BOR Papers in Press. Published on November 23, 2011 as DOI:10.1095/biolreprod.111.

097782

OF THE MANY SECRETS OF STEROIDOGENESIS Charles L. Chaffin1,3, Rebecca S. Brogan2 1 Department of OB/GYN & Reproductive Sciences, University of Maryland School of Medicine, Baltimore, MD 21210. email: cchaffin@upi.umaryland.edu. Phone (410) 706-3031. Fax: (410) 706-5747 2 Department of Biology, Loyola University Maryland, Baltimore, MD 21210 3 Correspondance So where is the excitement in the study of steroidogenesis? It is pretty well cut-anddried; after all, 17-estradiol is the same in mice, rats, cows, monkeys, and humans. The basic steroidogenic pathway of cholesterol to pregnenolone to progesterone and adrenal steroids, or to androgens and estrogens is the same in all mammalian taxa. The old debate about the rate limiting step in steroidogenesis has lost some traction as the role of the steroidogenic acute regulatory (STAR) protein has gained clarification. Far and away most work in the field of steroidogenesis currently revolves around gene regulation of specific players in the pathway to be sure, a critical and necessary aspect in understanding cell and tissue specificity. Finally, steroidogenesis does not pose a compelling clinical topic as steroids are easy to measure and easy to manipulate. Despite all this, the immensely important role steroids play in nearly all aspects of vertebrate biology underlies both the true complexity and elegance of steroidogenesis. Species are, in fact, dramatically different in how they regulate the steroidogenic pathway. For example, 17-hydroxyprogesterone (17OH-P) is a poor substrate for P450c17 (official symbol CYP17A1) in humans; because 17OH-P is a precursor for cortisol, this probably keeps substrate available for stress responses in the adrenal gland. In contrast, CYP17A1 efficiently metabolizes 17OH-P to androstenedione and, ultimately, to estrogen in species such as rats, perhaps allowing for a rapid resumption of follicle growth in short luteal phase species [1]. Another example of species variability lies in the compartmentalization of steroidogenesis. While arguably all mammalian species have the same basic relationship in the ovary of androgen producing theca cells and aromatizing granulosa cells, estrogen synthesis during pregnancy is wildly divergent, for example, fetal adrenal placenta in humans and placenta and maternal ovaries in rats. In this issue of Biology of Reproduction, Conley et al [4] hypothesize that compartmentalization of steroidogenic enzymes represents a functional aspect of estrogen synthesis. Implicit in this hypothesis is the concept of optimization of the rate of steroidogenesis, while not necessarily maximizing estrogen production. Conley et al. [4] tested their hypothesis using an insect cell line into which various combinations of CYP17A1, 3-HSD (official symbol HSD3B), and aromatase were transfected. Cells containing different constructs were co-cultured with pregnenolone as a substrate to determine the effect on estrogen synthesis. This is an exceptionally interesting idea considering the natural compartmentalization of enzymes leading to estrogen production. By co-expressing all three enzymes in the same cell, estrogen synthesis was higher than the condition of CYP17A1 alone and HSD3B plus aromatase, suggesting that isolating CYP17A1 limits the rate of estrogen synthesis. However,

Copyright 2011 by The Society for the Study of Reproduction.

consider the case of the ovary. The two-cell model predicts that theca cells generate androstenedione, thus requiring the co-expression of HSD3B and CYP17A1. While it is probably a bit bold to say this is definitively the case in bovids and humans / primates, there are substantial data supporting this hypothesis [2, 3]. The fact that Conley et al. report that this expression configuration actually results in the lowest level of estrogen synthesis strongly suggests for the selection of a compartmentalization system aimed at optimizing rather than maximizing androgen and estrogen synthesis. If, as in the case of the theca cell, CYP17A1 and HSD3B are co-expressed, then how can inhibitors of HSD3B result in the observed increase in androstendione levels? The answer may lay in the isoform and relative expression level of HSD3B. With high levels of either isoform, but especially type 1, inhibition of HSD3B actually promotes (or fails to suppress) androstenedione synthesis, while inhibition of HSD3B at low expression dose-dependently decreases androstenedione. These data also provide experimental evidence that in humans, delta-4 steroids (progestins) are poor substrates for CYP17A1, i.e., inhibiting HSD3B results in the expected increase in DHEA along with a corresponding loss of progesterone. Conley et al. thus suggest that hypoandrogenism could be alleviated by blocking HSD3B, a therapeutic intervention that would be preferred to systemic androgen treatment. What are the take home messages from the elegant studies by Conley et al [4] ? In simple terms, if you co-express CYP17A1 with HSD3B, estrogen synthesis is reduced because some of the substrate ends up in the delta-4 / progesterone pathway. However, because CYP17A1 and HSD3B often co-localize in nature, the relative expression levels become critical, with high HSD3B siphoning off substrate towards progesterone. The system does not seem to have evolved to maximize the rate of estrogen synthesis, but rather teleologically speaking - to maintain very strict control of the steroid environment. How this affects local steroid action and endocrine physiology really remains to be determined; for example, do the low levels of preovulatory progesterone presumably originating from the theca layer have a local function, or is this just a necessary evil associated with having theca-expressed HSD3B in oreder to make androsteneione? Conley and his research colleagues [4] have shown us, once again, the richness of the steroid system and by default, have posed many more questions than they have answered. If science should be about discovery over dollars, thoughtfulness over haste, creativity over translation, then perhaps the article by Conley et al does more than just test a fascinating hypothesis. Perhaps it reminds us of the fact that science should, quite simply, be its own reward. So where is the excitement in the study of steroidogenesis? It lies in all that is left to be learned and all the questions that remain unanswered. References 1. Conley AJ, Bird IM. The role of cytochrome P450 17 alpha-hydroxylase and 3 betahydroxysteroid dehydrogenase in the integration of gonadal and adrenal

steroidogenesis via the delta 5 and delta 4 pathways of steroidogenesis in mammals. Biol Reprod 1997; 56: 789-799. 2. Conley AJ, Kaminski MA, Dubowsky SA, Jablonka-Shariff A, Redmer DA, Reynolds LP. Immunohistochemical localization of 3 beta-hydroxysteroid dehydrogenase and P450 17 alpha-hydroxylase during follicular and luteal development in pigs, sheep, and cows. Biol Reprod 1995; 52: 1081-1094. 3. Galas J, Slomczynska M, Knapczyk-Stwora K, Durlej M, Starowicz A, Tabarowski Z, Rutka K, Szoltys M. Steroid levels and the spatiotemporal expression of steroidogenic enzymes and androgen receptor in developing ovaries of immature rats. Acta Histochem. 4. Conley AJ, Corbin CJ, Thomas JL, Gee NA, Lasley BL, Moeller BC, Stanley SD, Berger T. Costs and Consequences of Cellular Compartmentalization and Substrate Competition among Human Enzymes Involved in Androgen and Estrogen Synthesis. Biol Reprod 2012; (in press), published ahead of print 14 September 2011 as doi:10.1095/biolreprod.111.094706.