Rehma Saeed
Pronouns: She/Her
Research Mentor(s): Nadia Saadat
Research Mentor School/College/Department: Pediatrics / Medicine
Program:
Authors: Rehma Saeed, Arpita Vyas, Vasantha Padmanabhan, Nadia Saadat
Session: Session 4 1:40-3:30 p.m. Hussey Room
Poster:
Abstract
Background: Individuals with polycystic ovary syndrome (PCOS) have higher levels of testosterone (T), leading to adverse reproductive and metabolic outcomes observed in pregnant mothers and their offspring as a result of developmental programming. Previous studies in Padmanabhan lab established a prenatal T-treated sheep model of PCOS-phenotype, a precocial model of high translational relevance (1). Two decades of work with this model have established gestational T-treatment, in addition to developing a PCOS-phenotype female offspring, perturbs the maternal milieu manifested at the level of lipidome and oxidative stress and leads to fetal increases in T and estradiol. The steroidogenic pathways involved in T programming of the offspring PCOS-phenotype are not known. Objective: To obtain a detailed assessment of steroidogenic pathways perturbed in gestational T-treated sheep to relate to perturbations seen in the offspring. Methods: Time-mated pregnant Suffolk sheep were the subjects of this study with a T group (T, 100mg T-Propionate (Sigma-Aldrich, St. Louis, MO) in 2mL corn oil from days 30-90 of the 147-gestation period, n=12) and the control group (C, vehicle only, n=5). On day 127.12±5.32 of gestation, plasma samples were collected prior to comprehensive steroid panel analysis using LC-MS/MS. All animals were treated in accordance with the National Research Council’s Guide for the Care and Use of Laboratory Animals and were approved by the University of Michigan Animal Care and Use Committee. As steroids are interrelated and can have high multicollinearity, we used the multivariate approach to analyze all steroids simultaneously. The data was analyzed using unsupervised principal component analysis (PCA) using SIMCA. Overview PCA revealed an outlier that was removed from subsequent analysis. IBM SPSS 29.0, R 12.0, and GraphPad Prism 9.0 were utilized for univariate analysis to determine the normality, collinearity, and significance of the data (t-tests). Results: Unsupervised multivariate PCA showed clear separation between the C and T group, the differences were further investigated using a score plot and a loading plot that revealed steroids from both ?5 and ?4 pathways contributed to this separation. Univariate analysis revealed four ?5 pathway steroids, 17-hydroxypregnenolone, androsterone, allopregnanolone, and androstenediol, were significantly altered in the T group (p<0.05). Conclusion: We have previously observed disruptions in steroid hormone levels on day 90 of gestation, at the end of T-treatment (1). The disruptions found in ?4 and ?5 pathways evidenced over a month (37.12±5.32 days) after cessation of T-treatment, indicative of continued disruptions in the mother’s steroid milieu during late gestation, are suggestive of their role IUGR development (2, 3, 4) and offspring PCOS-phenotype (5). As sheep are translationally relevant to humans, future clinical studies may explore if similar steroidogenesis perturbations occur in humans. References 1. Abi Salloum, B., A. Veiga-Lopez, D. H. Abbott, C. F. Burant, and V. Padmanabhan. 2015. “Developmental Programming: Exposure to Testosterone Excess Disrupts Steroidal and Metabolic Environment in Pregnant Sheep.” Endocrinology 156(6):2323–37. doi: 10.1210/en.2014-2006. 2. Steckler, Teresa, Jinrong Wang, Frank F. Bartol, Shyamal K. Roy, and Vasantha Padmanabhan. 2005. “Fetal Programming: Prenatal Testosterone Treatment Causes Intrauterine Growth Retardation, Reduces Ovarian Reserve and Increases Ovarian Follicular Recruitment.” Endocrinology 146(7):3185–93. doi: 10.1210/en.2004-1444. 3. Manikkam, Mohan, Erica J. Crespi, Douglas D. Doop, Carol Herkimer, James S. Lee, Sunkyung Yu, Morton B. Brown, Douglas L. Foster, and Vasantha Padmanabhan. 2004. “Fetal Programming: Prenatal Testosterone Excess Leads to Fetal Growth Retardation and Postnatal Catch-up Growth in Sheep.” Endocrinology 145(2):790–98. doi: 10.1210/en.2003-0478. 4. Beckett, E. M., O. Astapova, T. L. Steckler, A. Veiga-Lopez, and V. Padmanabhan. 2014. “Developmental Programing: Impact of Testosterone on Placental Differentiation.” Reproduction (Cambridge, England) 148(2):199–209. doi: 10.1530/REP-14-0055. 5. Padmanabhan, V., and A. Veiga-Lopez. 2014. “Reproduction Symposium: Developmental Programming of Reproductive and Metabolic Health.” Journal of Animal Science 92(8):3199–3210. doi: 10.2527/jas.2014-7637.
