Maternal dysbiosis at birth as a model for increased risk of autism
Paul Ashwood, Ph.D. Dept. of Medical Microbiology and Immunology, UC Davis School of Medicine
Assess whether the alteration of maternal microbiota will result in long-term changes in microbiota composition in the offspring, altered local intestinal barrier function, increased neuroinﬂammation and altered behavioral outcomes.
Enhancement of tissue procurement from individuals with ASD
Thomas Blanchard, Ph.D. University of Maryland Brain and Tissue Bank
This 3-year proposal from the Maryland Brain and Tissue Bank is to identify additional medical examiners at national and state association meetings and to educate the public and medical examiners about the critical importance of tissue donations from individuals with ASD and matching controls for continued research into the etiology of autism.
Joint Hypermobility & Hypermobile Spectrum Disorders in Mothers of Children with Autism & ADHD
Emily Casanova. University of South Carolina
In this study on joint hypermobility, as well as related features, we plan to survey mothers of children with autism who are assessed by our clinic, which diagnoses and treats many hundreds of ASD children each year. In addition, we will assess the relationship of GJH features in mothers in association with: 1) estimates of cognitive impairment in the child, 2) maternal features associated with the Broader Autism Phenotype (BAP), and 3) maternally reported immune- (e.g., chronic respiratory allergies, asthma, hives, etc.) and hormone-mediated symptoms (e.g., polycystic ovary syndrome, menstrual disorders, etc.). Mothers of children with ADHD will be used as controls in this study.
To Determine the Minicolumnar Morphometry of Autistic, 15q dup and various Shank3 Mutant Mouse Models as Compared to Those in Control Tissue
Manuel F. Casanova, M.D. Endowed Chair in Translational Neurotherapeutics University of South Carolina Greenville, South Carolina
Examine differences in minicolumnar morphometry between the brains of 7 autistic individuals, 7 patients with 15q dup, 7 controls, all matched for age and sex (already obtained from the Autism Tissue Program), and the brains of various Shank3 mutant mice and wild type (n=15 total).
Role of Environmental Factors in Autism Spectrum Disorder
Ved and Abha Chauhan. The Research Foundation of Mental Hygiene
This project involves evaluating the effects of three environmental chemicals, i.e., bisphenol A (BPA), methylmercury (MeHg) and alcohol, alone and in combination, on development and behavior of Drosophila (fruit fly) larvae, along with behavior of flies. To understand the mechanism of developmental and behavioral alterations, they will also analyze the biochemical changes, gene expression, and protein expression.
Analysis of Common Factors in Genetic and Non-Genetic ASD Models
Andreas Grabrucker and Kieran McGourty. University of Limerickas
Using an original maternal zinc deficient mouse model to generate ASD behaviors in prenatal zinc deficient offspring, the investigators plan to use proteomic analysis of 4 brain regions to identify common biochemical pathways that are dysregulated in both genetic and non-genetic causes of ASD. Brain samples and behavioral analyses have previously been collected and this proposal will provide proteomic evaluation of these tissues and in addition provide an extended in silico comparison with other “omics” based ASD studies.
Metabolomics Analysis of Young Children with Autism Spectrum Disorders and Their Mothers Compared to Neurotypical Controls
Haiwei Gu and James Adams. Arizona State University
The overall goal of this proposal is to simultaneously analyze and compare urine samples from children with ASD and their mothers compared to NT controls using metabolomics technology. The investigators hypothesize that ASD induces a correlated reprogramming in children and their mothers which could be targeted for diagnosis and treatment before birth and in childhood.
Elevation of methionine sulfoxide in children with autism
Jill James, Ph.D. and Stepan Melnyk, Ph.D. University of Arkansas for Medical Sciences
The oxidized form of methionine, methionine sulfoxide was found to be elevated in children with autism relative to their siblings and unrelated controls. This proposal will extend and conﬁrm this ﬁnding with plasma previously collected from 50 ASD/sibling/control trios and determine the extent to which this marker of oxidative stress is related to other markers of protein damage. Positive results will provide a convenient marker for antioxidant treatment efficacy for protein damage.
Gender Dimorphism: Microbiome Analysis in Autistic Boys and Girls
Rafail Kushak, Ph.D., Dr. Sc., Massachusetts General Hospital Harvard Medical School
Test a new hypothesis suggesting that differences in the intestinal microbiota may underlie the well-established male preponderance in ASD.
Proteomic Analyses of Autistic Brain
Marvin Natowicz, M.D., Ph.D. Pathology & Laboratory Medicine, Genomic Medicine, Neurological and Pediatrics Institutes Cleveland Clinic, Cleveland, OH
This study, the ﬁrst proteomic analysis of ASD brains, deepens our understanding of the brain biology in autism. Conﬁrm earlier hypothesized dysregulated biological processes that were proposed on the basis of molecular genetic investigations such as disturbances of synapse biology. Provide new insights regarding the biology of ASD including, for example, insights regarding the dysregulation of key differentially expressed brain biochemical networks.
To study CREB and Akt after zinc therapy
A.J. Russo, PhD Hartwick College, Oneonta, New York
The PI has previously demonstrated that the transcription factor CREB and the protein kinase AKT are low in children with autism. In this proposal, the PI will determine or not zinc therapy in children with autism will signiﬁcantly raise both AKT and CREB levels in plasma and whether this increase will be associated with improved autism symptom severity in 50 children with autism and 50 age/ gender matched neurotypical children.
To assess MAPK pathway intracellular biomarkers in individuals with autism
A.J. Russo, PhD Hartwick College, Oneonta, New York
This research project is designed to determine whether or not dysrugulation of MAPK/ERK pathways, speciﬁcally over-activation and under-activation in lymphocytes, can be associated speciﬁc sub-populations of autism basedon symptom severity. The PI proposes to use a MAPK phosphorylation array to measure 17 pathway biomarkers and compare these marker concentrations with severity of 19 ASD symptoms.
Determination of Exosomal Biomarker Candidates in ASD
Judy Van de Water. UC Davis, MIND Institute
The study of exosomes is a relatively new and exciting area of research. Simply defined, exosomes are small extracellular vesicles that modulate important functions in cell physiology and development and are thought to have a pathologic function in certain conditions of the central nervous system (CNS). Exosomal contents include proteins, lipids and various RNA species that have been shown to be altered during disease. The fact that exosomes are released into the blood stream from blood cells and endothelial cells responding to CNS diseases as well as from the brain and spinal cord, and that they express markers that allow their tracking to the cell of origin make them appealing as a target metric for biomarker and diagnostic tool development. While the utilization of exosomes for biomarkers and diagnostics in diseases affecting the CNS are still in the early stages of discovery and development, the goal of this pilot project is to fill this knowledge gap and perform an unprecedented proof-of-principle study analyzing the exosomal RNA and protein cargo associated with autism spectrum disorder (ASD) aiming to identify potential biomarkers and to inform as to CNS status.
The role of the intestinal microbiome in children
Harland Winter, M.D. Rafail Kushak, Ph.D. Tim Buie, M.D. Harvard Medical School and Massachusetts General Hospital
This grant supports the continued enrollment of children with autism and neurotypical controls into the Harvard/ MassGeneral Hospital’s biorepository of blood and gastrointestinal tissue samples. Samples will be used for their own research and shared with other researchers who are pursuing novel and creative approaches to understand causes of autism and potential treatment options.
Identifying Metabolic Differences During Pregnancies in Mothers of Children Later Diagnosed with ASD
James B. Adams, David Haas, Haiwei Gu, and Juergen Hahn, Arizona State University
Prenatal supplements with folate are known to reduce the risk of neurological birth defects, and also seem to significantly reduce the risk of a child developing ASD, especially if taken preconception or early in the pregnancy. Our recent research on mothers of children with autism (several years after they gave birth) suggests that they have many metabolic differences from mothers of typically-developing children, and suggests that certain nutritional supplements including folate could be important in improving their metabolism. So, we propose to investigate levels of those metabolites in blood samples that were stored from a pregnancy cohort of 10,000 women, and compare the metabolite levels during pregnancy in the mothers whose children developed ASD vs. those who developed typically. Our hope is that this study will lead to the discovery of other nutrients that may improve the mother’s metabolism and reduce the risk and/or severity of ASD in their children, as well as co-occurring symptoms including seizures, intellectual disability, gastrointestinal problems, and sensory sensitivities.
The Domino-Drosophila Ensemble for the Functional Validation of New ASD Genes
Alexandros Kanellopoulos, University of Lausanne, Switzerland
Autism Spectrum Disorders (ASDs) are characterized by significant limitations in both intellectual functioning and adaptive behavior. Despite substantially different clinical phenotypes and genetic contributions, there seems to be a convergence on common cellular pathways, which ultimately affect brain wiring, function and behavior. A large list of potential ASDs risk genes has been compiled through analysis of copy number variants (CNVs) or single-nucleotide polymorphisms (SNPs). In addition, whole-exome sequencing (WES) and whole-genome sequencing (WGS) lead to the identification of de novo, rare recessive heritable mutations. In contrast to recessive conditions, identification of dominant mutations for ASDs is more difficult, due to the abundance of benign heterozygous variants that create massive background noise.
In our innovative project we plan to identify and experimentally validate new genes for Autism. Specifically, our approach is based on a machine-learning algorithm which predicts with high confidence, which genes may harbor autosomal-dominant mutations, distinguishing the true ASDs risk genes from false-positive associations. We have currently extracted 141 de-novo mutations from a public database that possibly cause ASDs, and 7 of these are novel putative genes. The validation will be performed using the fruit fly Drosophila melanogaster, a valuable animal model for a medium-throughput screening.
Single Session Intervention for Comorbid Internalizing Symptoms in ASD
Matthew Lerner, Stony Brook University
Teens with autism spectrum disorder (ASD) experience substantial internalizing (anxiety and depression) symptoms, as well as considerable social challenges, with direct effects on functional impairment and distress. Existing interventions for these symptoms are resource-intensive, with variable effects. Growth mindset single-session interventions (GM SSI) represent a promising, new, scalable approach to addressing these symptoms, which can be delivered via computer in just 30 minutes. GM SSIs have been shown to reduce internalizing symptoms in youth without ASD. This project is the first to examine a GM SSI in teens with ASD; it employs a randomized controlled design with a well-matched attention control (a supportive therapy [ST] SSI). This project will evaluate effects of GM SSI on internalizing symptoms and core social and executive functioning outcomes. Thirty youth with ASD will be randomly assigned to receive the GM SSI or ST SSI. They will complete a single assessment/intervention visit, with immediate pre-post assessment, then return after 3 months for follow-up. This approach allows for identification of whether GM SSIs can be effective for youth with ASD while controlling for nonspecific treatment factors, thereby enhancing the strength and interpretability of any obtained findings.
Using Zebrafish to Help Identify Which Environmental Toxicants Could Increase Autism Risk
Edward Levin, Duke University
In the proposed project, we will use the zebrafish model to investigate the behavioral consequences of environmental toxicants affecting retinoic acid receptors (RARs) during early neural development. Zebrafish provide an efficient model which bridges the gap between high throughput cell based in-vitro models which can quickly screen many compounds on a basic level and complex but slow and expensive investigations of humans in epidemiological studies and rodents in experimental studies. We will study RAR acting compounds because RAR signaling plays a critical role in proper formation of the neural tube/plate. Disruption of RAR signaling such as with the anticonvulsant drug valproic acid (VPA) has been shown to increase risk of Autism. We anticipate that even modest disruptions in RAR signaling will not result in dramatic alteration in neural organization until a later stage of neurodevelopment when the brainstem nuclei project rostrally to help organize later more rostral neurodevelopment.
Then, the full phenotypic expression of the early neurotoxic injury may manifest in behavioral dysfunction. Here we propose that exposure to environmental toxicants affecting RAR signaling contribute to the increasing rates of Autism. We have found the zebrafish tests are sensitive to VPA and hypervitaminosis A induced impairments in social will be used to screen environmental compounds exhibiting RAR agonist activity which have been identified through mining recent Tox21/ToxCast high throughput screening data. Zebrafish offer a higher throughput and economic model system enabling analysis neurobehavioral consequences as affected by toxicants impacting RAR signaling during development thereby contributing to the risk of developmental neurobehavioral toxicity.
Corneal Confocal Microscopy: A Surrogate Marker for Neurodegeneration in Autism
Rayaz Malik, Weill Cornell Medicine, Qatar
Major challenges in managing people with Autistic Spectrum Disorder (ASD) include its heterogeneity, poor knowledge of disease trajectories and an inability to accurately identify the extent and progression of underlying neurodegeneration. Neuroimaging studies confirm altered neural connectivity and trajectories for normal brain development, but lack precision. Whilst the focus of these studies has been on central brain pathology, recent studies show altered tactile discrimination and allodynia on the face, mouth, hands and feet of subjects with autism, indicating defects in peripheral somatosensory neurons. Indeed, a reduction in lower limb intraepidermal nerve fibre density has been reported in children with autism. We have pioneered the technique of corneal confocal microscopy (CCM) to rapidly image corneal nerve fibres and established it as a biomarker for peripheral and central neurodegenerative disease. In our pilot study of eight children with ASD, we show a significant reduction in the corneal nerve fiber length (P<0.01) and branch density (P<0.01) compared to healthy controls. We propose to assess if CCM can act as a rapid and reliable non-invasive biomarker of neurodegeneration in people with ASD.
Assessing Novel Impacts of RASopathies on Cortical GABAergic Neurons
Daniel Vogt, Michigan State University
Autism spectrum disorder (ASD) may be caused by multiple factors, including environmental impacts and gene mutations, the latter with several hundred genes implicated. It has been difficult to study ASD due to the shear enormity of the factors at play and multitude of symptoms that manifest. However, there are several monogenetic syndromes with high rates of ASD that may be the low hanging fruit (one gene that leads to ASD symptoms) that could uncover which biological mechanisms are altered, and in turn, may be targeted, to alleviate ASD symptoms. The RASopathies are syndromes caused by mutations in genes regulating the RAS/MAPK pathway, which is associated with high rates of ASD. While much work has uncovered genetic causes of these syndromes, the molecular and cellular changes, and their contribution to ASD symptoms, are poorly understood. We deleted the RASopathy gene, Nf1, in GABAergic interneurons, and uncovered a drastic loss of a key GABAergic control gene and loss of a specific types of interneuron, i.e. those expressing parvalbumin (PV). Since GABAergic neurons are the primary driver of brain inhibition, this discovery may uncover some mechanisms underlying the neuropsychiatric changes in ASD. This proposal will be the first to assess the core GABAergic molecular program in RASopathy models with low to high rates of ASD diagnoses as well as correlate our findings with rates of ASD diagnoses. Moreover, PV+ neurons have been implicated in human ASD cases and in many animal gene models of ASD, suggesting that understanding how these cells are impacted could provide new inroads into ASD.