Activin-Like Kinase 2 Functions in Peri-implantation Uterine Signaling in Mice and Humans


Implantation of a blastocyst in the uterus is a multistep process tightly controlled by an intricate regulatory network of interconnected ovarian, uterine, and embryonic factors. Bone morphogenetic protein (BMP) ligands and receptors are expressed in the uterus of pregnant mice, and BMP2 has been shown to be a key regulator of implantation. In this study, we investigated the roles of the BMP type 1 receptor, activin-like kinase 2 (ALK2), during mouse pregnancy by producing mice carrying a conditional ablation of Alk2 in the uterus (Alk2 cKO mice). In the absence of ALK2, embryos demonstrate delayed invasion into the uterine epithelium and stroma, and upon implantation, stromal cells fail to undergo uterine decidualization, resulting in sterility. Mechanistically, microarray analysis revealed that CCAAT/enhancer-binding protein β (Cebpb) expression is suppressed during decidualization in Alk2 cKO females. These findings and the similar phenotypes of Cebpb cKO and Alk2 cKO mice lead to the hypothesis that BMPs act upstream of CEBPB in the stroma to regulate decidualization. To test this hypothesis, we knocked down ALK2 in human uterine stromal cells (hESC) and discovered that ablation of ALK2 alters hESC decidualization and suppresses CEBPB mRNA and protein levels. Chromatin immunoprecipitation (ChIP) analysis of decidualizing hESC confirmed that BMP signaling proteins, SMAD1/5, directly regulate expression of CEBPB by binding a distinct regulatory sequence in the 3′ UTR of this gene; CEBPB, in turn, regulates the expression of progesterone receptor (PGR). Our work clarifies the conserved mechanisms through which BMPs regulate peri-implantation in rodents and primates and, for the first time, uncovers a linear pathway of BMP signaling through ALK2 to regulate CEBPB and, subsequently, PGR during decidualization.


Vyšlo v časopise: Activin-Like Kinase 2 Functions in Peri-implantation Uterine Signaling in Mice and Humans. PLoS Genet 9(11): e32767. doi:10.1371/journal.pgen.1003863
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003863

Souhrn

Implantation of a blastocyst in the uterus is a multistep process tightly controlled by an intricate regulatory network of interconnected ovarian, uterine, and embryonic factors. Bone morphogenetic protein (BMP) ligands and receptors are expressed in the uterus of pregnant mice, and BMP2 has been shown to be a key regulator of implantation. In this study, we investigated the roles of the BMP type 1 receptor, activin-like kinase 2 (ALK2), during mouse pregnancy by producing mice carrying a conditional ablation of Alk2 in the uterus (Alk2 cKO mice). In the absence of ALK2, embryos demonstrate delayed invasion into the uterine epithelium and stroma, and upon implantation, stromal cells fail to undergo uterine decidualization, resulting in sterility. Mechanistically, microarray analysis revealed that CCAAT/enhancer-binding protein β (Cebpb) expression is suppressed during decidualization in Alk2 cKO females. These findings and the similar phenotypes of Cebpb cKO and Alk2 cKO mice lead to the hypothesis that BMPs act upstream of CEBPB in the stroma to regulate decidualization. To test this hypothesis, we knocked down ALK2 in human uterine stromal cells (hESC) and discovered that ablation of ALK2 alters hESC decidualization and suppresses CEBPB mRNA and protein levels. Chromatin immunoprecipitation (ChIP) analysis of decidualizing hESC confirmed that BMP signaling proteins, SMAD1/5, directly regulate expression of CEBPB by binding a distinct regulatory sequence in the 3′ UTR of this gene; CEBPB, in turn, regulates the expression of progesterone receptor (PGR). Our work clarifies the conserved mechanisms through which BMPs regulate peri-implantation in rodents and primates and, for the first time, uncovers a linear pathway of BMP signaling through ALK2 to regulate CEBPB and, subsequently, PGR during decidualization.


Zdroje

1. ChaJ, SunX, DeySK (2012) Mechanisms of implantation: strategies for successful pregnancy. Nat Med 18: 1754–1767.

2. LargeMJ, DeMayoFJ (2012) The regulation of embryo implantation and endometrial decidualization by progesterone receptor signaling. Mol Cell Endocrinol 358: 155–165.

3. RamathalCY, BagchiIC, TaylorRN, BagchiMK (2010) Endometrial decidualization: of mice and men. Semin Reprod Med 28: 17–26.

4. BagchiIC, LiQ, CheonYP, MantenaSR, KannanA, et al. (2005) Use of the progesterone receptor antagonist RU 486 to identify novel progesterone receptor-regulated pathways in implantation. Semin Reprod Med 23: 38–45.

5. ZhangH, BradleyA (1996) Mice deficient for BMP2 are nonviable and have defects in amnion/chorion and cardiac development. Development 122: 2977–2986.

6. SoyalSM, MukherjeeA, LeeKY, LiJ, LiH, et al. (2005) Cre-mediated recombination in cell lineages that express the progesterone receptor. Genesis 41: 58–66.

7. Clementi C, Pangas SA, Matzuk MM (2013) In: Barbieri, Strauss, editors. Yen and Jaffe's Reproductive Endocrinology 7th ed: Elsevier Inc.

8. LeeKY, JeongJW, WangJ, MaL, MartinJF, et al. (2007) Bmp2 is critical for the murine uterine decidual response. Mol Cell Biol 27: 5468–5478.

9. LiQ, KannanA, WangW, DemayoFJ, TaylorRN, et al. (2007) Bone morphogenetic protein 2 functions via a conserved signaling pathway involving Wnt4 to regulate uterine decidualization in the mouse and the human. J Biol Chem 282: 31725–31732.

10. PariaBC, MaW, TanJ, RajaS, DasSK, et al. (2001) Cellular and molecular responses of the uterus to embryo implantation can be elicited by locally applied growth factors. Proc Natl Acad Sci U S A 98: 1047–1052.

11. YingY, ZhaoGQ (2000) Detection of multiple bone morphogenetic protein messenger ribonucleic acids and their signal transducer, Smad1, during mouse decidualization. Biol Reprod 63: 1781–1786.

12. EdsonMA, NalamRL, ClementiC, FrancoHL, DemayoFJ, et al. (2010) Granulosa cell-expressed BMPR1A and BMPR1B have unique functions in regulating fertility but act redundantly to suppress ovarian tumor development. Mol Endocrinol 24: 1251–1266.

13. GuZ, ReynoldsEM, SongJ, LeiH, FeijenA, et al. (1999) The type I serine/threonine kinase receptor ActRIA (ALK2) is required for gastrulation of the mouse embryo. Development 126: 2551–2561.

14. MishinaY, CrombieR, BradleyA, BehringerRR (1999) Multiple roles for activin-like kinase-2 signaling during mouse embryogenesis. Dev Biol 213: 314–326.

15. KaartinenV, NagyA (2001) Removal of the floxed neo gene from a conditional knockout allele by the adenoviral Cre recombinase in vivo. Genesis 31: 126–129.

16. TongW, PollardJW (1999) Progesterone inhibits estrogen-induced cyclin D1 and cdk4 nuclear translocation, cyclin E- and cyclin A-cdk2 kinase activation, and cell proliferation in uterine epithelial cells in mice. Mol Cell Biol 19: 2251–2264.

17. NagashimaT, LiQ, ClementiC, LydonJP, DeMayoFJ, MatzukMM (2013) BMPR2 is required for post-implantation uterine function and pregnancy maintenance. The Journal of Clinical Investigation 123 (6) 2539–50.

18. ChakrabortyI, DasSK, WangJ, DeySK (1996) Developmental expression of the cyclo-oxygenase-1 and cyclo-oxygenase-2 genes in the peri-implantation mouse uterus and their differential regulation by the blastocyst and ovarian steroids. J Mol Endocrinol 16: 107–122.

19. MantenaSR, KannanA, CheonYP, LiQ, JohnsonPF, et al. (2006) C/EBPbeta is a critical mediator of steroid hormone-regulated cell proliferation and differentiation in the uterine epithelium and stroma. Proc Natl Acad Sci U S A 103: 1870–1875.

20. DunnCL, KellyRW, CritchleyHO (2003) Decidualization of the human endometrial stromal cell: an enigmatic transformation. Reprod Biomed Online 7: 151–161.

21. ChristianM, PohnkeY, KempfR, GellersenB, BrosensJJ (2002) Functional association of PR and CCAAT/enhancer-binding protein beta isoforms: promoter-dependent cooperation between PR-B and liver-enriched inhibitory protein, or liver-enriched activatory protein and PR-A in human endometrial stromal cells. Mol Endocrinol 16: 141–154.

22. WangW, LiQ, BagchiIC, BagchiMK (2010) The CCAAT/enhancer binding protein beta is a critical regulator of steroid-induced mitotic expansion of uterine stromal cells during decidualization. Endocrinology 151: 3929–3940.

23. BagchiMK, MantenaSR, KannanA, BagchiIC (2006) Control of uterine cell proliferation and differentiation by C/EBPbeta: functional implications for establishment of early pregnancy. Cell Cycle 5: 922–925.

24. LydonJP, DeMayoFJ, ConneelyOM, O'MalleyBW (1996) Reproductive phenotpes of the progesterone receptor null mutant mouse. J Steroid Biochem Mol Biol 56: 67–77.

25. NairSC, RimermanRA, ToranEJ, ChenS, PrapapanichV, et al. (1997) Molecular cloning of human FKBP51 and comparisons of immunophilin interactions with Hsp90 and progesterone receptor. Mol Cell Biol 17: 594–603.

26. FinnCA, MartinL (1972) Endocrine control of the timing of endometrial sensitivity to a decidual stimulus. Biol Reprod 7: 82–86.

27. LiQ, KannanA, DeMayoFJ, LydonJP, CookePS, et al. (2011) The antiproliferative action of progesterone in uterine epithelium is mediated by Hand2. Science 331: 912–916.

28. FrancoHL, RubelCA, LargeMJ, WetendorfM, Fernandez-ValdiviaR, et al. (2012) Epithelial progesterone receptor exhibits pleiotropic roles in uterine development and function. FASEB J 26: 1218–1227.

29. LejeuneB, Van HoeckJ, LeroyF (1981) Transmitter role of the luminal uterine epithelium in the induction of decidualization in rats. J Reprod Fertil 61: 235–240.

30. GellersenB, BrosensIA, BrosensJJ (2007) Decidualization of the human endometrium: mechanisms, functions, and clinical perspectives. Semin Reprod Med 25: 445–453.

31. WilcoxAJ, WeinbergCR, O'ConnorJF, BairdDD, SchlattererJP, et al. (1988) Incidence of early loss of pregnancy. N Engl J Med 319: 189–194.

32. NorwitzER, SchustDJ, FisherSJ (2001) Implantation and the survival of early pregnancy. N Engl J Med 345: 1400–1408.

33. NorwitzER (2006) Defective implantation and placentation: laying the blueprint for pregnancy complications. Reprod Biomed Online 13: 591–599.

34. DiedrichK, FauserBC, DevroeyP, GriesingerG, Evian Annual Reproduction WorkshopG (2007) The role of the endometrium and embryo in human implantation. Hum Reprod Update 13: 365–377.

35. SimonC, LanderasJ, ZuzuarreguiJL, MartinJC, RemohiJ, et al. (1999) Early pregnancy losses in in vitro fertilization and oocyte donation. Fertil Steril 72: 1061–1065.

36. JonesRL, SalamonsenLA, FindlayJK (2002) Activin A promotes human endometrial stromal cell decidualization in vitro. J Clin Endocrinol Metab 87: 4001–4004.

37. TrenorCC (2011) Hormonal contraception and thrombotic risk: a multidisciplinary approach. Pediatrics 127: 347–357.

38. MueckAO (2010) Exogenous hormones, the risk of venous thromboembolism, and activated protein C resistance. Menopause 17: 1099–1103.

39. MueckAO, SeegerH, RabeT (2010) Hormonal contraception and risk of endometrial cancer: a systematic review. Endocr Relat Cancer 17: R263–271.

40. ChangH, BrownCW, MatzukMM (2002) Genetic analysis of the mammalian transforming growth factor-beta superfamily. Endocr Rev 23: 787–823.

41. MassagueJ (2008) TGFbeta in Cancer. Cell 134: 215–230.

42. PaduaD, MassagueJ (2009) Roles of TGFbeta in metastasis. Cell Res 19: 89–102.

43. TanCK, LeuenbergerN, TanMJ, YanYW, ChenY, et al. (2011) Smad3 deficiency in mice protects against insulin resistance and obesity induced by a high-fat diet. Diabetes 60: 464–476.

44. DabekJ, KulachA, Monastyrska-CupB, GasiorZ (2006) Transforming growth factor beta and cardiovascular diseases: the other facet of the ‘protective cytokine’. Pharmacol Rep 58: 799–805.

45. MargadantC, SonnenbergA (2010) Integrin-TGF-beta crosstalk in fibrosis, cancer and wound healing. EMBO Rep 11: 97–105.

46. WilkesJJ, LloydDJ, GekakisN (2009) Loss-of-function mutation in myostatin reduces tumor necrosis factor alpha production and protects liver against obesity-induced insulin resistance. Diabetes 58: 1133–1143.

47. SchleinitzD, KlotingN, BottcherY, WolfS, DietrichK, et al. (2011) Genetic and evolutionary analyses of the human bone morphogenetic protein receptor 2 (BMPR2) in the pathophysiology of obesity. PLoS One 6: e16155.

48. DudasM, SridurongritS, NagyA, OkazakiK, KaartinenV (2004) Craniofacial defects in mice lacking BMP type I receptor Alk2 in neural crest cells. Mech Dev 121: 173–182.

49. DebK, ReeseJ, PariaBC (2006) Methodologies to study implantation in mice. Methods Mol Med 121: 9–34.

50. FinnCA, HinchliffeJR (1964) Reaction of the Mouse Uterus during Implantation and Deciduoma Formation as Demonstrated by Changes in the Distribution of Alkaline Phosphatase. J Reprod Fertil 8: 331–338.

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Genetika Reprodukčná medicína

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