The Autism Research Institute (ARI) conducts, sponsors, and supports research on the underlying causes of, and treatments for, Autism Spectrum Disorders (ASDs). In order to provide parents and professionals with an independent, unbiased assessment of causal and treatment efficacy issues, ARI seeks no financial support from government agencies or drug manufacturers.
We therefore rely on the generosity of donors so that we may continue to advance autism research. Our founder Dr. Bernard Rimland would often say, ‘Research that makes a difference!’ to remind us of the need to focus on what might be beneficial here and now for people with ASDs.
MeHG/D-Penicillamine Experiment
(PI: Vas Aposhian, Ph.D., Dept. of Molecular & Cellular Biology, University of Arizona.)
This study involves two sets of experiments. The first set is designed to confirm a previous finding, that D-penicillamine reduces brain mercury, and to examine whether organic or inorganic mercury are excreted from an animal brain. The second set of experiments will be similar to the first set, except the animals will be autopsied at 1 day, 2 days, and 32 days of D-penicillamine treatment to determine if there is an initial increase in brain mercury, as is often observed with some chelating agents. In both sets of experiments, mercury levels will be measured in brain and kidney.
Autistic Enterocolitis: microscopic ileo-colitis in childhood and ileo-colonic Crohn’s disease in adulthood. Confirmation of endoscopical and histological evidence of a progressive inflammatory involvement of the bowel.
(PI: Federico Balzola M.D., Gastro-Epatologia Dipartimento Medico-Chirurgico delle Malattie dell’Apparato Digerente e della Nutrizione Azienda Ospedaliera Molinette C.so Bramante 86 10126 Torino, Italy.)
This two-year study aims: (1) to evaluate the prevalence of Autistic Enterocolitis with microscopic pattern in patients with regressive autism under 14 years of age, and to evaluate the prevalence of Autistic Enterocolitis with Crohn’s disease pattern in 40 consecutive patients over 14 years old with regressive autism; (2) to identify and evaluate a group of routine blood tests that could correlate with the intestinal disease and to evaluate the IBD serological and faecal markers in the two subgroups of patients; and (3) to evaluate any variations in the routine blood tests and IBD serological and faecal markers in the groups of patients who submitted to pharmacological therapy after 6 and 12 months of follow-up.
Improving the quality of life for people with autism and their families by integrating biomedical and behavioral approaches to address pain and illness.
(PI: Edward Carr, Ph.D. , State University of New York at Stony Brook and Developmental Disabilities Institute.)
Presently the field is undergoing a transformation to a “whole-body approach” to ASD in which biomedical factors, particularly those involving physical illness, are being closely examined for their impact on individuals’ functioning, and their families. Our research program at Developmental Disabilities Institute suggests that the integration of biomedical and behavioral approaches can result in benefits beyond those achieved by using either approach alone.
Markers of Inflammation and Oxidative Damage in Autism
(PI: Abha Chauhan, Ph.D., Developmental Neuroscience Laboratory, NYS Institute for Basic Research in Developmental Disabilities.)
The aim of this project is two-fold: (1) to study markers of oxidative stress in autism; and (2) to study whether anti-inflammatory response and/or immunological response is abnormal in autism.
To ascertain oxidative stress, we will analyze levels of ROS (free radicals) in the serum, levels of markers of oxidative damage to proteins and DNA in the plasma, and total antioxidant status in serum. With regard to anti-inflammatory responses, we will study inflammatory markers in autism by measuring (a) levels of acute phase proteins such as C-reactive protein, haptoglobin, alpha 1-antitrypsin, and also a2-macroglobulin in the serum from children with autism and controls; (b) complement cascade pathways of activation, i.e., classical, alternate and lectin pathways, and levels of different components of complement cascade (C3A, iC3b, C4a, C4d, C5A), membrane-attack complex (SC5b6789), and complement C1q- and C3d-containing circulating immune complexes will be studied in the serum of autism and controls; and (c) levels of different cytokines and chemokines in the plasma will be studied using Luminex technology. With regard to an imbalance in immune response, we will compare in the serum of autistic and non-autistic siblings the levels of different immunoglobulins and their subclasses: IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgA subclasses (IgA1, IgA2) and IgM.
We will also examine the relationship between oxidative stress/ inflammation/ immune response and severity, and/or behavior and language problems in low-functioning and high-functioning autistic children. Analysis of levels of ROS, oxidative markers, APP, immunoglobulins, complement proteins and cytokines in relation to severity and behavior/ language abnormalities in different cohorts of autism will suggest whether the measurement of these parameters can be of value in supporting the behavioral diagnosis of autism. The relationship, if any, between these abnormalities and low- or high-functioning autism groups, as well as severity of behavior deficits in autism, will also be studied.
Enhancement of Tissue Procurement from Autistic Individuals
(PI: Kathleen M. Currey, M.D. and H. Ronald Zielke, Ph.D., University of Maryland, Baltimore, NICHD Brain and Tissue Bank for Developmental Disorders.)
The focus of the NICHD Brain and Tissue Bank for Developmental Disorders at the University of Maryland is to retrieve brain tissue from individuals with developmental disorders, including autistic children, and matching controls. Our goal is to recruit four or five additional medical examiners. These tissue donations are critical to shorten the time until the causes of autism and other pervasive developmental disorders are known.
Measuring HCY Thiolactone in autism
(PI: Richard Deth, Ph.D. , Northeastern University.)
Homocysteine thiolactone (HcyTL) is a cyclic form of homocysteine (Hcy) that is produced as a “by-product” of the loading of amino acids onto transfer RNA (tRNA) during protein synthesis. There are two components of this research project: (1) develop a novel GC-MS or LC-MS-based assay for HcyTL; and (2) measurement of plasma HcyLT levels in autistic and normal control subjects.
Methionine Synthase Status as an Oxidative Stress Biomarker
(PI: Richard Deth, Ph.D. , Northeastern University.)
Initial studies will utilize cultured lymphoblasts to validate the utility of measuring methionine synthase modifications as indicators of oxidative stress status. Methionine synthase status in 8 pairs of lymphoblasts from autistic subjects and same-sex neurotypical siblings under normal growth conditions will be compared before and after induction of oxidative stress by blocking the synthesis of glutathione with buthionine sulfoxide (BSO) treatment. We will measure cellular levels of glutathione to confirm that they are lower after BSO treatment, and then use PCR to evaluate methionine synthase mRNA. We will utilize primer pairs directed against each of the five domains to determine if they are all present, or whether the Cap domain is absent. Primers directed against individual exons within the Cap domain will allow us to learn which specific exons are being removed by alternative splicing. Based upon our preliminary findings we hypothesize that the Cap domain will be partially spliced under normal growth conditions, but more extensively spliced under conditions of oxidative stress. In addition to providing information about alternative splicing of mRNA, the PCR assay will also indicate the overall amount of mRNA, a reflection of the level of transcription. If we did indeed observe changes in methionine synthase mRNA status, it would strongly suggest that alternative splicing of methionine synthase mRNA in blood cells might be a useful marker of oxidative stress status. In addition to mRNA analyses, we will assess the molecular size of methionine synthase in the 8 sib-pairs of lymphoblasts, to confirm absence of domains from alternative mRNA splicing, but also to determine whether additional domain deletion occurs at the level of the protein.
Pilot test of Actos in autistic children: A double-blinded, placebo-controlled, crossover study on the effects of Actos on behavioral and metabolic measures in children with ASD.
(PI: Douglas Feinstein, Ph.D., Department of Anesthesiology, University of Illinois.)
The goals of this study are: (1) to determine if treatment with Actos reduces behavioral abnormalities as assessed using the Aberrant Behavior Checklist (ABC) exam in children diagnosed with ASD; (2) to determine whether Actos will normalize plasma cytokine profiles; and (3) to determine if any measured improvements in behavior or cytokine levels are maintained for up to three months after stopping Actos treatment.
Impact of Innate Immunity on Regressive Autism
(PI: Harumi Jyonouchi, M.D., Department of Pediatrics, UMDNJ-New Jersey Medical School.)
This study focuses on the dissection of intrinsic abnormalities of innate immune cells, and their effects on adaptive immunity and tolerance induction. Identification of innate immune abnormalities that might be biomarkers for identifying ASD children in whom development of ASD is strongly associated with neuro-immune interactions. This will include (1) an assessment of responses to TLR stimulants by isolated innate immune cells from ASD children; and (2) an assessment of cellular immune reactivity to common luminal Ags in ASD children.
Rising Glutathione Levels in Children with Autism
(PI: Janet Kern, Ph.D. , Autism Treatment Center, Dallas, Texas.)
The proposed study will examine the use of two supplemental programs to determine their efficacy in raising GSH levels in children with autism. These are: (1) Oral Lipoceutical GSH; and (2) Transdermal N-acetyl L-cysteine (NAC) + oral Vitamin B6 + oral Vitamin C. The aim is (a) to measure the change in GSH levels after a 6-week trial, (b) to examine tolerance and side-effect, (c) to monitor any changes in the symptoms of autism while on the supplemental programs.
Neuronal Oxidative Stress in Autism; Characterization of CEP-modification as an oxidative stress marker in autistic brain
(PIs: Woody McGinnis, M.D. , Ashland, Oregon, and Xiongwei Zhu, Ph.D. , Dept of Pathology, Case Western University, Cleveland, Ohio.)
This study investigates whether episodic or sustained oxidative stress contributes to altered brain development and function, which then manifest as autistic behavior. The goal is: (1) to determine the differential content of CEP in various brain regions (both inside and outside of blood brain barrier) from autistic patients and to correlate alterations with morphological abnormalities, behavioral deficits or onset of autism; (2) to determine the association of CEP with abnormal systems in autism by double-staining for GABA-ergic, glutamatergic, cholinergic, and oxytocin receptors, as well as glutamic acid decarbosylase (GAD) and reelin; and (3) to determine the proteins modified by CEP in autistic brain.
Support for Several Studies
(PI: Elizabeth Mumper, M.D., The Rimland Center, Lynchburg, Virginia.)
These studies include: (1) a pilot study examining methylation abnormalities and oxidative stress in children undergoing hyperbaric therapy; (2) a study comparing metabolic organic acid profiles in urine of children with Autism Spectrum Disorder to 50 control (healthy, non-autistic) children; and (3) a study to compare the prevalence of Rh-negativity in mothers of children with neurodevelopmental disorders vs. children with normal development.
Brain region-specific oxidative stress in autism: standardization of tissue protocols and evaluation of potential artifacts
(PI: Elizabeth Sajdel-Sulkowska, D.Sc. , Assistant Professor of Biochemistry, Dept. of Psychiatry, Harvard Medical School, Boston, Mass.)
The proposed studies will test the hypothesis that elevated oxidative stress in autism is brain-region specific. By measuring oxidative-stress markers in diverse regions of brain, we will begin the first quantitative mapping of oxidative changes in autistic brains. The hypothesis will be tested using both postmortem human brain tissue from autistic and control cases, and rat brain tissue isolated from neonates exposed to neurotoxins (LPS, MSG, and Hg) during the critical developmental period (postnatal days 4-7) corresponding to the last trimester of human pregnancy. The specific aims are: (1) to standardize tissue protocols for measuring oxidative stress markers; (2) to evaluate potential artifacts in assessing oxidative stress markers in brain tissue; and (3) to compare the region-specific distribution of oxidative-stress markers in autistic and control brains, and in animals exposed to neurotoxins known to exert oxidative effects in brain.
Use of Probiotics to prevent the recurrence of autism in sibpairs
(PI: Gene Stubbs, M.D., Professor Emeritus of Psychiatry and Pediatrics, Oregon Health & Sciences University.)
This is a pilot study using a comparison of reported recurrence of autism in families (7-10%) versus those who have at least one child with autism whose newborn sibling gets treatment in our study with probiotics (VSL#3). Probiotics have been used safely in infants to prevent allergy onset. These products have been used safely anecdotally in children with autism who have abnormal bowel symptoms. VSL#3 has been studied in ulcerative colitis, Crohn’s Disease, and Irritable Bowel Disease, and shown to be safe and effective. VSL#3 has been shown to colonize the gut microflora during the period of time it is taken.
Efficacy of neuromodulation technique with children diagnosed with autism.
(PI: Robert H. Weiner, Ph.D. , Dallas, Texas.)
The purpose of this study is to determine whether neuromodulation technique (NMT) is effective in reducing maladaptive behaviors and increasing adaptive behaviors in children diagnosed with autism. This is a pilot, Phase I Study: Treatment, Randomized, Wait-list Control. This is project is part of an international (US, Canada, Mexico), multi-site and interdisciplinary study.
Clinical Trial of School Aged Children w/ASD using holistic approach through diet, lifestyle, and supportive supplement modification.
(PI: Marjie C. Andrejciw, MT (ASCP), MS, NC. Hawthorn University.)
The intent of the study is to improve behavior, self control, the ability to focus, social skills and the uncontrollable urges and tics in five to ten school age children with autism using an individualized diet (GFCF), lifestyle, and supportive supplements (e.g. Cod liver oil and Brain Child Nutritional spectrum supplement). A 12-week, pre and post-test, control group design will be employed. Pre and post-laboratory testing will be conducted for children in the experimental group. Parents/teachers/caregivers will fill out the Autistic Treatment Evaluation Checklist (ATEC) prior to beginning treatment and every four weeks thereafter for the twelve week study period. Control group parents will also fill out the ATEC on the same time schedule.
Evaluation of Tibial Bone Lead Levels in Autistic Children
(PI: Marvin Boris, M.D. Autism Associates of New York.)
Neurotoxicity of lead is a well-documented medical problem in children, and in the past several years many children with autism have been documented to have high levels of lead in urine after undergoing chelation therapy. The parameters for determining which children should undergo chelation are not clearly established. Developing a more precise measure of lead exposure and absorption may help evaluate lead exposure and chelation effectiveness. The study is designed to measure tibial lead content via the non-invasive X-Ray Fluorescence (XRF technique). This exploratory study will measure lead exposure through the XRF technique in a convenience sample of 20 patients from the medical practice.
Multidimensional Impact of Pain on Individual and Family Functioning in Autism Spectrum Disorder.
(PI: Edward G. Carr, Ph.D. Department of Psychology, State University of New York Stony Brook.)
Children with ASD frequently present with a wide array of health and illness issues. This study proposes to highlight the role of pain and discomfort as a final common pathway through which multiple medical conditions and physiological states exert their effects on individual behavior and family functioning. The relationship between pain and behavior will be examined through a series of measurements on 240 subjects. Parents/caregivers will fill out questionnaires and children will be examined. Instruments include but are not limited to: Non-communicating Children’s Pain Checklist-Revised, GI symptoms checklist, Aberrant Behavior checklist, Children’s Sleep Habits Questionnaire, Autism Treatment Evaluation Checklist, child Behavior Questionnaire, and Vineland (Communication subscales). Bivariate correlations between pain and dependent variables from the ABC and CSHQ will be conducted, followed by stepwise regression analysis to examine the effect of moderator variables such as communication skills, and autism treatment on the pain relationship. The study has implications for assessment, treatment, theoretical issues, and should pave the way for future research.
Markers of inflammation and oxidative damage in autism.
(PI: Abha Chauhan, Ph.D. New York State Institute for Basic Research in Developmental Disabilities/Research Foundation for Mental Hygiene, Inc.)
There is limited knowledge of the causative or secondary abnormalities in the biochemical pathways in autism. Increasing evidence suggests that autistic subjects have signs of oxidative stress, and that their immune response and/or inflammatory response may be compromised. Increase in oxidative stress in autism can be due to the imbalance between generations of free radicals i.e. reactive oxygen species (ROS) and defense mechanism against ROS. Anti-oxidant enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), catalase) play important roles in the biological redox system. The aims of the project are to study: the mechanism of oxidative stress in autism, and whether the anti-inflammatory and/or immunological response is abnormal in autism. The status of oxidative stress and antioxidant potential in lymphoblasts were compared in autistic subjects (n=11) and control subjects (n=11) by analyzing lipid peroxidation (LPO), generation of free radicals (ROS – reactive oxygen species), extent of membrane damage and activities of antioxidant enzymes.
Research Assistance for Clinical Biomedical Studies.
(PI: John Green, M.D. Evergreen Center, Oregon City, Oregon.)
This study will provide funding for research assistant to: review laboratory results, create charts and graphs, assist with bacteria and blood sample research studies, assist with laboratory quality assurance, develop presentation, conduct computer searches, education/outreach with physicians and parents, in conjunction with research efforts conducted at the Center.
Raising Glutathione Levels in Children with Autism.
(PI: Janet Kern, Ph.D., Autism Treatment Center, Dallas, Texas.)
The study is a continuation of the 2007 study that examines the use of two supplemental programs to determine their efficacy in raising GSH levels in children with autism. These are: (1) Oral Lipoceutical GSH; and (2) Transdermal N-acetyl L-cysteine (NAC) + oral Vitamin B6 + oral Vitamin C. The aim is (a) to measure the change in GSH levels after a 6-week trial, (b) to examine tolerance and side-effect and (c) to monitor any changes in the symptoms of autism while on the supplemental programs.
Measuring mercury body burden in three distinct groups to help elucidate the role of mercury in autism pathology.
(PI: Janet Kern, Ph.D. Autism Treatment Center, Dallas, Texas.)
Recent studies in autism found that children with ASD had significantly elevated urinary pentaporphyrin (5cxP), precoproporphyrin (prcP) and coproporphyrin (cP levels as compared to controls. These markers of mercury toxicity were reduced in children who had been chelated. The specifics aims of the study are to determine the mercury body burden differences using urinary porphyrins between three distinct groups: children in the US with autism who have not been chelated, children in the US who are typically developing, and children in France who are typically developing (thimerosal was banned in France in 2001.) Urine specimens will be collected from all children. All children will also be evaluated using the Childhood Autism Rating Scale (CARS). Typical children will be gender, age, and race matched to the children with autism. All porphyrins will be compared.
Autism: Analysis of Neocortical Lamination in Autism
(PI: Kathryn McFadden, M.D. University of Pittsburgh, Department of Pathology/Division of Neuropathology.)
Neuroanatomic and histologic studies of autistic subjects have consistently shown substantially altered postnatal brain growth trajectories associated with abnormal underlying cortical micro architecture. The changes have been noted to predominately affect the rostral cortex. The hypotheses for this study include: (1) there will be a disproportionate increase in neuronal density and laminar depth in superficial cortical layers derived from the SVZ relative to deeper layers derived from the VZ in autistic brains and (2) this divergence will be more pronounced in rostral cortical areas relative to more caudal regions. The proposal to test these hypotheses includes delineating cortical layers and certain neuronal subpopulations using “layer-specific” antibodies. We will compare various rostral to caudal cortical areas between autistic individual and age-matched controls. More specific definition of the cortical phenotype(s) in autism will allow us to better delineate which developmental processes are more impacted. This project was approved by the Autism Tissue Project and the investigators received samples from the Brain Atlas Celloidin Project on which to conduct preliminary studies.
Neuronal Oxidative Stress in Autism
(PI: Woody McGinnis, M.D. and Xiongwei Zhu, Ph.D. Case Western Reserve University, Department of Pathology.)
The pattern of CEP (carboxyethyl pyrrole (CEP)-protein adducts) appears to be a hallmark of the autistic brain, implications of this indicate that there is increased oxidative stress. The broad long term goals of the study are to: (1) determine the extent to which CEP and other oxidative stress parameters correlates with mental dysfunction; (2) determine the potential causes of oxidative stress and to understand the mechanism by which brain oxidative stress leads to autistic symptoms; (3) establish animal models of autism useful for further research and therapeutic intervention studies; (4) determine whether the reduction in oxidative stress correlates with neuronal survival and improvements in mental function; and (5) provide tools for measuring the efficacy of therapeutic treatments in reducing oxidative stress. The study will be completed using human postmortem fixed and frozen brain tissue from six autistic and six control subjects from the NICHD Brain and Tissue Bank at the University of Maryland.
The Involvement of Medullary Vagal Nuclei in Autism Spectrum Disorders.
(PI: Veronica Miller, Ph.D. New York State Department of Health, Wadsworth Centre for Laboratories and Research.)
Some patients with autism have dysfunctional heart rate, gastrointestinal, and inflammatory responses, which are suggestive of vagal nerve abnormalities. Exposure to environmental toxicants and pathogens are associated with the pathogenesis of autism. The hypothesis for this study is that medullary vagal nuclei may be particularly to toxicant or pathogen induced inflammation in autism for three different reasons. First, the dorsal nucleus of vagus (DMV) in the medulla is adjacent to the area postrema; a brain region lacking a complete blood-brain barrier. Secondly, vagal-sensory C fibers lack myelin and may be vulnerable to nerve damage, and thirdly, the DMV receives retrograde axonal input from sensory ganglia on the myenteric plexus and cardia tissues; providing a route for food-borne or blood-borne pathogens to the brain. The study will obtain brainstem tissues containing the DMV and additional medullary nuclei form Maryland Brain and Tissue Bank to determine if medullary inflammation is found in autism. Tissues from four patients with autism and four control patients will be used and pathological analysis of markers directed against activated neuralgia, macrophages, and ubiquitinated inclusions will be used to determine inflammation in the vagal nerve.
Brain region-specific oxidative stress in autism: standardization of tissue protocols and evaluation of potential artifacts.
(PI: Elizabeth Sajdel-Sulkowska, D.Sc. Department of Psychiatry, Harvard Medical School)
This study continues to test the hypothesis that elevated oxidative stress in autism is brain-specific. By measuring oxidative-stress markers in diverse regions of the brain, the study begins the first quantitative mapping of oxidative changes in autistic brains. The hypothesis is tested using both postmortem human brain tissue from autistic and control cases, and rat brain tissue isolated from neonates exposed to neurotoxins (LPS, MSG, and Hg) during the critical developmental period (postnatal 4-7 days) corresponding to the last trimester of human pregnancy. The specific aims are: (1) standardize tissue protocols for measuring oxidative stress markers; (2) evaluate potential artifacts in assessing oxidative stress markers in autistic and control brains, and in animals exposed to neurotoxins known to exert oxidative effects in brain.
Oxidative stress and changes in brain neurotrophin levels: relevance to autism.
(PI: Elizabeth Sajdel-Sulkowska, D.Sc. Department of Psychiatry, Harvard Medical School and Brigham and Women’s Hospital)
The study will test the hypothesis that oxidative stress implicated in autistic pathology is associated with altered levels of brain neurotrophins. The hypothesis will be tested using SHR rats that exhibit hypertension induced oxidative stress, assuming that these animals will exhibit increased oxidative stress when exposed to lipopolysaccharides (LPS) or thimerosal treatment during the developmental period corresponding to the fetal or early postnatal period. The study should relate the effect of environmental toxins to oxidative stress and its relationship to further changes in brain neurotrophins critical for brain development, as well as to suggest the possible use of neurotrophins in autism therapy. SHR rats will be exposed to either LPS or thimerosal, and tested for motor coordination, following testing the brain tissue will be moved for subsequent analysis 3-NT, neurotrophins, and Purkinje cell counting.
Split Sample Reproducibility Study
(PI: Elizabeth Mumper, MD. The Rimland Center, Virginia. Nancy O’Hara, MD, Optimal Health Initiatives, Wilton, CT.)
Many clinicians rely on lab results from specialty labs in making clinical decisions. It is important to know if these lab tests are accurate (correct answer), interpreted appropriately (normal reference range and correct commentaries), and specific (where the analysis pertains to the analyte being measured and only to that). As a first step in this process, we measured precision – the ability to test the same sample twice and get the same or similar results. It is critical to understand that our study only measured reproducibility, not reliability, accuracy, specificity, or the appropriateness of reference ranges or commentaries.
Blood and urine specimens were collected from the clinical practices of Elizabeth Mumper, Nancy O’Hara and Gail Szakacs, all physicians involved in the integrative care of children on the autism spectrum. The results were reported to the DAN! Think Tank. A list of labs that passed by our criteria for specific tests was distributed to clinicians and researchers. (Add link to split sample reproducibility results here.)
National randomized double-blind placebo controlled vitamin/mineral study.
(PI: Jim Adams, Ph.D. Arizona State University.)
The study will test a new liquid vitamin/mineral supplement for children and adults with autism in a 12-week double-blind, placebo-controlled trial. A total of 300 children and adults will be recruited into the study. Parents/guardians, and teachers/speech pathologists will fill out forms (ATEC, PDD-BI, Severity of Autism Scale) to collect data pre-and post-supplementation. A study nurse will collect data on dosing and symptoms every two weeks during the study.
CD8 + T lymphocyte function in autism.
(PI: Paul Ashwood, Ph.D. M.I.N.D. Institute, University of California Davis.)
Genomic profiling of children with autism has recently demonstrated alterations in genes specific to cytotoxic immune cell subsets namely, CD8+ cytotoxic T lymphocytes (CTL) and natural killer cells. The goal of this research is to assess the nature and extent of CD8+ cytotoxic T cell immune function in autism, in order to test the hypothesis that in autism there is a fundamental defect at the CD8+ CTL level that ultimately leads to abnormalities in antiviral and/or inflammatory immune function. The study will: (1) determine phenotypic and functional differences in peripheral blood CD8+ CTL cells between children with autism and typically-developing controls; (2) compare CD8+ CTL cell effector responses and proliferative capacities in response to stimulation; and (3) determine phenotypic and functional differences in CD8+ CTL cells that recirculate specifically to mucosal intestinal tissue.
Modulation of Neuronal Cysteine Update and Redox Status by Morphine, Gluten/Casein-derived Opiates and Naltrexone.
(PI: Richard Deth, Ph.D., Northeastern University School of Pharmacy, Boston, MA)
There is substantial evidence indicating that contemporary autism is associated with systemic oxidative stress and neuroinflammation. Using cultured human neuronal and glial cells, the project will investigate the dose-dependent ability of opiate receptor agonists (e.g. morphine and gluten/casein-derived opiod peptides) and antagonists (e.g. naltrexone and naloxone) to affect EAAT3-mediated cysteine uptake. The project will also assess the influence of these agents on intracellular levels of thiol metabolites as well as cellular redox status. Results from this project will provide a mechanistic link between the GF/CF diet, as well as LDN and the metabolic pathway whose dysfunction underlies autism.
Investigation of the Effect of 2 Treatments on ATP: NADH and Ribose.
(PI: Stuart Freedenfeld, Arizona State University.)
ATP is the primary fuel for the brain and body, so it is very important for normal functioning. Preliminary data found the 2/3 children with ASD have unusually low levels of ATP. Both NADH and Ribose are anticipated to increase ATP levels. In addition, since ribose uses adenosine to form the backbone for ATP, it may have an effect of lowering elevated adenosine. Elevated adenosine has been shown to interfere with the conversion of SAH to homocysteine and thereby has a negative effect on the methylation and sulfation pathway. We will investigate whether ribose lowers elevated adenosine levels and whether this improves the methylation and transulfuration pathways. The addition of guanosine and inosine will help to ascertain the mechanism of any effect on adenosine levels. The purpose of the study is to determine the biochemical effects of NADH and ribose in two groups of children. Each child will serve as their own control with baseline values and each child will be re- evaluated after treatment. Each child will receive the full test panel prior to initiation of intervention then they will be assigned randomly to either NADH or ribose. Evaluations will be repeated after two weeks of treatment. There will be ten children between the ages of 3 and 15 with a diagnosis of ASD in each group.
A Double Blind, Randomized Clinical Trial of Levocarnitine to Treat Autism Spectrum Disorders.
(PI: Mark Geier, MD, Ph.D. Institute of Chronic Illness, Silver Spring, MD.)
The project is a controlled study of oral, liquid L-carnitine (levocarnitine) administered to 20 children with ASD. Ten children without ASD (controls) will be given a placebo. Both will be given for 3 months. Children will be between the ages of 2 and 10 years of age. Children will be randomly assigned to treatment or placebo group. Groups will be blinded to caretakers and medical professionals. It is hypothesized that blood carnitine levels in children with ASDs have a significant impact on behavior, cognition, socialization, and health/physical traits. Outcome will be judged by behavioral and cognition metrics (ATEC, CARS, CGI) and muscle strength testing. Laboratory chemistry tests will also be performed at the beginning and end of the 90 day study period.
Impact of Innate Immunity on T and B cell differentiation in autistic children/Altered TLR response in a subset of children with regressive autism.
(PI: Harumi Jyonouchi, MD. Department of Pediatrics, UMDNJ- New Jersey Medical School, Newark, NJ.)
There is an urgent need for reliable (reproducible) biological markers along with genetic markers that distinguish ASD subsets, since therapeutic approaches likely differ in each subset. In this study, the ASD test group is categorized as regressive autism. They have markedly fluctuating behavioral symptoms and cognitive skills affected by benign immune insults such as common childhood microbial infection. Using blood samples, this study will determine mRNA expression in non-monocyte populations in the ASD test group, an ASD control group, and in non-ASD controls. The study will also determine whether mRNAs expressed in NK cells are up-regulated in ASD children as reported by others when RNA was extracted from whole blood. The study will also enumerate peripheral blood Treg, Th17, and memory B cell subsets in the ASD test, ASD control and non-ASD control groups.
Repository for Tissues from Children with and without Autism.
(PI: Rafail Kushak, Ph.D. Massachusetts General Hospital, Boston, MA.)
This project will assist with management and maintenance of the Digestive Function Tissue repository at the Massachusetts General Hospital. This repository was started in 1999 and contains tissues from individuals with autism and non-affected controls with gastrointestinal symptoms. Excess/residual tissues, as well as urine and stool samples, left after study completion are kept frozen in the repository for possible future research. An example of future research might be that blood samples from the repository may be used to develop an ELISA assay for casomorphin analysis in patients with ASD. Funding will be used to reduce shortages of storage space and equipment such as -20 and 80 degree freezers and purchase freezerworks software and freezerworks bar code accessories. Tissue samples kept in the repository represent a good source of material for current and future research.
Evaluation of altered fatty acid metabolism via Time-of-Flight Secondary Ion Mass Spectroscopy imaging in the propionic acid rat model of autism spectrum disorders.
(PI: Derrick F. MacFabe, MD. Kilee Patchell-Evans Autism Research Group, University of Western Ontario, Canada.)
Propionic acid (PPA) and related enteric short chain fatty acids (e.g. butyrate and acetate) are likely to be molecular candidates linking the disparate behavioral, dietary, gut, metabolic and immune factors implicated in ASD. Since PPA is metabolized through mitochondrial lipid oxidation pathways, it is proposed that excess exogenous PPA may overwhelm mitochondria-mediated lipid metabolism, thereby causing mitochondrial dysfunction and oxidative stress consistent with findings from ASD patients. PPA is proposed to play a key role in the dietary, gastrointestinal, neuropatholigcal, behavioral, metabolic and molecular disturbances seen in autism. The effects of PPA will be examined through a variety of routes (e.g. intra-cerebroventricular, systemic, intragastiric), and developmental time periods (pre-post natal, preadolescent, adult), on CNS/systemic lipid/fatty acid metabolism via traditional and novel lipid analysis techniques. The animal studies will provide further proof that behavioral, neuropatholigcal, electrophysiological, and biochemical abnormalities associated with PPA exposure can be reduced by treatment such as diet, costridial eradication, probiotics, carnitine, and MB12.
A Micro device for Immune Profiling of Children with Autism.
(PI: Alexander Revzin, Ph.D. UC Davis Department of Biomedical Engineering. Davis, CA.)
In connection with the requirement to predict how a given child will react to vaccination, there is an urgent need to develop technologies that enable multiplexed analysis of blood based on a small input of blood to ensure the compliance of small children and families. The project will design a miniature blood analysis system to distinguish early in life the immune patterns that could predict immune function and response to exogenous antigens, such as vaccines, using a small sample volume of whole blood. A microarray will be developed that is comprised of antibodies that will be used to capture and enumerate CD4 and CD8 T-cells, quantify T-cell secreted cytokines, and detect serum immunoglobulins. A miniature device will be designed that will allow researchers to perform differential multiparametric blood analysis on samples obtained from subjects compared to typically developing controls. Overall, the goal is to characterize and validate a novel technology for immune profiling of children with autism and healthy controls. In the future, the technology may be used prior to vaccination to determine immune competence of infants, and if necessary, to alter vaccination protocols.
Split Sample Reproducibility Study
(PI: Elizabeth Mumper, MD. The Rimland Center, Virginia. Nancy O’Hara, MD, Optimal Health Initiatives, Wilton, CT.)
Many clinicians rely on lab results from specialty labs in making clinical decisions. It is important to know if these lab tests are accurate (correct answer), interpreted appropriately (normal reference range and correct commentaries), and specific (where the analysis pertains to the analyte being measured and only to that). As a first step in this process, we measured precision – the ability to test the same sample twice and get the same or similar results. It is critical to understand that our study only measured reproducibility, not reliability, accuracy, specificity, or the appropriateness of reference ranges or commentaries.
Blood and urine specimens were collected from the clinical practices of Elizabeth Mumper, Nancy O’Hara and Gail Szakacs, all physicians involved in the integrative care of children on the autism spectrum. The results were reported to the DAN! Think Tank. A list of labs that passed by our criteria for specific tests was distributed to clinicians and researchers.
Is Low Serum Myeloperxidase (MPO) in Autistic Children with Severe GI Disease Acquired by Anti-Fungal Therapy or Inherited?
(PI: A.J. Russo, Ph.D. Health Research Institute/Pfeiffer Treatment Center, Warrenville, IL)
The purpose of the study is to assess serum myeloperoxidase (MPO) levels in autistic children with severe GI disease and its association with inflammatory GI disease, including ANCA and ASCA, as well as severity of disease, previously seen in a subgroup of autistic children, and to test the hypothesis that antifungal therapy may be associated with MPO deficiency in these patients. Fifty children with autism and GI disease will be studied for MPO concentration, before, during and after antifungal therapy and compared to MPO serum levels of 60 controls. Using PCR, determination will be made whether any observed MPO deficiency is inherited. The results of this study will help test the hypothesis that there is a relationship between low MPO levels and GI disease in ASD and whether this deficiency is acquired from antifungal therapy.