Alternative (backdoor) androgen production and masculinization in the human fetus

Peter J. O'Shaughnessy, Jean-Philippe Antignac, Bruno Le Bizec, Marie-Line Morvan, Konstantin Svechnikov, Olle Söder, Iuliia Savchuk, Ana Monteiro, Ugo Soffientini, Zoe C. Johnston, Michelle Bellingham (Corresponding Author), Denise Hough, Natasha Walker, Panagiotis Filis, Paul Fowler (Corresponding Author)

Research output: Contribution to journalArticle

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Abstract

Masculinization of the external genitalia in humans is dependent on formation of 5α-dihydrotestosterone (DHT) through both the canonical androgenic pathway and an alternative (backdoor) pathway. The fetal testes are essential for canonical androgen production, but little is known about the synthesis of backdoor androgens, despite their known critical role in masculinization. In this study, we have measured plasma and tissue levels of endogenous steroids in second trimester human fetuses using multidimensional and high-resolution mass spectrometry. Results show that androsterone is the principal backdoor androgen in the male fetal circulation and that DHT is undetectable (<1 ng/mL), while in female fetuses, there are significantly lower levels of androsterone and testosterone. In the male, intermediates in the backdoor pathway are found primarily in the placenta and fetal liver, with significant androsterone levels also in the fetal adrenal. Backdoor intermediates, including androsterone, are only present at very low levels in the fetal testes. This is consistent with transcript levels of enzymes involved in the alternate pathway (steroid 5α-reductase type 1 [SRD5A1], aldo-keto reductase type 1C2 [AKR1C2], aldo-keto reductase type 1C4 [AKR1C4], cytochrome P450 17A1 [CYP17A1]), as measured by quantitative PCR (qPCR). These data identify androsterone as the predominant backdoor androgen in the human fetus and show that circulating levels are sex dependent, but also that there is little de novo synthesis in the testis. Instead, the data indicate that placental progesterone acts as substrate for synthesis of backdoor androgens, which occurs across several tissues. Masculinization of the human fetus depends, therefore, on testosterone and androsterone synthesis by both the fetal testes and nongonadal tissues, leading to DHT formation at the genital tubercle. Our findings also provide a solid basis to explain why placental insufficiency is associated with disorders of sex development in humans.
Original languageEnglish
Article numbere3000002
JournalPLoS Biology
Volume17
Issue number2
DOIs
Publication statusPublished - 14 Feb 2019

Fingerprint

androsterone
Androsterone
masculinization
androgens
Androgens
fetus
Fetus
Testis
Dihydrotestosterone
testes
synthesis
Tissue
testosterone
genitalia
steroids
Testosterone
Steroids
Placental Insufficiency
Disorders of Sex Development
Genitalia

Keywords

  • androgen
  • alternative pathway
  • androsterone
  • masculinisation
  • human
  • fetal
  • testis
  • placenta
  • adrenal
  • liver
  • backdoor pathway
  • FETAL PLASMA
  • WALLABY POUCH YOUNG
  • 17,20-LYASE ACTIVITY
  • ENZYME EXPRESSION
  • MASS-SPECTROMETRY
  • PLASMA TESTOSTERONE
  • P450 OXIDOREDUCTASE
  • BIOSYNTHESIS
  • GENE-EXPRESSION
  • MATERNAL SMOKING

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

O'Shaughnessy, P. J., Antignac, J-P., Le Bizec, B., Morvan, M-L., Svechnikov, K., Söder, O., ... Fowler , P. (2019). Alternative (backdoor) androgen production and masculinization in the human fetus. PLoS Biology, 17(2), [e3000002]. https://doi.org/10.1371/journal.pbio.3000002

Alternative (backdoor) androgen production and masculinization in the human fetus. / O'Shaughnessy, Peter J.; Antignac, Jean-Philippe; Le Bizec, Bruno; Morvan, Marie-Line; Svechnikov, Konstantin ; Söder, Olle; Savchuk, Iuliia ; Monteiro, Ana; Soffientini, Ugo; Johnston, Zoe C.; Bellingham, Michelle (Corresponding Author); Hough, Denise; Walker, Natasha; Filis, Panagiotis; Fowler , Paul (Corresponding Author).

In: PLoS Biology, Vol. 17, No. 2, e3000002, 14.02.2019.

Research output: Contribution to journalArticle

O'Shaughnessy, PJ, Antignac, J-P, Le Bizec, B, Morvan, M-L, Svechnikov, K, Söder, O, Savchuk, I, Monteiro, A, Soffientini, U, Johnston, ZC, Bellingham, M, Hough, D, Walker, N, Filis, P & Fowler , P 2019, 'Alternative (backdoor) androgen production and masculinization in the human fetus', PLoS Biology, vol. 17, no. 2, e3000002. https://doi.org/10.1371/journal.pbio.3000002
O'Shaughnessy PJ, Antignac J-P, Le Bizec B, Morvan M-L, Svechnikov K, Söder O et al. Alternative (backdoor) androgen production and masculinization in the human fetus. PLoS Biology. 2019 Feb 14;17(2). e3000002. https://doi.org/10.1371/journal.pbio.3000002
O'Shaughnessy, Peter J. ; Antignac, Jean-Philippe ; Le Bizec, Bruno ; Morvan, Marie-Line ; Svechnikov, Konstantin ; Söder, Olle ; Savchuk, Iuliia ; Monteiro, Ana ; Soffientini, Ugo ; Johnston, Zoe C. ; Bellingham, Michelle ; Hough, Denise ; Walker, Natasha ; Filis, Panagiotis ; Fowler , Paul. / Alternative (backdoor) androgen production and masculinization in the human fetus. In: PLoS Biology. 2019 ; Vol. 17, No. 2.
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abstract = "Masculinization of the external genitalia in humans is dependent on formation of 5α-dihydrotestosterone (DHT) through both the canonical androgenic pathway and an alternative (backdoor) pathway. The fetal testes are essential for canonical androgen production, but little is known about the synthesis of backdoor androgens, despite their known critical role in masculinization. In this study, we have measured plasma and tissue levels of endogenous steroids in second trimester human fetuses using multidimensional and high-resolution mass spectrometry. Results show that androsterone is the principal backdoor androgen in the male fetal circulation and that DHT is undetectable (<1 ng/mL), while in female fetuses, there are significantly lower levels of androsterone and testosterone. In the male, intermediates in the backdoor pathway are found primarily in the placenta and fetal liver, with significant androsterone levels also in the fetal adrenal. Backdoor intermediates, including androsterone, are only present at very low levels in the fetal testes. This is consistent with transcript levels of enzymes involved in the alternate pathway (steroid 5α-reductase type 1 [SRD5A1], aldo-keto reductase type 1C2 [AKR1C2], aldo-keto reductase type 1C4 [AKR1C4], cytochrome P450 17A1 [CYP17A1]), as measured by quantitative PCR (qPCR). These data identify androsterone as the predominant backdoor androgen in the human fetus and show that circulating levels are sex dependent, but also that there is little de novo synthesis in the testis. Instead, the data indicate that placental progesterone acts as substrate for synthesis of backdoor androgens, which occurs across several tissues. Masculinization of the human fetus depends, therefore, on testosterone and androsterone synthesis by both the fetal testes and nongonadal tissues, leading to DHT formation at the genital tubercle. Our findings also provide a solid basis to explain why placental insufficiency is associated with disorders of sex development in humans.",
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AU - Le Bizec, Bruno

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AU - Soffientini, Ugo

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N2 - Masculinization of the external genitalia in humans is dependent on formation of 5α-dihydrotestosterone (DHT) through both the canonical androgenic pathway and an alternative (backdoor) pathway. The fetal testes are essential for canonical androgen production, but little is known about the synthesis of backdoor androgens, despite their known critical role in masculinization. In this study, we have measured plasma and tissue levels of endogenous steroids in second trimester human fetuses using multidimensional and high-resolution mass spectrometry. Results show that androsterone is the principal backdoor androgen in the male fetal circulation and that DHT is undetectable (<1 ng/mL), while in female fetuses, there are significantly lower levels of androsterone and testosterone. In the male, intermediates in the backdoor pathway are found primarily in the placenta and fetal liver, with significant androsterone levels also in the fetal adrenal. Backdoor intermediates, including androsterone, are only present at very low levels in the fetal testes. This is consistent with transcript levels of enzymes involved in the alternate pathway (steroid 5α-reductase type 1 [SRD5A1], aldo-keto reductase type 1C2 [AKR1C2], aldo-keto reductase type 1C4 [AKR1C4], cytochrome P450 17A1 [CYP17A1]), as measured by quantitative PCR (qPCR). These data identify androsterone as the predominant backdoor androgen in the human fetus and show that circulating levels are sex dependent, but also that there is little de novo synthesis in the testis. Instead, the data indicate that placental progesterone acts as substrate for synthesis of backdoor androgens, which occurs across several tissues. Masculinization of the human fetus depends, therefore, on testosterone and androsterone synthesis by both the fetal testes and nongonadal tissues, leading to DHT formation at the genital tubercle. Our findings also provide a solid basis to explain why placental insufficiency is associated with disorders of sex development in humans.

AB - Masculinization of the external genitalia in humans is dependent on formation of 5α-dihydrotestosterone (DHT) through both the canonical androgenic pathway and an alternative (backdoor) pathway. The fetal testes are essential for canonical androgen production, but little is known about the synthesis of backdoor androgens, despite their known critical role in masculinization. In this study, we have measured plasma and tissue levels of endogenous steroids in second trimester human fetuses using multidimensional and high-resolution mass spectrometry. Results show that androsterone is the principal backdoor androgen in the male fetal circulation and that DHT is undetectable (<1 ng/mL), while in female fetuses, there are significantly lower levels of androsterone and testosterone. In the male, intermediates in the backdoor pathway are found primarily in the placenta and fetal liver, with significant androsterone levels also in the fetal adrenal. Backdoor intermediates, including androsterone, are only present at very low levels in the fetal testes. This is consistent with transcript levels of enzymes involved in the alternate pathway (steroid 5α-reductase type 1 [SRD5A1], aldo-keto reductase type 1C2 [AKR1C2], aldo-keto reductase type 1C4 [AKR1C4], cytochrome P450 17A1 [CYP17A1]), as measured by quantitative PCR (qPCR). These data identify androsterone as the predominant backdoor androgen in the human fetus and show that circulating levels are sex dependent, but also that there is little de novo synthesis in the testis. Instead, the data indicate that placental progesterone acts as substrate for synthesis of backdoor androgens, which occurs across several tissues. Masculinization of the human fetus depends, therefore, on testosterone and androsterone synthesis by both the fetal testes and nongonadal tissues, leading to DHT formation at the genital tubercle. Our findings also provide a solid basis to explain why placental insufficiency is associated with disorders of sex development in humans.

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KW - alternative pathway

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KW - testis

KW - placenta

KW - adrenal

KW - liver

KW - backdoor pathway

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KW - WALLABY POUCH YOUNG

KW - 17,20-LYASE ACTIVITY

KW - ENZYME EXPRESSION

KW - MASS-SPECTROMETRY

KW - PLASMA TESTOSTERONE

KW - P450 OXIDOREDUCTASE

KW - BIOSYNTHESIS

KW - GENE-EXPRESSION

KW - MATERNAL SMOKING

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U2 - 10.1371/journal.pbio.3000002

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