USP11 regulates proliferation and apoptosis of human spermatogonial stem cells via HOXC5-mediated canonical WNT/β-catenin signaling pathway.
Humans
Male
Apoptosis
/ genetics
Cell Proliferation
/ genetics
Wnt Signaling Pathway
/ genetics
Homeodomain Proteins
/ metabolism
Azoospermia
/ metabolism
Spermatogonia
/ metabolism
Spermatogenesis
/ genetics
Adult Germline Stem Cells
/ metabolism
beta Catenin
/ metabolism
Testis
/ metabolism
Thiolester Hydrolases
HOXC5
Human spermatogonial stem cells
Proliferation and apoptosis
USP11
WNT/β-catenin pathway
Journal
Cellular and molecular life sciences : CMLS
ISSN: 1420-9071
Titre abrégé: Cell Mol Life Sci
Pays: Switzerland
ID NLM: 9705402
Informations de publication
Date de publication:
09 May 2024
09 May 2024
Historique:
received:
02
01
2024
accepted:
21
04
2024
revised:
23
03
2024
medline:
9
5
2024
pubmed:
9
5
2024
entrez:
9
5
2024
Statut:
epublish
Résumé
Spermatogonial stem cells (SSCs) are capable of transmitting genetic information to the next generations and they are the initial cells for spermatogenesis. Nevertheless, it remains largely unknown about key genes and signaling pathways that regulate fate determinations of human SSCs and male infertility. In this study, we explored the expression, function, and mechanism of USP11 in controlling the proliferation and apoptosis of human SSCs as well as the association between its abnormality and azoospermia. We found that USP11 was predominantly expressed in human SSCs as shown by database analysis and immunohistochemistry. USP11 silencing led to decreases in proliferation and DNA synthesis and an enhancement in apoptosis of human SSCs. RNA-sequencing identified HOXC5 as a target of USP11 in human SSCs. Double immunofluorescence, Co-immunoprecipitation (Co-IP), and molecular docking demonstrated an interaction between USP11 and HOXC5 in human SSCs. HOXC5 knockdown suppressed the growth of human SSCs and increased apoptosis via the classical WNT/β-catenin pathway. In contrast, HOXC5 overexpression reversed the effect of proliferation and apoptosis induced by USP11 silencing. Significantly, lower levels of USP11 expression were observed in the testicular tissues of patients with spermatogenic disorders. Collectively, these results implicate that USP11 regulates the fate decisions of human SSCs through the HOXC5/WNT/β-catenin pathway. This study thus provides novel insights into understanding molecular mechanisms underlying human spermatogenesis and the etiology of azoospermia and it offers new targets for gene therapy of male infertility.
Identifiants
pubmed: 38722330
doi: 10.1007/s00018-024-05248-6
pii: 10.1007/s00018-024-05248-6
doi:
Substances chimiques
Homeodomain Proteins
0
USP11 protein, human
0
beta Catenin
0
Thiolester Hydrolases
EC 3.1.2.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
211Subventions
Organisme : National Outstanding Youth Science Fund Project of National Natural Science Foundation of China
ID : 32170862
Organisme : National Outstanding Youth Science Fund Project of National Natural Science Foundation of China
ID : 31872845
Informations de copyright
© 2024. The Author(s).
Références
Agarwal A, Baskaran S, Parekh N et al (2021) Male infertility. The Lancet 397:319–333. https://doi.org/10.1016/S0140-6736(20)32667-2
doi: 10.1016/S0140-6736(20)32667-2
Ml E, Sc E, Dj L et al (2023) Male infertility. Nat Rev Disease Primers. https://doi.org/10.1038/s41572-023-00459-w
doi: 10.1038/s41572-023-00459-w
Krausz C, Riera-Escamilla A (2018) Genetics of male infertility. Nat Rev Urol 15:369–384. https://doi.org/10.1038/s41585-018-0003-3
doi: 10.1038/s41585-018-0003-3
Vn P, Tp K, As H (2020) Genetic mutations contributing to non-obstructive azoospermia. Best Pract Res Clin Endocrinol Metabol. https://doi.org/10.1016/j.beem.2020.101479
doi: 10.1016/j.beem.2020.101479
Halpern JA, Davis AM, Brannigan RE (2022) Diagnosis and treatment of infertility in men. JAMA 328:2056–2057. https://doi.org/10.1001/jama.2022.19294
doi: 10.1001/jama.2022.19294
Kasak L, Laan M (2021) Monogenic causes of non-obstructive azoospermia: challenges, established knowledge, limitations and perspectives. Hum Genet 140:135–154. https://doi.org/10.1007/s00439-020-02112-y
doi: 10.1007/s00439-020-02112-y
de Kretser DM, Loveland KL, Meinhardt A et al (1998) Spermatogenesis. Hum Reprod 13(Suppl 1):1–8. https://doi.org/10.1093/humrep/13.suppl_1.1
doi: 10.1093/humrep/13.suppl_1.1
Murat F, Mbengue N, Winge SB et al (2023) The molecular evolution of spermatogenesis across mammals. Nature 613:308–316. https://doi.org/10.1038/s41586-022-05547-7
doi: 10.1038/s41586-022-05547-7
Cheng H, Shang D, Zhou R (2022) Germline stem cells in human. Signal Transduct Target Ther 7:345. https://doi.org/10.1038/s41392-022-01197-3
doi: 10.1038/s41392-022-01197-3
Neto FTL, Bach PV, Najari BB et al (2016) Spermatogenesis in humans and its affecting factors. Semin Cell Dev Biol 59:10–26. https://doi.org/10.1016/j.semcdb.2016.04.009
doi: 10.1016/j.semcdb.2016.04.009
Phillips BT, Gassei K, Orwig KE (2010) Spermatogonial stem cell regulation and spermatogenesis. Philos Trans R Soc Lond B Biol Sci 365:1663–1678. https://doi.org/10.1098/rstb.2010.0026
doi: 10.1098/rstb.2010.0026
Diao L, Turek PJ, John CM et al (2022) Roles of spermatogonial stem cells in spermatogenesis and fertility restoration. Front Endocrinol (Lausanne) 13:895528. https://doi.org/10.3389/fendo.2022.895528
doi: 10.3389/fendo.2022.895528
Guo J, Sosa E, Chitiashvili T et al (2021) Single-cell analysis of the developing human testis reveals somatic niche cell specification and fetal germline stem cell establishment. Cell Stem Cell 28:764-778.e4. https://doi.org/10.1016/j.stem.2020.12.004
doi: 10.1016/j.stem.2020.12.004
Rajachandran S, Zhang X, Cao Q et al (2023) Dissecting the spermatogonial stem cell niche using spatial transcriptomics. Cell Rep 42:112737. https://doi.org/10.1016/j.celrep.2023.112737
doi: 10.1016/j.celrep.2023.112737
Di Persio S, Neuhaus N (2023) Human spermatogonial stem cells and their niche in male (in)fertility: novel concepts from single-cell RNA-sequencing. Hum Reprod 38:1–13. https://doi.org/10.1093/humrep/deac245
doi: 10.1093/humrep/deac245
Yang Q-E, Kim D, Kaucher A et al (2013) CXCL12–CXCR4 signaling is required for the maintenance of mouse spermatogonial stem cells. J Cell Sci 126:1009–1020. https://doi.org/10.1242/jcs.119826
doi: 10.1242/jcs.119826
Kubota H, Avarbock MR, Brinster RL (2004) Growth factors essential for self-renewal and expansion of mouse spermatogonial stem cells. Proc Natl Acad Sci U S A 101:16489–16494. https://doi.org/10.1073/pnas.0407063101
doi: 10.1073/pnas.0407063101
Caires KC, de Avila JM, Cupp AS, McLean DJ (2012) VEGFA family isoforms regulate spermatogonial stem cell homeostasis in vivo. Endocrinology 153:887–900. https://doi.org/10.1210/en.2011-1323
doi: 10.1210/en.2011-1323
Pellegrini M, Filipponi D, Gori M et al (2008) ATRA and KL promote differentiation toward the meiotic program of male germ cells. Cell Cycle 7:3878–3888. https://doi.org/10.4161/cc.7.24.7262
doi: 10.4161/cc.7.24.7262
Carlomagno G, van Bragt MPA, Korver CM et al (2010) BMP4-induced differentiation of a rat spermatogonial stem cell line causes changes in its cell adhesion properties. Biol Reprod 83:742–749. https://doi.org/10.1095/biolreprod.110.085456
doi: 10.1095/biolreprod.110.085456
Vij SC, Sabanegh E, Agarwal A (2018) Biological therapy for non-obstructive azoospermia. Expert Opin Biol Ther 18:19–23. https://doi.org/10.1080/14712598.2018.1380622
doi: 10.1080/14712598.2018.1380622
Abdelaal NE, Tanga BM, Abdelgawad M et al (2021) Cellular therapy via spermatogonial stem cells for treating impaired spermatogenesis non-obstructive azoospermia. Cells 10:1779. https://doi.org/10.3390/cells10071779
doi: 10.3390/cells10071779
Medrano JV, Rombaut C, Simon C et al (2016) Human spermatogonial stem cells display limited proliferation in vitro under mouse spermatogonial stem cell culture conditions. Fertil Steril 106:1539-1549.e8. https://doi.org/10.1016/j.fertnstert.2016.07.1065
doi: 10.1016/j.fertnstert.2016.07.1065
Hermann BP, Sukhwani M, Hansel MC, Orwig KE (2010) Spermatogonial stem cells in higher primates: are there differences from those in rodents? Reproduction 139:479–493. https://doi.org/10.1530/REP-09-0255
doi: 10.1530/REP-09-0255
Zhou D, Wang X, Liu Z et al (2020) The expression characteristics of FBXW7 in human testis suggest its function is different from that in mice. Tissue Cell 62:101315. https://doi.org/10.1016/j.tice.2019.101315
doi: 10.1016/j.tice.2019.101315
Hou J, Niu M, Liu L et al (2015) Establishment and characterization of human germline stem cell line with unlimited proliferation potentials and no tumor formation. Sci Rep 5:16922. https://doi.org/10.1038/srep16922
doi: 10.1038/srep16922
Hermann BP, Cheng K, Singh A et al (2018) The mammalian spermatogenesis single-cell transcriptome, from spermatogonial stem cells to spermatids. Cell Rep 25:1650-1667.e8. https://doi.org/10.1016/j.celrep.2018.10.026
doi: 10.1016/j.celrep.2018.10.026
Guo J, Grow EJ, Mlcochova H et al (2018) The adult human testis transcriptional cell atlas. Cell Res 28:1141–1157. https://doi.org/10.1038/s41422-018-0099-2
doi: 10.1038/s41422-018-0099-2
Guo J, Nie X, Giebler M et al (2020) The dynamic transcriptional cell atlas of testis development during human puberty. Cell Stem Cell 26:262-276.e4. https://doi.org/10.1016/j.stem.2019.12.005
doi: 10.1016/j.stem.2019.12.005
Nusse R, Clevers H (2017) Wnt/β-catenin signaling, disease, and emerging therapeutic modalities. Cell 169:985–999. https://doi.org/10.1016/j.cell.2017.05.016
doi: 10.1016/j.cell.2017.05.016
Hrycaj SM, Dye BR, Baker NC et al (2015) Hox5 genes regulate the Wnt2/2b-Bmp4 signaling axis during lung development. Cell Rep 12:903–912. https://doi.org/10.1016/j.celrep.2015.07.020
doi: 10.1016/j.celrep.2015.07.020
Guo T, Tang H, Yuan Z et al (2022) The dual role of USP11 in cancer. J Oncol 2022:9963905. https://doi.org/10.1155/2022/9963905
doi: 10.1155/2022/9963905
Zhang E, Shen B, Mu X et al (2016) Ubiquitin-specific protease 11 (USP11) functions as a tumor suppressor through deubiquitinating and stabilizing VGLL4 protein. Am J Cancer Res 6:2901–2909
doi: 10.1158/0008-5472.CAN-15-2120
Zhang C, Xie C, Wang X et al (2020) Aberrant USP11 expression regulates NF90 to promote proliferation and metastasis in hepatocellular carcinoma. Am J Cancer Res 10:1416–1428
Zhou Z, Luo A, Shrivastava I et al (2017) Regulation of XIAP turnover reveals a role for USP11 in promotion of tumorigenesis. EBioMedicine 15:48–61. https://doi.org/10.1016/j.ebiom.2016.12.014
doi: 10.1016/j.ebiom.2016.12.014
Deng T, Yan G, Song X et al (2018) Deubiquitylation and stabilization of p21 by USP11 is critical for cell-cycle progression and DNA damage responses. Proc Natl Acad Sci U S A 115:4678–4683. https://doi.org/10.1073/pnas.1714938115
doi: 10.1073/pnas.1714938115
Bijl JJ, van Oostveen JW, Walboomers JMM et al (1998) Differentiation and cell-type-restricted expression of HOXC4, HOXC5 and HOXC6 in myeloid leukemias and normal myeloid cells. Leukemia 12:1724–1732. https://doi.org/10.1038/sj.leu.2401106
doi: 10.1038/sj.leu.2401106
Alami Y, Castronovo V, Belotti D et al (1999) HOXC5 and HOXC8 expression are selectively turned on in human cervical cancer cells compared to normal keratinocytes. Biochem Biophys Res Commun 257:738–745. https://doi.org/10.1006/bbrc.1999.0516
doi: 10.1006/bbrc.1999.0516
Schmidt JA, Avarbock MR, Tobias JW, Brinster RL (2009) Identification of glial cell line-derived neurotrophic factor-regulated genes important for spermatogonial stem cell self-renewal in the rat. Biol Reprod 81:56–66. https://doi.org/10.1095/biolreprod.108.075358
doi: 10.1095/biolreprod.108.075358
Lin H, Cheng K, Kubota H et al (2022) Histone methyltransferase DOT1L is essential for self-renewal of germline stem cells. Genes Dev 36:752–763. https://doi.org/10.1101/gad.349550.122
doi: 10.1101/gad.349550.122
Zhao H, Ming T, Tang S et al (2022) Wnt signaling in colorectal cancer: pathogenic role and therapeutic target. Mol Cancer 21:144. https://doi.org/10.1186/s12943-022-01616-7
doi: 10.1186/s12943-022-01616-7
Xue X, Fan C, Wang L et al (2022) Ascorbic acid regulates mouse spermatogonial stem cell proliferation in a Wnt/β-catenin/ROS signaling dependent manner. Theriogenology 184:61–72. https://doi.org/10.1016/j.theriogenology.2022.02.028
doi: 10.1016/j.theriogenology.2022.02.028