Meconium androgens are correlated with ASD-related phenotypic traits in early childhood

Prenatal exposure to increased androgens has been suggested as a risk factor for autism spectrum disorder (ASD). This hypothesis has been examined by measurement of steroids in amniotic fluid and cord blood, with mixed results. To provide an orthogonal measure of fetal exposure, this study used meconium, the first stool of a newborn, to measure prenatal androgen exposure from infants in the Early Autism Risk Longitudinal Investigation (EARLI). EARLI is a familial-enriched risk cohort that enrolled pregnant mothers who already had a child with an ASD diagnosis. In the younger child, we investigated the association between meconium unconjugated (u) and total (t) concentrations of major androgens testosterone (T), dehydroepiandrosterone (DHEA) and androstenedione (A4), and ASD-related traits at 12 and 36 months of age. Autism traits were measured at 12 months with Autism Observation Scale for Infants (AOSI) and at 36 months with total score on the Social Responsiveness Scale (SRS). 137 children (61 males, 76 females) had data on both outcomes and meconium androgen measurements. Separate robust linear regressions between each of the log-transformed androgens and log-transformed AOSI and SRS scores revealed three-way interaction between sex of the child, sex of the proband, and androgen concentration. In the adjusted analyses; t-T, u-A4, and u-DHEA ( P ≤ 0.01) were positively associated with AOSI scores while u-T ( P =0.004) and u-DHEA ( P =0.007) were positively associated with SRS total score among females with female probands. Additionally, higher concentrations of u-T ( P =0.01) and t-T ( P =0.01) predicted higher SRS total score in males with male probands. Significance Using a prospective pregnancy cohort enriched for autism risk, we investigated prenatal androgen exposure measured from meconium as a risk factor for autism-related traits. Several meconium androgens were positively correlated with autism-related traits. In addition, we found a strong positive association between autism traits in the sub-group of individuals with an older female sibling with autism extending a previous finding based on cord blood measures in the same cohort. This study supports the utility of meconium for studies of endogenous fetal metabolism and suggests the sex of the proband should be considered as a biological variable in relevant studies. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement Data collection and analyses were supported by NIH R01ES016443, NIH R21ES02559, and Autism Speaks AS5938. BYP was supported by an Autism Speaks predoctoral fellowship (AS 8556). Meconium method development and analysis was supported by R21HD087866, R03CA211820, R01ES029336, and a NARSAD Young Investigator Brain and Behavior Foundation grant to NWS. ### Author Declarations All relevant ethical guidelines have been followed; any necessary IRB and/or ethics committee approvals have been obtained and details of the IRB/oversight body are included in the manuscript. Yes All necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines and uploaded the relevant EQUATOR Network research reporting checklist(s) and other pertinent material as supplementary files, if applicable. Yes Data is available via National Database for Autism Research (NDAR) and the Steering Committee of EARLI. The corresponding author remains blinded to potentially identifying information.