Nature Outlook Autismo by Javier Ortega

OUTLOOK

AUTISM

01 November 2012 / Vol 491 / Issue No 7422

OUTLOOK AUTISM

Produced with support from: Nancy Lurie Marks Family Foundation, the Simons Foundation, Roche, Autism Speaks, and The Autism Science Foundation

Illuminating a diverse disorder

Cover art: Mark Smith

Editorial Herb Brody, Apoorva Mandavilli, Michelle Grayson, Tony Scully, Aaron Fagan, Nick Haines Art & Design Wes Fernandes, Alisdair Macdonald, Andrea Duffy Production Donald McDonald, Yvonne Strong, Kelly Hopkins, Leonora Dawson-Bowling Sponsorship David Bagshaw, Yvette Smith, Reya Silao Marketing Elena Woodstock, Hannah Phipps Project Managers Claudia Deasy, Christian Manco Art Director Kelly Buckheit Krause Chief Magazine Editor Tim Appenzeller Editor-in-Chief Phil Campbell

magine you wake up one day and all the familiar signals of human communication are unintelligible. A smile no longer expresses mirth or happiness, a raised voice no longer reflects excitement or anger. You can exchange words with other people, but you’re talking through a heavy curtain of uncertainty. You find it difficult or impossible to behave the way people seem to expect. Welcome to the baffling world of autism, a range of disorders that is affecting a growing number of people and continues to perplex scientists searching for causes and cures. The degree to which autism is on the rise is a matter of some controversy. There’s no doubt that the number of children identified as having an autism-related disorder (often described as being on the autism spectrum) has surged in the past decade or so. What’s not so clear is whether this represents a true increase in prevalence or just greater awareness of the condition (page S2). Indeed, our understanding of the disorder is evolving — and the shifting definition could deny some children access to the educational and social services that give them a better chance to succeed (S12). This Outlook is an editorial collaboration between Nature and SFARI.org, the news website of the Simons Foundation Autism Research Initiative. SFARI.org operates with editorial independence from the Simons Foundation and did not participate in the editing or commissioning of articles about research funded by the Simons Foundation. We find that uncertainty still shrouds much of autism. Genetic analysis is beginning to yield candidate genes and some of the underlying physiology of autism (S4). Yet, there are very few treatments available (S14). And little is known about what happens when children with autism grow up (S10). We acknowledge the financial support of the Nancy Lurie Marks Family Foundation, the Simons Foundation, Roche, Autism Speaks and The Autism Science Foundation. As always, Nature retains responsibility for all editorial content. Herb Brody Supplements Editor

Nature Outlooks are sponsored supplements that aim to stimulate interest and debate around a subject of interest to the sponsor, while satisfying the editorial values of Nature and our readers’ expectations. The boundaries of sponsor involvement are clearly delineated in the Nature Outlook Editorial guidelines available at http://www. nature.com/advertising/resources/pdf/outlook_guidelines.pdf

CONTENTS S2 EPIDEMIOLOGY

Complex disorder To define autism we must first figure it out

S4 GENETICS

Searching for answers Collaborations tackle the complexity

S7 CHILD DEVELOPMENT

The first steps Study of siblings points to better detection

S10 ADULTHOOD

Life lessons Research is maturing along with its participants

S12 DIAGNOSIS

Redefining autism Draft guidelines a matter of contention

S14 TREATMENTS

In the waiting room Clinicians eager for targeted treatments

S17 PERSPECTIVE

Imaging autism Nicholas Lange cautions against using brain scans for diagnosis

S18 CULTURE

Diverse diagnostics Behaviour can only be assessed in context

S20 PERSPECTIVE

Brain scans need a rethink Evidence for a leading autism theory is flawed, say Ben Deen and Kevin Pelphrey

COLLECTION S21 Autistic-like behaviour in Scn1a+/– mice

and rescue by enhanced GABA-mediated neurotransmission Sung Han et al.

S27 Neural mechanisms of social risk for

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psychiatric disorders Andreas Meyer-Lindenberg & Heike Tost

S33 Using iPSC-derived neurons to uncover

cellular phenotypes associated with Timothy syndrome Sergiu P. Paşca et al.

S39 Exome sequencing in sporadic autism

spectrum disorders identifies severe de novo mutations Brian J. O’Roak et al.

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EPIDEMIOLOGY

Complex disorder Researchers are digging into the myriad causes of autism to refine its definition and find elusive biological signatures. BY VIRGINIA HUGHES

efining ‘autism’, a single word encompassing a wide spectrum of behaviours, is no easy task. Each person with autism has a different combination of symptoms, and the combination may change over that person’s lifetime. Some people with autism don’t speak at all, whereas others are fluent. Some are bothered by sounds, whereas others are musical prodigies. Some are intellectually disabled, whereas others are savants. No wonder then that many researchers have begun referring instead to ‘autisms’. It has become clear over the past few years that the biology underlying autism is similarly multifarious, with hundreds of potential genetic, developmental and environmental causes. Still, researchers are on the hunt for a biological signature of autism, one they say is the unmistakable core — a distinctive mix of social disinterest and repetitive behaviours — that marks every individual with the disorder. “That’s the big mystery,” says David Amaral, a neuroscientist at the MIND Institute at the University of California, Davis. “How could so many different aetiologies, so many different genetic risks, lead to a behavioural syndrome that a clinician could say, ‘Well, that person has autism’?”

Since Leo Kanner defined autism in 1943, its symptomatic profile has widened dramatically. It now includes not only social impairments and repetitive behaviours, but many associated ones, such as motor problems, hyperactivity, seizures and distinctive facial features. Autism’s prevalence has also grown. As recently as the mid-1990s it was thought to be rare, occurring in 1 in every 2,500 people. Estimates published in 2012 by the US Centers for Disease Control and Prevention put it at 1 in 88. Many theories attempt to explain this spike, although no single theory accounts for all of it. A substantial component of the risk is demonstrably genetic. When one identical twin has autism, for example, there is a roughly 70% chance that the other will also have the disorder. Many research teams have searched for genes that may be involved. They haven’t turned up any prime candidates yet, only dozens, even hundreds, of bit players. “I truthfully believed that sporadic autism, like Kanner had described it, might be caused by a handful of genes with maybe one predominant gene,” says Huda Zoghbi, a geneticist at Baylor College of Medicine in Houston, Texas, who in 1999 first identified the gene that causes an autismrelated condition called Rett syndrome. “Little did we know.”

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To complicate matters, many of the genetic blips associated with autism are found in the general population, too. Still, some themes are emerging. For example, many of the genes function at the synapse, the junction between neurons, which has led many pharmaceutical companies to begin early investigations of drugs that target this part of the cell. But the genetic findings are hardly simple. Other implicated genes are related to an array of cell functions, from the immune system, to gastrointestinal repair, and even the production of cellular energy. Part of the increase must be due to greater awareness. As more people learn about autism, it is likely that more people will spot early signs and seek help. This happens more quickly in some regions than others, depending on cultural and socioeconomic factors. A mix of demographic differences could help explain why, for example, there is a four-fold difference in autism prevalence between Utah, the US state with the highest rate, and Alabama, which has the lowest. Autism is also diagnosed differently these days. The widening criteria for diagnosis means that many high-functioning people with autism might have gone undiagnosed in the past. Conversely, some of those who have severe cognitive problems might have been diagnosed with mental retardation. The rapidly rising prevalence has also led scientists to look at environmental causes. A small part of the increase in autism seems to result from ageing fathers, for example. As men age, their sperm acquires harmful mutations that can be transmitted to the child. There are any number of other theories, some more plausible than others: the distribution of gut bacteria, autoimmune conditions in the mother, fetal exposure to drugs such as antidepressants, or even living near main roads. “There have been so many changes in lifestyles,” Zoghbi says. “Everything is game.” Despite this seemingly endless list of causes and consequences, researchers are looking for commonalities at every biological level, whether in the genome, a brain circuit or even a style of thinking. For example, in the 1980s, Uta Frith and colleagues proposed that people with autism lack ‘theory of mind’, the ability to grasp what others think or believe. Researchers have since localized this ability to a part of the brain called the temporoparietal junction. Pairing these kinds of cognitive studies with molecular ones will be crucial to understanding the disorder, says Frith, a developmental psychologist at the Institute of Cognitive Neuroscience at University College London. “You must look at the right level to pull all of the strings together and to talk about this unity of experience that I would call autism.” ■ Virginia Hughes is a freelance science writer based in Brooklyn, New York.

CHARLY FRANKLIN/GETTY IMAGES

OUTLOOK AUTISM

THE RISE OF AUTISM

Once thought of as a rare disease, autism’s prevalence, funding and research has risen dramatically over the past twenty years.

PREVALENCE OF AUTISM AND RELATED RESEARCH

EVOLVING DEFINITIONS 3,000

1 in every 88 children is diagnosed with autism

2,500

Scientific studies on autism have increased 12-fold since 1980.

2,000

In 1998, autism prevalence in a New Jersey town was found to be more than 16 times higher than estimates.

1,500

1,000

For just over a century, researchers have grappled with how to define autism — and what causes it.

Number of research publications

Prevalence of autism (x 1,000 people)

5, 318–326 (1995)/YEARGIN-ALLSOPP, M. J. AM MED ASS. 289, 49–55 (2003)/ BERTRAND, J. ET AL. PEDIATRICS 108, 1155–11561 (2001).

SOURCES: CENTER FOR DISEASE CONTROL AND PREVENTION/ THE INTERAGENCY AUTISM COORDINATING COMMITTEE./BURD, L. ET AL. J. AM. ACAD. CHILD. ADOLESC. PSY. 5, 700–703. (1987)/RITVO, E. R. ET AL. AM J PSYCHIATRY. 146, 1032–1036. (1989)./KIRBY, R. S J. DEV. BEHAV. PEDIATR.

AUTISM OUTLOOK

500

1984

1988

1992

1996

2000

2004

2008

BREAKDOWN OF AUTISM RESEARCH FUNDING IN THE USA, 2010: Infrastructure and surveillance ($50.8M) Lifespan issues ($6.6M)

Services ($64.8M)

Treatments and interventions ($68.1M)

84%

increase in research funding between 2008 and 2010

Diagnosis ($45.6M)

408.5M

PUBLIC AND PRIVATE FUNDING (US$)

Biology ($91.3M)

Risk factors ($81.2M)

1911

Swiss psychiatrist Eugen Bleuler coins the term ‘autism’ in describing self-absorbed adults with schizophrenia.

1943

US psychiatrist Leo Kanner publishes a report of 11 children with autism, defines disorder as “autistic disturbances of affective contact.”

1944

Austrian paediatrician Hans Asperger publishes a report of children with profound social problems, lack of empathy and clumsiness.

1967

Bruno Bettelheim’s The Empty Fortress claims that autism stems from social deprivation, adding fuel to the popular, though incorrect, theory that emotionally cold mothers were the cause.

1980

‘Infantile autism’ is added to the Diagnostic and Statistical Manual of Mental Disorders (DSM) III. Defined by 6 criteria, including a lack of responsiveness to others, gross language and resistance to change.

1992

Asperger’s syndrome becomes a distinct diagnosis when it’s included in the tenth edition of the World Health Organization’s diagnostic manual.

1994

The fourth edition of the DSM greatly expands the autism spectrum, outlining criteria for autistic disorder, Asperger’s syndrome and pervasive developmental disorder not otherwise specified (PDD-NOS). Genetic risk factors ($50.8M) Gene-environment ($20.5M) Environment ($4.4M) Epigenetics ($5.5M)

2013

The fifth edition of the DSM is likely to merge the various autism disorders into a single category called autism spectrum disorder.

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LUIS DE LA TORRE UBIETA

OUTLOOK AUTISM

Researchers are mutating autism-associated genes in brain cells in cell culture to see how the cellular biochemistry is altered.

G E NE T I CS

Searching for answers Solving the riddle of autism genetics will require looking beyond the growing list of candidate genes to epigenetics and personalized medicine. B Y S A R A H C . P. W I L L I A M S

t Rutgers University in New Brunswick, New Jersey, blood samples from 3,000 people with autism and their families have been carefully collected and tested over the past five years. The DNA from each of the samples — almost 13,000 in total — has been extracted, studied and shared with researchers around the world. And each person with autism whose genes are in the collection has been put through batteries of tests and examinations to characterize their condition. The Simons Simplex Collection, as the set of data is known, is one of the largest — and most comprehensive — of the handful of autism cohorts around the world. But even so, it hasn’t given up many of autism’s secrets. Each new study using the Simons data — or one of the other similar collections — adds to a growing list of gene variations that could relate to the disorder, or perhaps even be the cause in some cases. But on the whole, the expanding

catalogue of genes doesn’t explain what causes autism at a cellular or molecular level, what characteristics the mutations share, or how to treat the disorder. For researchers, autism is a riddle that is ever more complex. And for patients and their families, it’s a story of how science can lead to discovery after discovery but be slow to improve treatments and change lives. Now, however, a new research strategy that integrates protein and epigenetic studies into classic genetic screens might reveal some of the disease’s causes and potentially lead to successful, targeted treatments.

THE ROLE OF GENETICS

“From a genetics standpoint, it’s been clear for a while now that there isn’t going to be an autism gene,” says Daniel Geschwind, a neurologist and autism researcher at the University of California, Los Angeles. “But the idea now is that we’re moving from just looking under the proverbial street light to integrating many different approaches to solve this problem.”

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The latest estimates by the US Centers for Disease Control and Prevention state that 1 in 88 children in the United States has been identified with an autism spectrum disorder. Symptoms range from mild social impairment to severe developmental delays and challenges in language, communication, social interaction and behaviour. It’s been known for decades that the dis“There is just not order has a genetic enough postcomponent: studies mortem tissue of identical twins that fits our in the 1980s found criteria.” that if one twin has autism, the other has the disorder almost 100 per cent of the time1. More recent studies have found slightly lower numbers in twin and sibling studies, but they still estimate that well over half of all autism cases can be attributed to genetic, rather than environmental, factors2. And there’s another piece of evidence for inheritance: it’s become increasingly clear that

AUTISM OUTLOOK older parents — fathers in particular — are at increased risk of conceiving children with autism. A study published in August 2012 found that for every increasing year of a father’s age, from 20 to 65 years, the number of mutations passed along to his offspring increased3. The research suggests that a man’s genetic material accumulates mistakes over his lifetime, as sperm cells are continuously copied. But which of these genetic mutations lead to autism?

DAVID CRAUSBY/ GETTY IMAGES

SEARCHING THE GENOME

Many of the early studies hunting for autism genes were genome-wide association studies (GWAS). Such studies compare the DNA of people with and without a disorder using thousands or millions of markers scattered throughout their genomes. However, GWAS can only point to large chromosome regions rather than identifying particular genes or gene mutations. The first GWAS on autism, published in 2009, implicated two regions with mild effects: one on chromosome 5, and one on chromosome 20 (ref. 4). Subsequent GWAS on autism have failed to turn up any other parts of the genome with statistical significance. Many researchers were disappointed by these results, which suggested that the common genetic variants that GWAS detects explain only a small fraction of the symptoms of autism. “The commonvariant hypothesis was essentially tested,” says Geschwind. “And it doesn’t mean that common variants don’t contribute at all, but they have small effects. So let’s look at the rare variants.” But at the University of Southern California in Los Angeles, neurogeneticist Dan Campbell wanted to be sure that all the information possible was culled from the GWAS data before he moved on. “Our approach has been to go back to those genome-wide signals,” says Campbell. He was convinced that common variants played some role in autism because of studies that had found broad autistic traits in up to half of all children with autism. So Campbell’s lab started studying, in more depth than ever before, the two regions that had been located. They’ve already found a gene on chromosome 5 that could play a role. The surprise, and the reason it hadn’t been identified earlier, is that it’s a non-coding RNA. Instead of being the basis for a protein, the gene is translated into an RNA molecule that has activity in the cell in its own right. In this case, the RNA binds to a different gene — one that makes a protein called moesin, which is known to be involved in brain development. The non-coding RNA turns off the moesin gene, which is critical to early brain development. Without moesin, the neuronal projections (axons and dendrites) are shorter than normal. Brain samples from deceased autism patients, Campbell’s team reported in April 2012, have on average 12 times more of the non-coding RNA than usual, suggesting that levels of moesin might have been stunted at some time during development5. More work is needed to test whether

the altered moesin levels lead to altered brain morphology in people with autism. Like all the findings on genes implicated in autism, this non-coding RNA doesn’t explain all instances of the condition. And even in people who have the common variant and display an increase in these RNA levels, it might not be the full story. “Some cases of autism you can nail down to, ‘there’s a mutation in this gene’,” says Campbell. “Some cases will be due to people inheriting a common genetic variant.” And other cases, he said, could be the result of a combination of common genetic variants with an uncommon gene variant or an environmental factor. With the dropping price of genetic sequencing, many researchers have moved from GWAS to more in-depth sequencing of DNA from people with autism and their unaffected family members in an effort to unearth rarer mutations. In April 2012, three research papers reported the sequencing of exomes (the encoding portions of the genome) from a combined

Children born of older fathers are more likely to have autism.

total of more than 1,000 autism patients and hundreds of family members. Together, the studies turned up in excess of 1,000 genes from across the genome associated with autism, far surpassing previous estimates of how many rare genes could contribute to the disorder. The challenge, says geneticist Jay Shendure of the University of Washington in Seattle, is that the rarer a mutation is, the larger the sample size required to prove its association with autism. There are hundreds of thousands of people with autism spectrum disorder in the United States, for example, but only a tiny fraction of that population is part of any wellcharacterized study like the Simons Simplex Collection. “The cost of sequencing is dropping at such an incredible rate that I do think there will come a time in the next few years when all the good cohorts out there have been sequenced,” says Shendure. “But the sample sizes to implicate these genes may need to be much larger, and that means we need more cohorts.” The number of people with autism in the United States is large, but it takes time to enrol patients in studies — each person must have in-depth, standardized testing.

Finding the genetic variations is one mission; understanding what they mean is another. Shendure says that new analytical techniques are needed to put the growing list of autism genes into context. By using maps of protein–protein interactions throughout cells, for example, researchers can find networks of proteins affected by many different genetic mutations, suggesting a common mechanism for disease. Or, by relying on new catalogues of gene expression data, they can pin down which cells might be influenced by known mutations. “The more sorts of data we have to intersect our data with, the more we can get out of it,” says Shendure.

MOVING BEYOND GENES

The systems biology approach that Shendure proposes is one that Geschwind also endorses. The rare mutations that turn up in exome studies are not necessarily all causative of autism. So using data from the Simons Simplex Collection, Geschwind and colleagues recently profiled the transcriptomes (the set of all RNAs produced) of 244 people with autism and members of their family to get a sense of which genetic mutations led to real changes in gene expression. “Pathway convergence is likely to be quite important,” says Geschwind. “You can imagine that many distinct genes could end up leading to a similar circuit-level dysfunction in the brain.” Geschwind’s study found that many of these genes with altered expression patterns were clustered in pathways known to play important roles in neuron function, development and structure6. This discovery suggests common mechanisms through which seemingly different gene mutations could cause disease. Treatments targeting those pathways might help many patients who have different gene mutations playing into the same networks. Geschwind’s team analysed gene expression patterns in the brains of 19 people with autism and 17 without, using brain tissue collected after each person’s death7. For each brain, the scientists studied three different regions, all thought to be important in autism. They found that in people with autism, two regions of the brain that normally have distinct patterns of gene expression — the frontal and temporal lobes of the cerebral cortex — instead had almost identical patterns, with the same genes switched on or off. The altered genetics suggest a lack of specialization among some brain cells, which could lead to differences in how the brain processes information. Geschwind’s brain study was small. “There is just not a lot of post-mortem tissue that fits our criteria,” he says. “So you are stuck with small cohorts.” As tissue collections grow in the future, he hopes to repeat the studies and explore more of the unique brain morphology associated with autism. At the University of California, Davis, genetics and immunology researcher Janine LaSalle is also looking beyond straightforward genetic

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OUTLOOK AUTISM A CATALOGUE OF CANDIDATES

A sample of genetic suspects that may be responsible for causing autism Gene

Chromosome

Studies that have linked autism to genetic factors

Description

CACNA1H

Mutations in this calcium channel could change the way neurons function or the way the brain develops.

CNTNAP2

This gene’s protein associates with potassium channels on neurons and may play a role in the differentiation of a nerve cell’s axons.

CNTN4

Encodes a membrane protein that helps axons form in the developing nervous system.

FOXP2

Encodes a protein that regulates other genes, including CNTNAP2. Studies have shown that it is important to neural plasticity.

MECP2

X chromosome

The MECP2 protein is implicated in Rett syndrome and known to be involved in turning off other genes in nerve cells.

MET

Best known as a proto-oncogene, but signalling by MET has also been linked to the development of certain parts of the brain.

NRXN1

Encodes membrane proteins active at the synapse between two neurons.

OTXR

The OTXR protein helps control the levels in the brain of the neurotransmitter oxytocin.

RAI1

Mutations in RAI1 are associated with certain syndromic causes of autism.

SHANK3

Involved in the development of synapses between neurons. Mice lacking Shank3 show symptoms.

screens for autism variants, in the hope that she will tie together the hundreds of autism genes being discovered with a finding on another level. Instead of focusing on proteins, LaSalle is looking at epigenetics — changes to gene expression through chemical and structural modifications to DNA, such as the addition of a methyl group to DNA, rather than changes to the code itself. Epigenetic patterns in cells can be passed from parent to child or influenced by the environment, and can cause major changes to molecular pathways. If epigenetic alterations were discovered in people with autism, they could go so far as to explain the complexity of the disorder, says LaSalle. “Even a subtle hit to one of these pathways through changes in methylation could have the same impact as knocking out an entire gene,” she says. Already, Rett syndrome, an autism spectrum disorder, is providing hints. This neurodevelopmental disorder, which primarily affects females, leads to physical abnormalities as well as repetitive motions and a lack of verbal skills. The syndrome has been linked to a gene, MECP2, that controls the epigenetic silencing of other genes in neurons. LaSalle has shown that in mice, mutations in MECP2 change the way genes are turned on and off in response to a chemical found in flame retardants. Her results don’t prove that these chemicals are a cause of autism, but they suggest that MECP2’s epigenetic mechanisms could link genetic and environmental impacts in autism8. Researchers in the emerging field of environmental epigenetics have already found that many

environmental toxins and pollutants reduce overall methylation in cells, she says. But epigenetics is a young field and new techniques are needed to be able to fully understand how methylation is controlled and, in particular, how it might have unique roles in neurons. There’s evidence that in neurons, methylation is influenced not only by environmental and inherited factors, but also by neuronal activity itself, setting up a feedback loop. Moreover, neurons have been found to show different epi“ I see cancer as genetic patterns to9 the rest of the body . being about a decade ahead of Until recently, it was thought that most neuroscience.” me t hy l at ion w as concentrated in the promoters of genes, so most commercial tests for methylation are biased toward promoters. As the technology for studying these patterns improves, LaSalle says, additional epigenetic associations with autism will emerge. For example, epigenetic modifications could influence the same genes and pathways that genetic mutations alter. “The real key is overlaying the epigenetic data with data from genome-wide association studies and having people work together,” says LaSalle.

OPTIMISTIC HUNTERS

So far, no single gene has been discovered that accounts for more than a few per cent of autism cases. Still, the discovery over the past decade of more than 100 autism-related genes is progress of a sort. Indeed, according

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to Geschwind, known diseases and mutations now collectively explain almost one-fifth of autism cases. “If you asked me 10 years ago if I thought we’d be on the threshold of understanding some genetic risk in 20–25% of autism, I would have thought that would be very, very optimistic,” he says. “But the field has moved rapidly.” Most recently, a potential treatment for a rare form of autism that involves a mutation in the BCKDK gene was reported10. Experiments with BCKDK-deficient mice showed improvement when administered diets rich with branched-chain amino acids (BCAA). BCKDK mutations diminish BCAA levels, and in human patients the regimen helped restore plasma BCAA levels. These are promising developments that could lead to treatment by sequencing exomes in order to link specific genes to their physiological effects. Eventually, Geschwind says, treatments for autism may become personalized. Genetic tests could determine which treatments would work in a certain patient’s case. But first, studies on broader protein networks and the underlying causes of autism are likely to lead to more general treatments that work for at least some patients, although as these treatments will target broad cellular pathways, they might bring unwanted side effects. Geschwind cites examples from other clinical sciences. “I see cancer as being about a decade ahead of neuroscience,” he says. “In cancer they’ve been effective in some cases but have also seen offtarget effects. In many ways that field is now moving towards more targeted drugs.” The large databases of autism gene candidates that are now available make the quest to explain autism more complicated than researchers had hoped. But the complexity of the condition is stimulating the expansion of approaches taken and enticing scientists to look beyond straightforward genetic explanations for autism. “We’ve figured out that explaining autism is not simple,” says Geschwind. “But I have a pretty optimistic view. We’re going to continue to make progress — and a lot of it is because of great collaboration in the field and an influx of new people tackling autism.” ■ Sarah C. P. Williams is a freelance science writer based in Kailua, Hawaii. 1. Ritvo, E. R. et al. Am. J. Psychiatry 142, 74–77 (1985). 2. Ronald, A. et al. J. Am. Acad. Child Adol. Psy. 45, 691–699 (2006). 3. Kong, A. et al. Nature 488, 471–475 (2012). 4. Weiss, L. A. et al. Nature 461, 802–808 (2009). 5. Kerin, T. et al. Sci. Transl. Med. 4, 128ra40 (2012). 6. Luo, R. et al. Am. J. Hum. Genet. 91, 38–55 (2012). 7. Voineagu, I. et al. Nature 474, 380–384 (2011). 8. Woods, R. et al. Hum. Mol. Genet. 21, 2399–2411 (2012). 9. Iwamoto, K. et al. Genome Res. 21, 688–696 (2011). 10. Novarino, G. et al. Science doi: 10.1126/ science.1224631.

STACEY WIESNER

AUTISM OUTLOOK

A child at low risk for autism wears an electrode cap, which captures the faint electrical signals generated by neurons, as part of a study on autism development.

CH ILD D EVELO PMEN T

The first steps Because infants born into families with autism are more likely to develop the condition, studying them might lead to ways to diagnose people in the general population earlier. B Y K AT H E R I N E B O U R Z A C

t six months of age, Case 6 was a happy baby. She locked eyes with people and smiled back, reacted when her name was called, and enjoyed playing ‘peek-a-boo’. By twelve months, she babbled and knew three words. Six months later, she hadn’t learned any new words. She didn’t point at things to ask for them, as normally developing babies do at this age. She fell frequently and began hitting herself on the head. At 36 months of age, she was diagnosed with autism. Many parents whose children have been diagnosed with autism can tell a similar story. The individual details are different, but the global progress of the disorder is the same, says Lonnie Zwaigenbaum, now co-director of the Autism Research Centre at Glenrose Rehabilitation Hospital in Edmonton, Canada, one of the leaders of the study that included Case 6. The girl labelled ‘Case 6’ was part of a study that followed, from infancy to toddlerhood, the younger siblings of children diagnosed with autism. Baby sibs, as researchers

affectionately call them, are at a much higher risk of developing the disorder than the general population. One in five baby sibs will be diagnosed with autism1; the prevalence in the general population is 1 in 88. Because the population of baby sibs is almost 20 times richer in children who will go on to develop autism, this cohort is ideal for studying the origins of the disorder. In baby sib studies, researchers record the behaviour, cognitive ability and brain function of baby sibs throughout development, as well as infants the same age who do not have any siblings with autism. By comparing the developmental trajectory of the two groups, they hope to gain a better understanding of how autism unfolds. But more importantly, researchers are on the hunt for markers that might enable them to identify infants at high risk of autism in the general population, while developing earlier interventions. The story of babies like Case 6 does not have to be set in stone at birth. “There is no reason to accept the disabling consequences of autism,” says Mayada Elsabbagh, a psychiatrist at McGill

University in Montreal, Canada, who studies the brain function in autism. “If we start intervention earlier, when the brain is more malleable, we can prevent these consequences.”

EARLY SIGNS

Autism can be diagnosed at three years of age. However, owing to disparities in access to healthcare, the median age at diagnosis in the United States is more than five years. By the time most children with autism come into the clinic, they already have major problems with social engagement and language, as well as other difficulties, such as repetitive behaviours. Early intervention is important because autism can create a vicious cycle. Without the ability to draw people in and be drawn to them, a child will lose learning opportunities that shape the rapidly developing brain. But cause and effect are difficult to tease apart in autism, NATURE.COM and problems in brain Unlocking the brain’s function are likely to neural circuitry to precede social and lan- treat brain disorders: guage difficulties. As a go.nature.com/2npoxd

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result of this muddle, “by the time you see the end state, it’s hard to separate the core symptoms from their effects,” says Mark Johnson, who directs the Centre for Brain and Cognitive Development at Birkbeck, University of London. Prospective baby sib studies might fill in the gap. In the mid-1990s, researchers started the first prospective studies that followed both baby sibs and children with no siblings with autism. The earliest of these studies focused on behavioural markers. In 2005, having followed a group of infants (including Case 6) from six months of age, Zwaigenbaum co-authored one of the most influential baby sib studies to date2. Based on this investigation, Zwaigenbaum says, while there is a clear global progression of behaviours in baby sibs who go on to develop the disorder, when each infant is watched individually, there are exceptions. At six months, most of the babies who go on to develop autism are in line with their peers. Like Case 6, they smile when their parents smile, and when strangers smile. Most will perk up when someone says their name.

However, while most babies around this age develop the strength and coordination to hold up their heads and sit independently, baby sibs who go on to develop autism often do not. “They may be floppy — some can’t hold their head up,” says Zwaigenbaum. These early motor problems are one of the best-established early indicators of the disorder, but why they occur, or whether they might cause subsequent problems, is unknown. After six months, the development of baby sibs who go on to develop autism seems to plateau, whereas their peers become more social, more talkative, and more creative. By 12 months, the Canadian researchers found, many children who will eventually be diagnosed with autism have started to become withdrawn. They don’t cue into what other people are doing, but may be intensely interested in nonsocial stimuli. For example, they may stare at toys but not play with them in ways other children do. “They don’t smile as much, they don’t light up to peek-a-boo, and they show intense visual engagement with toys,” says Zwaigenbaum. “But it’s variable — you might find they

are reserved but not impaired, and they may score normally on behavioural assessments.” Children who will later be diagnosed with autism start to show more stable, recognizable signs after 18 months. “They make poor eye contact, they don’t play [normally], and they begin repetitive behaviour,” says Zwaigenbaum. At this age most normally developing children begin pretending with toys, but a child who goes on to develop autism is less likely to. Instead of playing imaginatively, the child may, for example, put the wheels of a toy car up to his or her face and stare intently as they spin. Although these behavioural and motor signs are clearly correlated with an autism outcome, says Zwaigenbaum, neither his group of children nor any others have demonstrated that they can be used to predict the disorder. Yet even though the results are confounding, they also offer hope. Baby sibs who do not go on to develop the disorder often score as atypical at some point, compared with infants who have no siblings with autism. This suggests that for some baby sibs — who are presumably at higher genetic risk for the disease — there may be protective factors at work that researchers could identify and encourage.

QUANTIFYING RISK

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A new crop of baby sib studies goes further and deeper. Ami Klin, chief of the Marcus Autism Center in Atlanta, Georgia, is trying to fill in autism’s developmental map with dense, quantitative data on behaviour. His approach is to measure early and often. He believes this is the only way to get at the richness of normal development in the first year of life, and how it goes awry in autism. In his group’s studies, the children are assessed monthly for their first 6 months, then every 3 months until they reach 24 months of age. Measuring once every 6 months, he says, is like “sending a spaceship to another planet, taking a picture from far away, and saying, ‘no water’,” — without ever sending a rover for on-the-ground exploration. Klin’s lab is developing technologies to quantify infant attention. One of these is an eye-tracking system that very densely samples where infants look, as often as 60 times a second, with software to analyse the data. Klin developed these tools and tested them in older children with autism. For example, Klin’s group reported3 an eye-tracking test that compared how attentive toddlers with and without autism were to social and non-social motion by tracking them while they watched simple animations. Toddlers without autism preferred to look at human motion, whereas those with autism showed no preference. Klin has now done similar vision-tracking experiments in baby sibs, although the results haven’t been published yet. He is waiting for the children to reach 36 months, when the autism diagnosis is considered stable, so he can compare baby sibs who did and did not develop the disorder with the control group.

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OUTLOOK AUTISM

HELEN MARIS

AUTISM OUTLOOK He’s also working with speech scientist Gordon Ramsay of the Yale Child Study Center in New Haven, Connecticut, to make monthly recordings of the babies’ vocal environment, to watch how they progress from babbling to language, and how they learn to modulate their voices to convey emotion.

THE CHICKEN AND THE EEG

Behavioural checklists and quantitative measures of cognition such as Zwaigenbaum’s and Klin’s may eventually lead to a test that paediatricians can use to identify high-risk babies in the general population. To complete the developmental map of the disease, other researchers are undertaking studies of brain function. But there’s still a chicken-and-egg problem in autism: do problems in brain function precede behavioural problems, or vice versa? And to what extent do these feed off one another? “Changes in brain development precede changes in behaviour,” states Charles Nelson, director of research for the Developmental Medicine Center at Boston Children’s Hospital in Massachusetts. Nelson’s is one of a handful of groups studying brain function in baby sibs. Since most behavioural signs of autism emerge after the first year, Nelson and others believe that the key will be to find the origins of autism in the brain during the critical first 12 months of life. Early results from a handful of these studies have come out this year, and many more will follow. One of the most powerful tools available to developmental neuroscientists such as Nelson is electroencephalography, or EEG. Researchers fit babies with electrode caps that capture faint electrical signals generated by neurons firing on the surface of the brain. In 2012, Nelson’s group showed that 6-month-old baby sibs, when resting, showed lower levels of all types of EEG activity compared with low-risk babies4. His group is still processing data from older ages. In the spring of 2012, researchers led by Johnson and Elsabbagh showed that brain function between 6–10 months of age could be used to predict which baby sibs will be diagnosed with autism at 36 months of age5. While monitoring them with EEG, the researchers showed the babies two images: one of a woman looking at the viewer, and one with her eyes averted. Most of those who did not develop autism showed a different EEG response depending whether the woman’s gaze shifted to or away from them. Babies who were later diagnosed with autism did not appear to differentiate. In these brain studies, as in the behavioural studies, the clearest differences were not between infants who developed autism and those who didn’t, but between high-risk infants — the baby sibs —and low-risk infants. “Most babies at risk are processing stimuli differently, but many compensate for that” and do not develop the disorder, says Johnson. Perhaps they compensate because the infant

You looking at me? Whether infants repond to a direct gaze is one of the tests for autism being evaluated.

brain is plastic, or perhaps they have a particularly supportive environment. Whatever it is, it’s giving researchers hope that there might be protective effects — and that early interventions could eventually alleviate autism. It may be a decade or more before behavioural and brain-function studies in baby sibs yield a reliable checklist that can be used to predict the risk of autism in the general population. For one thing, researchers can’t crunch the numbers until three years into the study, when the children reach an age at which a stable diagnosis of autism can be determined. And then, once the data from these prospective baby sib studies are sufficiently robust, researchers will then have to use them to test babies in the general population and follow them until 36 months as well. In the meantime, some researchers are testing infant interventions in baby sibs even without having a reliable prediction of autism. Because only one in five of these children will receive a diagnosis, any intervention has to be supportive and positive for both the infant and the parents, regardless of whether the child develops autism. Programmes in testing emphasize fostering the relationship between parent and baby — something that’s good for any family, and that may have particular benefits for children who will go on to develop autism, perhaps lessening their eventual symptoms, says Jean Kelly, co-director of the Center on Infant Mental Health and Development at the University of Washington in Seattle. Whether improving parent–baby relationships works to lower the risk of autism is the subject of baby sibs intervention studies at the University of Washington and elsewhere. Kelly’s group takes videos of parent–child interactions, and coaches parents — many of whom are stressed because they already have a child with autism. Kelly holds these sessions weekly for 3 months with baby sibs 8–15 months of age. Kelly’s group is trying to help teach, “How do I reach this child?” And, she says, “we are promoting trust, security and attachment on the side of the child.”

Rebecca Landa, director of the Center for Autism and Related Disorders at the Kennedy Krieger Institute in Baltimore, Maryland, has seen the power of interventions in high-risk toddlers from two years of age and is now trying to use these measures in babies. Some of the interventions her group is studying involve bringing together 12-month-old baby sibs who have shown troubling behavioural signs. The group then performs social tasks that take advantage of the peculiarities of repetitive autistic behaviours to encourage them to imitate one another — something these children normally won’t do. These children tend to move objects back and forth and stare at them, so if the researchers give them all the same object, they end up doing the same motion, and then they notice one another. In one exercise, the 1-year-olds wipe the table in front of them with wet wipes, and the researchers then get them to sing “This is the way we wash the table” and carry the wipe to the bin. After this exercise, says Landa, “the parents are in disbelief that the children are doing an activity they normally do at home, that they’re learning the words ‘wash table’, that they are socially engaged,” she says. The intervention studies are still in progress, but Landa is optimistic. “We want to stop these problems in their tracks,” she says. If these methods are shown to have benefits for baby sibs, Landa and Kelly both hope to develop a standardized program that could be widely implemented by paediatricians and child minders. “For some of these babies, autism doesn’t have to be their destiny,” says Johnson. Genes influence the path of development, but the course of development is not inevitable. ■ Katherine Bourzac is a freelance science writer based in San Francisco, California. 1. Ozonoff, S. et al. Pediatrics 128, 488–495 (2011). 2. Zwaigenbaum, L. et al. Int. J. Dev. Neurosci. 23, 143–152 (2005). 3. Klin, A. et al. Nature 459, 257–261 (2009). 4. Tierney, A. L. et al. PLoS ONE 7 (6), e39127 (2012). 5. Elsabbagh, M. et al. Curr. Biol. 22, 338–342 (2012).

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AD ULT HO O D

Life lessons We know little about autism past adolescence, but a wellstudied generation of children with autism will change that. B Y L I N D S AY B O R T H W I C K

he first child ever diagnosed with autism, by child psychiatrist Leo Kanner in 1943, was five-year-old Donald Gray Triplett. Described by his father as living “within himself ” with “no apparent affection” and a “mania for spinning blocks”, Donald would become the archetype for a developmental disorder that is now one of the most common in the United States — affecting an estimated 1 in 88 children, according to the US Centers for Disease Control and Prevention. These days, though, Donald lives much like any other American retiree. A 2010 profile in

The Atlantic magazine portrays him as a happy, independent, Cadillac-driving septuagenarian — albeit one with a few quirks, including a prodigious gift for numbers — who plays golf and travels the world. The contrast between Donald as a young boy and an old man is striking. The question is, how did he get there? We don’t know, says Joseph Piven, a psychiatrist at the University of North Carolina at Chapel Hill, who in 2010 spearheaded a working group on adults older than 50 years of age with the disorder. Older adults with autism, he says, “are virtually unstudied”. It is even unclear whether the disorder’s

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core symptoms — social and communication deficits, repetitive behaviours and restricted interests — persist during adulthood. Nor do we know much about the prevalence of autism among adults or about their mental state, brain activity or general health. “Early in my career, if you said this is a lifelong condition, people would get upset. They didn’t want to hear that,” says Piven. But that perception of autism as a childhood disorder has begun to change, largely due to shifting demographics. Autism prevalence has risen sharply since the 1990s, so much so that neuroscientist and psychiatrist Thomas Insel has likened it to a “huge wave moving through the system”. Insel is director of the US National Institute of Mental Health in Rockville, Maryland, and chairs the Interagency Autism Coordinating Committee (IACC), a group of scientists and advocates charged with guiding federal funding of autism research in the United States. Starting in 2009, the IACC made research on adults with the disorder a strategic priority. That same year, Autism Speaks, a United States– based autism science and advocacy organization, launched a major initiative for adults with the disorder that calls for legislation to increase access to services for these adults. A 2012 report funded by Autism Speaks estimates that the annual cost of caring for people with autism in the United States is US$137 billion, most of which is for adults. The organization is measuring the economic benefits of providing services to individuals with autism as they transition from adolescence to adulthood. Individual researchers, too, are turning their attention to adults with autism. For example, Catherine Lord, who developed the gold-standard screening tests for autism, is refining the tools used to diagnose the disorder in adults. Lord is a psychologist who directs the Center for Autism and the Developing Brain, a subsidiary of Weill Cornell Medical College in New York and New York Presbyterian Hospital in White Plains. She has been following 200 young adults since they were diagnosed with autism at 2 years of age. “We have a very good sense of who these kids were when they were little and now what they’re like at 20,” says Lord. “That’s been a huge source of information for us.” Another group, at the University of Wisconsin–Madison, is following more than 400 families with a history of autism. Their study, which started 12 years ago, is focusing on how symptoms of autism change over time and how the changes affect parents and siblings. (A handful of long-term studies were conducted in past decades, but they are difficult to interpret because the diagnostic criteria and techniques for assessing autism have changed over time.) In that context, Donald Triplett’s case raises important questions that we’re only beginning to answer. Can children diagnosed with autism expect to live a full life 30, 40 or 50 years from now? Can a subset of children with the disorder outgrow their symptoms, or at least learn to cope

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OUTLOOK AUTISM

OUT OF SIGHT

Adults are rarely included in studies of behavioural interventions for individuals with autism. 80 Number of participants

with them? And, perhaps most importantly, how can we improve the odds that they do? Most studies show that autism’s core deficits, including social and communication problems, persist into adulthood, and that most adults with the disorder remain dependent on family members for support and struggle to fulfil their basic needs such as housing and employment. But work over the past decade or so has offered a more hopeful scenario, partly because many of the children diagnosed today are less severely impaired than those in the past. In some cases, autism’s core symptoms do seem to diminish over time. And a 2009 study of 120 individuals first diagnosed with autism in adulthood found that one-quarter had a college or university degree, and more than 40% held a job or were studying1. About half of the participants aged over 23 were living independently. But the researchers also found higher rates of anxiety and depression in these adults, all of whom had average IQ scores. According to Julie Taylor, a developmental psychologist at the Vanderbilt Kennedy Center in Nashville, Tennessee, who studies the transition to adulthood for people with autism, only about 10% have independent employment in the community. (The numbers are slightly better — ranging from about 25% to 45% — if they include community-based jobs with support from someone like a job coach or jobs in sheltered workshops.) The crucial question arises because of this 10%: what determines whether a child with autism is able to graduate from college, hold down a job and live independently? Some trends are emerging. Faring best are children who score well on standardized intelligence tests, have early verbal skills and display fewer disruptive behaviours such as aggression and hyperactivity. These factors are hard to change, so the challenge to researchers is to find others that can be influenced more readily. “Knowing about IQ and early language as predictors of better outcomes is all fine and good, but when you have a 12-, 18- or 30-yearold, there’s just not a whole lot you can do,” says Taylor. So she is beginning to study societal and family-related factors, such as how connected families are, to better design interventions. The late teen years seem to be particularly critical. One recent study, for example, showed that although autism symptoms generally improve during high school, this improvement slows or ceases after graduation2. This is the time when access to specialized services through the school system dries up. A survey of nearly 1,000 young adults (aged 19–23) with autism published in 2011 found that nearly 40% receive no services after graduation3. “Parents of kids with autism often liken it to falling off a cliff,” says Geraldine Dawson, chief science officer at Autism Speaks. This abrupt loss of services is particularly true for individuals with average or aboveaverage IQ scores, Dawson says, because they don’t qualify for services intended for adults

Percentage of articles

EDWARDS, T. L. RES. AUT. SPECT. DIS. 6, 996–999 (2012).

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10 12 14 Age in years

with intellectual disabilities. Higher-functioning adults are three times more likely to have no regular vocational or educational activities than those with lower cognitive function4. That the adults who are best equipped to succeed are the ones slipping through the cracks was a real wake-up call, says Taylor. Whether therapies — vocational, educational, behavioural or drug-based — can help adults with autism meet those expectations is still unclear. Only 32 studies conducted so far of therapies for autism are aimed at adolescents or adults with the disorder (aged 13–30 years), most of which were of poor quality, according to “Young adults with autism can a report published in make enormous August 2012 by the US progress as they Agency for Healthcare5 Research and Quality . grow up.” Compare that with a previous report of 159 studies of treatments that enrolled children 12 years old and under with autism. Another 2012 review, by psychologist Al Poling of Western Michigan University in Kalamazoo and his colleagues, found that fewer than 2% of participants in studies of behavioural interventions are aged 20 or older6. That’s likely to change over the next ten years or so, says Taylor. And that’s important because therapies may affect adults and children differently. For example, a small placebo-controlled trial published in 2011 found that adults who take the antidepressant fluoxetine (marketed as Prozac) show improvements in the repetitive behaviours typical of autism. Neither fluoxetine nor other antidepressants have had much success in children with the disorder. Research on therapies for autism has focused on children because their brains are still developing (see ‘In the waiting room’, page S14). But there is reason to believe that adult brains also respond to treatment. A mouse study published in 2012 showed that a compound called CTEP reverses many of the neurochemical and behavioural symptoms of fragile X syndrome, an inherited form of intellectual disability that produces an autism-related disorder7. The mice were given

>20

CTEP at 4–5 weeks of age — the mouse equivalent of human adolescence — suggesting that it may offer benefits after the so-called ‘critical window’ of brain development has passed. Another drug, arbaclofen, has undergone phase II clinical testing for fragile X syndrome in children and adults, and is being tested on adults with autism. A new study shows that it alleviates some of the social impairments and behavioural problems that are characteristic of the syndrome. A third drug, GRN-529, may help people with autism8. It significantly improves social behaviour in a mouse model of the disorder compared with controls. “I think the outlook is extremely positive, even for adults,” says Randi Hagerman, who led the arbaclofen trial, at the University of California, Davis. “We are pushing the companies hard to do trials for autism with these drugs and they say they will.” Although research on adults with autism is still lagging behind their need for treatment, the fact that autism is now being viewed — and studied — as a lifelong disorder is encouraging. The 200 children with autism that Lord has followed for the past 20 years are a constant reminder that neural development doesn’t cease. “What’s been so striking is that young adults with autism can make enormous progress as they grow up,” she says. “It’s imperative that we look at adult development and how we can support it — from a humanitarian and an economic point of view.” ■ Lindsay Borthwick is a freelance writer based in New Haven, Connecticut. 1. Hofvander, B. et al. BMC Psychiatry 9, 35 (2009). 2. Taylor, J. L. & Seltzer, M. M. J. Autism Dev. Disord. 40, 1431–1446 (2010). 3. Shattuck, P. T. et al. Arch. Pediatr. Adolesc. Med. 165, 141–146 (2011). 4. Taylor, J. L. & Hodapp, R. M. Am. J. Intellect. Dev. Disabil. 117, 67–79 (2012). 5. Lounds Taylor, J. Comparative Effectiveness Review 65 (Agency for Healthcare Research and Quality, 2012). 6. Edwards, T. L.. et al. Res. Aut. Spec. Dis. 6, 996–996 (2012). 7. Michalon, A. et al. Neuron 12, 49–56 (2012). 8. Silverman, J. L. et al. Sci. Transl. Med. 4, 131ra51 (2012).

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controversy surrounds the goal to make the guidelines sensitive enough to diagnose those who have the disorder, and specific enough to exclude those who don’t.

CONSTANT FLUX

D IAGNO SIS

Redefining autism Draft diagnostic guidelines are raising concerns that mild forms of the disorder may no longer be recognized. B Y E M I LY S I N G E R

he word ‘autism’ once invoked the picture of someone with severe problems, serious challenges in communications and rigid behavioural patterns — perhaps someone like Dustin Hoffman’s character in the 1988 movie Rain Man. These days, the perception of autism tends to be one of a much milder condition — people may know someone with a child with some form of autism, many of whom go to regular schools. The definition of autism has changed many times since it was first described in the 1940s. It has evolved from so-called childhood

schizophrenia in the 1950s and 1960s, to infantile autism in the 1980s, to the broad autism spectrum disorder (ASD) we know today. The fifth and newest edition of psychiatry’s diagnostic bible, the Diagnostic and Statistical Manual of Mental Disorder (DSM-5), is due to be published in May 2013. Four years in the making, the DSM-5 has another stab at refining our concept of autism. The process is always closely watched because the guidelines have broad impact: they are used in the United States and many other countries, by clinicians, pharmaceutical companies, researchers and insurance companies to define psychiatric conditions. In this case,

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The two biggest changes in the DSM-5 are a shift from three classes of symptoms to two, and the collapse of a number of different autism-related disorders, including Asperger’s syndrome (which describes people who have some of the social-interaction deficits common in autism but no impairments in language) under the single heading of autism. The proposal to erase the boundaries between distinct autism-related disorders has raised fears that some children would no longer meet diagnostic criteria under the DSM-5 definition, making it difficult to get the help they need. “My worry is there will be a substantive change in the way we make diagnoses, and that will complicate the lives of researchers and families,” says Fred Volkmar, director of the Yale University Child Study Center in New Haven, Connecticut, and a critic of DSM-5. Supporters of the DSM-5’s autism definition counter that the changes are based on a better understanding of the condition’s symptoms. What’s more, they say, the preliminary studies indicate that few will be excluded. “To the public it seems like the changes are huge and capricious,” says Sally Ozonoff, an expert in autism diagnosis at the MIND Institute at the University of California, Davis, who was not involved in the revisions. “But it’s based on a very exhaustive review of all the research, and I think they got it right.” Adding fuel to the fire is that unlike previous versions, the DSM-5 guidelines have been open to public comment as they were developed. The openness of the process has made for heated — and sometimes emotional — reactions. “It hurt like hell that newspapers were accusing us of trying to hurt kids by denying them services,” Susan Swedo, chair of the working group charged with revising the guidelines, told a packed lecture hall at an autism conference in Toronto, Canada, in May 2012. Swedo, a specialist in behavioural paediatrics at the National Institute of Mental Health in Rockville, Maryland, says the new guidelines are very similar to the old ones. “What’s really different is there is more guidance for clinicians,” she says, such as how different symptoms might look in toddlers or young adults. The goal of expanding these descriptions, she adds, was to make the guidelines more sensitive to autism in groups that are often overlooked, such as girls and minorities.

A CRITERION FOR CRITERIA

One of the aims of the new guidelines was to help standardize diagnosis. A 2011 study by a team led by psychologist and autism researcher Catherine Lord of Weill Cornell Medical

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OUTLOOK AUTISM

COURTESY OF YALE SCHOOL OF MEDICINE

AUTISM OUTLOOK College in New York suggests that although clinicians give similar scores on autism screening tests, the label they attach to that score — such as classical autism or the milder Asperger’s syndrome — is hugely variable1. To develop the new autism criteria, researchers analysed data from several large data sets that included children with classical autism, autism-related disorders and other diagnoses, such as language delay, intellectual disabilities and attention-deficit hyperactivity disorder. The children had each been screened with two widely used tests: the Autism Diagnostic Observation Schedule (ADOS) and the Autism Diagnostic Interview-Revised (ADI-R). To determine what should be included in the criteria, the researchers first ran a statistical analysis designed to isolate the factors responsible for the diagnosis of an autismrelated disorder. They then created different versions of the DSM-5 and tested how accurately each identified children with autism of different ages, gender and language ability. “This allowed us to look at the proportion of children with various clinical diagnoses who would be included or excluded from ASD with the new criteria,” says Lord, a member of the DSM-5 working group and the creator of both the ADOS and ADI-R. The DSM-5 collapses the current triad of impairments as described in DSM-IV — deficits in social behaviour, language delay, and repetitive or restrictive behaviours — into just two main classes: social communication difficulties, and repetitive or restrictive behaviours. “It was really kind of random whether a particular behaviour was designated as social or non-verbal communication [in the DSM-IV],” says Lord. “When we looked at various large data sets, there were two separate factors in ASD.” DSM-5’s simplification, she says, more clearly reflects the reality of the condition. Two studies that analysed different sets of data support this change2,3. The conflation of communication and social reciprocity into one category fits very much into our clinical experience,” says David Skuse, a developmental neuropsychologist at University College London who led one of the studies. (Skuse is not a member of the DSM-5 group.) The more controversial change in the DSM-5 is to bring together a number of formerly distinct disorders under the single heading of autism. The DSM-IV recognized several disorders as being distinct, including classical autism, Asperger’s syndrome and pervasive development disorder not otherwise specified (PDD-NOS) — a category that applies to many children who meet some, but not all, of the criteria for autism. Some autism researchers say there is little scientific basis for defining these as different disorders. “Replicable research findings show that it’s almost impossible to discriminate people with autism from Asperger’s syndrome if you control for intelligence and language level,”

says Lord. As for PDD-NOS, “when you look at the research, it’s basically less severe autism,” she says. “No one has come up with characteristics that pull out that group.” One potential impact of the redefinition that its opponents have cited is that some people with Asperger’s syndrome and PDD-NOS will no longer qualify for an autism diagnosis and the support services that come with it. For example, Volkmar and his collaborators reanalysed data collected via questionnaires for the DSM-IV field trials4, linking it to criteria outlined in the DSM-5. Their 2012 analysis

Fred Volkmar is a critic of plans to redefine autism.

suggests that only about 60% of 1,000 people diagnosed with autism in 1994 under DSM-IV would meet the new diagnostic criteria. A handful of studies published over the past year support Volkmar’s concern5–8. “All show a big drop off in numbers of people diagnosed with ASD under the DSM-5,” says Johnny Matson, a psychologist at Louisiana State University in Baton Rouge and author of some of these studies. “I would rather see a tweak than a major change.” But critics of these studies point out that they are retrospective, meaning they used old data, often from limited questionnaires. Some of the symptoms in the new guidelines, such as sensory deficits, may simply not have been collected in the past. The result, they say, is that individuals who would meet the diagnostic criteria if assessed today would not be considered autistic based on the previously collected information. Retrospective studies are “always going to underestimate sensitivity, because they didn’t ask all the questions”, says Lord. The Volkmar study was “fatally flawed because it used data to do the analysis that couldn’t answer questions being asked”, says Swedo. “Just because you have an idea of how a child met criteria for the DSM-III or DSM-IV

doesn’t mean it can be applied to the DSM-5.” A new study, published this month in the American Journal of Psychiatry by Lord and collaborators, uses a much richer data set to show that the new guidelines are just as sensitive as the DSM-IV and unlikely to exclude many9. Experts on both sides agree that the real answer will come from prospective studies, meaning that new data will be collected as participants are simultaneously evaluated with the DSM-IV and the DSM-5. “We won’t know until [results from those] trials are published,” says Ozonoff. Such research is now underway. The research and advocacy organization Autism Speaks is funding a prospective study, as is the American Psychiatric Association (APA), which publishes the DSM. Preliminary results from the APA field trials, presented at the International Meeting for Autism Research in Toronto in May 2012, are in line with Lord’s study and suggest that the new guidelines are unlikely to exclude many people with an ASD. Ozonoff, who is broadly supportive of the DSM-5 guidelines, says one flaw is the creation of a diagnostic category called pragmatic, or social, communication disorder. This label will apply to children who show some of the social communication deficits of autism but not the repetitive and restrictive behaviours. These children are often diagnosed with PDD-NOS. Unpublished data from Skuse’s group suggest that about onethird of those people are likely to be shifted to social communication disorder. “It seems very similar to autism spectrum disorder,” says Ozonoff. “I wonder about the rationale for creating a new diagnosis unless we have good data that it’s different.” Just as Asperger’s syndrome came and went as a form of autism, the same trajectory may be in store for this latest subcategory. The only way to resolve the dispute is more research, says Volkmar, who in the 1980s helped evaluate the current criteria in the fourth edition. “Ultimately,” he says, “it’s not a point for opinion — it’s a point for data.” ■ Emily Singer is news editor at SFARI.org, the website of the Simons Foundation Autism Research Initiative in New York City. 1. Lord, C. et al. Arch. Gen. Psychiatry 69, 306–313 (2012). 2. Mandy, W. P. et al. J. Am. Acad. Child Adolesc. Psychiatry 51, 41–50 (2012). 3. Frazier, T. W. et al. J. Am. Acad. Child Adolesc. Psychiatry 51, 28–40 (2012). 4. McPartland, J. C. et al. J. Am. Acad. Child Adolesc. Psychiatry 51, 368–383 (2012). 5. Worley, J. A. & Matson, J. L. Autism Spectr. Disord. 6, 965–970 (2012). 6. Mandy, W. et al. Autism Res. 4, 121–131 (2011). 7. Taheri, A. & Perry, A. J. Autism Dev. Disord. 42, 1810–1817 (2012). 8. Matson, J. L. et al. Dev. Neurorehabil. 15, 185–190 (2012). 9. Huerta, M. et al. Am. J. Psychiatry 169, 1056–1064 (2012).

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have become good at stealing drugs developed for other disorders,” jokes Hardan. But even second-hand drugs are better than nothing, given how disruptive these behaviours can be.

STRIKING A BALANCE

A seven-year-old boy with autism is given oxytocin nasally to see if it alters his sociability.

T REAT MENTS

In the waiting room After years of making do with drugs developed for other conditions, doctors and scientists are eagerly pursuing drugs that target the social symptoms of autism. BY MICHAEL EISENSTEIN

hy would any parent feed their child worm eggs? In Stewart Johnson’s case, it was because he believed he had stumbled across a promising treatment for the symptoms of his teenage son’s autism after limited success with other treatments. Although he is not a professional scientist, his bold experiment with porcine whipworm (Trichuris suis) ova has culminated in a formal clinical trial spearheaded by a leading autism research centre. However, Johnson’s story also tells of the desperation felt by many parents to find something — anything — that might improve the lives of children with autism spectrum disorders (ASDs).

That desperation is borne of the limited success achieved in developing drugs to treat autism. “When I give a lecture on medication, I usually start by saying that I won’t be talking about the state of pharmacology in autism, but about the miserable state of pharmacology in autism,” says Antonio Hardan, a child psychiatrist at Stanford University School of Medicine in California. In fact, no drugs are approved by the US Food and Drug Administration (FDA) for autism per se, although a handful of medications approved for “Talking about other psychiatric indiimmune stuff cations have proven was considered useful in mitigating fringe and some of the sympfunky.” toms of ASDs. “We

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‘Irritability’ may sound like a minor failing, but the behaviours it encompasses — mood swings, aggression and self-harming — can be debilitating and dangerous to people with autism and their caregivers. That’s why irritability is a primary target for pharmaceuticals in ASDs. Two antipsychotic drugs, aripiprazole (marketed as Abilify by Otsuka Pharmaceuticals) and risperidone are now in widespread use in people with ASDs. “These drugs are effective in 60–70% of children and adolescents that have significant irritability,” says Christopher J. McDougle, director of the Lurie Center for Autism at Massachusetts General Hospital in Boston. Both drugs, however, come with side effects. Jeremy Veenstra-VanderWeele, a child psychiatrist and researcher at Vanderbilt University Medical Center in Nashville, Tennessee, has determined that aripiprazole and risperidone frequently induce drowsiness and weight gain, and he cautions against prescribing these drugs unless symptoms are severe. “When you’re using a medicine in a seven-year-old child that increases their body weight by 10% over the course of a couple of months, that potentially changes the way their metabolism is going to work down the line,” he says, noting the chance for increased risk of metabolic syndrome or cardiovascular disease. Many clinicians prescribe anti-depressants from the selective-serotonin reuptake inhibitor (SSRI) family, such as fluoxetine (Prozac) and citalopram, to treat ASD patients, despite scant evidence supporting their use. Randomized controlled trials have shown that SSRIs may reduce compulsive, repetitive behaviour in adults with autism, but have no observed effect on these symptoms in children and adolescents. But these trials did not examine social anxiety, a symptom for which some doctors have observed improvements in children with SSRIs. McDougle, who pioneered the study of SSRIs for ASDs while working at Yale University School of Medicine in the 1990s, believes factors related to ageassociated brain development may also limit their use. “One reason some children didn’t get better was that they couldn’t tolerate even low starting doses,” he says. “They became much more irritable, agitated and aggressive.” Since the value of both SSRIs and antipsychotics is based largely on the extent to which they enable children with autism to participate in school, caregivers must weigh the potential medical benefits and adverse effects carefully. “If we’re putting kids to sleep, even if their aggression is down, we haven’t actually facilitated their psycho-educational intervention,” says Evdokia Anagnostou, a child

UNC ASPIRE RESEARCH PROGRAM

OUTLOOK AUTISM

ILANIT GORDON, YALE CHILD STUDY CENTER

AUTISM OUTLOOK

A C T I VAT I N G T H E A U T I S T I C B R A I N Functional MRI images from a small, preliminary study (7 participants) show that the administration of oxytocin could potentially change brain patterns in a way that would reduce the symptoms of autism.

Medial prefrontal cortex (decision-making and social judgment)

O X YTO C I N

Cingulate cortex (cognitive and emotional behavior)

Precuneus (memory and consciousness)

Insula (self-awareness and perception of social cues)

Temporal parietal junction (attention, self-perception and theory of mind)

R Superior temporal sulci (responds to voice and gaze of others)

PLA C E B O

Inferior temporal sulci

neurologist at the Bloorview Research Institute in Toronto, Canada.

THE HEART OF THE MATTER

No medications target the core symptoms of autism, which include attention deficit, abnormal social behaviour and poor ability to communicate. Accordingly, when a potential treatment option for these symptoms is identified, it grabs the community’s attention. For example, extensive research in both animals and humans has identified a prominent role for the hormone oxytocin in behaviours associated with trust, empathy and attachment — emotions commonly impaired in people with ASDs. At least half-a-dozen clinical trials are underway to test whether oxytocin affects these traits in people with ASDs. The Swiss pharmaceutical company Novartis makes a nasal oxytocin spray, Syntocinon, and this is the primary mode of delivery in development. Veenstra-VanderWeele cautions that the benefits remain unclear. “Kids with ASD are having oxytocin squirted up their noses, and we need to know if that makes any sense,” he says. Early studies have shown modest improvements — for example, an increased tendency to make eye contact or elevated activity in social centres of the brain. But most of these investigations have looked at the impact of single doses in small numbers of patients. A new round of larger-scale initiatives should help clarify both the efficacy and the longterm safety of oxytocin use. For example, a treatment network funded by the US National Institutes of Health through its Autism Centers of Excellence programme is set to embark

on an ambitious five-year study of 300 children and adolescents to examine whether extended treatment with oxytocin confers measurable improvements in social function. One of the biggest obstacles confronting clinical studies of oxytocin — and, indeed, any treatment of ASD’s core symptoms — is the lack of robust, reproducible measurements of social deficits and improvements in those deficits. “Social behaviour is so complicated, and trying to measure changes in it is really tricky,” says McDougle. This is especially true for early-stage experimental drugs, which target neurological pathways with unclear contributions to ASD and in which the therapeutic effects are likely to be weaker than for approved drugs such as risperidone. Furthermore, the inclusivist definition of autism as a spectrum of disorders overlooks very real differences. “Autism is a heterogeneous disorder, and it’s very unlikely that you’re going to find a compound that’s effective in the treatment of social deficits in all kids with autism,” says Hardan.

FRAGILE X MARKS THE SPOT

Some of the most exciting progress has emerged from work focused on ASDs with a clearly defined genetic cause. For example, research by Mark Bear, a neuroscientist at the Massachusetts Institute of Technology in Cambridge, has shown that defects in brain signalling mediated by the neurotransmitter glutamate appear to be critical to the pathophysiology of fragile X syndrome (FXS), an inherited disability in which patients often manifest autistic-like social deficits.

FXS is caused by mutations in the gene encoding a protein that regulates the production of other proteins involved in neuron signalling. Bear and colleagues discovered that the protein produced by the mutated gene represses a glutamate receptor called mGluR5. Bear hypothesized that the neurological symptoms of FXS might arise from overactive mGluR5, so his group studied animal models of FXS to test this theory1. “We took fragile X mice and engineered them to have 50% of the normal level of mGluR5, with the prediction that this would correct multiple aspects of the disease, and it did,” says Bear. “That was a powerful proof of principle.” Since then, numerous drug candidates have been identified that can potentially correct mGluR5-associated deficits — Novartis, Swiss drug giant Roche, and Seaside Therapeutics, based in Cambridge, Massachusetts, and co-founded by Bear, all have agents in clinical trials. Mouse models of FXS have proven useful in testing these candidates, as they directly mimic the genetic and cellular-scale defects seen in FXS, even if they do not always recapitulate the full spectrum of behavioural and social symptoms. Most encouragingly, a collaboration between Bear’s team and Lothar Lindemann’s research group at Roche has discovered that mGluR5 inhibitors can even repair FXS-associated symptoms in grown mice2 — suggesting that the window for repairing cognitive damage may be far wider than expected. “That’s something we wouldn’t have imagined 10 years ago,” says Veenstra-VanderWeele, who was not involved in the study.

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OUTLOOK AUTISM Novartis has published data from a phase II trial of its mGluR5 inhibitor, AFQ056. The results suggest that the drug may have positive behavioural effects in a subset of adult FXS patients 3 , but the size of the cohort that benefited was too small to be conclusive. The company is now recruiting for a larger phase III trial. Seaside has also announced (but not published) encouraging phase II results in treating social symptoms of FXS with arbaclofen, a drug that inhibits glutamate release. This spate of trials announces the entry of the pharmaceutical industry into a sector that it had long shied away from, owing to issues of complexity and a lack of concrete targets. “I think they are getting more interested — it’s a huge market out there, and the science is getting intriguing,” says neuroscientist Gerald Fischbach, scientific director of the New York-based Simons Foundation Autism Research Initiative. There is also hope, but not yet supporting evidence, that the deficits seen in FXS may share common roots with some other ASDs. “We need to understand the pathophysiology of each of these models and look for points of convergence, but we should not make assumptions about what those might be,” says VeenstraVanderWeele. Nevertheless, the key role of glutamate signalling in the brain has given some scientists cause for optimism, and several trials are underway to examine the potential benefits of repurposing other known glutamatergic modulators. For example, Anagnostou and colleagues are conducting pilot studies with memantine (marketed as Namenda by Forest Laboratories) and riluzole (marketed as Rilutek by Sanofi-Aventis), which were approved by the FDA for Alzheimer’s disease and amyotrophic lateral sclerosis respectively. “This is a very rich area to look for molecular targets,” she says.

PICKING UP HINTS

All the same, there is a very real risk that treatments for patients with ASD subsyndromes such as FXS or the neurodevelopmental disorder Rett syndrome will benefit only a small subset of people with autism outside these categories. Some researchers further caution against drawing overly broad connections between these conditions that may be relatively superficial or even misleading. “Everybody says people with those syndromes also exhibit characteristics of autism,” says McDougle, “but I can tell you that if you know autism, then you can see that the impaired social relatedness in those disorders is quite different — it’s not autism.” Categorization of patients could be improved by the identification of biomarkers — molecular or physiological traits that correlate with specific symptoms or manifestations of ASD. Such markers could steer patient selection for clinical trials, as well

as illuminate autism’s still-mysterious aetiology. Several candidate genes have been identified, although they have not yet been woven into a coherent model. In April 2012, a large-scale gene sequencing study of 343 families revealed more than 300 potential susceptibility factors — and an unexpectedly strong link to FXS, with many genes encoding targets regulated by fragile X mental retardation protein (FMRP)4. “This will still need to be settled with empirical studies in humans,” says Bear. In some cases, causative mutations may be exceedingly rare or even limited to certain bloodlines. For example, one recent genetic study identified a link between a defect in a metabolic gene and autism in two related families, and subsequent animal studies suggested that autistic individuals with this mutation might benefit from dietary supplementation5. This finding is unlikely to help the vast majority of people with autism, however, and the search continues for broader aspects of ASD pathology. Some researchers are also begin“We have ning to investigate become good at the contributions stealing drugs of i n f l am m at or y developed immune responses for other and autoimmundisorders.” ity in causing, or at least exacerbating, autism. “Not long ago, talking about immune stuff was considered fringe and funky, and I’m not fringe or funky,” says McDougle. “But I think that immune dysfunction is going to prove to be important in a very meaningful subgroup of patients — maybe as many as 10–25%.” While the evidence for this correlation remains circumstantial, other scientists are taking the possibility seriously. Stewart Johnson’s maverick experiments with whipworm eggs were informed by research data suggesting that the pig parasitic worms, which do not permanently colonize the human gut, can suppress harmful inflammatory responses6. Johnson reported improvement in many of his son’s behavioural symptoms, and his work was sufficiently persuasive to convince psychiatrist and autism specialist Eric Hollander at the Albert Einstein College of Medicine in New York to embark on a formal clinical trial of this treatment people with ASD . Feedback from parents has led to other investigations of therapeutic modalities besides the whipworm. For example, input and financial support from parents led Hardan to conduct a clinical trial examining N-acetylcysteine, an antioxidant and modulator of glutamate signalling, as a potentially milder alternative to antipsychotic drugs for irritability symptoms7. Fischbach notes that parents of children with autism helped call attention to a surprising observation that could offer novel insights into ASD pathology.

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“One-quarter of the people in our study group of almost 3,000 families report that when their children have a mild fever, their social interaction improves — some start speaking or making more eye contact,” he says. “We are trying to formulate controlled trials to understand that phenomenon better.” Not every anecdotal finding pays dividends — Veenstra-VanderWeele cites the prominent example of secretin, a digestive hormone that was described as an effective treatment for core deficits in autism based on a case report8 of three children published by a research group in 1998, but it ultimately led researchers down a costly rabbit hole. “We figured out three or four years later that it does absolutely nothing,” says VeenstraVanderWeele. “People were spending a lot of time and effort pursuing that instead of something with more evidence behind it.” The problem is that parents, desperate for treatment options, are confronted by a staggering amount of ambiguous data — and misinformation — and clinicians must take care to keep parents informed of what has been scientifically confirmed. Anagnostou says her top priority is to steer families pursuing alternative approaches away from known hazards, such as excessively high doses of vitamin B6 or omega-3 fatty acids, or unsafe treatments such as chelation therapy, which entails the use of compounds intended to remove heavy metals from the body. “I take all of the non-evidence-based things that they’re interested in and tell them what’s dangerous, and then I hope that they will stay in the non-dangerous territory,” she says. “But the truth is, we haven’t given them that many options.” With luck, this will change as the current round of clinical studies concludes. Clinicians and researchers eagerly await news of the first drug to successfully chip away at autism’s core symptoms. However, VeenstraVanderWeele points out that even a runaway success on the pharmacological front will only be the starting point for effective therapy, enabling people with autism to receive the behavioural and educational training they need to function in society. “You can set the brain up to learn,” he says, “but you still need to teach.” ■ Michael Eisenstein is a freelance science writer based in Philadelphia, Pennsylvania. Dölen, G. et al. Neuron 56, 955–962 (2007). Michalon, A. et al. Neuron 74, 49–56 (2012). Jacquemont, S. et al. Sci. Transl. Med. 3, 64ra1 (2011). Iossifov, I. et al. Neuron 74, 284–299 (2012). Novarino, G. et al. Science doi: 10.1126/ science.1224631 (2012). 6. Summers, R. W. et al. Am. J. Gastroenterol. 98, 2034–2041 (2003). 7. Hardan, A. Y. et al. Biol. Psychiatry 71, 956–961 (2012). 8. Horvath, K. et al. J. Assoc. Acad. Minor Phys. 9, 9–15 (1998). 1. 2. 3. 4. 5.

AUTISM OUTLOOK

PERSPECTIVE Imaging autism

Several studies in the past two years have claimed that brain scans can diagnose autism, but this assertion is deeply flawed, says Nicholas Lange.

study published recently in the widely read clinical journal Radiology correctly identified 36 out of 39 children and adolescents with autism solely by the use of functional magnetic resonance imaging (fMRI)1. The researchers reported that as the subjects listened to their parents’ voices, those with autism had lower brain activity in the superior temporal gyrus, a brain region involved in language reception, compared with controls. Although this finding makes sense, we should be cautious about its interpretation. This is not an autism-detecting brain scan. Language deficits are a core and defining feature of the disorder; one doesn’t need a brain scan to show this. And there are many people with language difficulties who do not have autism. Therefore, a technology that detects language problems doesn’t move the ball towards a differential diagnosis. To diagnose autism reliably, we need to better understand what goes awry in people with the disorder. Until its solid biological basis is found, any attempt to use brain imaging to diagnose autism will be futile. I agree with my colleagues who have said that any candidate biomarker for any disorder is of no clinical use unless we first establish a stable and biologically valid concept of the illness2. During the 70 years since psychiatrist Leo Kanner originally observed and named autism, its diagnostic criteria (see ‘Redefining autism’, page S12) have not included any biological measurements. Elevated blood level of the neurotransmitter serotonin is its only candidate biological marker to date, but it is hardly specific to autism. For instance, tumours in the gastrointestinal tract and recreational drug use also increase blood serotonin. Given this absence of a simple physical test for autism, it’s tempting to think that a 5- or 10-minute brain scan can do the trick, by quickly and accurately identifying something distinctive in the brain of an individual with autism. But we don’t yet know its biological underpinnings, and we cannot expect brain imaging to diagnose autism without that a priori biology. To believe otherwise is to place false confidence in technology3, and potentially to mislead or even harm affected individuals, their parents, families and communities. Brain imaging does of course have a place in autism study. While it has little if any diagnostic utility at present, this technology has already helped us understand the disorder. Volumetric MRI has shown us that about one in five children with autism has an early abnormal enlargement of the brain that tends to stabilize during the first 18 months of life4,5. Functional MRI has helped us learn where people with autism focus during social interaction, or when watching films with intense social content. Positron emission tomography (PET) has begun to show us differences in the distribution of serotonin and dopamine receptors — and of the evolving roles of serotonin throughout life — in the brains of people with autism compared with typical controls. Indeed, if the underlying autism pathology puzzle is solved, PET may provide some guidance as a marker of drug effectiveness. Similarly, magnetic

resonance spectroscopy may eventually prove helpful in treatment studies. For instance, it may help quantify differences between the neurotransmitters glutamate and gamma-aminobutyric acid (GABA) in people with autism and those with schizophrenia. A correct diagnosis of autism often benefits an individual. A false diagnosis can do harm. It is therefore essential — but unfortunately rare — to publish the positive predictive value of a proposed test (the proportion of correct diagnoses among all diagnoses). One of my own autism studies offered an MRI-based classification algorithm (not diagnostic) with the highest overall ability to date, having 90% positive predictive value6. But, like other such studies, mine was limited by small sample and effect sizes and requires external replication. What individuals with autism and their parents urgently need is for us to conduct large, long-term, multicentre studies to identify physical features unique to the brains of individuals with autism. We should pay particular attention to differences in genetics, physiology, immune challenge profile, neurochemistry and brain connectivity between individuals with autism, those with other developmental disorders, and typically developing individuals. These studies need to enrol patients across the entire spectrum, and should use existing reference imaging databases of typical brain development. No one has conducted a study of this magnitude or potential authority, and reductions in research funding make it a tough sell. Yet because autism is an individually heterogeneous disorder, it is only in this way that we can achieve the goals of a future ‘stratified psychiatry’2. This term refers to a newly proposed system for all psychiatric disorders in which homogeneous subgroups of individuals are defined by dimensional combinations of biological and behavioural features — such as an altered neurotransmitter pattern and a history of major depression — rather than by categorical criteria. Autism prevalence worldwide is rising rapidly. In the United States, prevalence jumped by 78% between 2002 and 2008 (ref. 7). Larger numbers and solid biology may mean, however, that we can power our studies with enough participants to find reliable, sensitive and specific markers of the disorder with clinically acceptable predictive value, and improve the lives of people with autism.

UNTIL ITS SOLID BIOLOGICAL BASIS IS FOUND, ANY ATTEMPTS TO USE BRAIN IMAGING TO DIAGNOSE AUTISM WILL BE FUTILE

Nicholas Lange is a biostatistician at Harvard Medical School in Boston, Massachusetts. email: nlange@hms.harvard.edu 1. Lai, G. et al. Radiology 260, 521–530 (2011). 2. Kapur, S., Phillips, A. G. & Insel, T. R. Mol. Psychiatry doi: 10.1038/mp.2012.105 (2012). 3. Racine, E. et al. Nature Rev. Neurosci. 6, 159–164 (2005). 4. Nordahl, C. W. et al. Proc. Natl Acad. Sci. USA 108, 20195–20200 (2011). 5. Lainhart, J. E. & Lange, N. J. Am. Med. Assoc. 306, 2031–2032 (2011). 6. Lange, N. et al. Autism Res. 3, 350–358 (2010). 7. http://www.cdc.gov/ncbddd/autism/addm.html. 1 NOV E M B E R 2 0 1 2 | VO L 4 9 1 | NAT U R E | S 1 7

of London, who worked with children with autism from ethnically diverse backgrounds in east London1. “Other cultures might have very different expectations of how children behave.” This viewpoint makes it challenging to use behavioural diagnostic tests for autism in places where the disorder may look — and even be — different from in the West. But with growing interest in autism’s true prevalence worldwide and the need for autism services in poor countries, researchers are grappling with the best ways to objectively diagnose the disorder. Parent support groups for autism exist in more than 100 countries. “We know that autism is diagnosable and observable across cultures,” says Mayada Elsabbagh, a researcher at McGill University in Montreal, Canada, who is leading a group within the International Society for Autism Research on cross-cultural issues. “But the exact details of how different cultures or settings modify autism is unknown,” she says. For many years, the relevance of culture to autism was ignored. Some researchers believed that autism was intrinsically linked to modernity and Westernization, and was rare in other cultures. Others assumed that because autism is a neurobiological disorder, its expression should be the same everywhere2. But many researchers are beginning to take a subtler point of view. “While autism itself, the neuropathology of it, may not be culturally determined, our interpretation of those behaviours and our response to those behaviours is,” says David Mandell, associate director of the Center for Autism Research at the Children’s Hospital of Philadelphia in Pennsylvania.

DIAGNOSTIC DIFFERENCES

CULT URE

Diverse diagnostics The study of autism around the globe must account for a variety of behavioural norms in different societies. BY SARAH DEWEERDT

n rural South Africa, young children may look at adults’ faces while having a conversation, but they don’t usually make direct eye contact because it is considered disrespectful. Yet a lack of eye contact is a hallmark of social deficits in people with autism, and as such it is something Western clinicians look for when diagnosing the disorder.

There are other examples of children’s behaviour — such as finger pointing to draw attention to something, or conversing with adults as if they are peers — that are commonplace in the West and included in tests of autism. “Most autism research originates in the West, and we have a particular view of what autism is, a particular view about how children behave and interact with adults,” says Courtenay Norbury at Royal Holloway, University

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Before they can assess autism worldwide, researchers must measure how often various behaviours occur in different cultures and establish the norm, says Charles Zaroff, a psychology researcher at the University of Macau in China. They will also need to work with parents of children with autism to identify how it might manifest in a given culture. In many Asian cultures, for example, children are expected to express respect for their elders through their language and behaviour, but these aspects of social interaction can be tricky for children with autism to master. Such difficulties would hardly be noticed in much of the West. “Lacking that deference would appear completely appropriate in the United States, but that lack of attention to the strata in society based on age would appear very abnormal in places like China,” Zaroff says. Because the most widely used screening tests for autism were developed in the United States and the United Kingdom, researchers are finding that they have to adapt the tests to identify autism in other countries. For example, part of the Autism Diagnostic Observation Schedule (ADOS), one of the gold-standard diagnostic instruments for the disorder, involves observing a child having a pretend birthday party

RICHARD JONES/REX FEATURES

OUTLOOK AUTISM

FUSE/ GETTY IMAGES

AUTISM OUTLOOK — singing ‘Happy Birthday’, cutting and distributing slices of cake, and so on. But in rural areas of South Africa, birthdays often aren’t celebrated, so even typically developing children might be unfamiliar with this ritual. So, for a study of early autism diagnosis in KwaZulu-Natal province, researchers developed an alternative scenario of shared excitement, involving a traditional African song. “It’s finding the intention of what you’re trying to elicit, and then finding an alternative,” says Amy Wetherby, director of the Autism Institute in the College of Medicine at Florida State University in Tallahassee, who led the work. Wetherby developed a list of 22 early signs of autism based on studies of several hundred children in Florida. Unpublished data from 19 children show that many of the same red flags differentiate children with autism from their typically developing peers in South Africa as well, says Wetherby. “The amazing part to me is the unusual gestures,” says Wetherby. For example, rather than pointing or looking together at an object, children with autism in the United States may communicate by taking an adult’s hand and moving it to the object. “That’s an early sign of autism, and we see that [in South Africa] as well.” One reason the patterns of autism symptoms appear to be similar across cultures may be that the participants in this study are only 18–36 months of age. “The earlier we go,” Wetherby says, “the more similarities we will see.” More culturally specific or environmental symptoms of autism may emerge as children grow up. For example, according to one recent study, 5–12-year-olds with autism in the United States are more sensitive to sights and sounds than are children with autism in Israel — although the authors note that genetic, cultural and environmental factors might be an influence, as well as the parents’ reporting of their child’s behaviour3.

SUBTLE SPEECH

If cultural differences emerge later in childhood, this could further complicate the diagnosis of autism. The disorder tends to be diagnosed later outside the United States and Western Europe, in part because of a lack of awareness of both developmental norms and autism. A parent who notices that a child is withdrawn or has a language delay may not recognize that as a symptom of autism. In-depth interviews in Goa, India, show that the parents there aren’t attuned to early social and communication milestones, and they usually become alarmed only when a child starts preschool and has trouble connecting with peers4. “What really concerns the parents initially is, ‘he’s not fitting in with everybody else’,” says Gauri Divan, a paediatrician working with the child health organization Sangath in Goa. In some cultures, parents may notice symptoms that are not typically associated with autism. Among Latino migrant workers in

Florida, for example, “the first complaint seems to be that the child is a picky eater,” says Roy Richard Grinker, a George Washington University anthropologist in Washington DC, who is collaborating with Wetherby on a study of autism in this community. “But then, if you go into more detail, you start to see that these children the mothers are describing are probably going to fall on the autism spectrum.” Grinker speculates that these mothers are particularly aware of eating habits because they are poor and food is scarce. Some evidence suggests that doctors need to be trained to spot the signs of autism from

Picky eating might point to autism, but a parent noticing a child’s eating habits is cultural.

oblique comments made by parents. For example, Mandell says, white parents in the United States often emphasize a child’s lack of communication by saying, ‘my child doesn’t respond when I call his name’, while black parents tend to use phrases like ‘my child won’t mind me’. Doctors may be less apt to consider a diagnosis of autism when they think a parent is describing a disobedient child rather than a socially impaired one — possibly helping to explain why autism is diagnosed less frequently among black children.

RAISING RATES

If parents in different cultures developed the same sense of autism awareness as in the West, research suggests, autism prevalence around the world might look no different to — or may be even higher than — in the United States or the United Kingdom. Perhaps the most dramatic demonstration of this is a study of more than 55,000 children in South Korea, which estimated autism prevalence at 2.64% (ref. 5). That’s more than twice the autism prevalence in the United States estimated by the US Centers of Disease Control and Prevention, and more than 50 times higher than the South Korean government’s figure for autism prevalence of 0.046% (ref. 6). One reason for that higher estimate may be that the researchers screened children in the

general population for autism symptoms, rather than recruiting them only from clinics for autism and other developmental disorders. In South Korea, researchers suggest that one reason for the underdiagnosis in autism may be that the stigma attached to the disorder is particularly strong in that country7. The diagnosis of a Korean child with autism diminishes the marriage prospects of siblings, and it can even affect his or her parents’ careers. Parents often prefer that their child be labelled as having ‘reactive attachment disorder’, or ‘lack of love’ as it’s known in Korean, a diagnosis that affects the mother’s reputation. Still, about two-thirds of the children the South Korean study identified as having autism attended mainstream schools and were not receiving any autism-related services. This widespread mainstreaming raises the question of whether a Western-defined autism diagnosis is meaningful if children are able to function reasonably well in their cultural context. “That’s an interesting issue,” says Norbury, who questions “whether we should be worried about these kids, and whether we should be making families worried about them, if there wasn’t any kind of worry before.” Other children identified in the study may be what Koreans would call a ‘border child,’ a new term that is emerging to describe some who would probably be diagnosed with autism in the West7. “This is a child who is high-functioning enough to be in a mainstream school, but who has significant social impairment,” Grinker says. Parents prefer this label, he says, because it implies that the child is only impaired socially, not intellectually, and that the condition is temporary. In stigmatizing autism, South Korea is not unique. But Elsabbagh points out that attitudes towards a disorder often change as more resources and services become available. That, she says, is one powerful argument for more cross-cultural research on autism. Another is that it will advance understanding of the biology of autism. “We’ve constrained our participant pools to those of European ancestry, and we have also not considered very thoroughly some of the cultural determinants that may shape autism in different ways,” says Elsabbagh. “Taking a more global perspective would allow us to see that underlying commonality much more easily.” ■ Sarah DeWeerdt is a science writer based in Seattle, Washington. 1. Norbury, C. F. & Sparks, A. Dev. Psychol. (published online 5 March 2012). 2. Daley, T. C. Trans. Psychiatry 39, 531–550 (2002). 3. Caron, K. G. et al. Am. J. Occup. Ther. 66, e77–e80 (2012). 4. Divan, G. et al. Autism Res. 5, 190–200 (2012). 5. Kim, Y. S. et al. Am. J. Psychiatry 168, 904–912 (2011). 6. Kang-Yi, C. D. et al. J. Autism Dev. Disord, (published online 22 June 2012). 7. Grinker, R. et al. Autism Res. 5, 201–210 (2012).

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OUTLOOK AUTISM

PERSPECTIVE Brain scans need a rethink

Head movement can bias brain imaging results, undermining a leading theory on the cause of autism, say Ben Deen and Kevin Pelphrey.

Kevin Pelphrey

ne of the most popular and widely accepted theories on the cause of autism spectrum disorders attributes the condition to disrupted connectivity between different regions of the brain. This ‘connectivity hypothesis’ claims that the social and cognitive abnormalities in people with autism can be explained by a dearth of connections between distant regions of the brain1. Some flavours of this theory also predict more connections between nearby brain regions. Recent studies, however, have found that when a person moves their head while undergoing functional magnetic resonance imaging (fMRI) — a method that maps how different neuroanatomical structures of the brain interact in real time, its functional connectivity — it looks like the neural activity observed in autism. That’s a sobering discovery: it means that a major source of evidence for a leading hypothesis on autism, and one that several research teams have pursued for years, may arise from an artefact. Many studies have investigated functional connectivity in the brains of people with autism, and most have reported evidence supporting the connectivity hypothesis. These findings are consistent with results from some animal models of autism, and from studies using diffusion tensor imaging, which measures the bundles of fibres connecting parts of the brain. But three studies published in 2012 have come to the same conclusion: head motion leads to systematic biases in fMRI-based analyses of functional connectivity2–4. Specifically, motion makes it appear as if long-range connections are weaker than they really are, and that short-range connections are stronger than they really are. This bias affects all functional connectivity analyses, but it is particularly insidious for studies of autism. That’s because it would lead to precisely the patterns that have been observed in fMRI scans of children with autism, and because children with autism typically move more than unaffected children do. How can autism researchers overcome this bias? One approach would be to define a measure of head motion in each participant over the course of a scan — for instance, to compute the displacement of the head between consecutive time points, and average this over time. Researchers can then check that the autism and control groups are well matched on this measure, or include this value as a nuisance variable in regression analyses. The matching, however, would need to be precise: as one of the new studies showed, even a difference as small as 0.004 millimetre in average head motion across groups of patients can lead to significant differences in correlation strengths4. Furthermore, it is likely that motion artefacts can persist even when groups are matched on mean head motion. First, there is some evidence that head motion relates to functional connectivity measures in a nonlinear fashion2,4. If that’s true, it would not be sufficient to account for only linear effects of motion. Even if head motion doesn’t significantly differ across groups, a nonlinear function of head motion could.

Second, a given estimate of average motion can correspond to rather different scenarios — a few isolated but large movements, or constant small movements — which would have different effects on fMRI signals and on measures of functional connectivity. For instance, it has been shown that large, jerky movements lead to brief spikes in fMRI signal strength. Based on this observation, cognitive neuroscientist Steven Petersen and his colleagues at Washington University in St Louis, Missouri, propose a strategy for mitigating these head-motion artefacts2. They recommend removing periods of high motion. They have shown that this technique, which they call ‘scrubbing’, corrects at least some of the spurious correlations caused by head motion. This approach is promising but many questions remain. For instance, the extent of motion-induced correlations even after scrubbing is not well understood. Moreover, the optimal motion threshold for removing periods has not been studied in detail. Still, performing scrubbing in addition to group matching on average motion estimates and other standard noise-reduction methods represents current best practice in functional connectivity research. So far, only a few research groups have used scrubbing or similar techniques in fMRI studies on autism5,6. Given the importance of understanding whether previous results on functional connectivity in autism are real or merely motion-related artefacts, we hope that the authors of fMRI functional connectivity studies in autism (and particularly those reporting results consistent with the connectivity hypothesis) will reanalyse their data using these techniques. Scrubbing can be easily implemented using freely available software such as Artifact Detection Tools or by implementing custom modifications to the programs. Reanalysing data should be facilitated by tools provided by the US National Database for Autism Research, part of the National Institutes of Health. In addition, a trove of information has recently become publicly available in the Autism Brain Imaging Data Exchange — a collection of resting-state fMRI imaging data sets from 539 individuals with autism and 573 controls. Revisiting fMRI studies with these approaches would help establish whether there really is a connectivity deficit in the brains of people with autism.

A MAJOR SOURCE OF EVIDENCE FOR A LEADING HYPOTHESIS ARISES FROM AN ARTEFACT

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Ben Deen is a graduate student in brain and cognitive sciences at the Massachusetts Institute of Technology in Cambridge. Kevin Pelphrey is the Harris Associate Professor in the Child Study Center at Yale University in New Haven, Connecticut. email: kevin.pelphrey@yale.edu 1. 2. 3. 4. 5. 6.

Just, M. A. et al. Brain 127, 1811–1821 (2004). Power, J. D. et al. Neuroimage 59, 2142–2154 (2012). Satterthwaite, T. D. et al. Neuroimage 60, 623–632 (2012). Van Dijk, K. R. A. et al. Neuroimage 59, 431–438 (2012). Kennedy, D. P. et al. Neuroimage 39, 1877–1885 (2008). Deen, B. et al. International Meeting for Autism Research (Philadelphia, 2010).