioengineering of the Fachhochschule (FH) Campus Wien, partner of the Austrian Center of Industrial Biotechnology (ACIB), has been established in order to evaluate novel filtration solution for biotechnological applications.

ELISABETH GASSMANN

F

iltration is an essential operation included

production process may be designed with robust-

in every biotechnological process and

ness and some safety contingency.

therefore the most efficient and economic filtration is required to achieve the re-

quired purification and clarification of biotechnological products. Purification by filtration is a critical stage in complex biotechnological production processes: Usually, genetically engineered cells are cultivated to express active substances which are later separated and finally purified.

Filterability studies serve to evaluate the throughput and lifetime performance of a filter media as well as its ability to remove processrelated impurities and contaminants. Further-

The objectives of studies are to

• evaluate filtration media • undergo economic assessment of production processes

• assess target biological product safety An interview with Prof Dr Michael Maurer from the FH Campus in Vienna, partner of 3M, provides an insight in the exciting work at the research centre. The interview was held in November 2015.

more, the scalability of the production process from laboratory to pilot scale can be investigated.

Elisabeth Gassmann: What kind of research centre

By testing at pilot-scale, the biopharmaceutical

is the FH Campus Wien and what is its position and integration in Vienna? How would you describe the collaboration with other institutes in Austria, such as the University of Vienna?

About FH Campus Wien

Michael Maurer: The FH Campus Wien is responsible for teaching and applied research in biotech-

About 5,400 students make the FH Campus Wien the biggest technical college

nology. It is partner of the ACIB and maintains col-

of Austria. Numerous R&D projects commissioned by study programmes or

laborations with other universities and research

contract research are running. FH Campus Wien is networked with enterprises,

institutes worldwide. In Austria, cooperation exists

associations, schools and public institutions. Moreover, recognized certificates

with the Technical Universities of Vienna and Graz,

confirm the college’s high-quality management standard.

as well as the BOKU (department for soil culture) from the University of Natural Resources and Life

Contact: FH Campus Wien • Fachbereich Bioengineering FH-Prof DI Dr

Sciences Vienna.

Michael Maurer • Lehre und Forschung • +43 1 606 68 77-3620 michael.maurer@gh-campuswien.ac.at • www.fh-campuswien.ac.at • www.acib.at

When did you start working for the institute and what are your main tasks?

Lifesciences plus 01 I 2016

47

L A B & P R O C E S S I S y s t e m s B i oLl A o gBy & P R O C E S S I U n t e r r u b r i k

I started as a professor of the FH in Vienna in 2016. Besides lectureship, I am leading and supervising different research projects.

How did the cooperation with 3M come about and what does it involve?

It was during a workshop, when ACIB, the FH Campus Wien and 3M first met and the collaboration resulted. The ACIB and 3M agreed upon a research programme of 5 years, during which the ACIB is performing independent tests with 3M’s filters. In this context, 3M is providing free filter material for the research projects of the ACIB. How would you describe the collaboration with 3M? Does it meet your expectations?

The collaboration with 3M allows us to remain up-to-date on the latest techniques and to work with cutting-edge 3M products for filtration applications even before they are available on the market.

3M in turn profits from FH Campus Wien’s network, the complete equipment and the broad range of expression systems. Even a pro-

01

Dr Michael Maurer, group leader at FH Campus

tein-A-column is available.

Wien, gives an insight in the exciting work on filter evaluation for biotechnologically-produced substances. (Pictures: FH Campus Wien)

Which filtration systems and application of 3M have you tested so far?

We evaluated the entire product line Zeta Plus for deep-bed filtration (depth filtration), LifeASSURE PDA010 (a membrane filter for sterile filtration), the latest product line of Emphaze AEX Hybrid Purifier as well as filter disks together with the filter case Ligaster. We are testing filter sizes for laboratory and production scale (pilot plant for 180 l).

What projects are you currently working on and which products of 3M are you evaluating in this context?

The filtration performance of the Emphaze AEX Hybrid Purifier is currently being compared to the filter capacity of the Zeta Plus filter for depth filtration. For this purpose, we use monoclonal antibodies (CHO).

Which research results could you achieve? Were the studies successful?

We were able to verify the great potential of protein enrichment with Emphaze compared to Zeta Plus. Moreover, we can prove the scalability from laboratory to pilot scale, which means that we were able to achieve comparable results. This leads to process optimisation by minimising the filter area, better yield and less production loss.

Were these results published and presented to the (Austrian) public?

Of course. Different posters showing the research on 3M’s Emphaze

02

Pilot system: Zeta Plus and Emphaze AEX Hybrid Purifier are compared in the laboratory.

AEX Hybrid Purifier product line have already been presented at conferences in Scotland, France and Austria.

What are your future plans for the collaboration with 3M? The cooperation agreement between 3M and FH Campus Wien expires in 2019. Further projects are planned in order to test the Emphaze AEX Hybrid Purifier for the reduction of endotoxins when using E. coli bacteria in biotechnology. In addition, samples of the AEX-ST (salt tolerant) which expand the Emphaze product line will

AUTHOR

be tested soon.

Elisabeth Gassmann. www.3M.com

Marketing Manager at 3M Purification.

48

Lifesciences plus 01 I 2016 © istockphoto.com

F I LT R AT E

the time from October 2015 to January 2016 – a large number. These babies suffer from an abnormally small skull which affects growth of the brain leading to mental and physical disabilities. Since the increase coincides with an increase of registered Zika infections, experts suppose that Zika virus is responsible for the cranial malformations. This hypothesis is supported by the fact that Zika virus RNA was detected in the amniotic fluid of several pregnant women. Their fetuses showed

ZIKA VIRUS

microcephaly which indicates that the virus had crossed the placenta and could have caused a mother-

Small Bite – Big Impact

to-child infection.

Apparently many pregnant women in Brazil got infected by Zika – most likely via bites of the mosquito Aedes aegypti, the very same mosquito which also transmits diseases such as Dengue fever and Chikungunya.

But there is also good news: Zika infection itself seems to be uncomplicated in healthy adults and children. Actually, in 80 % of the cases infection passes even

Human diseases that are spread in a mosquito– human–mosquito cycle are not uncommon.

The recent Zika outbreak, however, is alarming.

Researchers, health ministries as well as travelers and inhabitants of affected areas are worried about the fast spread. Whereas the acute infection is quite harmless, its late effects can be terrible.

There is urgent need in strategies to stop the spread worldwide.

unnoticed. The other 20 % of infections usually cause moderate symptoms which include mild illness with fever and rash. Sometimes conjunctivitis as well as muscle and joint pain and fatigue may occur. Gastrointestinal problems are seldom and the symptoms disappear after 2 to 7 days, without special medical treatment. The disease itself seems to be much less threatening than Ebola which is, in comparison, highly contagious and often lethal. Zika is transmitted by the mosquito and it is very likely that it may be passed on via direct blood contact (which includes sexual intercourse).

The bad news is that complications during or after infections may occur. The first outbreak of Zika fever was noticed in 2013 – 2014 in French Polynesia indirectly

SONJA BICHSEL-KÄSER

by noticing an increase in patients with Guillain-Barré

T

syndrome. The same happened in Brazil in 2015, when hese days, World Health Organisation as well

the first outbreak was noted there. Guillain-Barré syn-

as other health ministries worldwide are kept

drome is an autoimmune disease, where immune cells

busy again: Brazil has seen an unusual rise in

attack the neurons and may lead to death by respiratory

the number of patients with Zika virus infec-

paralysis in the worst cases.

tion over the past two years (possibly after the virus ar-

To date, there is no cure against acute Zika virus in-

rived with World Cup travellers in 2014). Last year, more

fection available. The symptoms of an infection are

than 1,5 million people were affected. Although the vi-

treated with common pain and fever medicines. More-

rus is known in Africa, the Americas, Asia and Western

over patients have to avoid dehydration. Only if condi-

Pacific, the current outbreak in Brazil is worrying, since

tions worsen should patients seek medical advice.

the infection may cause severe side-effects.

Zika fever can be diagnosed via symptoms and in the laboratory with a blood test. The virus is a member of

Mild Illness but Severe Consequences

the virus family Flaviviridae and the genus Flavivirus.

Recently, the Brazilian Ministry of Health registered

Zika is related to the dengue, yellow fever, Japanese en-

about 5,000 cases of new-borns with microcephaly in

cephalitis, and West Nile viruses.

An illustration of the Zika virus, whose symptoms include mild headaches, maculopapular rash, fever, malaise, conjunctivitis, and arthralgia. (© istockphoto.com)

Lifesciences plus 01 I 2016

49

F I LT R AT E I Z i k a Vi r u s

Regarding its shape, Zika virus is

it on as they bite again. Unfortunately

icosahedral. Its genome consists of

besides Aedes aegypti which survives

nonsegmented, single-stranded, posi-

only in tropical and subtropical regions,

tive-sense RNA. To date, two Zika virus

also Aedes albopictus, another tiger

lineages – African and Asian – are

mosquito, transmits the virus. The latter

known, whereas the virus in America is

hibernates and survives in cooler cli-

probably very closely related to the

mates and has been registered also in

Asian strain. The complete genome se-

the south of Switzerland (Tessin).

quence of the Zika virus has been pub-

Fortunately, tiger mosquitos are not the

lished. Researchers assume that a re-

strongest flyers. It only flies about 400

cent change in Zika virus codon led to

meters. Travelling

the current outbreak with drammatical

country therefore is slow. In Switzer-

increase in infections. But this is not

land, the Alps build a natural barrier.

confirmed yet.

Nevertheless, the mosquito can be un-

from

country

The primary vertebrate hosts of the

intentionally transported by humans in

virus were monkeys and transmission

planes, truckss etc. If the climate of the

to humans occurred only occasionally.

destination is suitable, Aedes might re-

Before the current pandemic began in

produce and thereby introduce the Zika

2007, Zika virus rarely caused infections

virus into new regions.

in humans. Other arboviruses (arthro-

Zika fever rash on arm. (Picture: Wikimedia)

to

pod-borne viruses, which are transmit-

Precautions for Europe

ted via arthropods) have become estab-

The situation in Switzerland and Eu-

lished as a human disease though, and

rope is not alarming yet. During winter,

spread in a mosquito–human–mosquito

mosquitos are inactive. But what will

cycle (yellow fever, dengue fever and

happen in summer, especially with cli-

Transmission Electron Micrograph (TEM) picture of Zika

chikungunya fever).

mate warming and heat waves hitting

virus. Virus particles are 40 nm in diameter, with an outer

moderate climates, is not predictable.

envelope, and an inner dense core. (Picture: Wikimedia)

Spread Zika virus is transmitted via two vectors: man and mosquito, whereas the

Theoretically, Zika as well as Dengue spread is possible.

Another

problem

might

be

the

Aedes aegypti mosquito biting

latter is more complicated to handle. Fe-

scheduled 2016 Olympic and Paralympic

and sucking blood from human.

male mosquitos bite more than one per-

games in Rio de Janeiro. Besides Brazil-

(© istockphoto.com)

son and pick up the virus with the blood

ian residents, also attendees from all

they suck and pass it on to the next vic-

over the world might get in contact with

tim. Three days after biting, they lay

the virus and bring it along to their

eggs that survive up to 1 year, even

homes on their return.

without water! In small quantities of

Travellers and inhabitants of affected

water, the larvae hatch and develop to

areas are advised to use insect repellents.

adult mosquitos. These get infected

Wearing clothes to cover as much of the

from people carrying the virus and pass

body as possible is recommended as well

50

Lifesciences plus 01 I 2016

F I LT R AT E I Z i k a Vi r u s

as using mosquito nets, screens and clos-

University in Denmark is executive

also member of the ESCMID study

ing doors and windows. In addition, trav-

commitee member of an ESCMID study

group speculates that “the next stage

ellers returning from affected area to

group and explains: “Research into

for the virus might be to move from

Switzerland should not donate blood for

rapid diagnostics, treatments and vac-

Cabo Verde to Guinea-Bissau, and from

an interval of 28 days and women are ad-

cines are urgently needed and under-

vised to avoid pregnancy during a period

way.” Moreover, he brings into consid-

of 28 days, states the Swiss Federal Office

eration: “The emergence of Zika virus

for Public Health (BAG).

soon after the Ebola outbreak is anoth-

The European Society of Clinical

er reminder for the need for a coordi-

Microbiology and Infectious Diseases

nated global effort to have sufficiently

(ESCMID) is currently developing tools

resourced rapid response groups for

to monitor the spread of the Zika virus.

proactive surveillance and conduct of

The goal is to gather data in order to

priority research in emergency situa-

better assess the risks of the disease.

tions.” Dr Nick Beeching from the Liv-

Professor Eskild Petersen from Aarhus

erpool School of Tropical Medicine and

Cornelia Staehelin MD

5 Questions to Cornelia Staehelin MD, Senior Physician at the Department of Infectious Diseases, Inselspital Berne Lifesciences plus: Aedes mosquitos are endemic in Switzerland. In addition, Zika virus causes mild symptoms and is thought to be passed on via sexual intercourse.

Thus, people might infect their partners without even knowing they carry the virus. Dr Staehelin, taken these facts into account, how likely is a Zika virus spread in Europe in your opinion?

transmission seems to be negligible compared to the efficiency of transmission through mosquito bites. However, the Federal Commission for Travel Medicine (EKRM) recommends practicing safe sex for at least one month after returning from an affected region.

Women should avoid pregnancy during two menstrual cycles after returning from an affected region.

Currently, there is no explicit phase of high or low risk known. In other words, the entire pregnancy is a high-risk phase.

Which measure to stop the epidemic Would you travel to a Zika-affected is more favourable: a vaccination country like Brazil at present? or the elimination of the vector?

Ideally, both should be possible.

But the development of a vaccination lasts up to 10 years or more.

Thus, in the current situation, Cornelia Staehelin: Since the Is re-infection possible, mosquito control should be pushed mosquito Aedes albopticus is or can immunity be acquired? endemic in the Mediterranean forward. This is already being region (including parts of the Canton This is not known yet. Flaviviruses practised in the affected countries. of Ticino), a possible spread of related to Zika with similar Brazil for instance deploys its Zika is theoretically possible. The epidemics are known to need troops in the combat against Aedes. main vector for the current Zika a “naïve population” (not yet From a historical point of view, this epidemic is however Aedes aegypti, infected individuals) for a large Zika epidemic is very interesting which is not found in Europe. spread. In countries with an and yet tragic. In the 1970ies, tiger Nevertheless, similar diseases established Zika virus circulation mosquitos were eradicated in South like Dengue fever or Chikungunya for decades, such huge epidemics America, except for the countries fever showed only very limited affecting all segments of the Venezuela, Guyana and Surinam. outbreaks in the south of Europe population are not seen, suggesting After the use of DDT as an insec(217 cases of Chikungunya in that people are infected earlier ticide was banned due to the Italy in 2007). Therefore, the risk in life resulting in some form of accumulation in the human body, of a possible spread is rated minilasting immunity. However, in case the tiger mosquitos reconquered mal according to the European of Zika, a final assessment is not the whole continent during the Centre for Disease Prevention and possible yet and investigations are following decades. In parallel, Control (ECDC). Sexual transmisin progress. during the 1970es, Dengue was sion though is very uncommon for only present in Venezuela and the a virus which is mainly transmitted Pregnant women are worried adjacent countries. With the return by a mosquito. So far only two about the health of their unborn of the tiger mosquito, all of the cases are documented. But the children. Is there a permanent risk four types of Dengue spread in virus could be detected in sperm for microcephaly during 40 weeks South and Central America. In 2013 3 – 4 days after disease outbreak. of pregnancy or is the risk limited and 2014, tiger mosquitos spread For the current epidemic, sexual to a certain trimester?

Chikungunya, and in 2015s, Zika

Lifesciences plus 01 I 2016

51

is on the rise. Thus, control of Aedes aegypti and related mosquitos is certainly the crucial strategy and will have a wider impact than only controlling the Zika virus.

Yes. The absolute number of travellers returning with Dengue or Chikungunya remains low.

Even if the number of infected returnees rises during epidemics, these numbers are still very limited.

(86 cases of Chikungunya and 127 cases of Dengue in 2015 in Switzerland according to the BAG).

Currently pregnant women are strongly recommended to postpone or cancel journeys to the high-risk regions. If travelling is absolutely required, they should be pedantic regarding the precautions against mosquito bites. We strongly advise to wear long and loose clothing during the day (Aedes mosquitos are daytime biters) and to impregnate their clothes with a Permethin-containing spray.

Additionally, insect repellents should be frequently and thoroughly applied to the uncovered skin. We recommend to only use repellents that show the sign of quality testing by the Swiss Tropical and Public Health Institute.

F I LT R AT E I Z i k a Vi r u s

there to neighbouring countries

of Aedes’ and hence Zika’s next

in West Africa. It could also be

destination – depending on the cli-

exported to Madeira, which is part

mate and weather conditions of

of Europe.” If the virus reaches

summer 2016.

Southern

Europe,

the

situation

could become alarming. Countries like France, Spain and Italy which

REFERENCES

in the past have registered cases of Dengue transmission would then

www.wikipedia.com • www.who.int, www.bag.adim.ch

be at risk. Under these circum-

www.spiegelonline.de • www.ninds.nih.gov

stances, Switzerland could be one

www.escmid.org • www.infektiologie.insel.ch

Guillain-Barré Syndrome Guillain-Barré syndrome was first described by G. Guillain, J.-A. Barré and A. Strohl in 1916 and can affect anybody like many autoimmune disorders.

Symptoms and Reconvalescence First symptoms include a weakness or tingling in the legs. Later, the sensations spread to the arms and upper body.

During further progression of the disorder, the symptoms increase in intensity until finally some muscles are paralyzed. In severe cases this can be lethal, since breathing becomes impossible and cardiac arrhythmia occur. Fortunately, most patients recover fully.

The syndrome is rare (1:100,000), men are slightly more often concerned than women. Usually, Guillain-Barré occurs a few days after an infection (respiratory or gastrointestinal). Typically, after 2– 4 weeks symptoms are most intense and begin to subside after this peak phase.

Recovery can take up to 4 months, whereby about two third of affected people recover fully. However, 30 percent of the patients feel residual weakness for up to 3 years. A relapse of muscle weakness and tingling sensations after many years after the initial attack occur in about 3 percent of the patients.

Mechanisms Guillain-Barré is not contagious, since it is known to be an autoimmune disease. In case of Guillain-Barré, the immune system starts to destroy the myelin sheath surrounding the axons of peripheral nerves. Sometimes also the axons are attacked. Axons are the extensions of nerve cells which contact other cells (neural or muscular) and transmit signals. Myelin surrounds the axons and work as a signal enhancer like a kind of electrical insulator. If the

52

myelin sheath of afferent and efferent neurons is degraded, signals are transmitted inefficiently: muscles cannot adequately respond to the brain’s command. In turn, the brain doesn’t get enough information about sensations in the periphery, which ends up in feelings like tingling, pain or heat/cold.

Trigger Very often, Guillain-Barré forms after an infection with virus or bacteria or very rarely after vaccinations. Regarding bacteria, Campylobacter jejuni – a pathogen which causes gastroenteritis and stomach ulcer – is one of the most common malefactors. Antibodies against Campylobacter are formed and attack the myelin. In all cases of GuillainBarré, the immune cells are re-programmed and start treating myelin as foreign, triggered by the stimulus with a virus or a bacterium.

Diagnose and Treatment Syndroms are difficult to diagnose, since they show symptoms which may be similar to other diseases. Therefore, only differential diagnose and the analysis of cerebrospinal fluid (which bathes brain and spinal cord) bring the issue to light. However, there is no cure for Guillain-Barré available. Sometimes plasmapheresis (exchange of plasma) or high-dose immunoglobulin therapy are successful. Since there is no standard cure available, the disease has to be waited out and patients have to be monitored accurately in order to keep respiration stable.

Lifesciences plus 01 I 2016

NEWS

What does she think to herself? The motives behind our behaviour normally remain hidden. (© istockphoto.com)

Eastern Water Frogs “conquer” Canton of Aargau

In the past

two decades, water frogs have spread rapidly in Central Europe. Using a new statistical model, researchers from the University of Basel were now able to show that local species such as the Yellow-bellied Toad and the Common Midwife Toad are suffering from the more dominant water frogs. So-called invasive species may either hunt other species or replace them in their natural habitat. Some of the most invasive amphibians belong to the water frog species complex. The Eurasian Marsh Frog, native to Eastern Europe, has become particularly dominant in Switzerland in the past years. Humans are to blame for this development, as they began to import the frog species to Central Europe for consumption beginning of the 1960s.

Hidden Motives Become “Visible“

To understand

human behaviors, it is crucial to understand the motives behind them. So far, there was no direct way to identify motives. Simply observing behavior or eliciting explanations from individuals for their actions will not give reliable results. Psychologist and neuroscientist Grit Hein and Ernst Fehr from the Department of Economics, University of Zurich, teamed up with Yosuke Morishima, Susanne Leiberg, Sunhae Sul and found that the way relevant brain regions communicate with each other is altered depending on the motives driving a specific behavioral choice. This interplay between brain regions allowed them to identify the underlying motives.

The Basel zoologists Tobias Roth, Christoph Bühler and Valentin Amrhein have now used the data of over 1,000 bodies of water in the Swiss Canton of Aargau in order to study the impact of water frogs on native amphibians using a new statistical model. The cantonal Amphibian Monitoring Aargau supplied the necessary observational data. However, such data is often difficult to interpret: If, for example, there are only few observations of a species, does that mean that the species is actually rare or is it only observed rarely? The statistical model used by the Swiss researchers takes into account the different detection probabilities of species as well as other environmental factors. The results show that Yellow-bellied Toads and Common Midwife Toads have smaller communities whenever water frogs are also present at the same body of water. “Based on our analysis we estimate that the populations of both species would be up to five times bigger without the water frogs’ presence,” says Christoph Bühler, Head of Amphibian Monitoring Aargau. # www.unibas.ch

During the study, participants were placed in an fMRI scanner and made altruistic decisions driven by an empathy motive (the desire to help a person for whom one feels empathy) or a reciprocity motive (the desire to reciprocate an individual’s previous kindness). Simply looking at the functional activity of specific regions of the brain couldn’t reveal the motive underlying the decisions. Broadly speaking, the same areas in the brain lit up in both settings. “However, using Dynamic Causal Modeling (DCM) analyses, we could investigate the interplay between these brain regions and found marked differences between empathybased and reciprocity-based decisions^,” explains Grit Hein. “The impact of the motives on the interplay between different brain regions was so fundamentally different that it could be used to classify the motive of a person with high accuracy,” she continues.

A further important result was that motives are processed differently in selfish and prosocial people. In selfish people, the empathy but not the reciprocity motive increased the number of altruistic decisions. In contrast, prosocial people behaved even more altruistically after activating the reciprocity, but not the empathy motive. # www.uzh.ch

Invasive species: The Eurasian Marsh Frog has been imported since the 1960s for human consumption and was released to nature.

(Picture: Christoph Buehler)

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Lifesciences plus 01 I 2016

F I LT R AT E I N e w s

New Tobacco-Based Influenza Vaccine

A new tobacco-based

seasonal influenza vaccine being developed by Mitsubishi Tanabe Pharma and currently in Phase III studies could potentially rival traditional chicken egg-based vaccines, as it aims to launch in the US for the 2018 – 19 flu season, according to an analyst with research and consulting firm GlobalData. The technology involved in the new vaccine, which can be produced in four weeks, six times faster than egg-based methods, involves implanting influenza genetic material into tobacco leaves, a manufacturing process originally developed by Medicago, a Canadian company acquired by Mitsubishi Tanabe in 2013. Achilleas Livieratos, PhD, GlobalData’s Analyst covering Infectious Diseases, states that there are a number of serious limitations that come with the use of egg-based vaccines, leaving a substantial need for alternatives: “As well as taking six months to work, during which time minor genetic mutations can decrease vaccine efficacy, individuals with egg allergies cannot safely receive vaccines, leaving them vulnerable to infection. A number of vaccine giants including Sanofi, GlaxoSmithKline and MedImmune / Astra Zeneca are also developing their vaccine portfolios.

However, their current egg-based, quadrivalent, inactivated (split virus) seasonal influenza vaccines lack the manufacturing efficiency of tobacco plant-derived vaccines that can also generate virus-like particles (VLPs),” says Mr Livieratos.

Mitsubishi Tanabe will need to demonstrate strong safety data and yearly production consistency of its tobacco-based vaccine.

If the company’s product, or one like it, is approved, GlobalData expects a novel vaccine that boasts a rapid, plant-based manufacturing process to have a significant impact on the seasonal influenza vaccine landscape. # www.globaldata.ch A new Influenza vaccine is produced through tabacco leaves.

(Picture: Wikipedia)

Broccoli Ingredient has Positive Influence on Drug Efficacy

Colon cancer cells

that are pretreated with an ingredient found in cruciferous vegetables are more likely to be killed by a cancer drug that is currently in development, found ETH scientists. Certain foods can alter the activity of endogenous enzymes and thus influence the efficacy of drugs. It is well known, for example, that grapefruit has an adverse effect on a number of anti-arrhythmic and cholesterol-lowering drugs: it contains ingredients that inhibit an endogenous enzyme responsible for the degradation of certain medications in the liver. Consumption of grapefruit thus increases the side-effects associated with these drugs. Until now, only a few examples existed of food ingredients that influence the efficacy of the drug to the benefit of the patient through nutrition.

Scientists from ETH Zurich and the University of Zurich have recently discovered a new example of such a correlation. The researchers struck gold when they investigated the effects of sulforaphane on human intestinal cells. Sulforaphane is a naturally occurring ingredient in a number of cruciferous vegetables, such as broccoli. The scientists, led by Shana Sturla, Professor at the Department of Health Sciences and Technology at ETH Zurich, treated various types of colon cancer cells and intestinal cells from healthy subjects with this substance in the laboratory. The concentration used was approximately equivalent to that which reaches the intestines after consuming a meal with broccoli, and importantly, at a dose that did not itself act in killing the cells. The researchers discovered that sulforaphane increases the concentration of a number of enzymes in the colon cancer cells, including those of an enzyme with the abbreviation AKR1C3. Interestingly, sulforaphane does not have this effect in all cases: in colon cancer cells that already exhibit a significantly elevated concentration of AKR1C3 as a result of the cancer, the broccoli substance caused a further increase in the concentration of the enzyme. But sulforaphane had no influence in colon cancer cells with an initially very low concentration of AKR1C3. The same was found with intestinal cells unaffected by cancer. AKR1C3 is central in the efficacy of a cancer drug (PR-104A) that is currently still in development and clinical testing. # www.ethz.ch

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