BNIP3-mediated Autophagy Via the MTOR/ULK1 Pathway Induces Primordial Follicle Loss After Ovarian Tissue Transplantation

Overview

Journal J Assist Reprod Genet

Publisher Springer

Specialties Genetics
Reproductive Medicine

Date 2023 Mar 3

PMID 36869237

Authors

Fengxia Liu

Mujun Li

Affiliations

Soon will be listed here.

Abstract

Purpose: To explore the underlying mechanism of primordial follicle loss in the early period following ovarian tissue transplantation (OTT).

Methods: BNIP3 was selected through bioinformatic protocols, as the hub gene related to autophagy during OTT. BNIP3 and autophagy in mice ovarian grafts and in hypoxia-mimicking KGN cells were detected using immunohistochemistry, transmission electron microscopy (TEM), western blotting, qPCR, and fluorescence staining. The regulatory role played by BNIP3 overexpression and the silencing of KGN cells in autophagy via the mTOR/ULK1 pathway was investigated.

Results: Ultrastructure examination showed that autophagic vacuoles increased after mice ovarian auto-transplantation. The BNIP3 and autophagy-related proteins (Beclin-1, LC3B, and SQSTM1/p62) in mice ovarian granulosa cells of primordial follicle from ovarian grafts were altered compared with the control. Administration of an autophagy inhibitor in mice decreased the depletion of primordial follicles. In vitro experiments indicated that BNIP3 and autophagy activity were upregulated in KGN cells treated with cobalt chloride (CoCl). The overexpression of BNIP3 activated autophagy, whereas the silencing of BNIP3 suppressed it and reversed the autophagy induced by CoCl in KGN cells. Western blotting analysis showed the inhibition of mTOR and activation of ULK1 in KGN cells treated with CoCl and in the overexpression of BNIP3, and the opposite results following BNIP3 silencing. The activation of mTOR reversed the autophagy induced by BNIP3 overexpression.

Conclusions: BNIP3-induced autophagy is crucial in primordial follicle loss during OTT procedure, and BNIP3 is a potential therapeutic target for primordial follicle loss after OTT.

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References

Bhardwaj J, Paliwal A, Saraf P, Sachdeva S . Role of autophagy in follicular development and maintenance of primordial follicular pool in the ovary. J Cell Physiol. 2021; 237(2):1157-1170. DOI: 10.1002/jcp.30613. View

Wang L, Hao H, Wang J, Wang X, Zhang S, Du Y . Decreased autophagy: a major factor for cardiomyocyte death induced by β1-adrenoceptor autoantibodies. Cell Death Dis. 2015; 6:e1862. PMC: 4558518. DOI: 10.1038/cddis.2015.237. View

Fransolet M, Noel L, Henry L, Labied S, Blacher S, Nisolle M . Evaluation of Z-VAD-FMK as an anti-apoptotic drug to prevent granulosa cell apoptosis and follicular death after human ovarian tissue transplantation. J Assist Reprod Genet. 2018; 36(2):349-359. PMC: 6420548. DOI: 10.1007/s10815-018-1353-8. View

Lee S, Cho H, Kim B, Lee J, Kim T . The Effectiveness of Anti-Apoptotic Agents to Preserve Primordial Follicles and Prevent Tissue Damage during Ovarian Tissue Cryopreservation and Xenotransplantation. Int J Mol Sci. 2021; 22(5). PMC: 7961474. DOI: 10.3390/ijms22052534. View

Gawriluk T, Hale A, Flaws J, Dillon C, Green D, Rucker 3rd E . Autophagy is a cell survival program for female germ cells in the murine ovary. Reproduction. 2011; 141(6):759-65. DOI: 10.1530/REP-10-0489. View

Zheng Y, Ma L, Liu N, Tang X, Guo S, Zhang B . Autophagy and Apoptosis of Porcine Ovarian Granulosa Cells During Follicular Development. Animals (Basel). 2019; 9(12). PMC: 6940823. DOI: 10.3390/ani9121111. View

Dolmans M, Falcone T, Patrizio P . Importance of patient selection to analyze in vitro fertilization outcome with transplanted cryopreserved ovarian tissue. Fertil Steril. 2020; 114(2):279-280. DOI: 10.1016/j.fertnstert.2020.04.050. View

Liu J, Van der Elst J, Van den Broecke R, Dhont M . Early massive follicle loss and apoptosis in heterotopically grafted newborn mouse ovaries. Hum Reprod. 2002; 17(3):605-11. DOI: 10.1093/humrep/17.3.605. View

Gorbunova A, Yapryntseva M, Denisenko T, Zhivotovsky B . BNIP3 in Lung Cancer: To Kill or Rescue?. Cancers (Basel). 2020; 12(11). PMC: 7697772. DOI: 10.3390/cancers12113390. View

10.

Delcour C, Amazit L, Patino L, Magnin F, Fagart J, Delemer B . ATG7 and ATG9A loss-of-function variants trigger autophagy impairment and ovarian failure. Genet Med. 2018; 21(4):930-938. DOI: 10.1038/s41436-018-0287-y. View

11.

Yadav A, Yadav P, Chaudhary G, Tiwari M, Gupta A, Sharma A . Autophagy in hypoxic ovary. Cell Mol Life Sci. 2019; 76(17):3311-3322. PMC: 11105528. DOI: 10.1007/s00018-019-03122-4. View

12.

He M, Zhang T, Zhu Z, Qin S, Wang H, Zhao L . LSD1 contributes to programmed oocyte death by regulating the transcription of autophagy adaptor SQSTM1/p62. Aging Cell. 2020; 19(3):e13102. PMC: 7059144. DOI: 10.1111/acel.13102. View

13.

Xu Z . Autophagy phenomenon in mice ovaries following transplantation. Theriogenology. 2022; 195:40-45. DOI: 10.1016/j.theriogenology.2022.10.018. View

14.

Yang Y, Cheung H, Law W, Zhang C, Chan W, Pei X . New Insights into the Role of Autophagy in Ovarian Cryopreservation by Vitrification. Biol Reprod. 2016; 94(6):137. DOI: 10.1095/biolreprod.115.136374. View

15.

Bellot G, Garcia-Medina R, Gounon P, Chiche J, Roux D, Pouyssegur J . Hypoxia-induced autophagy is mediated through hypoxia-inducible factor induction of BNIP3 and BNIP3L via their BH3 domains. Mol Cell Biol. 2009; 29(10):2570-81. PMC: 2682037. DOI: 10.1128/MCB.00166-09. View

16.

Yan Z, Li Q, Zhang L, Kang B, Fan W, Deng T . The growth and development conditions in mouse offspring derived from ovarian tissue cryopreservation and orthotopic transplantation. J Assist Reprod Genet. 2020; 37(4):923-932. PMC: 7183038. DOI: 10.1007/s10815-020-01734-5. View

17.

Kim J, Kundu M, Viollet B, Guan K . AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol. 2011; 13(2):132-41. PMC: 3987946. DOI: 10.1038/ncb2152. View

18.

Munoz-Sanchez J, Chanez-Cardenas M . The use of cobalt chloride as a chemical hypoxia model. J Appl Toxicol. 2018; 39(4):556-570. DOI: 10.1002/jat.3749. View

19.

Zhang J, Ney P . Role of BNIP3 and NIX in cell death, autophagy, and mitophagy. Cell Death Differ. 2009; 16(7):939-46. PMC: 2768230. DOI: 10.1038/cdd.2009.16. View

20.

Zhou X, Zhang J, Li Y, Chen Y, Wu X, Li X . Advanced Oxidation Protein Products Induce G1/G0-Phase Arrest in Ovarian Granulosa Cells via the ROS-JNK/p38 MAPK-p21 Pathway in Premature Ovarian Insufficiency. Oxid Med Cell Longev. 2021; 2021:6634718. PMC: 8337115. DOI: 10.1155/2021/6634718. View