Diagnostic for childhood risk of autism spectrum disorder
Provided herein are methods of obtaining and applying measurements of metabolites to diagnosing ASD in a subject, particularly children, with high specificity and sensitivity. The metabolites can be measured in urine, serum, and whole blood samples.
1 . A method of treating Autism Spectrum Disorder (ASD) in a subject suspected of having or at risk of having ASD, the method comprising:
(a) in a biological sample obtained from the subject, measuring or having measured a level of each of at least two metabolites in combination wherein the at least two metabolites in combination are selected from the group consisting of: 4-Hydroxy-3-methylbenzoic acid and Tryptamine; and 4-Hydroxy-3-methylbenzoic acid and 5-Hydroxytryptophan;
(b) comparing the measured level of each metabolite in the selected combination against a control metabolite level for each metabolite, wherein the control level for each metabolite is determined by measuring a metabolite level in a control, typically developing (TD) subject, wherein the comparing comprises calculating the Type I false positive rate (FPR) and Type II false negative rate (FNR) errors for the selected combination of metabolites using Fisher's Discriminant Analysis (FDA) or logistic regression;
(c) indicating the subject as having or at risk of ASD when, for the selected combination, the Type I error is about or below 20%, and the Type II error is about or below 20%; and
(d) administering to the subject identified as having or at risk of ASD, a treatment selected from the group consisting of:
(i) a medication selected from an antipsychotic drug, a selective serotonin re-uptake inhibitor (SSRI), a tricyclic, a psychoactive or anti-psychotic medication, a stimulant, an anti-anxiety medication, or an anticonvulsant;
(ii) a nutritional supplementation; and
(iii) a composition comprising fecal microbiota from a healthy neurotypical human donor.
2 . The method of claim 1 , wherein step (a) comprises preparing a sample extract from the biological sample and obtaining the level of each metabolite in the combination of at least two metabolites in the sample extract using Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectroscopy (UPLC-MS/MS).
3 . The method of claim 1 , wherein the method determines the subject as having or at risk of ASD at birth or pre-birth.
4 . The method of claim 1 , wherein the biological sample is a urine sample.
5 . The method of claim 4 , wherein the two metabolites are 4-Hydroxy-3-methylbenzoic acid and Tryptamine.
6 . The method of claim 1 , wherein the biological sample is serum.
7 . The method of claim 1 , wherein the treatment is further personalized for the subject determined as having or at risk of having ASD.
8 . The method of claim 1 , further comprising measuring a level of each of at least two metabolites in combination wherein the at least two metabolites in combination are selected from the group consisting of: 4-Hydroxy-3-methylbenzoic acid and Picolinic acid; 6-Hydroxynicotinic acid and Picolinic acid; 4-Hydroxy-3-methylbenzoic acid and Ethylmalonic acid; Mannose and 4-Hydroxy-3-methylbenzoic acid; Acetylglucosamine and 4-Hydroxy-3-methylbenzoic acid; Glutaric acid and 4-Hydroxy-3-methylbenzoic acid; Tyrosine and 4-Hydroxy-3-methylbenzoic acid; Nicotinamide and 4-Hydroxy-3-methylbenzoic acid; 4-Hydroxy-3-methylbenzoic acid and 2-Aminoadipic acid; Glutamine and 4-Hydroxy-3-methylbenzoic acid; Tyrosine and Nicotinamide; Kynurenic acid and 4-Hydroxy-3-methylbenzoic acid; Glutaric acid and Tryptamine; and Gentisic acid and 4-Hydroxy-3-methylbenzoic acid.
9 . A method of treating Autism Spectrum Disorder (ASD) in a subject suspected of having or at risk of having ASD, the method comprising: administering to the subject a treatment, wherein the subject has been determined as having or at risk of ASD by having a Type I error of about or below 20%, and a Type II error of about or below 20% for a level of each of two metabolite in a selected combination of at least two metabolites in a biological sample obtained from the subject, wherein two metabolites in combination are selected from the group consisting of: 4-Hydroxy-3-methylbenzoic acid and Tryptamine; and 4-Hydroxy-3-methylbenzoic acid and 5-Hydroxytryptophan; and wherein the treatment is selected from:
(i) a medication selected from an antipsychotic drug, a selective serotonin re-uptake inhibitor (SSRI), a tricyclic, a psychoactive or anti-psychotic medication, a stimulant, an anti-anxiety medication, or an anticonvulsant;
(ii) a nutritional supplementation; and
(iii) a composition comprising fecal microbiota from a healthy neurotypical human donor.
10 . The method of claim 9 , wherein the level of each metabolite is obtained from a sample extract prepared from the biological sample using Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectroscopy (UPLC-MS/MS).
11 . The method of claim 9 , wherein the subject is determined as having or at risk of ASD at birth or pre-birth.
12 . The method of claim 9 , further comprising measuring a level of each of at least two metabolites in combination wherein the at least two metabolites in combination are selected from the group consisting of: 4-Hydroxy-3-methylbenzoic acid and Picolinic acid; 6-Hydroxynicotinic acid and Picolinic acid; 4-Hydroxy-3-methylbenzoic acid and Ethylmalonic acid; Mannose and 4-Hydroxy-3-methylbenzoic acid; Acetylglucosamine and 4-Hydroxy-3-methylbenzoic acid; Glutaric acid and 4-Hydroxy-3-methylbenzoic acid; Tyrosine and 4-Hydroxy-3-methylbenzoic acid; Nicotinamide and 4-Hydroxy-3-methylbenzoic acid; 4-Hydroxy-3-methylbenzoic acid and 2-Aminoadipic acid; Glutamine and 4-Hydroxy-3-methylbenzoic acid; Tyrosine and Nicotinamide; Kynurenic acid and 4-Hydroxy-3-methylbenzoic acid; Glutaric acid and Tryptamine; and Gentisic acid and 4-Hydroxy-3-methylbenzoic acid.