Methods and systems for determining autism spectrum disorder risk
In certain embodiments, the invention stems from the discovery that analysis of population distribution curves of metabolite levels in blood can be used to facilitate predicting risk of autism spectrum disorder (ASD) and/or to differentiate between ASD and non-ASD developmental delay (DD) in a subject. In certain aspects, information from assessment of the presence, absence, and/or direction (upper or lower) of a tail effect in a metabolite distribution curve is utilized to predict risk of ASD and/or to differentiate between ASD and DD.
1. A method of differentiating between autism spectrum disorder (ASD) and non-ASD developmental delay (DD) in a subject, the method comprising:
(i) measuring levels of a plurality of metabolites in a sample obtained from the subject, wherein the plurality of metabolites comprise hydroxychlorothalonil and a second metabolite,
(ii) determining the hydroxychlorothalonil level in the sample is within an ASD tail effect with a right tail if the level of hydroxychlorothalonil in the sample is greater than 2.645 ng/ml, and
determining the second metabolite level in the sample is within an ASD tail effect,
wherein the second metabolite is selected from the group consisting of: 3-carboxy-4-methyl-5-propyl-2-furanpropanoate (CMPF), 3-indoxyl sulfate, 4-ethylphenyl sulfate, 5-hydroxyindoleacetate, 8-hydroxyoctanoate, gamma-CEHC, hydroxyisovaleroylcarnitine (C5), indoleacetate, isovalerylglycine, lactate, N1-Methyl-2-pyridone-5-carboxamide, p-cresol sulfate, pantothenate (Vitamin B5), phenylacetylglutamine, pipecolate, xanthine, octenoylcarnitine, and 1,5-anhydroglucitol (1,5-AG);
wherein the level of 3-carboxy-4-methyl-5-propyl-2-furanpropanoate (CMPF) is less than 0.396 ng/ml,
wherein the level of 3-indoxyl sulfate is less than 0.584 ng/ml,
wherein the level of 4-ethylphenyl sulfate is less than 0.281 ng/ml,
wherein the level of 5-hydroxyindoleacetate is greater than 2.027 ng/ml,
wherein the level of 8-hydroxyoctanoate is less than 0.711 ng/ml,
wherein the level of gamma-CEHC is less than 0.505 ng/ml,
wherein the level of hydroxyisovaleroylcarnitine (C5) is less than 0.619 ng/ml,
wherein the level of indoleacetate is less than 0.707 ng/ml,
wherein the level of isovalerylglycine is less than 0.438 ng/ml,
wherein the level of lactate is greater than 1.288 ng/ml,
wherein the level of N1-Methyl-2-pyridone-5-carboxamide is less than 0.554 ng/ml,
wherein the level of p-cresol sulfate is less than 0.378 ng/ml,
wherein the level of pantothenate (Vitamin B5) is great than 1.980 ng/ml,
wherein the level of phenylacetylglutamine is less than 0.498 ng/ml,
wherein the level of pipecolate is greater than 1.711 ng/ml,
wherein the level of xanthine is greater than 1.507 ng/ml,
wherein the level of octenoylcarnitine is less than 0.479 ng/ml, and
wherein the level of 1,5-anhydroglucitol (1,5-AG) is less than 0.680 ng/ml;
iii) determining the subject has ASD and not DD based on the identified ASD tails as determined in step (i) and step (ii).
2. The method of claim 1 , wherein the second metabolite is selected from the group consisting of 5-hydroxyindoleacetate (5-HIAA), 1,5-anhydroglucitol (1,5-AG), 3-(3-hydroxyphenyl)propionate, 3-carboxy-4-methyl-5-propyl-2-furanpropanoate (CMPF), 3-indoxyl sulfate, 4-ethylphenyl sulfate, 8-hydroxyoctanoate, gamma-CEHC, hydroxyisovaleroylcarnitine (C5), indoleacetate, isovalerylglycine, lactate, N1-Methyl-2-pyridone-5-carboxamide, p-cresol sulfate, pantothenate (Vitamin B5), phenylacetylglutamine, pipecolate, xanthine, hydroxy-chlorothalonil, octenoylcarnitine, and 3-hydroxyhippurate.
3. The method of claim 1 , wherein the second metabolite is selected from the group consisting of phenylacetylglutamine, xanthine, octenoylcarnitine, p-cresol sulfate, isovalerylglycine, gamma-CEHC, indoleacetate, pipecolate, 1,5-anhydroglucitol (1,5-AG), lactate, 3-(3-hydroxyphenyl)propionate, 3-indoxyl sulfate, and pantothenate (Vitamin B5).
4. The method of claim 1 , wherein the plurality of metabolites comprises at least three metabolites selected from the group consisting of phenylacetylglutamine, xanthine, octenoylcarnitine, p-cresol sulfate, isovalerylglycine, gamma-CEHC, indoleacetate, pipecolate, 1,5-anhydroglucitol (1,5-AG), lactate, 3-(3-hydroxyphenyl)propionate, 3-indoxyl sulfate, and pantothenate (Vitamin B5).
5. The method of claim 1 , wherein the plurality of metabolites comprise at least one pair of metabolites of hydroxy-chlorothalonil and p-cresol sulfate.
6. The method of claim 1 , wherein the plurality of metabolites comprises at least one triplet of metabolites of indoleacetate, hydroxy-chlorothalonil, and p-cresol sulfate.
7. The method of claim 1 , wherein the plurality of metabolites comprise at least one pair of metabolites that, combined together as a set of two metabolites, provides an AUC that is higher than the AUC of each individual metabolite of the pair of metabolites, where AUC is an area under a ROC curve that plots false positive rate (1-specificity) against true positive rate (sensitivity) for a classifier based only on the set of two metabolites.
8. The method of claim 1 , wherein the plurality of metabolites comprises at least one triplet of metabolites that, combined together as a set of three metabolites, provide an AUC that is higher than the AUC of any pair of metabolites of the triplet, where AUC is an area under a ROC curve that plots false positive rate (1-specificity) against true positive rate (sensitivity) for a classifier based only on the set of three metabolites.
9. The method of claim 1 , wherein the sample is a plasma sample.
10. The method of claim 1 , wherein the metabolites are measured by mass spectrometry.
11. The method of claim 1 , wherein the subject is no greater than about 54 months of age.
12. The method of claim 1 , wherein the subject is no greater than about 36 months of age.
13. A method of differentiating between autism spectrum disorder (ASD) and non-ASD developmental delay (DD) in a subject, the method comprising:
(i) measuring levels of a plurality of metabolites in a sample obtained from the subject, wherein the plurality of metabolites comprise hydroxychlorothalonil and a second metabolite,
(ii) determining if one or more of the metabolites are present in the sample in a level within a DD tail, wherein one or more metabolites are selected from the group consisting of: 3-(3-hydroxyphenyl)propionate, 3-indoxyl sulfate, isovalerylglycine, p-cresol sulfate, phenylacetylglutamine, pipecolate, xanthine, 3-hydroxyhippurate;
wherein the level of 3-(3-hydroxyphenyl)propionate is less than 0.270 ng/ml,
wherein the level of 3-indoxyl sulfate is greater than 1.601 ng/ml,
wherein the level of isovalerylglycine is greater than 3.182 ng/ml,
wherein the level of p-cresol sulfate is greater than 2.231 ng/ml,
wherein the level of phenylacetylglutamine is greater than 2.305 ng/ml,
wherein the level of pipecolate is less than 0.651 ng/ml,
wherein the level of xanthine is less than 0.731 ng/ml,
wherein the level of 3-hydroxyhippurate is less than 0.375 ng/ml;
(iii) determining that the subject has DD and not ASD based on the identified DD tails as determined in step (i) and step (ii).
14. The method of claim 13 , wherein the sample is a plasma sample.
15. The method of claim 13 , wherein the metabolites are measured by mass spectrometry.
16. The method of claim 13 , wherein the subject is no greater than about 54 months of age.
17. The method of claim 13 , wherein the subject is no greater than about 36 months of age.