Infertility has become one of the most serious diseases worldwide,and 50% of this disease can be attributed to male-related factors.Spermatogenesis,by definition,is a complex process by which spermatogonial stem cells...Infertility has become one of the most serious diseases worldwide,and 50% of this disease can be attributed to male-related factors.Spermatogenesis,by definition,is a complex process by which spermatogonial stem cells(SSCs)self-renew to maintain stem cell population within the testes and differentiate into mature spermatids.It is of great significance to uncover gene regulation and signaling pathways that are involved in the fate determinations of SSCs with aims to better understand molecular mechanisms underlying human spermatogenesis and identify novel targets for gene therapy of male infertility.Significant achievement has recently been made in demonstrating the signaling molecules and pathways mediating the fate decisions of mammalian SSCs.In this review,we address key gene regulation and crucial signaling transduction pathways in controlling the self-renewal,differentiation,and apoptosis of SSCs,and we illustrate the networks of genes and signaling pathways in SSC fate determinations.We also highlight perspectives and future directions in SSC regulation by genes and their signaling pathways.This review could provide novel insights into the genetic regulation of normal and abnormal spermatogenesis and offer molecular targets to develop new approaches for gene therapy of male infertility.展开更多
Icariin is a pure compound derived from Epimedium brevicornu Maxim,and it helps the regulation of male reproduction.Nevertheless,the role and underlying mechanisms of Icariin in mediating male germ cell development re...Icariin is a pure compound derived from Epimedium brevicornu Maxim,and it helps the regulation of male reproduction.Nevertheless,the role and underlying mechanisms of Icariin in mediating male germ cell development remain to be clarified.Here,we have demonstrated that Icariin promoted proliferation and DNA synthesis of mouse spermatogonial stem cells(SSCs).Furthermore,surface plasmon resonance iron(SPRi)and molecular docking(MOE)assays revealed that phosphodiesterase 5A(PDE5A)was an important target of Icariin in mouse SSCs.Mechanically,Icariin decreased the expression level of PDE5A.Interestingly,hydrogen peroxides(H2O2)enhanced the expression level of phosphorylation H2A.X(p-H2A.X),whereas Icariin diminished the expression level of p-H2A.X and DNA damage caused by H2O2 in mouse SSCs.Finally,our in vivo animal study indicated that Icariin protected male reproduction.Collectively,these results implicate that Icariin targets PDE5A to regulate mouse SSC viability and DNA damage and improves male reproductive capacity.This study thus sheds new insights into molecular mechanisms underlying the fate decisions of mammalian SSCs and offers a scientific basis for the clinical application of Icariin in male reproduction.展开更多
This study was designed to explore the regulatory effects of male germ cell secreting factor NODAL on Sertoli cell fate decisions from obstructive azoospermia （OA） and nonobstructive azoospermia （NOA） patients. Hu...This study was designed to explore the regulatory effects of male germ cell secreting factor NODAL on Sertoli cell fate decisions from obstructive azoospermia （OA） and nonobstructive azoospermia （NOA） patients. Human Sertoli cells and male germ cells were isolated using two-step enzymatic digestion and SATPUT from testes of azoospermia patients. Expression of NODAL and its multiple receptors in human Sertoli cells and male germ cells were characterized by reverse transcription-polymerase chain reaction （RT-PCRI and immunochemistry. Human recombinant NODAL and its receptor inhibitor SB431542 were employed to probe their effect on the proliferation of Sertoli cells using the CCK-8 assay. Quantitative PCR and Western blots were utilized to assess the expression of Sertoli cell functional genes and proteins. NODAL was found to be expressed in male germ cells but not in Sertoli cells, whereas its receptors ALK4, ALK7, and ACTR-IIB were detected in Sertoli cells and germ cells, suggesting that NODAL plays a regulatory role in Sertoli cells and germ cells via a paracrine and autocrine pathway, respectively. Human recombinant NODAL could promote the proliferation of human Sertoli cells. The expression of cell cycle regulators, including CYCLIN A, CYCLIN D1 and CYCLIN E, was not remarkably affected by NODAL signaling. NODAL enhanced the expression of essential growth factors, including GDNF, SCF, and BMP4, whereas SB431542 decreased their levels. There was not homogeneity of genes changes by NODAL treatment in Sertoli cells from OA and Sertoli cell-only syndrome （SCO） patients. Collectively, this study demonstrates that NODAL produced by human male germ cells regulates proliferation and numerous gene expression of Sertoli cells.展开更多
Spermatogonial stem cells （SSCs）, also known as male germline stem cells, are a small subpopulation of type A spermatogonia with the potential of self-renewal to maintain stem cell pool and differentiation into sper...Spermatogonial stem cells （SSCs）, also known as male germline stem cells, are a small subpopulation of type A spermatogonia with the potential of self-renewal to maintain stem cell pool and differentiation into spermatids in mammalian testis. SSCs are previously regarded as the unipotent stem cells since they can only give rise to sperm within the seminiferous tubules. However, this concept has recently been challenged because numerous studies have demonstrated that SSCs cultured with growth factors can acquire pluripotency to become embryonic stem-like cells. The in vivo and in vitro studies from peers and us have clearly revealed that SSCs can directly transdifferentiate into morphologic, phenotypic, and functional cells of other lineages. Direct conversion to the cells of other tissues has important significance for regenerative medicine. SSCs from azoospermia patients could be induced to differentiate into spermatids with fertilization and developmental potentials. As such, SSCs could have significant applications in both reproductive and regenerative medicine due to their unique and great potentials. In this review, we address the important plasticity of SSCs, with focuses on their self-renewal, differentiation, dedifferentiation, transdifferentiation, and translational medicine studies.展开更多
Infertility has become a serious disease since it affects 10%–15%of couples worldwide,and male infertility contributes to about 50%of the cases.Notably,a significant decrease occurs in the newborn population by 7.82 ...Infertility has become a serious disease since it affects 10%–15%of couples worldwide,and male infertility contributes to about 50%of the cases.Notably,a significant decrease occurs in the newborn population by 7.82 million in 2020 compared to 2016 in China.As such,it is essential to explore the effective methods of obtaining functional male gametes for restoring male fertility.Stem cells,including embryonic stem cells(ESCs),induced pluripotent stem cells(iPSCs),spermatogonial stem cells(SSCs),and mesenchymal stem cells(MSCs),possess the abilities of both self-renewal and differentiation into germ cells.Significantly,much progress has recently been achieved in the generation of male germ cells in vitro from various kinds of stem cells under the specified conditions,e.g.,the coculturing with Sertoli cells,three-dimensional culture system,the addition of growth factors and cytokines,and/or the overexpression of germ cell-related genes.In this review,we address the current advance in the derivation of male germ cells in vitro from stem cells based on the studies of the peers and us,and we highlight the perspectives and potential application of stem cell-derived male gametes in reproductive medicine.展开更多
Dear Editor,An interesting article from Zhao et al.recently published in Asian Journal of Andrology,has shown the association of a novel loss-of function(LOF)variant in PARN-like ribonuclease domain-containing exonucl...Dear Editor,An interesting article from Zhao et al.recently published in Asian Journal of Andrology,has shown the association of a novel loss-of function(LOF)variant in PARN-like ribonuclease domain-containing exonuclease 1(PNLDCI)and male infertility,and we would like to contribute a commentary on this article.展开更多
Yes-associated protein 1(YAP1)is a downstream effector of the Hippo signaling pathway,and it is involved in tumorigenesis,tissue repair,growth,and development.In this review,the biological roles and the mechanisms of ...Yes-associated protein 1(YAP1)is a downstream effector of the Hippo signaling pathway,and it is involved in tumorigenesis,tissue repair,growth,and development.In this review,the biological roles and the mechanisms of YAP1 in mediating stem cell fate decisions are discussed,including cell proliferation,differentiation,and apoptosis.In general,YAP1 promotes the proliferation and differentiation of stem cells,including embryonic stem cells and adult stem cells.It inhibits apoptosis by binding to the transcription factors,e.g.,transcriptional enhanced associate domain(TEAD),Smad,runt-related transcription factor 1/2,p73,p63,and Erb84,to maintain tissue homeostasis.The translocalization of YAP1 in cellular nuclei and the phosphorylation in the cytoplasm work as important and unusual events for the activation of YAP1.Moreover,YAP1 serves as the crosstalk for the Hippo pathway and other signaling pathways,including the Wnt and Notch pathways.It is highlighted in this review that YAP1 is an essential regulator for stem cells that have significant applications in regenerative medicine and reproductive medicine.展开更多
Recent studies have reported that induced pluripotent stem （iPS） cells from mice and humans can differentiate into primordial germ cells. However, whether iPS cells are capable of producing male germ cells is not kn...Recent studies have reported that induced pluripotent stem （iPS） cells from mice and humans can differentiate into primordial germ cells. However, whether iPS cells are capable of producing male germ cells is not known. The objective of this study was to investigate the differentiation potential of mouse iPS cells into spermatogonial stem cells and late-stage male germ cells. We used an approach that combines in vitrodifferentiation and in vivotransplantation. Embryoid bodies （EBs） were obtained from iPS cells using leukaemia inhibitor factor （LIF）-free medium. Quantitative PCR revealed a decrease in Oct4 expression and an increase in StraSand Vasa mRNA in the EBs derived from iPS cells, iPS cell-derived EBs were induced by retinoic acid to differentiate into spermatogonial stem cells （SSCs）, as evidenced by their expression of VASA, as well as CDH1 and GFRal, which are markers of SSCs. Furthermore, these germ cells derived from iPS cells were transplanted into recipient testes of mice that had been pre-treated with busulfan. Notably, iPS cell-derived SSCs were able to differentiate into male germ cells ranging from spermatogonia to round spermatids, as shown by VASA and SCP3 expression. This study demonstrates that iPS cells have the potential to differentiate into late-stage male germ cells. The derivation of male germ cells from iPS cells has potential applications in the treatment of male infertility and provides a model for uncovering the molecular mechanisms underlying male germ cell development.展开更多
Spermatogonial stem cells(SSCs)have great applications in both reproductive and regenerative medicine.Primates including monkeys are very similar to humans with regard to physiology and pathology.Nevertheless,little i...Spermatogonial stem cells(SSCs)have great applications in both reproductive and regenerative medicine.Primates including monkeys are very similar to humans with regard to physiology and pathology.Nevertheless,little is known about the isolation,the characteristics,and the culture of primate SSCs.This study was designed to identify,isolate,and culture monkey SSCs.Immunocytochemistry was used to identify markers for monkey SSCs.Glial cell line-derived neurotrophic factor family receptor alpha-1(GFRAl)-enriched spermatogonia were isolated from monkeys,namely Macaca fascicularis(M.fascicularis),by two-step enzymatic digestion and magnetic-activated cell sorting,and they were cultured on precoated plates in the conditioned medium.Reverse transcription-polymerase chain reaction(RT-PCR),immunocytochemistry,and RNA sequencing were used to compare phenotype and transcriptomes in GFRAl-enriched spermatogonia between 0 day and 14 days of culture,and xenotransplantation was performed to evaluate the function of GFRAl-enriched spermatogonia.SSCs shared some phenotypes with rodent and human SSCs.GFRAl-enriched spermatogonia with high purity and viability were isolated from M.fascicularis testes.The freshly isolated cells expressed numerous markers for rodent SSCs,and they were cultured for 14 days.The expression of numerous SSC markers was maintained during the cultivation of GFRAl-enriched spermatogonia.RNA sequencing reflected a 97.3%similarity in global gene profiles between 0 day and 14 days of culture.The xenotransplantation assay indicated that the GFRAl-enriched spermatogonia formed colonies and proliferated in vivo in the recipient c-Kitw/w(W)mutant mice.Collectively,GFRAl-enriched spermatogonia are monkey SSCs phenotypically both in vitro and in vivo.This study suggests that monkey might provide an alternative to human SSCs for basic research and application in human diseases.展开更多
One of the most significant findings in recent stem cell research is the establishment of the induced pluripotent stem （iPS） cells, because they could have critical implications in both regenerative and repro- ducti...One of the most significant findings in recent stem cell research is the establishment of the induced pluripotent stem （iPS） cells, because they could have critical implications in both regenerative and repro- ductive medicine. Male gametes play a crucial role in transmitting genetic information to subsequent generations, and notably there are more and more patients with azoospermia, due to genetic and environmental factors. Recent advancements on generation of male gametes from human iPS cells would bring great promise to produce patient own male gametes for treating male infertility and provide an excellent platform for unveiling molecular mechanisms of male germ cell development.展开更多
Exosomes are extracellular vesicles secreted by most eukaryotic cells and participate in intercellular communication.The components of exosomes,including proteins,DNA,mRNA,microRNA,long noncoding RNA,circular RNA,etc....Exosomes are extracellular vesicles secreted by most eukaryotic cells and participate in intercellular communication.The components of exosomes,including proteins,DNA,mRNA,microRNA,long noncoding RNA,circular RNA,etc.,which play a crucial role in regulating tumor growth,metastasis,and angiogenesis in the process of cancer development,and can be used as a prognostic marker and/or grading basis for tumor patients.Hereby,we mainly summarized as followed:the role of exosome contents in cancer,focusing on proteins and noncoding RNA;the interaction between exosomes and tumor microenvironment;the mechanisms that epithelial-mesenchymal transition,invasion and migration of tumor affected by exosomes;and tumor suppression strategies based on exosomes.Finally,the application potential of exosomes in clinical tumor diagnosis and therapy is prospected,which providing theoretical supports for using exosomes to serve precise tumor treatment in the clinic.展开更多
Prostate cancer(PCa)is a common malignant tumor with high morbidity and mortality worldwide.The prostate cancer stem cell(PCSC)model provides novel insights into the pathogenesis of PCa and its therapeutic response.Ho...Prostate cancer(PCa)is a common malignant tumor with high morbidity and mortality worldwide.The prostate cancer stem cell(PCSC)model provides novel insights into the pathogenesis of PCa and its therapeutic response.However,the roles and molecular mechanisms of specific genes in mediating fate decisions of PCSCs and carcinogenesis of PCa remain to be elusive.In this study,we have explored the expression,function,and mechanism of AZGP1P2,a pseudogene of AZGP1,in regulating the stemness and apoptosis of PCSCs and treatment resistance of docetaxel in castration-resistant prostate cancer(CRPC).We revealed that AZGP1P2 was downregulated in CRPC cell lines and PCSCs,while it was positively associated with progression-free interval.Upregulation of the AZGP1P2 enhanced the sensitivity of docetaxel treatment in CRPCs via inhibiting their stemness.RNA pull-down associated with mass spectrometry analysis,co-immunoprecipitation assay,and RNA immunoprecipitation assay demonstrated that AZGP1P2 could bind to UBA1 and RBM15 as a“writer”of methyltransferase to form a compound.UBA1,an E1 ubiquitin-activating enzyme,contributed to RBM15 protein degradation via ubiquitination modification.Methylated RNA immunoprecipitation assay displayed that RBM15 controlled the mRNA decay of TPM1 in m6A methylation.Furthermore,a xenograft mouse model and patient-derived organoids showed that the therapeutic effect of docetaxel in CRPC was increased by AZGP1P2 in vivo.Collectively,these results imply that AZGP1P2 mediates the stemness and apoptosis of PCSCs and promotes docetaxel therapeutic effect by suppressing tumor growth and metastasis via UBA1/RBM15-mediated TPM1 mRNA decay in CRPC.展开更多
Spermatogenesis is a sophisticated biological process by which spermatogonial stem cells(SSCs)undergo self-renewal and differentiation into spermatozoa.Molecular mechanisms underlying fate determinations of human SSCs...Spermatogenesis is a sophisticated biological process by which spermatogonial stem cells(SSCs)undergo self-renewal and differentiation into spermatozoa.Molecular mechanisms underlying fate determinations of human SSCs by key genes and signaling pathways remain elusive.Here,we report for the first time that Yes1-associated transcriptional regulator(YAP1)is required for fate determinations of SSCs and male fertility by interacting with RAD21 and targeting NEDD4 in humans and mice.YAP1 was mainly located at cell nuclei of human SSCs.YAP1 silencing resulted in the decreases in proliferation and DNA synthesis as well as an enhancement in apoptosis of human SSCs both in vivo and in vitro.RNA sequencing and real-time polymerase chain reaction assays identified NEDD4 as a target of YAP1,and NEDD4 knockdown inhibited the proliferation of human SSCs and increased their apoptosis.Furthermore,YAP1 interacted with RAD21 to regulate NEDD4 transcription in human SSCs.Importantly,YAP1 abnormalities were found to be associated with non-obstructive azoospermia(NOA)as manifested as lower expression level of YAP1 in testicular tissues of NOA patients and YAP1 single-nucleotide variants(SNVs)in 777 NOA patients.Finally,Yap1 germline conditional knockout(cKO)mice assumed mitotic arrest,low sperm count,and motility.Collectively,these results highlight a critical role of YAP1 in determining the fate determinations of human SSCs and male infertility through the YAP1/RAD21/NEDD4 pathway.This study provides new insights into the genetic regulatory mechanisms underlying human spermatogenesis and the pathogenesis of NOA,and it offers new targets for gene therapy of male infertility.展开更多
We are now well entering the exciting era of stem cells.Potential stem cell therapy holds great promise for the treatment of many diseases such as stroke,traumatic brain injury,Alzheimer’s disease,Parkinson’s diseas...We are now well entering the exciting era of stem cells.Potential stem cell therapy holds great promise for the treatment of many diseases such as stroke,traumatic brain injury,Alzheimer’s disease,Parkinson’s disease,amyotrophic lateral-sclerosis,myocardial infarction,muscular dystrophy,diabetes,and etc..It is generally believed that transplantation of specific stem cells into the injured tissue to replace the lost cells is an effective way to repair the tissue.In fact,organ transplantation has been successfully practiced in clinics for liver or kidney failure.However,the severe shortage of donor organs has been a major obstacle for the expansion of organ transplantation programs.Toward that direction,generation of transplantable organs using stem cells is a desirable approach for organ replacement and would be of great interest for both basic and clinical scientists.Here we review recent progress in the field of organ generation using various methods including single adult tissue stem cells,a blastocyst complementation system,tissue decellularization/recellularization and a combination of stem cells and tissue engineering.展开更多
Spermatogonial stem cells(SSCs)have important applications in both reproduction and regenerative medicine.Nevertheless,specific genes and signaling transduction pathways in mediating fate decisions of human SSCs remai...Spermatogonial stem cells(SSCs)have important applications in both reproduction and regenerative medicine.Nevertheless,specific genes and signaling transduction pathways in mediating fate decisions of human SSCs remain elusive.Here,we have demonstrated for the first time that OIP5(Opa interacting protein 5)controlled the self-renewal and apoptosis of human SSCs.RNA sequencing identified that NCK2 was a target for OIP5 in human SSCs,and interestingly,OIP5 could interact with NCK2 as shown by Co-IP(co-immunoprecipitation),IP-MS(mass spectrometry),and GST pulldown assays.NCK2 silencing decreased human SSC proliferation and DNA synthesis but enhanced their apoptosis.Notably,NCK2 knockdown reversed the influence of OIP5 overexpression on human SSCs.Moreover,OIP5 inhibition decreased the numbers of human SSCs at S and G2/M phases,while the levels of numerous cell cycle proteins,including cyclins A2,B1,D1,E1 and H,especially cyclin D1,were remarkably reduced.Significantly,whole-exome sequencing of 777 patients with nonobstructive azoospermia(NOA)revealed 54 singlenucleotide polymorphism mutations of the OIP5 gene(6.95%),while the level of OIP5 protein was obviously lower in testes of NOA patients compared to fertile men.Collectively,these results implicate that OIP5 interacts with NCK2 to modulate human SSC self-renewal and apoptosis via cell cyclins and cell cycle progression and that its mutation and/or lower expression is correlated with azoospermia.As such,this study offers novel insights into molecular mechanisms underlying the fate determinations of human SSCs and the pathogenesis of NOA,and it provides new targets for treating male infertility.展开更多
Spermatogonial stem cells(SSCs)are essential for initiating and maintaining normal spermatogenesis,and notably,they have important applications in both reproduction and regenerative medicine.Nevertheless,the molecular...Spermatogonial stem cells(SSCs)are essential for initiating and maintaining normal spermatogenesis,and notably,they have important applications in both reproduction and regenerative medicine.Nevertheless,the molecular mechanisms controlling the fate determinations of human SSCs remain elusive.In this study,we identified a selective expression of APBB1 in dormant human SSCs.We demonstrated for the first time that APBB1 interacted with KAT5,which led to the suppression of GDF15 expression and consequent inhibition of human SSC proliferation.Intriguingly,Apbb1^(-/-)mice assumed the disrupted spermatogenesis and markedly reduced fertility.SSC transplantation assays revealed that Apbb1 silencing enhanced SSC colonization and impeded their differentiation,which resulted in the impaired spermatogenesis.Notably,4 deleterious APBB1 mutation sites were identified in 2,047 patients with non-obstructive azoospermia(NOA),and patients with the c.1940C>G mutation had a similar testicular phenotype with Apbb1^(-/-)mice.Additionally,we observed lower expression levels of APBB1 in NOA patients with spermatogenic arrest than in obstructive azoospermia patients with normal spermatogenesis.Collectively,our findings highlight an essential role of APBB1/KAT5/GDF15 in governing human SSC fate decisions and maintaining normal spermatogenesis and underscore them as therapeutic targets for treating male infertility.展开更多
基金supported by the grants from the National Nature Science Foundation of China(No.32170862)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province(No.2019SK1012)+1 种基金the Research Team for Reproduction Health and Translational Medicine of Hunan Normal University(No.2023JC101)Graduate Scientific Research Innovation Project of Hunan Province,China(No.CX2022520).
文摘Infertility has become one of the most serious diseases worldwide,and 50% of this disease can be attributed to male-related factors.Spermatogenesis,by definition,is a complex process by which spermatogonial stem cells(SSCs)self-renew to maintain stem cell population within the testes and differentiate into mature spermatids.It is of great significance to uncover gene regulation and signaling pathways that are involved in the fate determinations of SSCs with aims to better understand molecular mechanisms underlying human spermatogenesis and identify novel targets for gene therapy of male infertility.Significant achievement has recently been made in demonstrating the signaling molecules and pathways mediating the fate decisions of mammalian SSCs.In this review,we address key gene regulation and crucial signaling transduction pathways in controlling the self-renewal,differentiation,and apoptosis of SSCs,and we illustrate the networks of genes and signaling pathways in SSC fate determinations.We also highlight perspectives and future directions in SSC regulation by genes and their signaling pathways.This review could provide novel insights into the genetic regulation of normal and abnormal spermatogenesis and offer molecular targets to develop new approaches for gene therapy of male infertility.
基金supported by the grants from the National Nature Science Foundation of China(No.32170862)Developmental Biology and Breeding(No.2022XKQ0205)+2 种基金the Research Team for Reproduction Health and Translational Medicine of Hunan Normal University(No.2023JC101)Graduate Scientific Research Innovation Project of Hunan Province(No.CX2022520)Shanghai Key Laboratory of Reproductive Medicine(2022SKLRM01).
文摘Icariin is a pure compound derived from Epimedium brevicornu Maxim,and it helps the regulation of male reproduction.Nevertheless,the role and underlying mechanisms of Icariin in mediating male germ cell development remain to be clarified.Here,we have demonstrated that Icariin promoted proliferation and DNA synthesis of mouse spermatogonial stem cells(SSCs).Furthermore,surface plasmon resonance iron(SPRi)and molecular docking(MOE)assays revealed that phosphodiesterase 5A(PDE5A)was an important target of Icariin in mouse SSCs.Mechanically,Icariin decreased the expression level of PDE5A.Interestingly,hydrogen peroxides(H2O2)enhanced the expression level of phosphorylation H2A.X(p-H2A.X),whereas Icariin diminished the expression level of p-H2A.X and DNA damage caused by H2O2 in mouse SSCs.Finally,our in vivo animal study indicated that Icariin protected male reproduction.Collectively,these results implicate that Icariin targets PDE5A to regulate mouse SSC viability and DNA damage and improves male reproductive capacity.This study thus sheds new insights into molecular mechanisms underlying the fate decisions of mammalian SSCs and offers a scientific basis for the clinical application of Icariin in male reproduction.
文摘This study was designed to explore the regulatory effects of male germ cell secreting factor NODAL on Sertoli cell fate decisions from obstructive azoospermia （OA） and nonobstructive azoospermia （NOA） patients. Human Sertoli cells and male germ cells were isolated using two-step enzymatic digestion and SATPUT from testes of azoospermia patients. Expression of NODAL and its multiple receptors in human Sertoli cells and male germ cells were characterized by reverse transcription-polymerase chain reaction （RT-PCRI and immunochemistry. Human recombinant NODAL and its receptor inhibitor SB431542 were employed to probe their effect on the proliferation of Sertoli cells using the CCK-8 assay. Quantitative PCR and Western blots were utilized to assess the expression of Sertoli cell functional genes and proteins. NODAL was found to be expressed in male germ cells but not in Sertoli cells, whereas its receptors ALK4, ALK7, and ACTR-IIB were detected in Sertoli cells and germ cells, suggesting that NODAL plays a regulatory role in Sertoli cells and germ cells via a paracrine and autocrine pathway, respectively. Human recombinant NODAL could promote the proliferation of human Sertoli cells. The expression of cell cycle regulators, including CYCLIN A, CYCLIN D1 and CYCLIN E, was not remarkably affected by NODAL signaling. NODAL enhanced the expression of essential growth factors, including GDNF, SCF, and BMP4, whereas SB431542 decreased their levels. There was not homogeneity of genes changes by NODAL treatment in Sertoli cells from OA and Sertoli cell-only syndrome （SCO） patients. Collectively, this study demonstrates that NODAL produced by human male germ cells regulates proliferation and numerous gene expression of Sertoli cells.
文摘Spermatogonial stem cells （SSCs）, also known as male germline stem cells, are a small subpopulation of type A spermatogonia with the potential of self-renewal to maintain stem cell pool and differentiation into spermatids in mammalian testis. SSCs are previously regarded as the unipotent stem cells since they can only give rise to sperm within the seminiferous tubules. However, this concept has recently been challenged because numerous studies have demonstrated that SSCs cultured with growth factors can acquire pluripotency to become embryonic stem-like cells. The in vivo and in vitro studies from peers and us have clearly revealed that SSCs can directly transdifferentiate into morphologic, phenotypic, and functional cells of other lineages. Direct conversion to the cells of other tissues has important significance for regenerative medicine. SSCs from azoospermia patients could be induced to differentiate into spermatids with fertilization and developmental potentials. As such, SSCs could have significant applications in both reproductive and regenerative medicine due to their unique and great potentials. In this review, we address the important plasticity of SSCs, with focuses on their self-renewal, differentiation, dedifferentiation, transdifferentiation, and translational medicine studies.
基金supported by the grants from the National Nature Science Foundation of China (32170862 and 31872845)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province (2019SK1012)+2 种基金Key Grant of Research and Development in Hunan Province (2020DK2002)High-Level Talent Gathering Project in Hunan Province (2018RS3066)Natural Science Foundation of Hunan Province of China (2020J5380 and 2020JJ5383).
文摘Infertility has become a serious disease since it affects 10%–15%of couples worldwide,and male infertility contributes to about 50%of the cases.Notably,a significant decrease occurs in the newborn population by 7.82 million in 2020 compared to 2016 in China.As such,it is essential to explore the effective methods of obtaining functional male gametes for restoring male fertility.Stem cells,including embryonic stem cells(ESCs),induced pluripotent stem cells(iPSCs),spermatogonial stem cells(SSCs),and mesenchymal stem cells(MSCs),possess the abilities of both self-renewal and differentiation into germ cells.Significantly,much progress has recently been achieved in the generation of male germ cells in vitro from various kinds of stem cells under the specified conditions,e.g.,the coculturing with Sertoli cells,three-dimensional culture system,the addition of growth factors and cytokines,and/or the overexpression of germ cell-related genes.In this review,we address the current advance in the derivation of male germ cells in vitro from stem cells based on the studies of the peers and us,and we highlight the perspectives and potential application of stem cell-derived male gametes in reproductive medicine.
基金This work was funded by the grant from the National Natural Science Foundation of China(No.32170862).
文摘Dear Editor,An interesting article from Zhao et al.recently published in Asian Journal of Andrology,has shown the association of a novel loss-of function(LOF)variant in PARN-like ribonuclease domain-containing exonuclease 1(PNLDCI)and male infertility,and we would like to contribute a commentary on this article.
基金This work was supported by grants from the National Nature Science Foundation of China(32170862,31872845)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province(2019SK1012)+4 种基金Key Grant of Research and Development in Hunan Province(2020DK2002)High-Level Talent Gathering Project in Hunan Province(2018RS3066)Natural Science Foundation of Hunan Province(2020JJ5383,2021JJ40365)Health Commission Foundation of Hunan Province(202104052273,202102050927)Hunan Province College Student Research Learning and Innovative Experiment Project(S202010542084).
文摘Yes-associated protein 1(YAP1)is a downstream effector of the Hippo signaling pathway,and it is involved in tumorigenesis,tissue repair,growth,and development.In this review,the biological roles and the mechanisms of YAP1 in mediating stem cell fate decisions are discussed,including cell proliferation,differentiation,and apoptosis.In general,YAP1 promotes the proliferation and differentiation of stem cells,including embryonic stem cells and adult stem cells.It inhibits apoptosis by binding to the transcription factors,e.g.,transcriptional enhanced associate domain(TEAD),Smad,runt-related transcription factor 1/2,p73,p63,and Erb84,to maintain tissue homeostasis.The translocalization of YAP1 in cellular nuclei and the phosphorylation in the cytoplasm work as important and unusual events for the activation of YAP1.Moreover,YAP1 serves as the crosstalk for the Hippo pathway and other signaling pathways,including the Wnt and Notch pathways.It is highlighted in this review that YAP1 is an essential regulator for stem cells that have significant applications in regenerative medicine and reproductive medicine.
文摘Recent studies have reported that induced pluripotent stem （iPS） cells from mice and humans can differentiate into primordial germ cells. However, whether iPS cells are capable of producing male germ cells is not known. The objective of this study was to investigate the differentiation potential of mouse iPS cells into spermatogonial stem cells and late-stage male germ cells. We used an approach that combines in vitrodifferentiation and in vivotransplantation. Embryoid bodies （EBs） were obtained from iPS cells using leukaemia inhibitor factor （LIF）-free medium. Quantitative PCR revealed a decrease in Oct4 expression and an increase in StraSand Vasa mRNA in the EBs derived from iPS cells, iPS cell-derived EBs were induced by retinoic acid to differentiate into spermatogonial stem cells （SSCs）, as evidenced by their expression of VASA, as well as CDH1 and GFRal, which are markers of SSCs. Furthermore, these germ cells derived from iPS cells were transplanted into recipient testes of mice that had been pre-treated with busulfan. Notably, iPS cell-derived SSCs were able to differentiate into male germ cells ranging from spermatogonia to round spermatids, as shown by VASA and SCP3 expression. This study demonstrates that iPS cells have the potential to differentiate into late-stage male germ cells. The derivation of male germ cells from iPS cells has potential applications in the treatment of male infertility and provides a model for uncovering the molecular mechanisms underlying male germ cell development.
基金the National Natural Science Foundation of China(31671550,31872845)National Key R&D Project(2016YFC1000606)+3 种基金High Level Talent Gathering Project in Hunan Province(2018RS3066)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province(2019SK1012)Key Grant of Research and Development in Hunan Province(2020DK2002)The Open Fund of the NHC Key Laboratory of Male Reproduction and Genetics(KF201802).
文摘Spermatogonial stem cells(SSCs)have great applications in both reproductive and regenerative medicine.Primates including monkeys are very similar to humans with regard to physiology and pathology.Nevertheless,little is known about the isolation,the characteristics,and the culture of primate SSCs.This study was designed to identify,isolate,and culture monkey SSCs.Immunocytochemistry was used to identify markers for monkey SSCs.Glial cell line-derived neurotrophic factor family receptor alpha-1(GFRAl)-enriched spermatogonia were isolated from monkeys,namely Macaca fascicularis(M.fascicularis),by two-step enzymatic digestion and magnetic-activated cell sorting,and they were cultured on precoated plates in the conditioned medium.Reverse transcription-polymerase chain reaction(RT-PCR),immunocytochemistry,and RNA sequencing were used to compare phenotype and transcriptomes in GFRAl-enriched spermatogonia between 0 day and 14 days of culture,and xenotransplantation was performed to evaluate the function of GFRAl-enriched spermatogonia.SSCs shared some phenotypes with rodent and human SSCs.GFRAl-enriched spermatogonia with high purity and viability were isolated from M.fascicularis testes.The freshly isolated cells expressed numerous markers for rodent SSCs,and they were cultured for 14 days.The expression of numerous SSC markers was maintained during the cultivation of GFRAl-enriched spermatogonia.RNA sequencing reflected a 97.3%similarity in global gene profiles between 0 day and 14 days of culture.The xenotransplantation assay indicated that the GFRAl-enriched spermatogonia formed colonies and proliferated in vivo in the recipient c-Kitw/w(W)mutant mice.Collectively,GFRAl-enriched spermatogonia are monkey SSCs phenotypically both in vitro and in vivo.This study suggests that monkey might provide an alternative to human SSCs for basic research and application in human diseases.
文摘One of the most significant findings in recent stem cell research is the establishment of the induced pluripotent stem （iPS） cells, because they could have critical implications in both regenerative and repro- ductive medicine. Male gametes play a crucial role in transmitting genetic information to subsequent generations, and notably there are more and more patients with azoospermia, due to genetic and environmental factors. Recent advancements on generation of male gametes from human iPS cells would bring great promise to produce patient own male gametes for treating male infertility and provide an excellent platform for unveiling molecular mechanisms of male germ cell development.
基金supported by the National Natural Science Foundation of China(81802785[Y.J.])Hunan Provincial Natural Science Foundation of China(2020JJ5382[Y.J.],2020JJ5381[L.C.]).
文摘Exosomes are extracellular vesicles secreted by most eukaryotic cells and participate in intercellular communication.The components of exosomes,including proteins,DNA,mRNA,microRNA,long noncoding RNA,circular RNA,etc.,which play a crucial role in regulating tumor growth,metastasis,and angiogenesis in the process of cancer development,and can be used as a prognostic marker and/or grading basis for tumor patients.Hereby,we mainly summarized as followed:the role of exosome contents in cancer,focusing on proteins and noncoding RNA;the interaction between exosomes and tumor microenvironment;the mechanisms that epithelial-mesenchymal transition,invasion and migration of tumor affected by exosomes;and tumor suppression strategies based on exosomes.Finally,the application potential of exosomes in clinical tumor diagnosis and therapy is prospected,which providing theoretical supports for using exosomes to serve precise tumor treatment in the clinic.
基金grants from the National Natural Science Foundation of China(81802554 and 32170862)the Experimental Animal Fund of Shanghai Science and Technology Commission(22140903800)a Project Funded by China Postdoctoral Science Foundation(2021M692432).
文摘Prostate cancer(PCa)is a common malignant tumor with high morbidity and mortality worldwide.The prostate cancer stem cell(PCSC)model provides novel insights into the pathogenesis of PCa and its therapeutic response.However,the roles and molecular mechanisms of specific genes in mediating fate decisions of PCSCs and carcinogenesis of PCa remain to be elusive.In this study,we have explored the expression,function,and mechanism of AZGP1P2,a pseudogene of AZGP1,in regulating the stemness and apoptosis of PCSCs and treatment resistance of docetaxel in castration-resistant prostate cancer(CRPC).We revealed that AZGP1P2 was downregulated in CRPC cell lines and PCSCs,while it was positively associated with progression-free interval.Upregulation of the AZGP1P2 enhanced the sensitivity of docetaxel treatment in CRPCs via inhibiting their stemness.RNA pull-down associated with mass spectrometry analysis,co-immunoprecipitation assay,and RNA immunoprecipitation assay demonstrated that AZGP1P2 could bind to UBA1 and RBM15 as a“writer”of methyltransferase to form a compound.UBA1,an E1 ubiquitin-activating enzyme,contributed to RBM15 protein degradation via ubiquitination modification.Methylated RNA immunoprecipitation assay displayed that RBM15 controlled the mRNA decay of TPM1 in m6A methylation.Furthermore,a xenograft mouse model and patient-derived organoids showed that the therapeutic effect of docetaxel in CRPC was increased by AZGP1P2 in vivo.Collectively,these results imply that AZGP1P2 mediates the stemness and apoptosis of PCSCs and promotes docetaxel therapeutic effect by suppressing tumor growth and metastasis via UBA1/RBM15-mediated TPM1 mRNA decay in CRPC.
基金funded by the grants from National Nature Science Foundation of China(32470904 and 32170862)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province(2019SK1012)+3 种基金Key Grant of Research and Development in Hunan Province(2020DK2002)Developmental Biology and Breeding(2022XKQ0205)Shanghai Key Laboratory of Reproductive Medicine,Natural Science Foundation of Hunan Province of China(2024JJ5284,2023JJ30424,and 2024JJ5282)Research Foundation of Education Bureau of Hunan Province for Outstanding Young(23B0064).
文摘Spermatogenesis is a sophisticated biological process by which spermatogonial stem cells(SSCs)undergo self-renewal and differentiation into spermatozoa.Molecular mechanisms underlying fate determinations of human SSCs by key genes and signaling pathways remain elusive.Here,we report for the first time that Yes1-associated transcriptional regulator(YAP1)is required for fate determinations of SSCs and male fertility by interacting with RAD21 and targeting NEDD4 in humans and mice.YAP1 was mainly located at cell nuclei of human SSCs.YAP1 silencing resulted in the decreases in proliferation and DNA synthesis as well as an enhancement in apoptosis of human SSCs both in vivo and in vitro.RNA sequencing and real-time polymerase chain reaction assays identified NEDD4 as a target of YAP1,and NEDD4 knockdown inhibited the proliferation of human SSCs and increased their apoptosis.Furthermore,YAP1 interacted with RAD21 to regulate NEDD4 transcription in human SSCs.Importantly,YAP1 abnormalities were found to be associated with non-obstructive azoospermia(NOA)as manifested as lower expression level of YAP1 in testicular tissues of NOA patients and YAP1 single-nucleotide variants(SNVs)in 777 NOA patients.Finally,Yap1 germline conditional knockout(cKO)mice assumed mitotic arrest,low sperm count,and motility.Collectively,these results highlight a critical role of YAP1 in determining the fate determinations of human SSCs and male infertility through the YAP1/RAD21/NEDD4 pathway.This study provides new insights into the genetic regulatory mechanisms underlying human spermatogenesis and the pathogenesis of NOA,and it offers new targets for gene therapy of male infertility.
基金The study is supported by funds from the Chinese Ministry of Science and Technology(2012CB966800 and 2013CB945600 to WQG and RY and 2012CB967900)the National Natural Science Foundation of China(81130038 to WQG,31171422 and 31230048 to ZH)+2 种基金Science and Technology Commission of Shanghai Municipality(Pujiang program to WQG,11PJ1406400 to ZH)Shanghai Education Committee Key Disciplines and Specialties Foundation(J50208 to WQG)Shanghai Health Bureau Key Disciplines and Specialties Foundation(to WQG),KC Wong foundation(to WQG)and the China Postdoctoral Science Foundation(2012M510835 to YYL).
文摘We are now well entering the exciting era of stem cells.Potential stem cell therapy holds great promise for the treatment of many diseases such as stroke,traumatic brain injury,Alzheimer’s disease,Parkinson’s disease,amyotrophic lateral-sclerosis,myocardial infarction,muscular dystrophy,diabetes,and etc..It is generally believed that transplantation of specific stem cells into the injured tissue to replace the lost cells is an effective way to repair the tissue.In fact,organ transplantation has been successfully practiced in clinics for liver or kidney failure.However,the severe shortage of donor organs has been a major obstacle for the expansion of organ transplantation programs.Toward that direction,generation of transplantable organs using stem cells is a desirable approach for organ replacement and would be of great interest for both basic and clinical scientists.Here we review recent progress in the field of organ generation using various methods including single adult tissue stem cells,a blastocyst complementation system,tissue decellularization/recellularization and a combination of stem cells and tissue engineering.
基金the grants from the National Nature Science Foundation of China(32170862 and 31872845)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province(2019SK1012)+3 种基金Key Grant of Research and Development in Hunan Province(2020DK2002)Developmental Biology and Breeding(2022XKQ0205)Natural Science Foundation of Hunan Province of China(2020JJ5380,2020JJ5383,and 2021JJ40365)a grant from the Shanghai Key Laboratory of Reproductive Medicine.
文摘Spermatogonial stem cells(SSCs)have important applications in both reproduction and regenerative medicine.Nevertheless,specific genes and signaling transduction pathways in mediating fate decisions of human SSCs remain elusive.Here,we have demonstrated for the first time that OIP5(Opa interacting protein 5)controlled the self-renewal and apoptosis of human SSCs.RNA sequencing identified that NCK2 was a target for OIP5 in human SSCs,and interestingly,OIP5 could interact with NCK2 as shown by Co-IP(co-immunoprecipitation),IP-MS(mass spectrometry),and GST pulldown assays.NCK2 silencing decreased human SSC proliferation and DNA synthesis but enhanced their apoptosis.Notably,NCK2 knockdown reversed the influence of OIP5 overexpression on human SSCs.Moreover,OIP5 inhibition decreased the numbers of human SSCs at S and G2/M phases,while the levels of numerous cell cycle proteins,including cyclins A2,B1,D1,E1 and H,especially cyclin D1,were remarkably reduced.Significantly,whole-exome sequencing of 777 patients with nonobstructive azoospermia(NOA)revealed 54 singlenucleotide polymorphism mutations of the OIP5 gene(6.95%),while the level of OIP5 protein was obviously lower in testes of NOA patients compared to fertile men.Collectively,these results implicate that OIP5 interacts with NCK2 to modulate human SSC self-renewal and apoptosis via cell cyclins and cell cycle progression and that its mutation and/or lower expression is correlated with azoospermia.As such,this study offers novel insights into molecular mechanisms underlying the fate determinations of human SSCs and the pathogenesis of NOA,and it provides new targets for treating male infertility.
基金supported by grants from the National Natural Science Foundation of China(nos.82201771,32270912,and 32170862)Natural Science Foundation of Hunan Province(nos.2024JJ6083 and 2023JJ31018)+4 种基金Health Research Project of Hunan Provincial Health Commission(nos.W20243143 and 20231769)Natural Science Foundation of Changsha(nos.kq2202491 and kq2502312)Science and Technology Innovation Project of Hunan Province(no.2021SK53204)Clinical Medical Technology Demonstration Base for Genetic Research of Fetal Congenital Heart Disease in Hunan Province(no.2021SK4036)Hunan Province Children’s Safe Medication Clinical Medical Technology Demonstration Base(no.2023SK4083).
文摘Spermatogonial stem cells(SSCs)are essential for initiating and maintaining normal spermatogenesis,and notably,they have important applications in both reproduction and regenerative medicine.Nevertheless,the molecular mechanisms controlling the fate determinations of human SSCs remain elusive.In this study,we identified a selective expression of APBB1 in dormant human SSCs.We demonstrated for the first time that APBB1 interacted with KAT5,which led to the suppression of GDF15 expression and consequent inhibition of human SSC proliferation.Intriguingly,Apbb1^(-/-)mice assumed the disrupted spermatogenesis and markedly reduced fertility.SSC transplantation assays revealed that Apbb1 silencing enhanced SSC colonization and impeded their differentiation,which resulted in the impaired spermatogenesis.Notably,4 deleterious APBB1 mutation sites were identified in 2,047 patients with non-obstructive azoospermia(NOA),and patients with the c.1940C>G mutation had a similar testicular phenotype with Apbb1^(-/-)mice.Additionally,we observed lower expression levels of APBB1 in NOA patients with spermatogenic arrest than in obstructive azoospermia patients with normal spermatogenesis.Collectively,our findings highlight an essential role of APBB1/KAT5/GDF15 in governing human SSC fate decisions and maintaining normal spermatogenesis and underscore them as therapeutic targets for treating male infertility.
