Comparison of Three Clinical Trial Treatments for Autism Spectrum Disorder Through Multivariate Analysis of Changes in Metabolic Profiles and Adaptive Behavior

Several studies associate autism spectrum disorder (ASD) pathophysiology with metabolic abnormalities related to DNA methylation and intracellular redox homeostasis. In this regard, three completed clinical trials are reexamined in this work: treatment with (i) methylcobalamin (MeCbl) in combination with low-dose folinic acid (LDFA), (ii) tetrahydrobiopterin, and (iii) high-dose folinic acid (HDFA) for counteracting abnormalities in the folate-dependent one-carbon metabolism (FOCM) and transsulfuration (TS) pathways and also for improving ASD-related symptoms and behaviors. Although effects of treatment on individual metabolites and behavioral measures have previously been investigated, this study is the first to consider the effect of interventions on a set of metabolites of the FOCM/TS pathways and to correlate FOCM/TS metabolic changes with behavioral improvements across several studies. To do so, this work uses data from one case–control study and the three clinical trials to develop multivariate models for considering these aspects of treatment. Fisher discriminant analysis (FDA) is first used to establish a model for distinguishing individuals with ASD from typically developing (TD) controls, which is subsequently evaluated on the three treatment data sets, along with one data set for a placebo, to characterize the shift of FOCM/TS metabolism toward that of the TD population. Treatment with MeCbl plus LDFA and, separately, treatment with tetrahydrobiopterin significantly shifted the metabolites toward the values of the control group. Contrary to this, treatment with HDFA had a lesser, though still noticeable, effect whilst the placebo group showed marginal, but not insignificant, variations in metabolites. A second analysis is then performed with non-linear kernel partial least squares (KPLS) regression to predict changes in adaptive behavior, quantified by the Vineland Adaptive Behavior Composite, from changes in FOCM/TS biochemical measurements provided by treatment. Incorporating the 74 samples receiving any treatment, including placebo, into the regression analysis yields an R2 of 0.471 after cross-validation when using changes in six metabolic measurements as predictors. These results are suggestive of an ability to effectively improve pathway-wide FOCM/TS metabolic and behavioral abnormalities in ASD with clinical treatment

Advanced glycation endproducts, dityrosine and arginine transporter dysfunction in autism - a source of biomarkers for clinical diagnosis.

Background: Clinical chemistry tests for autism spectrum disorder (ASD) are currently unavailable. The aim of this study was to explore the diagnostic utility of proteotoxic biomarkers in plasma and urine, plasma protein glycation, oxidation, and nitration adducts, and related glycated, oxidized, and nitrated amino acids (free adducts), for the clinical diagnosis of ASD. Methods: Thirty-eight children with ASD (29 male, 9 female; age 7.6 \ub1 2.0 years) and 31 age-matched healthy controls (23 males, 8 females; 8.6 \ub1 2.0 years) were recruited for this study. Plasma protein glycation, oxidation, and nitration adducts and amino acid metabolome in plasma and urine were determined by stable isotopic dilution analysis liquid chromatography-tandem mass spectrometry. Machine learning methods were then employed to explore and optimize combinations of analyte data for ASD diagnosis. Results: We found that children with ASD had increased advanced glycation endproducts (AGEs), N\u3b5-carboxymethyllysine (CML) and N\u3c9-carboxymethylarginine (CMA), and increased oxidation damage marker, dityrosine (DT), in plasma protein, with respect to healthy controls. We also found that children with ASD had increased CMA free adduct in plasma ultrafiltrate and increased urinary excretion of oxidation free adducts, alpha-aminoadipic semialdehyde and glutamic semialdehyde. From study of renal handling of amino acids, we found that children with ASD had decreased renal clearance of arginine and CMA with respect to healthy controls. Algorithms to discriminate between ASD and healthy controls gave strong diagnostic performance with features: plasma protein AGEs\u2014CML, CMA\u2014and 3-deoxyglucosonederived hydroimidazolone, and oxidative damage marker, DT. The sensitivity, specificity, and receiver operating characteristic area-under-the-curve were 92%, 84%, and 0.94, respectively. Conclusions: Changes in plasma AGEs were likely indicative of dysfunctional metabolism of dicarbonyl metabolite precursors of AGEs, glyoxal and 3-deoxyglucosone. DT is formed enzymatically by dual oxidase (DUOX); selective increase of DT as an oxidative damage marker implicates increased DUOX activity in ASD possibly linked to impaired gutmucosal immunity. Decreased renal clearance of arginine and CMA in ASD is indicative of increased arginine transporter activity which may be a surrogate marker of disturbance of neuronal availability of amino acids. Data driven combination of these biomarkers perturbed by proteotoxic stress, plasma protein AGEs and DT, gave diagnostic algorithms of high sensitivity and specificity for ASD

Identification and Characterization of Genetic Components in Autism Spectrum Disorders 2020

The Identification of the Genetic Components of Autism Spectrum Disorders 2020 will be a useful resource for laboratory and clinical scientists, translational-based researchers, primary healthcare providers and physicians, psychologists/psychiatrists, neurologists, developmental pediatricians, clinical geneticists, teachers, special educators, and caregivers involved with individuals who have autism spectrum disorders (ASD), with the goal to translate information directly to the clinical, education and home settings. Other professionals, students at all levels, and families who are interested in this important neurodevelopmental disorder will find this textbook of value by obtaining a better awareness of the causes, testing, and understanding of genetic components leading to autism, and research that may open avenues for treatment with new approaches. This textbook includes nine chapters divided into three sections (clinical, genetics, other) written by experts in the field dedicated to genetics research and clinical care, description, and treatment by generating reviews for ASD and related disorders. These chapters include information on discoveries, risk factors, causation, diagnosis, treatment, and phenotyping with characterization of genomic or genetic factors and the environment, as genetics play an important role in up to 90% of individuals with autism via over 800 currently recognized genes

Nutrition and biomarkers in psychiatry : research on micronutrient deficiencies in schizophrenia, the role of the intestine in the hyperserotonemia of autism, and a method for non-hypothesis driven discovery of biomarkers in urine

This thesis describes the study of markers of nutrition and intestinal motility in mental disorders with a focus on schizophrenia and autism, and the development, evaluation and application of a biomarker discovery method for urine. The aim of the thesis is to investigate the role of long-chain polyunsaturated fatty acids (LCPUFA), B-vitamins and platelet (PLT) serotonin (5-HT) in schizophrenia and autism. The thesis proposes also that biomarker research in psychiatric disease is of great relevance and describes a biomarker discovery method in urine using a non-hypothesis driven ‘-omics’-like approach. The thesis ends by summarizing its contents and putting biomarker research in psychiatric disease and its implications in a broader perspective. In the Introduction the complex etiology and potential role of non-hypothesis driven biomarker research in psychiatric disease is reviewed, with an accent on schizophrenia and autism. The enormous economic and psychosocial global burden of mental disorders is described as well as their epidemiology, clinical presentation and classification/diagnosis. Hypothesized etiological factors are discussed to create a framework in which biomarkers and the research thereof can be positioned. Furthermore, advances in the field of biomarker research in psychiatry are discussed in the context of epigenetics, proteomics and metabolomics. The first part (Part I) of this thesis describes a study of LCPUFA and B-vitamins in schizophrenia, of PLT 5-HT and intestinal permeability in autism, and of the value of PLT 5-HT as marker of intestinal motility in newborns. The chapters, in which these studies are described, are preceded by a review (Chapter 1), which gives an overview of the role of LCPUFA and folate in the etiology and severity of psychiatric diseases such as depression, bipolar disorders, schizophrenia and autism. Pregnancy complications and folate-substrated carbon-1 metabolism are considered and their possible epigenetic effect on the etiology of mental disorders is described. Other nutritional factors, such as LCPUFA, that are important for brain development, physico-chemical properties of membranes, signal transduction and DNA-transcription, and that have been used in supplementation trials, are suggested to be important factors in the origin and severity of schizophrenia. In Chapter 2 we describe the results from a study concerning the essential fatty acid (EFA) and functional B-vitamin status in patients with schizophrenia. Aberrant EFA-status and increased homocysteine (Hcy; a marker of functional B-vitamin deficiency), have been reported before in subgroups of patients with schizophrenia. We describe the characteristics of large subgroups with marginal to severe deficiencies of LCPUFA and B-vitamins, notably folate and vitamin B12. Deficiencies proved easily correctable in the most severely deficient patients upon supplementation with ω3 fatty acids and B-vitamins. Chapter 3 attempts to integrate and link previously reported findings of increased intestinal permeability and increased PLT 5-HT levels in subgroups of children with pervasive developmental disorders (PDD). Platelet 5-HT and intestinal permeability were assessed in children with PDD in Curaçao. Differential urinary excretion of inert sugars after ingestion of a sugar solution was used as marker of intestinal permeability. In Chapter 4 we examined the potential of PLT 5-HT as marker of intestinal motility. For this we studied whole blood and PLT 5-HT in mothers (normal motility) and their newborns (developing intestinal motility) at birth. The course of PLT 5-HT in relation to changes in feeding mode (i.e. parenteral/enteral) was investigated in a small group of preterm born infants to see whether PLT 5-HT was responsive to changes in intestinal motility. Chapters 2, 3 and 4 describe hypothesis-driven research in mental disorders. However, the advance of knowledge about mental disorders is slow and it is likely to benefit from complementation by information that is generated through non-hypothesis driven research with state-of-the art techniques that profile proteins (i.e. proteomics) and metabolites (i.e. metabolomics). These ‘-omics’ techniques are likely to deliver a multitude of candidate diagnostic and prognostic markers as well as therapeutic targets, compared to hypothesis driven research. Part II is thus devoted to the development, evaluation and application of such a non-hypothesis driven method. In Chapter 5 we describe the comparative analysis of low molecular weight urinary components using LC-MS and subsequent multivariate statistical analysis of the processed LC-MS data. This chapter deals with the development, evaluation and preliminary application of the method to proteinuria in humans. The potential and pitfalls of the method are contemplated upon. Chapter 6 describes an advanced proof-of-principle of the method through the comparison of urinary profiles from pregnant and non-pregnant females using the improved methodology described in Chapter 5. The methodology is significantly optimized with respect to data processing and multivariate statistical analysis. More focus is put on the selection of discriminatory peaks. --------------------------------------------------------------- Éénderde van de patiënten met schizofrenie blijkt matige tot ernstige tekorten aan bepaalde B-vitaminen en ω3- en ω6-vetzuren te hebben. Deze tekorten zijn eenvoudig op te heffen met voedingsupplementen (o.a. foliumzuur en visolie). Een lage status van deze micronutriënten speelt waarschijnlijk een rol in het ontstaan en de ernst van diverse psychiatrische ziekten en het ontstaan van hart- en vaatziekten. De behandelende artsen vermoedden het bestaan van deze tekorten niet. Ongeveer een kwart van de patiënten met autisme heeft een verhoogd serotoninegehalte in bloedplaatjes (hyperserotonemie: “biomarker”). Serotonine is een neurotransmitter in onze hersenen en darmen. Autisten zouden ook vaker maagdarmstoornissen hebben. We vonden geen relatie tussen hyperserotonemie (26%) en de darmdoorlaatbaarheid (in 0% verhoogd) in kinderen met autisme. Het idee dat een verhoogde darmmotiliteit hyperserotonemie veroorzaakt, werd ondersteund door een twee maal hoger bloedplaatjesserotonine van moeders (actieve darm) t.o.v. hun pasgeboren baby’s (inactieve darm). Tevens bleek starten en staken van enterale voeding in pasgeborenen gecorreleerd aan respectievelijk stijgingen en dalingen van hun bloedplaatjesserotonine. Meer onderzoek naar het maagdarmstelsel (onze “second brain”) bij autisme is gewenst. Niet-hypothesegedreven onderzoek als aanvulling op het veelal hypothesegedreven onderzoek kan helpen bij het vinden van diagnostische en therapeutische biomarkers voor psychiatrische stoornissen. De hiertoe opgezette methode onderzoekt urinemonsters met vloeistofchromatografie-massaspectrometrie, waarna de 3D-data multivariaat geanalyseerd worden om biomarkers te ontdekken. Experimenten met urinemonsters van patiënten met eiwit in hun urine (vanwege een nierziekte) en controles, en van zwangere en niet-zwangere vrouwen, lieten duidelijke groepsverschillen zien (“proof-of-principle”). Dit geeft hoop dat de ontwikkelde methode in de toekomst gebruikt kan worden voor het opsporen van biomarkers bij psychiatrische stoornissen en andere ziektes.

A Personalized Medicine Approach to the Diagnosis and Management of Autism Spectrum Disorder

This collection of articles provides an overview of the current and future methods for applying a personalized medicine approach to the diagnosis, management, and treatment of autism spectrum disorder

Early Disruption of the Microbiome Leading to Decreased Antioxidant Capacity and Epigenetic Changes: Implications for the Rise in Autism

Currently, 1 out of every 59 children in the United States is diagnosed with autism. While initial research to find the possible causes for autism were mostly focused on the genome, more recent studies indicate a significant role for epigenetic regulation of gene expression and the microbiome. In this review article, we examine the connections between early disruption of the developing microbiome and gastrointestinal tract function, with particular regard to susceptibility to autism. The biological mechanisms that accompany individuals with autism are reviewed in this manuscript including immune system dysregulation, inflammation, oxidative stress, metabolic and methylation abnormalities as well as gastrointestinal distress. We propose that these autism-associated biological mechanisms may be caused and/or sustained by dysbiosis, an alteration to the composition of resident commensal communities relative to the community found in healthy individuals and its redox and epigenetic consequences, changes that in part can be due to early use and over-use of antibiotics across generations. Further studies are warranted to clarify the contribution of oxidative stress and gut microbiome in the pathophysiology of autism. A better understanding of the microbiome and gastrointestinal tract in relation to autism will provide promising new opportunities to develop novel treatment modalities

Evaluating the role of social attention in the causal path to Autism Spectrum Disorder

This thesis evaluated the evidence for the hypothesis that early disruptions in social attention are involved in the causal pathway to Autism Spectrum Disorder (ASD). The sample included infants at high and low familial risk for neurodevelopmental disorders participating in a prospective longitudinal study, and their family members. Five studies were conducted to test whether social attention atypicalities precede the onset of behavioural symptoms and whether they are related to familial, genetic and epigenetic burden for ASD. Chapter 2 examined neural correlates of attention measured with multi-channel electroencephalography in 8-month-old infants attending to faces and non-social stimuli, in relation to outcomes at age 3. Chapter 3 used structural equation modelling to investigate whether disruptions in neural response have cascading effects on learning from the environment via looking behaviour. Next, to further understand whether disruptions in social attention lie between genetic risk and ASD phenotype, Chapter 4 examined the association between ability to detect eye-gaze direction in a familial sample, severity of ASD symptoms and polygenic risk for ASD. Chapter 5 explored these patterns earlier in development, looking at the relationship between social attention at 14 months of age and familial burden, polygenic risk and parentreport traits of ASD and ADHD. Finally, Chapter 6, leveraging DNA methylation data, explored whether epigenetic signals were associated with early neural and behavioural correlates of social attention as well as developmental change leading to atypical outcome. Taken together, this work examined in depth the multifaceted nature of social attention, pointing to neural and behavioural atypicalities at critical time points as promising targets for cognitive and affective interventions. Furthermore, it pioneers future work integrating genetics, epigenetics and early neurocognitive measures of social attention in large prospective longitudinal studies of individuals at increased vulnerability for neurodevelopmental disorders, to shed light on the developmental mechanisms underlying the emergence of ASD