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.
T cell polarization and candida reactivity in autistic children with food allergy
Harumi Jyonouchi, M.D., University of Medicine & Dentistry of New Jersey
GI symptoms are frequently observed in ASD children. Non-IgE mediated, delayed-type food allergy (NFA) might have a role in GI symptoms observed in ASD children on the basis of our previous studies; we reported previously that FPIES accounts for the GI symptoms experienced in many young ASD children (Jyonouchi et al., 2005). We have also observed that resolution of GI symptoms in ASD/FPIES children following implementation of avoidance of offending food was associated with improvement of certain behavioral symptoms.
However, in our observation, even after recovering from NFA, some ASD children continue to suffer from recurrent or persistent GI symptoms. We also observed that flare ups of GI symptoms in these ASD/NFA children are usually triggered by GI insults such as microbial gastroenteritis, and prolonged oral antibiosis. ASD/NFA children with persistent GI symptoms in our observation also often exhibit other co-morbid conditions such as recurrent respiratory infection, multiple adverse reactions to medications, and seizure disorders.
Thus we hypothesized that 1) oral tolerance to FP and commensal microbiota is fragile and more easily broken in ASD/NFA children with persistent GI symptoms, and 2) this is associated with altered innate/adoptive immune responses in these ‘treatment-resistant’ ASD/NFA children. This study assessed innate immune responses, Th cell polarization, and T cell functions in ASD/NFA, ASD/non-NFA, non-ASD/NFA, and normal control children.
Our results indicate that altered adaptive and innate immune responses are observed in both ASD/NFA as compared to 3 control groups. In addition, among ASD/NFA children, a small numbers of children who exhibit fluctuating behavioral symptoms and cognitive skills with immune insults (we categorized this subgroup as ASD-inflammatory subtype; ASD-IS) exhibited significantly different innate immune responses as compared to other ASD/NFA children. Our findings thus indicate ASD/NFA and ASD-IS children are immunologically different compared to ASD/non-NFA or non-ASD/NFA children.
Our results also indicate that changes in innate/adaptive immune processes observed in ASD-IS children, and in some degree even in ASD/NFA children, are not likely to be attributed to NFA; they might be associated with aberrant innate immune responses affecting neuroimmune network.
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
Derrick MacFabe M.D., University of Western Ontario, London, Canada
Autism Research Institute-Funded Research: Metabolic Products of Autism-Associated Gut Bacteria Produce Autism-Like Behavioral and Brain Changes Through Altering Brain Fatty Acid Metabolism
Dietary factors, gastrointestinal symptoms, antibiotic exposure, unique bacterial populations, and immune and metabolic alterations are common in patients with autism spectrum disorders. Many patients have dysregulation in fatty acid metabolism and abnormalities in mitochondria, the energy storehouse of the cell. Mitochondrial dysfunction has been suggested to explain the complex medical and physiological abnormalities found in some children with autism. Some have suggested that the systemic abnormalities seen in autism may arise from environmental triggers in sensitive subpopulations. Can a common environmental factor link the disparate symptoms of autism? Researchers, led by Dr. Derrick MacFabe, Director of the Kilee Patchell-Evans Autism Research Group at the University of Western Ontario, Canada have found that enteric short-chain fatty acids, such as propionic acid (PPA), produce many of the behavioral (repetitive, antisocial) and brain (immunological, metabolic) changes consistent with autistic patients, when given to laboratory rodents. PPA is interesting in that can be produced by opportunistic gut bacteria that have been implicated in autism (e.g. Clostridia, Desulfovibrio and Bacterioidetes), isolated by Dr. Sidney Finegold (UCLA). These bacteria produce PPA, particularly following ingestion of the dietary carbohydrates that many autistic children crave. Autism Research Institute-funded work done by postdoctoral fellow Dr. Raymond Thomas has found PPA exposure in rats leads to alterations in brain fat metabolism which show much similarity to findings in autistic patients. These include increased oxidative stress markers, abnormalities in brain omega-3 and -6 profiles, reductions in cholesterol and alterations in brain carnitine metabolism. Interestingly, some of these effects may be transient and related to behavior. Similar alterations have been found by the Group using a novel brain-imaging technique (Time-of-Flight-Secondary Ion Mass Spectroscopy) in brain regions affected in autism, particularly brain white matter, enriched in many brain fatty acids. As the brain is largely fat, these lipid changes have major effects on brain development, energy and function, particularly as PPA inhibits brain mitochondrial energy production. This animal model is useful as it allows multi-disciplinary ethical examination of many biological processes implicated in autism. Furthermore it also might help to explain how many proposed treatments (carbohydrate reduction, omega-3 fatty acids, carnitine, Methyl B12, eradication of gut bacteria), may work in autism, as all are implicated in the metabolism of PPA.
Collaborative work with Dr. Richard Frye, Director of Autism Research, Arkansas Children’s Hospital Research Institute, is further examining these lipid alterations– found in the animal model– in autistic children, to attempt to find similar biomarkers to identify patients and develop rational therapies.
Human intestinal microbial ecology and its relationship to autism
Rosa Krajmalnik-Brown, Ph.D., Biodesign Institute, Arizona State University, Tempe, Arizona
Unusually high proportions of autistic children suffer from gastrointestinal problems, suggesting a link between autism and abnormalities in gut microbial function. Disturbances in gut microbial ecosystem not only cause gastrointestinal problems that can exacerbate autistic symptoms, but might also affect the immune and neurological systems. Previous studies have focused primarily on autism-related presence of pathogenic bacteria. Hence, we investigated autism-related alteration of intestinal bacterial composition by comparing non-autistic and autistic children with or without gastrointestinal symptoms. Interestingly, autistic children tend to have less diverse bacterial communities in their intestines, regardless of noticeable gastrointestinal symptoms. In addition, extensive bioinformatics and statistical analyses discovered that a specific microbial genus was mostly absent in autistic children, which in contrast was found in a large fraction of non-autistic children. Intriguingly, these microbes are known to function as a primary degrader of polysaccharides in gut, suggesting a possible influence of daily diets. Therefore, our findings suggest that the less diverse and imbalanced gut microbial profiles of autistic children make them more susceptible to frequent gastrointestinal disorders.
Do vagal and circumventricular inflammation contribute to the etiology of regressive autism?
Veronica Miller, Ph.D., Wadsworth Ctr. For Laboratories & Research, State of New York Dept. of Health, Albany, NY
It is important to find out whether vagal and circumventricular organs (CVO) inflammation is found in regressive autism, because damage therein may trigger autonomic symptoms in some children. Therefore we are quantifying inflammatory lesions in brainstem CVO and vagal tissues from regressive autism cases and controls. We are also determining whether the inflammation is specific to the brain or is actually systemic, by quantifying inflammatory and stress-related changes in target organs innervated by the vagus nerve, in tandem with analysis of extracranial vagal-nerve specimens, from regressive autism cases and controls.
Research Project About a Potential Infectious Origin of Autism
Luc Montagnier, Institut de Recherche Luc Montagnier, Paris France
The goal of this trial is the detection of pathogens in peripheral blood using a physics-technology approach when undetectable or ambiguous with classical techniques. The hypothesis is that different factors may cause intestinal mucosa inflammation, and facilitate overpassing of bacterial products in the blood stream. This may cause bacterial coating in capillary brain vessels leading to ischemia and neuronal death. However, the identification of bacterial species is of first importance. The technology presented above does not yet allow identification of the bacteria, or the gene responsible for the production of EMS. Nevertheless, Luc Montagnier’s team recently set up a proven PCR technology that allows identification of responsible germs.
Equipment Grant
Martha Herbert, M.D., Ph.D., TRANSCEND Research Program, Massachusetts General Hospital, Boston, MA
Purchase tissue freezer and coil to allow for phophorous magnetic resonance spectroscopy.
Survey on treatment for children with autism with and without seizures
Richard Frye, M.D., Arkansas Children’s Hospital Research Institute, University of Arkansas for Medical Sciences
This grant supported a national survey on treatments for seizures in children with autism spectrum disorder. Approximately 25% of children and adults with autism have seizures, and another 50% might have sub-clinical seizures that are undetected, but might affect their intellectual function. This national survey asked over 1,000 parents with children with autism (733 with seizures and 290 without seizures) to rate the therapeutic effects and side effects of common traditional and non-traditional treatments used to treat seizures and autism, including medical, dietary, and nutritional treatments. This study identified specific antiepileptic drugs which improve seizures and, on average, had minimal adverse effects, and certain non-anti-epileptic drug treatments, particularly the ketogenic diet, that improved both seizures and symptoms associated with autism. The results of this study were published in BMC
Pediatrics https://www.biomedcentral.com/1471-2431/11/37 with a highly accessed distinction.
Elevated urinary p-cresol: intestinal causes and behavioral consequences
Antonio M. Persico, M.D., Laboratory of Molecular Psychiatry & Neurogenetics University Campus Bio-Medico Rome, Italy
“Elevated urinary p-cresol in small autistic children: origin and consequences”: This application aims at continuing and possibly completing the three above-mentioned studies, as follows: [a] assessing gut permeability, urinary p-cresol, fecal c. difficile/toxin A/calprotectin and urinary cotinine in approximately 50-60 individuals should suffice, if our results continue to show no correlation between these three parameters. We shall also try to implement a fecal metabolomic approach to assess gut microbiota beyond c.difficile, in order to exclude infection by other cresol-producing bacteria and to point convincingly toward environmental exposure as the cause of elevated urinary p-cresol in autism; complete our replication study by recruiting at least 13 other case-control pairs, reaching a final N of at least 30 autistic individuals and 30 matched controls, equally split between age < 8 and age > 8; [c] performing a complete battery of tests assessing motor (Open Field Test, Rotarod Test and Grooming activity), cognitive (Object Recognition Test, Spatial Novelty Test), and emotional (Porsolt’s Test, Plus Maze, Social Novelty Test) responses to acute administration of p-cresol and p-cresylsulfate in BTBR mice, which are regarded as one of the most reliable rodent models of autism. Neurochemical analyses will be performed on the brains of these mice, to assess brain region-specific imbalances in major neurotransmitters.
Creating a specimen bank of neurotypical individuals
Anthony J. Russo, Ph.D., Health Research Institute/Pfeiffer Treatment Center, Warrenville, IL
The aim of this grant is to establish a center for control samples at minimal cost. The center will collect, transport and house plasma/serum, red blood cells, buffy coat, and urine from age-appropriate non-autistic/neurotypical children.
Study of anti-neuronal autoantibodies in behavioral and movement disorders
Madeleine W. Cunningham, University of Oklahoma Health Sciences Center, Oklahoma City, OK
Our hypothesis is that autoantibody-mediated neuronal-cell signaling might be present in sera, and play a role in the development of PANDAS, Sydenham chorea and related disorders. Our project aims to determine the presence of, and to understand mechanisms by which, anti-neuronal autoantibodies(Ab) affect neuronal cells in vitro as well as in the brain, and consequently behavior in vivo. We plan to test for and correlate crossreactive antibody specificities for dopamine receptors, tubulin, and lysoganglioside with antibody-mediated neuronal-cell signaling mechanisms(CaMKinase) with symptoms in patients. Ultimately, Sydenham chorea and related disorders may in part be due to altered signaling patterns following immune stimulation in the brain. Our findings will contribute significantly to an understanding of autoimmune and signaling mechanisms which might be linked to infection and/or autoimmunity in movement and behavioral disorders.
Molecular pathways involved in oxidative stress and leaky-gut impairment in autism spectrum disorders.
Nicola Antonucci, M.D., Dept. of Experimental Medicine, University of Naples, Naples, Italy
The main aim of this research is to investigate the relationship between the gastrointestinal tract and autism and to understand how gut, brain, nutritional, and toxic status are consistent with greater oxidative stress in autistic spectrum disorders (ASDs).
Researchers will perform detailed analyses on several biological aspects of peripheral blood mononuclear cells (PBMCs), and of the intestinal mucosa of ASD patients, in order to identify several proteins and genes involved in inflammatory and cellular stress response, as well as the innate and adaptive immunological response. In particular, we will study the molecular pathways responsible for apoptosis, inflammation and cell damage or suffering of ASD-PBMCs, identifying the proteases involved in the activation pathways as possible diagnostic and/or therapeutic tools.
(PI: Dario Siniscalco, Chem.D., Ph.D., Anna Sapone, M.D., Ph.D., Nicola Antonucci, M.D.; Second University of Naples, Italy)
Enhanced tissue procurement from autistic individuals
H. Ronald Zielke, Ph.D., NICHD Brain and Tissue Bank, Dept. of Pediatrics, University of Maryland, Baltimore, MD
Grant to support efforts to obtain tissues for the NICHD/University of Maryland Brain and Tissue Bank.
Multidimensional impact of pain on individuals and family functioning in ASD
Arthur Samuel, Ph.D., Edward Carr, Ph.D., The Research Foundation of SUNY, Stony Brook, NY
This study will examine the impact of pain on multiple aspects of functioning in ASD. A meaningful exploration of this issue requires attention to three aspects of the problem: (1) identifying appropriate measures of pain; (2) identifying plausible dependent variables that are likely to be impacted by pain; (3) identifying plausible moderator variables that alter the impact that pain has on the selected dependent variables.
To study why increased copper in individuals with autism normalizes post zinc therapy in individuals with concurrent GI disease
Anthony J. Russo, Ph.D., Health Research Institute/Pfeiffer Treatment Center, Warrenville, IL
To study why increased copper in individuals with autism normalizes post zinc therapy in individuals with concurrent GI disease
This study is assessing patients with autism and evaluating GABA, norepinephrine and dopamine levels, as they are associated with Cu and Zn levels, particularly in patients with concurrent GI disease.
Investigating the Efficacy of the Hane Face Window
Chris Bentley, Fraser Center, Minneapolis, MN
Research grant to conduct a pilot study on the efficacy of the Hane Face Window. The Hane Face Window is designed to improve visual processing of faces as well as reduce stress.
To study the relationship between decreased hepatocyte growth factor (HGF) and glutamate excitotoxicity in autistic children
Anthony J. Russo, Ph.D., Health Research Institute/Pfeiffer Treatment Center, Warrenville, IL
The MET gene variant exists in the genome of a significant number of autistic individuals. It facilitates the signaling of its ligand, hepatocyte growth factor (HGF)/scatter factor, and is involved in peripheral organ development and repair, immune function and gastrointestinal repair. In this study we are measuring HGF serum levels in autistic children, particularly those with GI disease.
Autism Severity and Muscle Strength: A Correlation Analysis
Janet Kern, Ph.D., University of Texas Southwestern Medical Center, Dallas, TX
The autism severity and muscle strength study examined the relationship between muscle strength, as measured by handgrip strength, and autism severity, as measured by the Childhood Autism Rating Scale (CARS). Thirty-seven (37) children with a diagnosis of autism spectrum disorder (ASD) were evaluated using the CARS and then tested for hand muscle strength using a handgrip dynamometer. Statistical analysis was then conducted to examine the relationship between autism severity and hand muscle strength. The model generated in the present study showed that the CARS score is a significant predictor of Max Hand Muscle Score after adjustment for age, race, gender, year of birth, and a history of prior chelation therapy. Evidence suggests that handgrip strength in children with ASD is related to the severity of the disorder. Further research is needed to determine the extent and consistency of the muscle weakness, and possible treatments.
Medical Etiologies of Neurodevelopmental Disorders: Cerebral Folate Deficiency
Richard Frye, M.D., Arkansas Children’s Hospital Research Institute, University of Arkansas for Medical Sciences
This grant supported research into the prevalence of cerebral folate deficiency and folate receptor autoantibodies (FRAs) in children with autism spectrum disorders (ASD) and the effect of folinic acid treatment in children with ASD and FRAs. The study measured serum FRAs concentrations in 93 children with ASD. A high prevalence rate (75.3 percent) of FRAs was found in the children participating in the study. Children with FRAs were treated with oral leucovorin calcium (folinic acid), and treatment response was measured and compared with a wait-list control group. Compared with controls, significantly higher improvement ratings were observed in treated children over a mean period of 4 months, in the areas of verbal communication, receptive and expressive language, attention, and stereotypical behavior.
The results of this study were published in Molecular Psychiatry and can be viewed at https://www.nature.com/mp/journal/vaop/ncurrent/full/mp2011175a.html.
The role of intestinal microbiome in children with autism
Harland Winter, M.D., Harvard Medical School, MassGeneral Hospital, Boston, MA
The purpose of the grant was to help the Digestive Function Laboratory (DFL) manage and extend its repository for tissues from children with and without autism. The award received from ARI should be used for: 1) buying capital equipment such as -80oC freezer; 2) buying Freezerworks software and Freezerworks Bar Code Accessories for the cryogenic-safe bar code labeling; and 3) salary for technician managing and entering data in the repository.
Further studies on the role of desulfovibrio in regressive autism
Sydney M. Feingold, M.D., VA Medical Center, Los Angeles, CA
The study that we are proposing herewith is designed to examine stool specimens for evidence of Desulfovibrio species from 20 additional regressive autism children and from 20 age- and sex-matched control children with gastrointestinal symptoms similar to those seen in the autistic children, but without autism.
Investigation of IL-9, IL-33 and TSLP in serum of autistic children
Theoharis Theoharides, M.D., Ph.D., Tufts University School of Medicine, Boston, MA
Levels of pro-inflammatory cytokines (IL-6, TNF) in the serum of autistic children have been inconclusive. However, recent findings indicate that chemokines (eotaxin, MCP-1, RANTES) are elevated in this population. These chemokines are responsible for accumulation of mast cells and mast cell activation, and they have been shown to be associated with autism. In this research project, levels of IL-9, IL-33, and TSLP, new cytokines recently considered “alarmins” through mast cell activation, will be measured in the serum of autistic children along with age and sex-matched neurotypical controls. Interactions among these key molecules may prove useful for both potential biomarkers and novel targets for treatment.
To study the relationship between Myeloperoxidase (MPO) deficiency and probiotic therapy in autistic children
Anthony J. Russo, Ph.D., Health Research Institute/Pfeiffer Treatment Center, Warrenville, IL
This study assessed serum myeloperoxidase levels in autistic children with severe GI disease and associated 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 might be associated with MPO deficiency in these patients.
Possible Protective Effects of Oxytocin and its Receptor on Gut Function
Martha Welch, M.D., Departments of Psychiatry and Pathology & Cell Biology, Columbia University, New York, NY
The correlation between gastrointestinal symptoms and ASD is well established. As well, oxytocin (OT) and oxytocin receptor (OTR) abnormalities have also been documented in ASD. Oxytocin (OT) is a hormone that is found in breast milk and is expressed by enteric neurons. We discovered the expression and developmental regulation of OTR in the enteric nervous system and gastrointestinal epithelium. We also showed that combined administration of secretin and OT inhibits chronic colitis and associated activation of forebrain neurons. In this work, we are further elucidating the roles of OT and its receptor (OTR) in gut function generally, and in mucosal barrier function specifically. By comparing OTRnull (OTR knock-out) mice with wild-type mice, we are testing the hypothesis that OT/OTR signaling provides protection to the bowel and enteric neurons. If correct, this could provide a link between OTR gene polymorphisms in autistic patients and abnormalities in their gut function. The exciting aspect of this work is that the elucidation of the mechanism by which OT and OTR signaling is mucosal- and neuro-protective could lead to a better understanding of the developmental role of OT/OTR and its possible protective effects against adverse behavioral and physiological outcomes in ASD patients, especially high-risk infants, such as preterm infants.
Behavioral and psycho-physiological study of attentional, perceptual, and emotional processing after treatment with ambient prism lenses and visuomotor exercises in children with autism spectrum disorder
Estate Sokhadze, Ph.D., University of Louisville, Louisville, KY
The proposed study aims to understand the neural and functional mechanisms underlying visual distortion in autism by incorporating neurophysiologic studies, behavioral studies, and measurements of sustained attention and spatial attention. The study will use ambient prism lenses shown to improve ambient vision in children with autism. It is proposed that wearing ambient lenses and performing daily visuomotor exercises for six months will result in better performance on attention and perception tasks and will lower anxiety as indexed by a profile of autonomic measures.