Scientia 2021-2022

SCIENTIA A

JOURNAL

BY

THE

TRIPLE

HELIX

AT

THE

2021-2022

https://www.peakpx.com/en/hd-wallpaper-desktop-vnrvd

UNIVERSITY

OF

CHICAGO

Scientia

TABLE OF CONTENTS 4 Investigating the Phase Separation Behavior of the Transcriptional Repressor Yan in Drosophila Melanogaster Daisy Maslan is a second-year in the college. After taking the winter quarter advanced biology class with Professor Rebay, she became interested in fly genetics. In the spring of her first year, she joined the Rebay lab and was mentored by Saman Tabatabae who now attends Stanford as a graduate student. In addition to her biological work, she is also a history major and involved in MUN and UChi Votes.

5 The Impact of Mutations in Wolframin on Psychiatric Disorders Saira Munshani

12 Activation of Host Kinase Vps34 by Murine Norovirus Non-structural Protein NS4 Jessica Oros is a third-year student at the University of Chicago pursuing a major in Biological Sciences with a specialization in microbiology, as well as a minor in Philosophy. After graduation, she plans to attend graduate school to study virology.

16 Long Lasting Effects of Early Life Stress on the Brain and Epigenome Omar Kassem is a 4th year student at Uchicago. He is interested in studying the role of synaptic and intrinsic plasticity in memory formation and learning. He is currently developing his thesis on the structural spine plasticity in SK2 KO mice somatosensory cortex.

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2021-2022

ABOUT SCIENTIA Dear Reader, We at The Triple Helix have been thrilled to resume in-person activities this year. During the Fall quarter, we were particularly happy to host a fascinating panel discussion with UChicago Professors Paul Serrano and Luke Joyner on the intersection of art and science. During the Winter quarter, we were excited to be joined by Kristin McWharter, a professor and artist at the School of the Art Institute of Chicago, and UChicago historian Chris Carloy to discuss the history and development of video game design, and ways in which the pandemic has shaped our aesthetic and social experience of virtual media.

We are excited to bring you this latest edition of Scientia, featuring a research abstract which provides insight into how researchers use ongoing scientific work to pose new research questions, along with three full-length research articles which provide an in-depth look at how researchers investigate those questions, analyze the results, and communicate their conclusions to the Scientific community and to you, our reader. Read ahead to learn about the hand UChicago undergrads have had in exploring the behavior of the Yan gene in drosophila transcription, the role of Wolframin mutations in psychiatric disorders, the effect of Murine Norovirus on host enzymes, and the effects of early-life stress on the brain and epigenome.

We are incredibly grateful to the students who contribute to the Triple Helix as writers, editors, and members of the event and design teams. Without the passion with which they approach their work, and their dedication to share their love of scientific inquiry with others, this journal would not be possible.

Scientia is always looking to broaden our scope and expand the reach of our publication. If you are working on a research project you want to see in print, or if there is a profesor performing eye-opening research you would like to share, please consider writing for us! We encourage all interested writers to contact a member of our team. In the meantime, please enjoy this edition of Scientia, presented by The Triple Helix.

Sincerely, John Naughton and Thea Applebaum Licht Editors-in-Chief of Scientia

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Scientia

ABSTRACT

Investigating the Phase Separation Be havior of the Transcriptional Repressor Yan in Drosophila Melanogaster Daisy Maslan Rebay Lab, University of Chicago Abstract Transcription factors are essential to genetic regulation. However, we do not have a good understanding of how transcriptional factors localize at sufficient concentrations to organize complexes at enhancer regions to affect genetic output. My work studies phase separation as a model that may answer this question. Broadly, phase separation can be understood as the organization of cellular components into a membraneless organelle-like condensate that exists at a different phase than the rest of its environment. Previous work has found that some transcription factors form aggregates, now understood as phase-separated bodies, which I will refer to as condensates.

My work studies this model as it relates to the transcriptional repressor Yan in Drosophila Melanogaster; Yan is an evolutionarily conserved member of the E26 (ETS) transcription factor family, with its human paralogue referred to as Tel. Previous work from the Rebay lab has identified that Yan can form liquid-liquid phase separated condensates when overexpressed in the salivary glands of late third instar larva. My project aims to identify the mechanisms by which these phaseseparated condensates form, and, in the long term, to examine how the condensates influence Yan’s repressive function. My summer project had three goals: to assess the requirement for Yan DNA binding in condensate formation by generating a Yan mutant that can’t bind DNA; to ask whether the corepressor Groucho colocalizes to Yan condensates; and to identify tissues in which endogenous (rather than overexpressed) Yan forms condensates.

Hanson, MA. “Drosophilia female clip art. Specifically drosophilia melanogaster.” 11 Feb 2021.

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2021-2022 IN-DEPTH

The Impact of Mutations in Wolframin on Psychiatric Disorders Saira Munshani 1, Eiman Y. Ibrahim 2*, Ilaria Domenicano 2, and Barbara E. Ehrlich 2*

Key words: Wolfram syndrome, schizophrenia, bipolar disease, WFS1 mutation, mood disorders.

1 University of Chicago, Chicago, IL, United States 2. Yale University, New Haven, CT, United States

Abstract: Wolfram Syndrome is a rare autosomal recessive disease characterized by early-onset diabetes

mellitus,

neurodegeneration,

and

psychological disorders. Mutations in the gene WFS1, coding for the protein wolframin, cause Wolfram Syndrome and are associated with bipolar disorder and schizophrenia. This report aims to connect WFS1 mutations to their impact on protein expression and structure, which ultimately translates to altered cell function and behavioral alterations of an individual.

Methods: Published data were used to compile WFS1 mutations associated with psychiatric symptoms, both in homozygous patients and heterozygous carriers of WFS1 mutations. These mutations were evaluated in silico using SNAP2, PolyPhen-2, and PROVEAN to predict the effects of sequence variants. Statistical analysis was performed to assess the correlation between the locations of the mutations and the damage prediction scores.

Results: Several mutations, clustering in the center and C-terminus of the WFS1 polypeptide, such as A559T and R558C, are found in individuals with psychiatric diseases and appear particularly impactful on protein structure. Our analysis showed that mutations in all regions of wolframin were present in patients with schizophrenia whereas only cytoplasmic and ER luminal mutations were reported in patients with manic episodes and bipolar disorders.

According to Poly-Phen-2 predictions, 82.4% of the ER lumen mutations and 85.7% of the membrane mutations are damaging.

Conclusion: We propose mood disorders in Wolfram Syndrome and heterozygous carriers of WFS1 mutations are the consequence of specific

mutations in WFS1 that alter the structure of

Wolfram Syndrome affects nearly 30,000 people

wolframin, resulting in intracellular calcium

worldwide and is categorized as an orphan

dysregulations and impaired cell signaling,

disease. The pathophysiological changes that

Understanding the effect of WFS1 mutations

patients experience have been associated with

on bipolar disorder and schizoprenia is integral

various mutations in the WFS1 gene (classified

to designing clinically targeted treatments for

by Online Mendelian Inheritance in Man (OMIM

both diseases, which need more specialized

#222300). WFS1 encodes for wolframin, an

treatments.

endoplasmic reticulum (ER) associated protein, that is highly expressed in the heart, brain, lungs,

Introduction

inner ear, and pancreas (1, 5). Patients typically

Wolfram Syndrome Etiology and Diagnostic

are first diagnosed around age six with diabetes

Criteria

mellitus (1). Progressive loss of peripheral vision, Wolfram Syndrome is an autosomal

color perception, and sensorineural hearing loss

recessive progressive disease in which patients

usually occurs 4-5 years later (1, 6). Starting in

experience altered physical and psychological

early adulthood, more than half of the patients

functions. The cardinal manifestations include

develop neurological disorders, most commonly

diabetes mellitus coexisting with diabetes

manifested as ataxia, seizures, organic brain

insipidus, bilateral optic atrophy, hearing

syndrome, and peripheral neuropathy (1). During

and vision loss along with progressive motor,

this period, psychiatric symptoms, such as

autonomic and psychiatric abnormalities. (1,

depression, mania, and suicidal behavior also

2) Wolfram Syndrome is also known by the

occur (7). Brain stem atrophy is also a prominent

acronym DIDMOAD (diabetes insipidus, insulin-

feature and it is this effect that leads to an early

deficient diabetes mellitus, optic atrophy and

death, secondary to respiratory failure and

deafness). (3, 4) The course of Wolfram Syndrome

central apnea (8). Wolfram Syndrome is an

is progressive, and the prognosis is poor and

aggressive and fatal disease, with the majority of

typically terminal between 30 and 40 years of age.

patients with the disease dying before the age of

Genetic testing enables reliable diagnosis. (1, 3)

40.

5

Scientia Current Treatments Currently,

there

is

no

disease-

modifying therapy for Wolfram Syndrome.

Management of this disorder is limited to treatment and control of symptoms. Patients typically receive daily insulin injections and dietary

modifications

to

control

diabetes

mellitus (9). Several other approaches have been used for the management of Wolfram Syndrome such as annual screening for diabetes mellitus, fundus examination, and urodynamic testing (1, 10). There are ongoing clinical trials for disease specific treatments. There is a phase 2 double-blind, placebo-controlled drug directed to evaluate valproic acid (Clinical Trial Number: NCT03717909), a well-known mood stabilizer (11). A second trial is testing dantrolene (Clinical Trial Number: NCT02829268), a drug used to

Figure 2: A schematic of WFS1 with mutations that have been shown to be associated with mood disorder. FIgure was adapted from 2014

treat malignant hyperthermia (12). The efficacy

Matsunaga et. al mutation map (1).

and safety of these agents for Wolfram Syndrome with the endoplasmic reticulum (17). Although full manic episodes (19). Depressive episodes the function of wolframin is unclear, several often become the more prominent aspect of remain to be established.

WFS1

Mutations

and

Neuropsychiatric

Disorders Mutations in the WFS1 gene have been associated with cognitive abnormalities, in particular psychiatric disorders. These psychiatric disorders include bipolar disorder, schizophrenia,

depression,

and

suicide

tendency. Subjects who are either homozygous or heterozygous carriers for mutations in WFS1 have presented with psychiatric symptoms, ranging from mild to severe (13). The finding of WFS1 mutations in psychiatric subjects who do not have Wolfram Syndrome led to the suggestion that these mutations are associated with psychiatric symptoms (14) and that mutations in the gene WFS1 can be biomarkers and predictors for mood disorder (14, 15). Studies have shown that heterozygous carriers of WFS1 mutations (estimated to be 1% of US population) have a 26fold increased risk of having a mood disorder (16).

Pathogenic mutations of WFS1 are found distributed across the entire gene, which encodes an 890-amino acid polypeptide associated

cellular pathways necessary for normal cell the disorder as patients age. Although the signaling appear to be altered by mutations in pathophysiology of bipolar disorder is not known, WFS1 including intracellular calcium signaling there are environmental, behavioral, cellular, and and its downstream effects, ER stress, and molecular components that contribute to the mitochondrial

health,

neurotransmitter risk factors (20). Evidence that bipolar disorder

production and release (17, 18). Studies of the has a genetic basis was shown by the increased role of genetics in psychiatric disorders are susceptibility if other family members have been challenging because these disorders appear diagnosed with bipolar disease, where there is to entail a network of overlapping genes. a tenfold increased risk of bipolar disease if the Similarly, it should be noted that there are both patient has an affected parent (21). Twin studies misdiagnoses, nuances, and overlap with other have shown a 70-85% concordance of disease diseases in the characterization of psychiatric expression, regardless whether the twins were disorders. Here we focus on two distinct raised separately or together (21).

Schizophrenia

psychological disorders, bipolar disorder and

impairments

in

is

characterized

schizophrenia.

by

high-level

cognitive

Bipolar Disorder and Schizophrenia Etiology

functions such as memory, executive function,

Bipolar disorder is a mood disorder and attention along with irregular thought that affects approximately 2% of the human processes, social interaction, and decreased population and is typically characterized emotional responsiveness. Additional features of by frequent episodes that include mania, schizophrenia include delusions, hallucinations, hypomania, major depression, and mixed and difficulties in maintaining long-term episodes. Bipolar I disorder usually involves relationships. Although little is known about the a manic/mixed episode, whereas bipolar II genetics and molecular bases, genetic studies have must involve a major depressive episode and shown that schizophrenia and bipolar disorder hypomanic episode, with no requirement for share a number of the same genes related to the risk of disease (21). Thus, identifying genetic Figure 3: Distribution of mutations associated with psychiatric symptoms. WFS1 mutations associated

factors provides a guide to potential mechanisms

with mood disorder were categorized into two groups:

of action.

mutations identified in Wolfram Syndrome patients and

Study Aim

mutations identified in heterozygous carriers of WFS1 mutations.

In this report, we examined the relationship

between

reported

pathogenic

mutations in WFS1 and psychiatric disorders.

We propose that mutations in WFS1 predict

6

2021-2022 Syndrome.

Published papers that ranged from small patient studies to in depth genomic analyses were gathered. The literature was mined for relevant patient data and identification of mutations in WFS1. In addition, the references of each paper were examined for any other relevant papers. We excluded any study that does not address the full protein sequence of WFS1.

However, the paucity of studies that examined mutations in the N-terminus of WFS1 highlights that mutations in the first 310 residues of the protein are underrepresented.

Data synthesis through in-silico evaluation software: In order to select the relevant Figure 4: the SNAP2 predicted effect of mutations in WFS1 in with the SNAP2 predicted accuracy for each mutation’s impact. The closer to 100 a particular mutation is, the greater impact it has on wolframin structure and function.

mutations, the WFS1 amino acid sequence was run through SNAP2 (screening for nonacceptable

Note how the pattern for predicted effect and percent accuracy mirror each other.

polymorphisms),

PolyPhen-2

(Polymorphism Phenotyping v2), and PROVEAN (Protein Variation Effect Analyzer), all of which are software that predict the functional effects of sequence variants using statistical methods (22, 23).

It is difficult to characterize a certain psychiatric symptom as a definite indicator of a particular disorder. Thus, our investigation of mutations in the gene WFS1 incorporated data that include psychiatric symptoms associated with bipolar disorder and schizophrenia, even if those patients were not formally diagnosed with one of the disorders. To take into consideration lack of diagnosis and misdiagnosis, all patients with the following symptoms were considered: suicide tendency, suicide, depression, major Figure 5: The impact of WFS1 mutations in protein structuree and function based on SNAP2, PolyPhen-2, and PROVEAN data (data points were normalized to highest value in each data set).

an increased risk of bipolar disorder and regarding Wolfram Syndrome and heterozygous schizophrenia. We also note that there are carriers of WFS1 mutations with psychiatric virtually no published studies examining disorders (Figure 1). Databases searched included mutations in the N-terminus of WFS1 which PubMed Central, MEDLINE, Google Scholar, encodes for the primary region of wolframin and Bookshelf. Within all databases, the search residing in the cytoplasmic compartment terms “Wolfram Syndrome and Mood Disorders,” of cells and represents over 20% of the “N-terminus mutations in WFS1,” and “WFS1 protein. Combining results from all these mutations” AND [bipolar disorder, mood disorder, studies will assist in the understanding of the depression, or psychiatric disease] were used. pathophysiology of both Wolfram Syndrome and Papers (ranging from 1998-2020) were selected psychiatric disorders.

after reading the abstract and determining relevancy to the objective. The aim was to

Methods:

determine if particular WFS1 mutations found in

Searching strategy and study selection:

Wolfram Syndrome patients were associated with

Comprehensive literature searches psychiatric symptoms regardless of whether the and analyses were done to compile available data patient exhibited classic symptoms of Wolfram

7

depressive disorder, schizophrenia, bipolar disorder, manic episodes, hallucinations, and general psychiatric illness. As little is known about the cellular and neurological function of either wolframin or WFS1, there is not an abundance of data surrounding its connection to psychiatric disorders, especially as psychiatric disorders are not a defining characteristic of the disease (1). In total, approximately 2,392 patients were included in these collective studies.

Statistical analysis: Fisher’s Exact Tests were performed to investigate: i) the correlation between mutation location and the predicted PolyPhen-2 score and ii) the correlation between mutation location and psychiatric disorders. A P-value lower than 0.05 was considered statistically significant, and correlation indices such as

Scientia Cramer’s V and Contigency Coefficient were computed.

In order to illustrate the correlation of psychiatric phenotypes with the different locations, we categorized the psychiatric disorders into 5 main groups: i. Aggression, hallucinations, general psychiatric illness and psychiatric problems ii. Depression, depression with therapy and major depression iii. Manic episodes and bipolar disorder iv. Schizophrenia v. Suicide Results: The first round of literature search yielded over one hundred possible WFS1 mutations that were associated with mood

Figure 6: The potential progression of mood disorder ina given patient due ot mutations in wolframin.

disorder. In order to refine this list, SNAP2

confirm the location of mutational hotspots in Syndrome but had psychiatric symptoms (34). The

was used to determine the predicted effect and

the protein. From the present information, we degree of alteration in protein structure appears

accuracy on the protein structure and amino

identified regions of interest which included to correlate with the impact on functions related

acid sequence (UniProtKB - O76024 [WFS1_

mutations that had multiple reports in the to Wolfram Syndrome. The mutations to WFS1,

HUMAN]). As the experimental validation of the

C-terminus (residues 672-717 and 771-776) and at especially those to W700, should be examined

wolframin structure is not yet established, it was

the interface between the membrane and the ER further for effects on wolframin structure that

necessary to utilize this molecular prediction to

lumen (A326, L432, P533, G576).

identify the likely location of the mutations on the protein.

Wolframin

We then sought to determine which events. mutations found in our search were associated

is

a

lead to both medical and psychiatric downstream We then aligned the magnitude of

multi-pass

with Wolfram Syndrome patients and which every mutation’s impact along the protein (Figure

transmembrane protein associated with the

mutations were associated with heterozygous 5). It is important to note that there are no data

endoplasmic reticulum (ER) (17, 18, 27), where the

carriers of WFS1 mutations. A Venn diagram regarding the N-terminal mutations in WFS1 and

N-terminus is cytoplasmic and the C-terminus

was generated in order to visualize the overlap mood disorder, thus our analysis of mutations

resides in the lumen of the ER. The identified

(Figure 3). Note that two mutations in wolframin begins after the N-terminus, and the X axis starts

mutations were highlighted on the proposed

(A559T and A684V) were present in both Wolfram at residue position 300. The mutations which

folding structure of wolframin (Figure 2). This

Syndrome and heterozygous carriers of WFS1 were detected across all three platforms to be

location and orientation are appropriate for a

mutations in multiple cases. A559T is associated particularly impactful (in order from most to

protein that regulates calcium signaling (18).

with manic episodes and major depression (16, least impactful) were W700C, P553S, and R558C.

This mutation map demonstrates the many

28, 29), and A684V is associated with depression W700C was given a PROVEAN of -12.937, which is

reported C-terminal mutations, but lack of

and hallucinations (30-32) in both populations.

known mutations in the N-terminus. The

Subsequently, the SNAP2 predicted effect (-7.923 and -7.134, respectfully). Note that P533S

absence of published reports on mutations in

values were plotted in comparison to the and R558C mutations have only been detected in

the N-terminus was unexpected. Although the

SNAP2 predicted accuracy values (Figure 4). heterozygous carriers of WFS1 mutations. Thus,

mutations associated with Wolfram Syndrome

The SNAP2 results were most accurate when these mutations (P533S and R558C) may be points

appear to span the entire protein, the most

considering mutations with extreme impact on of interest for psychiatric disease because of their

frequently identified mutations associated with

protein structure and function. Patients with location in the middle of the protein, not the N-

psychiatric symptoms are A559T and A684V,

mutations at W700 have severe manifestations or C- termini (Table 1). This central location may

both of which are predicted to be localized in

of Wolfram Syndrome because the mutation allow wolframin to function relatively normally

extra-membrane regions of the protein. Several

W700X causes truncation of the majority of and cause solely psychiatric symptoms as opposed

mutations in non-Wolfram Syndrome patients

the C-terminus (10). Also, patients with the to physical symptoms like those associated with

(R772C, E717K, and M312R) occurred four unique

mutation W700C have been found to have Wolfram Syndrome patients.

times in different data sets, suggesting that

severe mood disorder with the mutation (9,

these are important positions for the function

33). The psychiatric symptoms associated with 4 mutations occurred in the cytoplasm, 17 in the

of wolframin. When a defined structure for

W700X were not reported. Patients with M312 ER lumen, and 7 in the membrane. We found a

wolframin is available, it will be possible to

mutations had milder symptoms of Wolfram statistically significant correlation between the

notably larger than the ratings of P533S and R558C

Over 29 mutations, we observed that

mutation location and the predicted PolyPhen-2

8

2021-2022 score (Cramer’s V = 0.40, Contigency The analysis did not show a statistically Coefficient = 0.49, P-value in Fisher’s significant

correlation

between

a

Exact Test = 0.02). The data showed certain psychiatric disorders group and that 82.4% (14/17) of the ER lumen mutation location (Cramer’s V = 0.29, mutations and 85.7% (6/7) of the Contigency Coefficient = 0.38, p-value membrane mutations are predicted as reported by the Fisher’s Exact Test = “probably damaging” according to Poly- 0.77). The majority of the mutations, Phen whereas 60% (3/4) of cytoplasmic indeed, occurred in the ER lumen, mutations are predicted to be “benign” regardless of the psychiatric disorders (Table 2). This finding suggests that if a groups. In patients with schizophrenia mutation occurs in the cytoplasm it is there are two mutations in cytoplasm, 4 more likely to be benign; conversely if in the ER lumen and 2 in the membrane. the mutation occurs in the lumen or in In patients with manic episodes and the membrane, then it is more likely to bipolar disorders one cytoplasmic and be damaging.

2 ER lumen mutations were reported (Table 3).

Table 2: Relationship between mutation locations and PolyPhen-2 predicted effect

Table 3: Distribution of the mutations associated to the psychiatric disorder groups across different locations

Table 1: Mutations in wolframin listed with their associated symptoms, SNAP2 predicted effect score/ SNAP2 predicted percent accuracy, PolyPhen-2 rating, and PROVEAN rating/ PROVEAN numerical score.

Discussion and Conclusion: Wolfram Syndrome is a rare multisystem disease characterized by juvenile onset diabetes mellitus, vision and hearing loss, and psychiatric disorders. There is no definitive diagnostic tool nor disease-modifying tool for Wolfram Syndrome-related mood disorders. Presently, there is little known about the protein wolframin and gene WFS1. Similarly, a coherent molecular or genetic understanding of mood disorders is still needed. The goal of this literature analysis was to determine whether there are hotspots in wolframin that could link mutations in the protein associated with Wolfram Syndrome to mood disorders such as bipolar disorder and schizophrenia.

Relevant to this work, there is a growing interest in the field of genetic mutations related to psychiatric diseases. For example, evidence suggests the existence of mutations that are causative to autism spectrum disorder (35), Tourette’s Syndrome (36), attention-deficit/hyperactivity disorder (37), and Huntington’s Disease (38). The correlation we observed between WFS1 mutations and psychiatric symptoms is consistent with these previous findings in other diseases.

Our findings support the proposal that specific mutations in WFS1 are associated with mood related disorders in both Wolfram Syndrome patients and heterozygous carriers of WFS1 mutations. When the location of these mutations is considered (Table 1), there is evidence that there are hot spots even though mutations were found throughout the protein.

9

Studies that have examined the N-terminal region of WFS1 have indicated that this region includes many “loss of function” mutations (1, 3). These mutations are often truncations or frameshifts which completely alter the remainder of the protein and are associated with the most detrimental forms of Wolfram Syndrome (1). More information describing the consequences of these N-terminus mutations is needed. We expect that there is a high incidence of mood disorders in individuals who carry mutations in this region, in both Wolfram Syndrome and heterozygous carriers of WFS1 mutations.

Mutations in heterozygous carriers of WFS1 such as R772C, E717K, and M312R, occurred four unique times in different data sets throughout this search, which suggests that they are important positions. A next step is to perform mutagenesis of wolframin in cell and animal models of Wolfram Syndrome to assess how the mutations affect mRNA and protein stability in vitro and in vivo. R772C and E717K are both in the C-terminus, which is associated with severe Wolfram Syndrome and psychiatric illness. It ise important to learn if these residues are localized near W700 in the 3D folded structure of wolfram, another

Scientia

residue associated with severe disease. In contrast, M312R is closer to disorder, patients are typically seen when their disease has progressed to an the N-terminus on a transmembrane loop predicted to be close to the unmanageable point. Although Wolfram Syndrome is usually detected early, cell cytoplasm. Interestingly, this location at the interface between the disease progresses quickly, and leaves little time to reverse the damage membrane and cytoplasm may be relevant as other mutations associated at the cellular level. As mood disorders are complex, management may need with psychiatric symptoms were found there (A559T, E394V and R558C). to be personally tailored, and thus there is a demand for a targetable and In addition, the mutation R558C has been found to cause a mild form druggable pathway for these conditions. Further research on the pathways of Wolfram Syndrome and is associated with risk for type 2 diabetes in and mutations underlying Wolfram Syndrome, particularly the N-terminus the Ashkenazi Jewish population (39). This group of individuals provides of WFS1, is crucial to provide early diagnoses and effective therapeutics.

In conclusion, WFS1 mutations have shown impactful change in

a controlled group to study the effects of the low impact mutation both

physically and psychologically. Once the wolframin structure is known, the structure of wolframin, leading to impaired calcium signaling and cell interactions among these sites will be determined. Future efforts and survival (18). These changes are expected to result in mood related disorders experiments will extend these findings and assist our understanding about in Wolfram Syndrome and heterozygous carriers of WFS1 mutations (Figure 6). Understanding the effect of WFS1 mutations on psychiatric disease is an

Wolfram Syndrome and its related psychiatric symptoms.

The findings of this report should be interpreted considering important challenge that will aid to development of targeted treatments for several limitations. Most importantly, knowledge of N-terminus mutations both Wolfram Syndrome and mood disorders. in Wolfram Syndrome patients is still limited and the availability of literature discussing this specific topic is sparse. In addition, information Abbreviations: about heterozygous carriers of WFS1 mutations is also under reported. DIDMOAD (diabetes insipidus, insulin-deficient diabetes mellitus, optic These limitations establish significant obstacles to find and select relevant atrophy and deafness) studies to substantiate our findings. Therefore, we formulated our search ER (endoplasmic reticulum) with a focus on schizophrenia and bipolar disease, as opposed to generally searching for every term associated with mental illness. We consider this report as an exploratory study intended to lay the groundwork for a more comprehensive research study in the future, which could References: hopefully include further information regarding psychiatric symptoms and N-terminal WFS1 mutations. In addition, with a debilitating disease such as Wolfram Syndrome, there is a chance that some of the psychiatric disorders identified in patients were an effect of living with a complex and incurable disease and not caused directly by the gene mutated in Wolfram Syndrome. Although we completed the review with these ambiguities in mind, understanding the distinction between cause and effect of mood disorders in particular is an area of research which needs further study.

Even though it is hypothesized that the location of WFS1 mutations would correlate with psychiatric disorders, the results of our statistical analysis failed to confirm this hypothesis. There are several explanations for these results. For example, the wide spectrum of psychiatric symptoms reported in Wolfram Syndrome would require larger sample sizes and more evident phenotypes to adequately test for a correlation. Furthermore, there is variation in both the prevalence and the clinical assessment of psychiatric symptoms in different regions the world. Additionally, psychiatric manifestations are known to be influenced by environmental factors.

When we examined Poly-Phen 2 predictive scores, we found that most of membrane and ER lumen mutations are damaging. As ER function is intimately related to the DNA damage response, and given the accumulating data supporting the hypothesis that ER and membranal wolframin mutations significantly induce psychiatric illness, we suggest that altered calcium homeostasis due to decreased NCS1 protein abundance is a contributing factor for the impaired cell signaling leading to abnormal neuronal function.

Current treatments for both Wolfram Syndrome and mood disorders are limited. Often, the treatments are centered around symptom management as opposed to addressing the underlying mechanism responsible for the disease. In the case of Wolfram Syndrome related-mood

1.

Urano F. Wolfram Syndrome: Diagnosis, Management, and Treatment. Current diabetes reports. 2016;16(1):6-.

2.

Maleki N, Bashardoust B, Zakeri A, Salehifar A, Tavosi Z. Diabetes mellitus, diabetes insipidus, optic atrophy, and deafness: A case of Wolfram (DIDMOAD) syndrome. J Curr Ophthalmol. 2015;27(3-4):132-5.

3.

Rohayem J, Ehlers C, Wiedemann B, Holl R, Oexle K, Kordonouri O, et al. Diabetes and neurodegeneration in Wolfram syndrome: a multicenter study of phenotype and genotype. Diabetes Care. 2011;34(7):1503-10.

4.

Barrett TG, Bundey SE, Macleod AF. Neurodegeneration and diabetes: UK nationwide study of Wolfram (DIDMOAD) syndrome. Lancet (London, England). 1995;346(8988):1458-63.

5.

Swift RG, Polymeropoulos MH, Torres R, Swift M. Predisposition of Wolfram syndrome heterozygotes to psychiatric illness. Mol Psychiatry.

1998;3(1):86-91.

6.

Schmidt-Kastner R, Kreczmanski P, Preising M, Diederen R, Schmitz C, Reis D, et al. Expression of the diabetes risk gene wolframin (WFS1) in the human retina. Exp Eye Res. 2009;89(4):568-74.

7.

Bischoff AN, Reiersen AM, Buttlaire A, Al-lozi A, Doty T, Marshall BA, et al. Selective cognitive and psychiatric manifestations in Wolfram Syndrome.

Orphanet Journal of Rare Diseases. 2015;10(1):66.

8.

Pallotta MT, Tascini G, Crispoldi R, Orabona C, Mondanelli G, Grohmann U, et al. Wolfram syndrome, a rare neurodegenerative disease: from pathogenesis to future treatment perspectives. J Transl Med. 2019;17(1):238-.

9.

Hofmann S, Philbrook C, Gerbitz KD, Bauer MF. Wolfram syndrome: structural and functional analyses of mutant and wild-type wolframin, the WFS1 gene product. Hum Mol Genet. 2003;12(16):2003-12.

10.

Hofmann S, Bauer MF. Wolfram syndrome-associated mutations lead to instability and proteasomal degradation of wolframin. FEBS Letters.

2006;580(16):4000-4.

11.

Kakiuchi C, Ishigaki S, Oslowski CM, Fonseca SG, Kato T, Urano F. Valproate, a Mood Stabilizer, Induces WFS1 Expression and Modulates Its Interaction with ER Stress Protein GRP94. PLOS ONE. 2009;4(1):e4134.

12.

Abreu D, Asada R, Revilla JMP, Lavagnino Z, Kries K, Piston DW, et al.

Wolfram syndrome 1 gene regulates pathways maintaining beta-cell health and survival. Lab Invest. 2020;100(6):849-62.

13.

Swift RG, Polymeropoulos MH, Torres R, Swift M. Predisposition of Wolfram syndrome heterozygotes to psychiatric illness. Molecular Psychiatry.

1998;3(1):86-91.

14.

Sequeira A, Kim C, Seguin M, Lesage A, Chawky N, Desautels A, et al. Wolfram syndrome and suicide: Evidence for a role of WFS1 in suicidal and impulsive behavior. Am J Med Genet B Neuropsychiatr Genet. 2003;119b(1):108-13.

15.

Koido K, Kõks S, Nikopensius T, Maron E, Altmäe S, Heinaste E, et al.

Polymorphisms in wolframin (WFS1) gene are possibly related to increased risk

10

2021-2022 for mood disorders. The international journal of neuropsychopharmacology / official scientific journal of the Collegium Internationale Neuropsychopharmacologicum (CINP). 2005;8:235-44.

16.

Swift M, Swift RG. Wolframin mutations and hospitalization for psychiatric illness. Mol Psychiatry. 2005;10(8):799-803.

17.

Angebault C, Fauconnier J, Patergnani S, Rieusset J, Danese A, Affortit CA, et al. ER-mitochondria cross-talk is regulated by the Ca(2+) sensor NCS1 and is impaired in Wolfram syndrome. Sci Signal. 2018;11(553).

18.

Nguyen L, Fischer TT, Abreu D, Arroyo A, Urano F, Ehrlich BE. Calpain Inhibitor and Ibudilast Rescue β-Cell Function in a Cellular Model of Wolfram Syndrome. PNAS. 2020;117(29):17389-98.

19.

Charney AW, Ruderfer DM, Stahl EA, Moran JL, Chambert K, Belliveau RA, et al. Evidence for genetic heterogeneity between clinical subtypes of bipolar disorder. Transl Psychiatry. 2017;7(1):e993.

20.

Rowland TA, Marwaha S. Epidemiology and risk factors for bipolar disorder. Ther Adv Psychopharmacol. 2018;8(9):251-69.

21.

Smeland OB, Bahrami S, Frei O, Shadrin A, O’Connell K, Savage J, et al. Genome-wide analysis reveals extensive genetic overlap between schizophrenia, bipolar disorder, and intelligence. Mol Psychiatry. 2020;25(4):844-53.

22.

Adzhubei I, Jordan DM, Sunyaev SR. Predicting functional effect of human missense mutations using PolyPhen-2. Curr Protoc Hum Genet.

2013;Chapter 7:Unit7 20.

23.

Choi Y, Sims GE, Murphy S, Miller JR, Chan AP. Predicting the functional effect of amino acid substitutions and indels. PLoS One. 2012;7(10):e46688.

24.

Bromberg Y, Rost B. SNAP: predict effect of non-synonymous polymorphisms on function. Nucleic Acids Research. 2007;35(11):3823-35.

25.

Saleh MA, Solayman M, Paul S, Saha M, Khalil MI, Gan SH. Impacts of Nonsynonymous Single Nucleotide Polymorphisms of Adiponectin Receptor 1 Gene on Corresponding Protein Stability: A Computational Approach. Biomed Res Int. 2016;2016:9142190.

26.

Roy A, Kucukural A, Zhang Y. I-TASSER: a unified platform for automated protein structure and function prediction. Nat Protoc. 2010;5(4):72538.

27.

Philbrook C, Fritz E, Weiher H. Expressional and functional studies of Wolframin, the gene function deficient in Wolfram syndrome, in mice and patient cells. Exp Gerontol. 2005;40(8-9):671-8.

28.

Khanim F, Kirk J, Latif F, Barrett TG. WFS1/wolframin mutations, Wolfram syndrome, and associated diseases. Hum Mutat. 2001;17(5):357-67.

29.

Smith CJ, Crock PA, King BR, Meldrum CJ, Scott RJ. Phenotype-genotype correlations in a series of wolfram syndrome families. Diabetes Care. 2004;27(8):2003-9.

30.

Cryns K, Sivakumaran TA, Van den Ouweland JM, Pennings RJ, Cremers CW, Flothmann K, et al. Mutational spectrum of the WFS1 gene in Wolfram syndrome, nonsyndromic hearing impairment, diabetes mellitus, and psychiatric disease. Hum Mutat. 2003;22(4):275-87.

31.

Kobayashi M, Miyagawa M, Nishio SY, Moteki H, Fujikawa T, Ohyama K, et al. WFS1 mutation screening in a large series of Japanese hearing loss patients: Massively parallel DNA sequencing-based analysis. PLoS One. 2018;13(3):e0193359.

32.

Rendtorff N, Lodahl M, Boulahbel H, Johansen I, Pandya A, Welch K, et al. Identification of p.A684V Missense Mutation in the WFS1 Gene as a Frequent Cause of Autosomal Dominant Optic Atrophy and Hearing Impairment. American journal of medical genetics Part A. 2011;155A:1298-313.

33.

Qian X, Qin L, Xing G, Cao X. Phenotype Prediction of Pathogenic Nonsynonymous Single Nucleotide Polymorphisms in WFS1. Sci Rep. 2015;5:14731.

34.

Hansen L, Eiberg H, Barrett T, Bek T, Kjaersgaard P, Tranebjaerg L, et al. Mutation analysis of the WFS1 gene in seven Danish Wolfram syndrome families; four new mutations identified. Eur J Hum Genet. 2005;13(12):1275-84.

35.

De Rubeis S, He X, Goldberg AP, Poultney CS, Samocha K, Cicek AE, et al. Synaptic, transcriptional and chromatin genes disrupted in autism.

Nature. 2014;515(7526):209-15.

36.

Pauls DL, Leckman JF, Cohen DJ. Familial relationship between Gilles de la Tourette’s syndrome, attention deficit disorder, learning disabilities, speech disorders, and stuttering. J Am Acad Child Adolesc Psychiatry.

1993;32(5):1044-50.

37.

Thapar A. Discoveries on the Genetics of ADHD in the 21st Century: New Findings and Their Implications. Am J Psychiatry. 2018;175(10):943-50.

38.

McColgan P, Tabrizi SJ. Huntington’s disease: a clinical review. Eur J Neurol. 2018;25(1):24-34.

39.

Bansal V, Boehm BO, Darvasi A. Identification of a missense variant in the WFS1 gene that causes a mild form of Wolfram syndrome and is associated with risk for type 2 diabetes in Ashkenazi Jewish individuals.

Diabetologia. 2018;61(10):2180-8.

40.

Fischer TT, Nguyen LD, Ehrlich BE. Neuronal Calcium Sensor 1 (NCS1) as a Potential Drug Target for Treatment of Wolfram Syndrome. The FASEB Journal. 2020;34(S1):1-.

41.

Koh PO, Undie AS, Kabbani N, Levenson R, Goldman-Rakic PS, Lidow MS. Up-regulation of neuronal calcium sensor-1 (NCS-1) in the prefrontal cortex of schizophrenic and bipolar patients. Proc Natl Acad Sci U S A. 2003;100(1):313-7.

42.

Won E, Kim YK. An Oldie but Goodie: Lithium in the Treatment of Bipolar Disorder through Neuroprotective and Neurotrophic Mechanisms. Int J Mol Sci. 2017;18(12).

43.

Matsunaga K, Tanabe K, Inoue H, Okuya S, Ohta Y, Akiyama M, et al.

Wolfram syndrome in the Japanese population; molecular analysis of WFS1 gene and characterization of clinical features. PLoS One. 2014;9(9):e106906.

11

Scientia

Activation of Host Kinase Vps34 by Murine Norovirus Non-structural Protein NS4 Introduction:

IN-DEPTH Jessica Oros Randall Lab, University of Chicago

that other viruses such as dengue virus (DENV),

Despite Vps34 being used in the host

and encephalomyocarditis virus (EMCV) cannot

for autophagy, MNV may be utilizing Vps34

Each year, millions of individuals

replicate when the TAG-dependent antiviral

to aid the process of viral replication. Thus,

are infected with human norovirus, securing

pathway is knocked down. If the TAG-dependent

we hypothesize that Vps34 may be a point of

norovirus as the number one worldwide cause

antiviral pathway was the only means through

control by MNV during infection as it may

of gastroenteritis and causing about 200,000

which viral replication was controlled, DENV

support viral replication in an unknown way.

deaths each year. Tens of thousands of those

and EMCV would be able to replicate in TAG-

Furthermore, we aim to characterize which

deaths occurring in children in developing

knockout conditions. Since this is not observed,

viral non-structural protein is responsible for

countries. There are presently no norovirus

it indicates that there is some other TAG-

the change in Vps34 activation level. Out of the

vaccines or cures, making it a pressing issue to

independent antiviral pathway present that is

6 non-structural proteins of MNV, the most

understand the biology of norovirus infection.

still blocking viral replication. Interestingly,

likely candidates are NS4, NS5, and NS6 because

Unfortunately, human norovirus cannot be

MNV can still replicate in this TAG-deficient

of their previously elucidated roles in immune

easily grown in laboratory cultures. However,

model, meaning there is some mechanism by

evasion. NS4 has been shown to localize to host

murine norovirus (MNV) can serve as a model.

which MNV is evading the immune response.

intracellular membranes and thus may play a

MNV is used in the lab to understand the

Evasion may be possible at the viral replication

role in replication complex formation, but its

mechanisms behind infection in murine cell

stage via the replication complex or through

function is largely uncharacterized. The Randall

lines. MNV is a positive-sense RNA virus. Like

virus-derived factors such as interactions

Lab has previously shown that NS5 can produce

other viruses within the same class, MNV creates

between viral and host proteins. The aim of this

an immune evasion phenotype upon infection

a replication complex upon entry and uncoating

project is to elucidate the mechanisms by which

with DEMV and EMCV of BV2 cells transduced

inside the cell using host-cell membranes . The

MNV presents a phenotype of immune evasion

with NS5. Upon entry and uncoating, the viral

purpose of this replication complex seems to be

through interactions with host proteins.

genome is translated into a polyprotein, and

protecting viral genome replication from the

One possible player in MNV immune

NS6 is the protease responsible for the cleavage

hostile environment of the host cell cytoplasm.

evasion and replication could be the cellular class

and release of individual MNV non-structural

Luckily for host organisms, these replication

III phosphatidylinositol-3 kinase (PI3K) vacuolar

proteins .

complexes can be targetted via an interferon-

protein sorting 34 (Vps34), which plays a role in

In this study, we have shown that

gamma (IFNG) mediated antiviral pathway,

cellular trafficking via initiation of autophagy.

infection with MNV results in a decrease in

allowing for lysosomal degradation of the

Vps34

phosphatidylinositol

inhibitory Vps34 acetylation, indicating an

replicating virus. This pathway uses proteins

to produce phosphatidylinositol-3-phosphate

increase in activity. Out of the viral non-

involved in autophagy, a cellular process by

(PI3P), which acts as a membrane marker to

structural proteins tested, only non-structural

which foreign particles or faulty proteins are

begin the formation of the autophagosome

protein 4 (NS4) was able to reproduce the

engulfed and degraded through lysosomal

. In healthy cells, Vps34 is maintained in an

phenotype, indicating that it is responsible for

fusion. This recognition by the host cell is done

inhibitory state by the nutrient-sensing kinase

the release of Vps34 from its inhibitory state.

by the Atg12-Atg5-Atg16L1 complex, which is

mTOR. Under cellular stress or starvation, mTOR

comprised of autophagy-related (Atg) proteins

phosphorylates UVRAG, a protein complex

Materials and Methods:

that interact with host membranes to form an

involved in endocytosis, which will activate

Tissue Culture:

autophagosome . Conjugation of the ubiquitin-

Vps34 to initiate autophagy. The Randall lab

Murine norovirus (MNV) was used as

like microtubule associated protein light-chain

has previously shown that knockdown of Vps34

a model to study norovirus activity. Cell culture

3 (LC3) with Atg-5 allows interferon-inducible

prevents MNV replication, indicating that there

was done with murine microglial BV2 cells using

GTPases to localize to LC3

phosphorylates

and target the

may be interactions between the virus and

standard tissue culture protocol, which involves

replication complex, inhibiting viral replication

the host lipid kinase. Vps34 activity is tightly

growing the adherent cells in flasks with Gibco

. This pathway does not operate via degradative

regulated by the host cell through acetylation

Dulbecco’s Modified Eagle Medium (DMEM)

canonical autophagy, but it uses autophagy

by the acetyltransferase p300, which itself

supplemented with 10% fetal bovine serum

proteins to prevent MNV from replicating inside

is activated by autoacetylation . Acetylation

(FBS) and 1% each of non-essential amino acids,

the cell . The pathway is called the Targeting by

of Vps34 is inhibitory, and activation by

hydroxyethyl

AutophaGy protein (TAG) – dependent antiviral

deacetylation contributes to the formation of

(HEPES), and PIS. Cells were seeded and used for

pathway. Prior work in the Randall lab shows

autophagosomes.

experiments. In the case of interferon gamma

12

piperazineethanesulfonic

acid

2021-2022 (IFNG) treatment, 100µL/mL were used for a 24hour period. For infections, MNV strain CW3 was diluted in complete DMEM media and added to cells at 0.05 MOI. Transductions: Transductions of viral non-structural proteins or wild-type Vps34 were done using plasmids previously generated by the Randall lab. Third-generation lentiviral constructs were used, and transfection was prepared for HEK293T cells using CaCl2, HEPES buffered saline solution, sterilized water, lentivirus packaging plasmids, pseudotyping envelope plasmids, and the previously described plasmids containing V5-

Figure 1: MNV Infection decreases levels of acetylp-300 and acetyl Vps-34 Western blots of control BV2 and control BV2+wtVps34 alongside infected BV2 and infected BV2+wtVps34. Infection was done with 0.05 MOI of MNV for 24 hours. (A) Infected cell lysates are shown in lane 2. Acetylp-300 and p300 bands were flanked by two

tagged Vps34 or Flag-tagged MNV non-structural

unspecific bands on the top and bottom. (B) Immunoprecipitations of cell lysates captured Vps34, with the middle column showing

proteins NS4, NS5, or NS6, with the tags being used

infected lysates. Input Vps34 and actin bands are taken from input samples collected from supernatant of lysates with precipitated

for immunoprecipitation and immunoblotting.

Supernatant was collected and filtered to isolate lentivirus particles 48 hours post transfection, which were added to BV2 cells. Selection for successfully transduced BV2 cells was done with Thermo Fisher Scientific Hygromycin B at a concentration of 250µg/mL. Gel Electrophoresis and Transfer: After 24 hours post infection (hpi) or post seeding cells transduced with MNV nonstructural proteins, cells were lysed using 2x Laemmli sample buffer. Samples were then heated at 95ºC for 10 minutes before gel electrophoresis was done on an SDS-PAGE to separate samples by size alongside Thermo Fisher Scientific PageRuler 10-250 kDa. Protein samples were transferred from the SDS-PAGE gel to 0.2μm PVDF membranes via semi-dry transfer. Membranes were then blocked in 5% skim milk blocking buffer before immunoblotting

with

polyclonal

antibodies

targeting the desired proteins.

Immunoprecipitation: In the case of immunoprecipitation, after 24hpi, cells were treated with cold coimmunoprecipitation lysis buffer was used and comprised 2.5mL 1M Tris at pH 7.5, 1.5mL 5M NaCl, 5mL 10% Triton X-100, proteinase inhibitor cocktail, and 41mL distilled water. Lysed cells were briefly frozen at -80ºC and thawed to complete lysis. Cell lysates were incubated overnight at 4ºC on a rotator with Cytiva Protein A Sepharose beads and mouse anti-V5 antibody. Precipitated protein was washed using Co-IP wash buffer before 2x Laemmli sample buffer was added, and samples were heated at 95ºC for 10 minutes before an SDS-PAGE and semi-dry transfer was done following the same protocol as whole cell lysates described above.

Vps34

Antibodies:

this phenotype. This indicates that MNV the

infection results in a decrease of acetylation

immunoprecipitation of Vps34 were Cell Signaling

Antibodies

for

of the acetyltransferase p300 such that the

Technology Mouse V5-tag (E9H80). Detection

levels of active p300 are at a lower level. Levels

of Vps34 and acetyl-Vps34 via immunoblot was

of acetylated Vps34 were measured (Figure 1b)

done with Cell Signaling Technology Rabbit

after infection using an immunoprecipitation,

V5-tag (D3H80) and Cell Signaling Technology

as endogenous levels of Vps34 are below

Rabbit Acetylated-Lysine, respectively. Imaging

detection threshold. When compared to

p300 in whole cell lysate was done with Santa

BV2 cells transduced with wild-type Vps34

Cruz Biotechnology Mouse monoclonal IgG p300

(BV2+wtVps34), infected cells showed a marked

(F-4), while acetyl-p300 was captured with Cell

decrease in the levels of acetylated Vps34.

Signaling Technology Rabbit Acetyl-CBP (K1535)/

Levels of Vps34 were consistent across infected

p300 (K1499). To normalize Western blot bands,

and control cells, meaning the decrease in

actin was detected in each sample using Santa

acetyl-Vps34 is notable.

Cruz Biotechnology Mouse monoclonal IgG Beta-

MNV NS4 decreases expression of acetyl-

actin (C4) HRP. Detection of viral non-structural

Vps34

proteins was done with Sigma-Aldrich Rabbit

BV2+wtVps34 cells were transduced

polyclonal Anti-FLAG antibodies. Secondary

using lentivirus with NS4, NS5, or NS6, and an

antibodies used were Thermo Fischer Scientific

immunoprecipitation was run on cell lysates

goat anti-Rabbit IgG secondary antibody, HRP

to precipitate Vps34. Immunoprecipitation

and Thermo Fischer Scientific goat anti-Mouse

products were subject to gel electrophoresis

IgG secondary antibody, HRP.

and immunoblotting to detect levels of Vps34 and acetyl-Vps34. Immunoblotting was done

Results:

to detect the presence of the transduced viral

MNV Infection decreases levels of acetyl-p300

non-structural protein and endogenous actin

and acetyl-Vps34

for normalizing measurements. Additionally,

BV2 cells were infected with MNV at a

input lanes were included to ensure the efficacy

multiplicity of infection (MOI) of 0.05 for 24 hours,

of the immunoprecipitation. Transduction

after which cells were lysed and the lysate was run

with NS6 did not replicate the major decrease

on an SDS-PAGE gel. Western blots using anti-

in acetyl-Vps34 levels (Figure 2a). However,

acetyl-CBP rabbit polyclonal antibodies and anti-

due to problems with either transfer or

CBP mouse polyclonal antibodies were used to

immunoblotting, repeated instances of noise

detect levels of acetyl-p300 and p300, respectively

on the membrane obstructed any detailed view

(Figure 1a). Cells infected with MNV showed a

of the acetyl-Vps34 bands. Transductions with

decrease in the fraction of p300 acetylated, as

NS5 were performed and the level of Vps34

control cells showed a greater increase from p300

and acetyl-Vps34 were measured without

to acetyl-p300 while infected cells did not mirror

obstruction from noise. NS5 transduction

13

Scientia resulted in a minor increase in the levels of Vps34 (Figure 2b), but as compared to the BV2+wtVps34 control, the ratio of acetyl-Vps34 to Vps34 was lower, indicating that NS5 is not responsible for the release of Vps34 from its acetylated inhibitory state. NS4 was transduced into BV2+wtVps34 cells, and a decrease in the level of acetyl-Vps34 was observed (Figure 2c). Because problems with the gel in the case of NS6 resulted in obstruction of acetyl-Vps34 bands, densitometry data was measured and graphed comparing levels of acetylVps34 in NS4 and NS5 transduced cells (Figure 2d). As seen in Figure 2b, an increase in acetyl-Vps34 was present in NS5 transduced cells, but NS4 transduced cells showed a decrease in expression of acetyl-Vps34 when compared to BV2+Vps34 cells without viral protein. This indicates that MNV protein NS4 is responsible for the decrease in Vps34 acetylation observed in infection.

MNV NS4 decreases p300 acetylation Cells were transduced with NS4 and NS6, and lysates were run through gel electrophoresis and levels of p300 and acetyl-p300 were detected via immunoblot. NS4 transduction resulted in a notable decrease in not only acetyl-p300 levels, but overall p300 expression as well Figure 2: MNVNS5 and NS6 do not produce dramatic decrease in acetyl-Vps34, NS4 decreases acetyl-Vps34 levels Western blots of control BV2 and control BV2-wtVps34 alongside BV2 cells transduced with NS6 (A), NS5 (B), and NS4 (C)

(Figure 3a). NS6, however, did not show a change in the

are shown with immunoblotting for Vps34 and acetyl-Vps34. Viral proteins were imaged using rabbit anti-Flag antibodies

acetyl-p300 levels as compared to BV2 controls (Figure

to indicate transduction was successful. Input lanes were included to ensure the efficacy of immunoprecipitation.

3b), indicating that NS6 cannot reproduce the phenotype

Densinometry calculations for expression levels of acetyl-Vps34 were done using data fron NS4-transduced and NS5transduced cells (D).

observed in MNV infection. When compared, it is evident that expression levels of p300 are much lower during NS4

Figure 3: MNV NS4 decreases

transduction than NS6 transduction (Figure 3c). These

acetyl-p300 levels while NS6 does not

results further point to NS4 as being responsible for the

Western blots of control BV2 cells and

phenotype in MNV infection whereby acetylation levels of

BV2 cells transduced with NS4 (A) and NS6 (B) are shown with immunoblotting for p300 and acetyl-p300. Densinometry calculations for expression level of acetyl-Vps34 were done (C).

p300 and Vps34 are decreased.

Role of NS4 interaction with Vps34 is not IFNG-dependent To test whether the deacetylation of Vps34 plays a role in viral immune evasion in the presence of interferon gamma (IFNG), cells transduced with NS4 were treated with IFNG at a concentration of 100µL/mL. Immunoprecipitated Vps34 was immunoblotted via Western blot and levels of Vps34 and acetyl-Vps34 were captured (Figure 4). The levels of both Vps34 and acetyl-Vps34 are constant across samples from NS4-transduced BV2 cells incubated with and without IFNG. This indicates there was no noticeable change in the acetylation state of Vps34 during IFNG treatment. Because IFNG does not alter Vps34 activation in NS4 conditions, NS4 does not appear to play a role in the IFNG antiviral pathway and may instead be involved in viral replication.

Discussion: The mechanism of MNV replication and immune evasion are not fully characterized, as it is relatively

Figure 4: Acetyl-Vps34 levels do not change in NS4 cells treated with IFNG Western blots of control BV2+wtVps34 cells and NS4-transduced cells in response to treatment with IFNG (2uL/mL). Input Vps34 shown for success of immunoprecipitation and NS4 shown for success of transduction.

unknown what purposes many non-structural proteins serve upon entry and uncoating. We have shown that activation of the lipid kinase Vps34 is increased during MNV infection through a release of inhibition on Vps34.

Furthermore, it was shown that the levels of the active

14

2021-2022 upstream regulator of Vps34, acetyltransferase p300, decreased during infection. These results suggest that Vps34 does play a role in viral replication and therefore is a point of control in MNV infection. These results were replicated in cells transduced with MNV non-structural protein NS4, while the large decrease in acetylation levels of Vps34 and p300 was not observed in NS5 or NS6 transduced cells.

This indicates that, during MNV infection, the phenotype wherein p300 expresses decreased activation and Vps34 expressess increased activation is being brought on by NS4.

It is also notable that the interaction between NS4 and Vps34 is not affected by the presence of IFNG. Had there been changes in Vps34 activation in the presence of NS4 and IFNG treatment, it may have led to the belief that Vps34 aids MNV in evading antiviral pathways, as IFNG is responsible for the detection and disruption of the viral replication complex.

Instead, it is likely that Vps34 may be involved with aiding viral replication. The Randall lab will perform additional trials of the experiments described in this paper to verify the data and evaluate its statistical significance.

It is still unknown exactly what NS4 does in the host cell upon infection. Studies have found it to localize to the Golgi apparatus and endosomesix, leading to a tentative belief that it may be involved in recruiting host membranes for the formation of the viral replication complex.

Downstream effects of Vps34 will be studied to further understand how Vps34 creates a pro-viral state within the cell for viral replication. Upon autophagosome initiation, Vps34 functions to produce phosphatidylinositol-3-phosphate

References O’Donnell, T. B., Hyde, J. L., Mintern, J. D., & Mackenzie, J. M. (2016). Mouse Norovirus infection promotes autophagy induction to facilitate replication but prevents final autophagosome maturation. Virology, 492, 130-139.

Lopman, B. A. (2015). Global burden of norovirus and prospects for vaccine development.

Hyde, J. L., Sosnovtsev, S. V., Green, K. Y., Wobus, C., Virgin, H. W., & Mackenzie, J. M. (2009). Mouse norovirus replication is associated with virus-induced vesicle clusters originating from membranes derived from the secretory pathway. Journal of virology, 83(19), 9709-9719.

Walczak, M., & Martens, S. (2013). Dissecting the role of the Atg12–Atg5-Atg16 complex during autophagosome formation. Autophagy, 9(3), 424-425.

Furlong K, Hwang S. Autophagy and Noroviruses. Viruses. 2019; 11(3):244. https://doi. org/10.3390/v11030244 Biering, S. B., Choi, J., Halstrom, R. A., Brown, H. M., Beatty, W. L., Lee, S., ... & Hwang, S. (2017). Viral replication complexes are targeted by LC3-guided interferon-inducible GTPases. Cell host & microbe, 22(1), 74-85.

Biering, S. B., Choi, J., Halstrom, R. A., Brown, H. M., Beatty, W. L., Lee, S., ... & Hwang, S. (2017). Viral replication complexes are targeted by LC3-guided interferon-inducible GTPases. Cell host & microbe, 22(1), 74-85.

Hwang, S., Maloney, N. S., Bruinsma, M. W., Goel, G., Duan, E., Zhang, L., ... & Virgin, H. W. (2012). Nondegradative role of Atg5-Atg12/Atg16L1 autophagy protein complex in antiviral activity of interferon gamma. Cell host & microbe, 11(4), 397-409.

Kim, J., Kim, Y. C., Fang, C., Russell, R. C., Kim, J. H., Fan, W., ... & Guan, K. L. (2013). Differential regulation of distinct Vps34 complexes by AMPK in nutrient stress and autophagy. Cell, 152(1-2), 290-303.

Munson, M. J., Allen, G. F., Toth, R., Campbell, D. G., Lucocq, J. M., & Ganley, I. G. (2015). mTOR activates the VPS34-UVRAG complex to regulate autolysosomal tubulation and cell survival. The EMBO journal, 34(17), 2272–2290. https://doi.org/10.15252/embj.201590992 Su, H., Yang, F., Wang, Q., Shen, Q., Huang, J., Peng, C., ... & Liu, W. (2017). VPS34 acetylation controls its lipid kinase activity and the initiation of canonical and non-canonical autophagy. Molecular cell, 67(6), 907-921.

Hyde, J. L., & Mackenzie, J. M. (2010). Subcellular localization of the MNV-1 ORF1 proteins and their potential roles in the formation of the MNV-1 replication complex. Virology, 406(1), 138-148.

Herod, M. R., Salim, O., Skilton, R. J., Prince, C. A., Ward, V. K., Lambden, P. R., & Clarke, I. N. (2014). Expression of the murine norovirus (MNV) ORF1 polyprotein is sufficient to induce apoptosis in a virus-free cell model. PLoS One, 9(3), e90679.

Boukhalfa, A., Nascimbeni, A.C., Ramel, D. et al. PI3KC2α-dependent and VPS34-independent generation of PI3P controls primary cilium-mediated autophagy in response to shear stress. Nat Commun 11, 294 (2020). https://doi.org/10.1038/s41467-019-14086-1

(PI3P) at the source of autophagosome membranes . To test whether Vps34 is functioning to form the viral replication

Acknowledgements:

complex during infection, immunofluorescence assays

University of Chicago

are currently underway to see the localization of PI3P in

Randall Lab

response to MNV infection. Large amounts of colocalization

Biological Sciences Collegiate Division (BSCD) Summer

of PI3P with sites of autophagosome membrane sources

Fellowship Program

during infection would indicate that MNV is utilizing the host cell’s autophagy pathway for the construction of the viral replication complex.

Additionally, the mechanisms by which Vps34 is aiding viral replication remain unknown. Further work will be done to understand differences in activity of upstream regulators of Vps34 during MNV infection and NS4 transduction, as it is unclear how NS4 is decreasing levels of Vps34 acetylation. It is unlikely that NS4 functions directly as a deacetylase, so the pathway by which Vps34 is deacetylated indirectly during NS4 transduction is yet to be elucidated. Further progress will examine other regulators of Vps34 that install post-translational modifications other than acetylation, such as phosphorylation or ubiquitination.

15

Scientia

IN-DEPTH

Long-lasting Effects of Early Life Stress on the Brain and Epigenome Omar Kassem Abstract Stable caregiving in early life is critical for normal social and emotional development. Exposure to chronic or extreme stress early in life has been associated with increased risk for psychiatric disorders, especially affective disorders. Early life stress (ELS) has long-lasting impacts on the development and function of diverse brain regions that are involved in emotional processing, cognitive functions, and social behavior. Despite the adverse effects of ELS on brain development, the neural underpinnings of those effects are not yet well characterized. This review highlights ELS-provoked changes in networks wiring, the cellular impacts on neuron excitability and myelination, and the epigenetic modifications that contribute to those changes.

Understanding those neurobiological aberrations aids in understanding the increased risk for psychopathology that has been reported in children exposed to ELS and provides insights into potential Introduction

have revealed that ELS can provoke changes conducted on adolescents years after exposure

Brain development and maturation on various levels including changes in neuron but the impact of ELS was still significant. Further are modulated by an interaction between excitability

and

myelination,

rewiring

of studies in mice have provided more insight into

environmental inputs and genes. It is well networks, and epigenetic modifications that are the underlying neural changes in those regions established that exposure to stress at different in some cases transgenerational [2,9,10,11]. Thus, in response to stress. The main physiological ages leads to different outcomes, with early stages integrating findings from circuitry, cellular, and model of memory formation in the brain is of development being associated with more severe genetic studies is essential to fully understand through adjusting synaptic weights in networks and long-lasting effects [1]. Early childhood is this

process.

Understanding

the

neural to encode memories. Several studies have

a critical period of brain development, which underpinnings of the relationship between ELS demonstrated that ELS interferes with synaptic renders it highly vulnerable to disorganizing and the mental health of the victims is of great plasticity and induces dendritic as well as spine environmental influences. Sustained or adverse value in the pursuit of possible treatments and atrophy [13,16,17]. The cumulative effect of these early-life stress (ELS), such as childhood trauma or interventions. Moreover, a better understanding alterations causes the loss of functional synapses, caregiver deprivation, modifies the expression of of caregiver-related stress effects on child which contributes to cognitive deficits. These multiple neurotransmitters and other molecules development provides many useful insights for influences on neural networks are mediated by that affect neuron and network development improving orphanage systems.

multiple molecules including stress-associated

in specific brain regions, potentially causing

glucocorticoid hormones, neurotransmitters,

long-term structural and functional changes [2]. Impact of ELS through disruption of the and neuropeptides. Better characterization of Brain regions involved in emotional processing, maturation of brain networks and circuitry such as the amygdala, cognitive regions such as

the stress toolbox has enhanced our ability to

The long-term effects of ELS stem pharmacologically intervene and manage stress

the hippocampus and prefrontal cortex, as well from the fact that developmental processes that symptoms [18,19]. as networks involved in social behaviors seem organize circuits and connectivity patterns are

Similarly, ELS, especially in the form

to be particularly responsive and vulnerable still occurring. In humans, the development of of caregiver deprivation, has been shown to to the impact of stress [1,3,4,5,6]. ELS-induced the hippocampus and limbic circuit, which are affect the development of the amygdala, which is long-lasting changes in these regions would be involved in cognitive and emotional functions, highly involved in emotion processing, affective expected to increase susceptibility to emotional takes place largely shortly after birth and valuation, and learning [20]. One key study did disorders and psychiatric illnesses, which has continues for years into adolescence. Therefore, an fMRI scan of brain activity of children adopted been reported in many epidemiological human unlike the effects of stress on adult cognitive from orphanages in a behavioral paradigm where studies [7,8]. Although the associations that functions that are reversible, ELS can often they were instructed to “go” to a neutral cue, and were statistically inferred from epidemiological permanently impact these processes and systems, “not go” to a rare threat cue [3]. Results showed studies are highly indicative of the impact of ELS, yet behavioral and pharmacological interventions that ELS-exposed preadolescents took longer it is difficult to prove causality and elucidate the can still be effective [12,13,14].. An epidemiological to approach a cue when anticipating a potential biological mechanisms underlying this influence study

conducted

on

post-institutionalized threat, indicating that the disorganized care of

in humans. Therefore, many subsequent studies adolescents showed that chronic early life the orphanage experience can alter emotional have used animal models to investigate the stress, being raised in orphanages, is associated and behavioral regulation. FMRI data showed biological underpinnings of ELS consequences. with a reduced volume of prefrontal cortex and that only activity in the amygdala differentiated Studies in animal models, especially rodents, hippocampus [15]. Remarkably, this study was the ELS-exposed children from the control group

16

2021-2022 when anticipating an aversive stimulus. These findings help explain the

adversity expedites their maturation.

epidemiological association between the institutional rearing of children

ELS effects on the epigenome

and psychiatric disorders, especially internalizing ones such as anxiety and

Epigenetics refers to the regulation of gene expression without

disruptive disorders [21,22]. Moreover, these results gesture towards the

altering the genetic code itself. This change in regulation is achieved by

critical importance of providing stable caregiver affection in orphanages,

changing the 3D structure of chromatin. By making the DNA more tightly

and families, to ensure the healthy mental development of children.

or loosely packed, epigenetic machinery can regulate the accessibility

ELS neuron-level effects

of the genome, thus, regulating the expression of genes. Epigenetic

Many studies have attempted to understand the neuron-

modifications involve adding a chemical modification to the DNA or

level impacts of ELS that mediate the observed cortical activity changes

histone proteins, which the DNA is wrapped around, to change its chemical

and behavioral phenotype. The two main influences that have been

properties and 3D shape in the cell. These modifications are added by a

characterized are changes in the electrical properties and myelination of

writer protein and removed by an eraser. The study of epigenetic changes

neurons [23,24,25]. One study used adolescent social isolation of rats to

in the nervous system, called Neuroepigenetics, has provided a lot of

investigate the electrical properties of basolateral amygdala pyramidal

insight into the mechanisms of gene regulation in neurons, especially

cells. This study revealed that the stressed group showed a significant

during development.

increase in neuron excitability, meaning that neurons were more easily

One key study by Kronman et al found that early-life stress in mice

and frequently firing, in comparison to control rats. This effect was

induces epigenetic changes, dimethylation of lysine 79 of histone H3

mediated by a decrease in SK potassium channel activity. Using a drug to

(H3K79me2), in D2-type medium spiny neurons in the depression-

enhance SK channel activity to normalize neuron excitability was found

associated region nucleus accumbens, renders mice more vulnerable

to attenuate anxiety-related behaviors, which provides insights into the

to stress later in life [11]. When ELS-exposed mice faced a second stress

potential of pharmacological interventions that could elevate symptoms.

event in adulthood they exhibited more depression-like behaviors than

Another study by Tanti et al [27] attempted to investigate the

control mice, including lower social interaction, decreased exploration,

effect of altered brain activity of the brain during early development,

and greater immobility in a swim test. They then used a viral vector to

and how that could induce long-term changes similar to those observed

genetically interfere with the function of the writer (Dot1l) and eraser

in children that face early childhood adversity. This study showed that

(Kdm2b) enzymes of the implicated epigenetic modification. Inducing

pharmacological reduction of the activity of the prefrontal cortex of

increased modification through overexpression of Dot1lor knockdown of

mice during development reproduces the cellular and behavioral defects

Kdm2b induced depression-like behaviors in control mice, in other words

induced by repeated ELS: increased behaviors associated with anxiety

inducing an ELS-like phenotype. Conversely, knocking down Dot1l or

and depression, a persistent decrease in neuronal activity, and early

overexpressing Kdm2b reversed the depression-like behaviors in response

differentiation of oligodendrocyte precursors (OPCs), cells that will be

to stress, showing that interventions on an epigenetic level could be a

responsible for producing the myelin surrounding neurons. Conversely,

potential therapeutic approach to remedy the long term effects of ELS.

restoring normal neuronal activity in the prefrontal cortex in mice

Although the effects of ELS could be reversed with genetic manipulation

undergoing early maternal separation leads to the disappearance of

in mice, such an intervention is not very practical in humans. Drugs that

short-term memory deficits, behavioral alterations associated with the

inhibit protein functions are much more feasible for human applications.

depressive state, and oligodendrocyte development. However, behavioral

The authors tested the effect of using a drug that inhibits Dotl1 enzyme to

disorders associated with anxiety persisted. This difference in effect on

interfere with ELS disruptions. Treated mice showed reduced modification

anxiety disorders and depressive disorders could be due to a spatial bias of

levels in their genome and improved behavioral performance, social

the viral injection they used to modulate activity, as it could have modified

interaction scores. These results provided important insights into the

activity in only a subset of emotional circuits involved. Furthermore, the

potential of using epigenetic drugs to remedy disruptions in the brain and

pharmacogenetic technique used to modulate neuronal activity does not

reverse some of those long-lasting effects.

reproduce the physiological state of developing neural networks. These Conclusion and future directions

differences could contribute to the persistence of anxiety behaviors.

The same study revealed that ELS is associated with changes in myelination

Overall, these studies demonstrate that early life stress has

and oligodendrocyte maturation. ELS induces premature differentiation of

profound effects on the developing brain including disrupting critical

oligodendrocyte precursors into mature oligodendrocytes. These results

networks, disrupted electrical properties, and myelination of neurons.

suggest that early emotional stress could lead to expedited maturation

These effects are at least partly mediated by epigenetic modifications that

in brain regions associated with emotion processing, which comes with

change the 3D shape of DNA, thus regulating gene expression in neurons.

a short-term advantage, that being processing the immediate emotional

These ELS effects are often long-lasting and increase susceptibility to

disturbances, and a long-term deficit. The decrease in OPCs could lead

mental and psychiatric disorders, which has been repeatedly shown

to a decrease in quantity and plasticity of myelination later in life. This

to be a significant influence on institutionally reared children. A better

falls in line with human studies associating institutionalized rearing with

understanding of the biological underpinnings of ELS disruptions has

a decline in white matter volume [26], which is related to the volume of

provided significant insights into prospective therapeutic approaches

the fatty myelin layer surrounding axons. Additionally, early maturation

that could reverse those long-lasting effects, such as epigenetic and

of OPCs has been reported in young adults who were sexually assaulted

pharmacological intervention. Moreover, studies on early-life caregiver

in childhood [27], which further supports the hypothesis that early life

deprivation and social isolation emphasize the importance of providing a

17

stable and consistent social environment for children at a young age, which should be incorporated into institutional rearing organizations. There are many future research directions that would provide a more comprehensive understanding of this phenomenon and provide insights into minimizing its impact on children. Elucidating the nature and underlying biology of sex-specific consequences of early-life stress is one important question that has been posed by epidemiological studies that show higher prevalence in females. Many behavioral cognitive therapies have been shown to promote rectifying changes in the brain, the potential of such treatments for reversing the ELS-induced biological and neurological changes is also one issue that hasn’t been studied. Also, many epigenetic studies have revealed that some epigenetic modifications could be transgenerational, potentially making future generations more vulnerable to disruptions that parents experienced. The question of whether epigenetic changes that happen in response to early life adversity could be inherited is another interesting perspective that could have implications for parents with a history of ELS. Finally, the studies discussed here provide evidence for the accelerated maturation of emotion circuitry following ELS. Whether this altered developmental pattern comes at the cost of lagged development of other systems is still unknown and is of great importance for improving the lives of individuals with histories of ELS.

References 1.

Lupien SJ, McEwen BS, Gunnar MR, Heim C (2009). Effects of stress throughout the lifespan on the brain, behaviour and cognition. Nat Rev Neurosci 10: 434–445.

2.

Chen, Y. & Baram, T. Z (2016). Toward Understanding How Early-Life Stress Reprograms Cognitive and Emotional Brain Networks. Neuropsychopharmacology 41, 197–206 3.

Malter Cohen M, Jing D, Yang RR, Tottenham N, Lee FS, Casey BJ (2013). Early-life stress has persistent effects on amygdala function and development in mice and humans. Proc Natl Acad Sci USA 110: 18274–18278. Sandi C, Haller J (2015). Stress and the social brain: behavioural effects and neurobiological mechanisms. Nat Rev Neurosci 16: 290–304 4.

Sandi C, Haller J (2015). Stress and the social brain: behavioural effects and neurobiological mechanisms. Nat Rev Neurosci 16: 290–304 5.

Kim JJ, Diamond DM (2002). The stressed hippocampus, synaptic plasticity and lost memories. Nat Rev Neurosci 3: 453–462.

6.

Joëls M, Baram TZ (2009). The neuro-symphony of stress. Nat Rev Neurosci 10: 459–466.

7.

Brown AS, Susser ES, Lin SP, Neugebauer R, Gorman JM (1995). Increased risk of affective disorders in males after second trimester prenatal exposure to the Dutch hunger winter of 1944-45.

Br J Psychiatry 166: 601–606 8.

Bremner JD, Southwick SM, Johnson DR, Yehuda R, Charney DS (1993). Childhood physical abuse and combat-related posttraumatic stress disorder in Vietnam veterans. Am J Psychiatry 150: 235–239 9.

Lewis, C. R. & Olive, M. F(2014). Early life stress interactions with the epigenome: potential mechanisms driving vulnerability towards psychiatric illness. Behav Pharmacol 25, 341–351 10. Herzberg, M. P. & Gunnar, M. R (2020). Early life stress and brain function: Activity and connectivity associated with processing emotion and reward. Neuroimage 209, 116493 11. Kronman, H. et al (2021). Long-term behavioral and cell-type-specific molecular effects of early life stress are mediated by H3K79me2 dynamics in medium spiny neurons. Nature Neuroscience 24, 667–676 12. Brunson KL, Chen Y, Avishai-Eliner S, Baram TZ (2003). Stress and the developing hippocampus: a double-edged sword? Mol Neurobiol 27: 121–136 13. Ivy AS, Rex CS, Chen Y, Dubé C, Maras PM, Grigoriadis DE et al (2010). Hippocampal dysfunction and cognitive impairments provoked by chronic earlylife stress involve excessive activation of CRH receptors. J Neurosci 30: 13005–13015.

14. Shonkoff JP (2011). Protecting brains, not simply stimulating minds. Science 333: 982–983.

15. Hodel AS, Hunt RH, Cowell RA, Van Den Heuvel SE, Gunnar MR, Thomas KM (2015). Duration of early adversity and structural brain development in postinstitutionalized adolescents. Neuroimage 105: 112–119.

16. Wang, X.-D. et al (2011). Forebrain CRF₁ modulates early-life stress-programmed cognitive deficits. J Neurosci 31, 13625–13634 17. Brunson KL, Kramár E, Lin B, Chen Y, Colgin LL, Yanagihara TK, Lynch G, Baram TZ et al (2005).

Mechanisms of late-onset cognitive decline after early-life stress. J Neurosci 25: 9328–9338.

18. Nemeroff, C. & Dunlop, B. 688 (2017). The CRH-1 Receptor Antagonist Saga in Mood and Anxiety Disorders: The Good, the Bad and the Ugly. Biological Psychiatry 81, S279 Kumar, A., Rinwa, P., Kaur, G. & Machawal, L (2013). Stress: Neurobiology, consequences and management. J Pharm Bioallied Sci 5, 91–97 19. Fareri, D. S. & Tottenham, N (2016). Effects of early life stress on amygdala and striatal development. Dev Cogn Neurosci 19, 233–247 20. Cohen, M. M. et al (2013). Early-life stress has persistent effects on amygdala function and development in mice and humans. PNAS 110, 18274–18278 21. Ellis, B.H., Fisher, P.A., Zaharie, S (2004). Predictors of disruptive behavior, developmental delays anxiety, and affective symptomatology among institutionally reared romanian children. J. Am. Acad.

Child Adolesc. Psychiatry43, 1283–1292 22. Zeanah, C.H., Egger, H.L., Smyke, A.T., Nelson, C.A., Fox, N.A., Marshall, P.J. (2009).Institutional rearing and psychiatric disorders in romanian preschool children.Am. J. Psychiatry 166, 777–785.

23. Teissier, A. et al (2020). Early-life stress impairs postnatal oligodendrogenesis and adult emotional behaviour through activity dependent mechanisms. Molecular Psychiatry 25, 1159–1174 24. Rau, A. R., Chappell, A. M., Butler, T. R., Ariwodola, O. J. & Weiner, J. L (2015). Increased Basolateral Amygdala Pyramidal Cell Excitability May Contribute to the Anxiogenic Phenotype Induced by Chronic Early-Life Stress. J Neurosci 35, 9730–9740 25. Teissier, A. & Gaspar, P. Comment le stress précoce altère le comportement à l’âge adulte (2020).

Med Sci (Paris) 36, 218–221 26. Nelson, C. A (2017). Hazards to Early Development: The Biological Embedding of Early Life Adversity. Neuron 96, 262–266 27. Tanti, A. et al (2018). Child abuse associates with an imbalance of oligodendrocyte-lineage cells in ventromedial prefrontal white matter. Mol Psychiatry 23, 2018–2028

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