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Dissecting the cell microenvironment of ovarian endometrioma through single-cell RNA sequencing

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Science China Life Sciences 2025年第1期68卷 116-129页

作者： Jiangpeng Wu Siyu Xia Wenting Ye Yan Sun Jing Cai Fubing Yu Haiping Wen Xiuwei Yi Taikang Li Mingwei Chen Jiayun Chen Ge Song Chuanbin Yang Yali Song Jigang Wang Department of reproductive medicine Dongguan Maternal and Child Health Care Hospital Department of Urology and Shenzhen Clinical Research Centre for GeriatricsShenzhen People's HospitalSouthern University of Science and Technology State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs Artemisinin Research Centerand Institute of Chinese Materia MedicaChina Academy of Chinese Medical Sciences reproductive medicine Center Foshan Women and Children Hospital State Key Laboratory of Antiviral Drugs School of PharmacyHenan University

Ovarian endometrioma（OE）, also known as “chocolate cysts,” is a cystic mass that develops in the ovaries due to endometriosis and is a common gynecological condition characterized by the growth of endometrial tissue outside the uterus, leading to symptoms such as dysmenorrhea, pelvic pain, and infertility. However, the precise molecular and cellular mechanisms driving this pathophysiology remain largely unknown, posing challenges for diagnosis and treatment. Here, we employed integrated single-cell transcriptomic profiling of over 52,000 individual cells from endometrial tissues of OE patients and healthy donors and identified twelve major cell populations. We identified notable alterations in cell type-specific proportions and molecular signatures associated with OE. Notably, the activation of IGFBP5+macrophages with pro-inflammatory properties, NK cell exhaustion, and aberrant proliferation of IQCG+and KLF2+epithelium are key features and may be the potential mechanisms underlying the pathogenesis of OE. Collectively, our data contribute to a better understanding of OE at the single cell level and may pave the way for the development of novel therapeutic strategies.

关键词： ovary high-throughput sequencing transcriptomics gene expression analysis ligand-receptor analysis

Bringing epigenetics into the diagnostics of the andrology laboratory： challenges and perspectives

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Asian Journal of Andrology 2014年第5期16卷 669-674,I0006页

作者： Ruth Klaver Jorg Gromoll centre of reproductive medicine and Andrology University Clinics of MOnster MOnster Germany

Recent studies have shown significant associations of aberrant DNA methylation in spermatozoa with idiopathic male infertility, increased frequency of spontaneous abortions and imprinting disorders. Thus, the analysis of DNA methylation of specific genes in spermatozoa has the potential to become a new valuable diagnostic marker in clinical andrology. This perspective article discusses the current state and value of DNA methylation analysis in the diagnostic setup of infertile men and outlines challenges and perspectives. It highlights the potential of DNA methylation in andrological diagnostics and its putative benefit in the examination of hitherto idiopathic infertile patients is described.

关键词： DNA methylation male infertility spermatogenesis

The epididymis, cytoplasmic droplets and male fertility

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Asian Journal of Andrology 2011年第1期13卷 130-138页

作者： Trevor G Cooper centre of reproductive medicine and Andrology University of MOnster MOnster D-48149 Germany

The potential of spermatozoa to become motile during post-testicular maturation, and the relationship between the cytoplasmic droplet and fertilizing capacity are reviewed. Post-testicular maturation of spermatozoa involves the autonomous induction of motility, which can occur in vivo in testes with occluded excurrent ducts and in vitro in testicular explants, and artefactual changes in morphology that appear to occur in the testis in vitro. Both modifications may reflect time-dependent oxidation of disulphide bonds of head and tail proteins. Regulatory volume decrease （RVD）, which counters sperm swelling at ejaculation, is discussed in relation to loss of cytoplasmic droplets and consequences for fertility. It is postulated that：（i） fertile males possess spermatozoa with sufficient osmolytes to drive RVD at ejaculation, permitting the droplet to round up and pinch off without membrane rupture; and （ii） infertile males possess spermatozoa with insufficient osmolytes so that RVD is inadequate, the droplet swells and the resulting flagellar angulation prevents droplet loss. Droplet retention at ejaculation is a harbinger of infertility caused by failure of the spermatozoon to negotiate the uterotubal junction or mucous and reach the egg. In this hypothesis, the epididymis regulates fertility indirectly by the extent of osmolyte provision to spermatozoa, which influences RVD and therefore droplet loss. Man is an exception, because ejaculated human spermatozoa retain their droplets. This may reflect their short midpiece, approximating head length, permitting a swollen droplet to extend along the entire midpiece; this not only obviates droplet migration and flagellar angulation but also hampers droplet loss.

关键词： epididymis fertility infertility sperm maturation

Maturation of sperm volume regulation in the rat epididymis

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Asian Journal of Andrology 2010年第4期12卷 578-590页

作者： Oliver S. Damm Trevor G. Cooper centre of reproductive medicine and Andrology of the University Miinster D-48149 Miinster Germany

Sperm maturation in the epididymis may involve differences between mature and immature spermatozoa in their volume regulatory osmolyte response. Spermatozoa obtained from the rat caput and cauda epididymidis were examined for their ability to regulate volume after transfer from in situ epididymal osmolality （measured to be 343 ± 13 and 365 ± 19 mmol kg^-1, respectively） to that of the female tract in single- and multiple-step protocols. Cells withstood the single-step treatment better than the multistep protocol. Sperm volume estimates by flow cytometric measure- ments of forward scatter of cells with intact head membranes was more sensitive than those by assessing cell coiling microscopically. At osmolalites below 210 mmol kg l both caput and cauda cells ruptured, limiting the use of flow cytometry. Above this critical value, the use of quinine showed that both caput and cauda cells could regulate volume, but cauda cells were the more effective. Of several organic osmolytes studied, myo-inositol, glutamate and KCl caused only temporary and slight swelling of spermatozoa cells in hypotonic medium. Spermatozoa of both maturities seemed to use potassium as the preferred osmolyte for regulating volume.

关键词： rat sperm sperm maturation volume regulation

Aquaporins in spermatozoa and testicular germ cells： identification and potential role

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Asian Journal of Andrology 2010年第4期12卷 490-499页

作者： Ching-Hei Yeung centre of reproductive medicine and Andrology University Clinic Muenster D48149 Germany

Mammalian spermatozoa have relatively high water permeability and swell readily, as in the hypo-osmotic swelling test used in the andrology clinic. Physiologically, spermatozoa experience changes in the osmolality of the surrounding fluids in both the male and the female tracts on their journey from the testis to the ovum. Sperm volume regulation in response to such osmotic challenges is important to maintain a stable cell size for the normal shape and function of the sperm tail. Alongside ion channels for the fluxes of osmolytes, water channels would be crucial for sperm volume regulation. In contrast to the deep knowledge and numerous studies on somatic cell aquaporins （AQPs）, the understanding of sperm AQPs is limited. Among the 13 AQPs, convincing evidence for their presence in spermatozoa has been confined to AQP7, AQP8 and AQP 11. Overall, current findings indicate a major role of AQP8 in water influx and efltux for sperm volume regulation, which is required for natural fertilization. The preliminary data suggestive of a role for AQP7 in sperm glycerol metabolism needs further substantiation. The association of AQP 11 with the residual cytoplasm of elongated spermatids and the distal tail of spermatozoa supports the hypothesis of more than just a role in conferring water permeability and also in the turnover and recycling of surplus cellular components made redundant during spermiogenesis and spermiation. This would be crucial for the maintenance of a germinal epithelium functioning efficiently in the production of spermatozoa.

关键词： fertility sperm volume regulation spermatid residual cytoplasm spermiogenesis water channels

Sperm chromatin structure assay （SCSA）： a tool in diagnosis and treatment of infertility

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Asian Journal of Andrology 2011年第1期13卷 69-75页

作者： Mona Bungum Leif Bungum Aleksander Giwercman reproductive medicine centre （RMC） Skane University Hospital SE - 205 02 Maimo Sweden

Diagnosis of male infertility has mainly been based on the World Health Organization （WHO） manual-based semen parameter＇s concentration, motility and morphology. It has, however, become apparent that none of these parameters are reliable markers for evaluation of the fertility potential of a couple. A search for better markers has led to an increased focus on sperm chromatin integrity testing in fertility work-up and assisted reproductive techniques. During the last couple of decades, numerous sperm DNA integrity tests have been developed. These are claimed to be characterized by a lower intraindividual variation, less intralaboratory and interlaboratory variation and thus less subjective than the conventional sperm analysis. However, not all the sperm chromatin integrity tests have yet been shown to be of clinical value. So far, the test that has been found to have the most stable clinical threshold values in relation to fertility is the sperm chromatin structure assay （SCSA）, a flow cytometric test that measures the susceptibility of sperm DNA to acid-induced DNA denaturation in situ. Sperm DNA fragmentation as measured by SCSA has shown to be an independent predictor of successful pregnancy in first pregnancy planners as well as in couples undergoing intrauterine insemination, and can be used as a tool in investigation, counseling and treatment of involuntary childlessness. More conflicting data exist regarding the role of sperm DNA fragmentation in relation to fertilization, pre-embryo development and pregnancy outcome in in vitro fertilization and intracytoplasmic sperm injection （ICSI）.

关键词： assisted reproductive techniques infertility sperm chromatin structure assay sperm DNA

Persistence and intergenerational transmission of differentially expressed genes in the testes of intracytoplasmic sperm injection conceived mice

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2013年第5期14卷 372-381页

作者： Li-ya WANG Ning WANG Fang LE Lei LI Le-jun LI Xiao-zhen LIU Ying-ming ZHENG Hang-ying LOU Xiang-rong XU Xiao-ming ZHU Yi-min ZHU He-feng HUANG Fan JIN Key Laboratory of reproductive Genetics (Zhejiang) Ministry of Educationand Centre of Reproductive Medicine Women's HospitalSchool of MedicineZhejiang University Department of Gynecologic Oncology Henan Cancer Hospital

Intracytoplasmic sperm injection (ICSI) is commonly used to solve male infertility problems. Previous studies showed that early environmental exposure of an embryo may influence postnatal development. To detect whether ICSI operations affect the reproductive health of a male or his offspring, we established assisted reproductive technologies (ART) conceived mouse models, and analyzed gene expression profiles in the testes of both ICSI and naturally conceived (NC) newborn F1 mice using micro-array analysis. Among the differentially expressed genes, we focused on the expression of eight male reproduction-related genes. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was used to analyze the expression of these genes in the testes of both adult and old F1 generation mice and adult F2 generation mice. Our results showed that down-regulated and somatic cell-expressed genes in newborn mice retained their differential expression patterns in adult and old F1 generation individuals, implying the persistence and fetal origin of the alteration in the expression of these genes. The intergenerational transmission of differential gene expression was observed, but most changes tended to be reduced in adult F2 generations. Controlled ovarian hyperstimulation (COH) and in vitro fertilization (IVF) mice models were added to explore the precise factors contributing to the differences in ICSI offspring. The data demonstrated that superovulation, in vitro culture, and mechanical stimulation involved in ICSI had a cumulative effect on the differential expression of these male reproductive genes.

关键词： Intracytoplasmic sperm injection Testis Intergenerational transmission

Improving precision in the assessment of round cell numbers in human semen

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Asian Journal of Andrology 2010年第1期12卷 111-114页

作者： Trevor G. Cooper Barbara Hellenkemper centre of reproductive medicine and Andrology University of Miinster Domagkstrasse 11 D-48149 Miinster Germany

The aim of this study was to increase the precision of assessment of the number of round cells observed in the per- oxidase test for detection of seminal leukocytes （granulocytes）. The dilution of semen was reduced and the volume of suspension examined was increased for semen samples containing between 0.6 and 6 million round cells per mL. A 1 ＋ 5 （1：6） dilution was compatible with measurable peroxidase activity and a sufficiently clear background for cell assessment. At this dilution, and with assessment of all 18 grids on both sides of the Neubauer-improved counting chamber, only three of the 10 samples （nominal cell concentrations of 1.9 ×10^-6-3.3×10^-6 mL-1） presented 400 round cells or more. As lower seminal dilutions were incompatible with easy detection of round cells or their peroxidase reaction product, it was not pos- sible to provide precise measurements （sampling error 5%） of the suggested lower reference limit of 1 ×10^-6 cells per mL. The results indicate that this poor precision of measuring 1×10^-6 round cells per mL could explain the discrepant reports on the acceptability of the cut-off values for leukocytospermia. Such reference limits need to be established with statistically sound methods.

关键词： cell number peroxidase precision round cells semen

Sperm banking for male reproductive preservation： a 6-year retrospective multi-centre study in China

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Asian Journal of Andrology 2010年第3期12卷 356-362,I0009页

作者： Ping Ping Wen-Bing Zhu Xin-Zong Zhang Kang-Shou Yao Peng Xu Yi-Ran Huang Zheng Li Department of Urology Shanghai Human Sperm Bank Shanghai Institute of Andrology Renji Hospital Shanghai Jiao Tong University School of Medicine Shanghai 200001 China Instinrte of Human Reproduction and Stem Cell Engineering Human Sperm Bank Central South University Changsha 410078 China Zhejiang Institute of Planned Parenthood Research ＆ Zhejiang Human Sperm Bank Hangzhou 310012 China jinghua Hospital Shenyang East Group Shenyang Reproductive Medicine Centre Shenyang 110005 China

Sperm banking can preserve male fertility effectively, but the current conditions of sperm cryopreservation in China have not been investigated. This retrospective investigation was based on data collected at multiple centres in China from January 2003 to December 2008. The collected data included urogenital history, indication for cryopreservation, semen parameters, use rate, type of assisted reproductive technique （ART） treatment and pregnancy outcome. The study population included 1 548 males who had banked their semen during the study period at one of the clinics indicated above. Approximately 1.9% （30/1 548） of the cryopreserved semen samples were collected from cancer patients; about 88.8% （1 374/1 548） of the patients had banked their semen for ART and 8.6% （134/1 548） had a male infertility disease （such as anejaculation, severe oligozoospermia and obstructive azoospermia）. The total use rate of cryopreserved semen was 22.7% （352/1 548）, with 119 live births. The cancer group use rate was 6.7% （2/30）, with one live birth by intracytoplasmic single sperm injection （ICSI）. The ART group use rate was 23.2% （319/1 374）, with 106 live births. The reproductive disease group use rate was 23.1% （31/134）, with 12 live births. The semen parameters in each category varied; the cancer patient and infertility disease groups had poor semen quality. In vitro fertilization （IVF） and ICSI were the most common ART treatments for cryopreserved sperm. Semen cryopreservation as a salvage method is effective, but in many conditions it is underutilized, especially in cancer patients. Lack of awareness, urgency of cancer treatment and financial constraints are the main causes of the low access rate. The concept of fertility preservation should be popularized to make better use of this medical service in China.

关键词： cancer cryopreservation infertility sperm