Decoding Microscopic Life Promotes Advances in Clinical Disciplines

1Jie Qiao

1Peking University

Humanity's exploration of the destiny of life is deepening with technological advancements and the continuous maturation of research tools. Among these, the decoding of microscopic life from genetic, epigenetic has played a transformative role in unraveling the fundamental mysteries of life and advancing clinical medicine development. This report focuses on analyzing and elaborating on three key questions within the field of human gamete and embryonic development research: Utilizing high-throughput polar body sequencing results to precisely reconstruct maternal genomic information lays the foundation for elucidating the laws of chromosomal recombination and genetic disease linkage analysis. Simultaneously, revealing the patterns of the two rounds of epigenetic reprogramming during gamete and embryonic development from the epigenetic level has led to the proposal of a novel theory regarding the differential contributions of parental epigenetic information, thereby further driving the advancement of non-invasive embryo diagnostic technologies based on epigenetic modifications. Focusing on the clinical issue of Klinefelter syndrome, which causes male infertility, research found that the lack of X-chromosome inactivation in the fetal germ cells of Klinefelter syndrome patients leads to germ cell developmental arrest, and that TGFβ inhibitors represent a potential new strategy for clinical intervention. Addressing the macro-level issue of environmental impacts on healthy reproduction, research on the etiology of Polycystic Ovary Syndrome (PCOS) using the gut microbiome as an entry point, along with the proposal of intervention strategies, provides a reference paradigm for the 'bench-to-bedside' research-translation pathway. The aforementioned research delineates an innovative practice loop extending from fundamental decoding to clinical application, vividly demonstrating that the power to change human destiny fundamentally depends on humanity's own scientific exploration.

Genetic Risk Assessment and Source Prevention of Chronic Diseases

1Hefeng Huang

1Institute of Genetic Medicine and Development，Zhejiang University

The U.S. CDC defines chronic diseases as conditions that last 1 year or more and require ongoing medical attention, or limit activities of daily living, or both. That makes a wide range of diseases, from heart diseases, cancer, chronic lung disease, stroke, Alzheimer’s disease, diabetes, to chronic kidney diseases. The rapidly increasing burden of chronic diseases is a key determinant of global public health. Chronic diseases now account for 71% of global mortality according to WHO data. The most effective measure for the prevention of chronic disease is to identify patients before they are obviously sick. Genetic information enables physicians to proactively conduct more precise and comprehensive personalized disease management. The occurrence of hereditary traits/diseases especially chronic disease is influenced not only by multiple microefficacy genes, but also by environmental factors. Polygenic risk scores (PRSs), Developed on the basis of GWAS, can integrate the effects of multiple susceptibility loci to build tumor risk prediction models, assessing an individual’s risk of developing cancer, cardiovascular disease and diabetes, enabling early prevention and intervention of chronic disease. The world’s first PGT-P baby was born in Aurea in 2020. In 2022, The first PRS+ model for diabetes based on family members was developed by us. The model, which enable diabetes prevention pre-pregnancy by PGT-P, was validated and optimized based on family members and 27,497 individuals with type 2 diabetes in the UKBB. We then constructed a PRS model for breast cancer using genetic and sample information from 18,483 female breast cancer patients and 246,222 control samples in UK Biobank. Based on the corrected site effect values of Asian population, the AUC was 0.63, which was better than that of European population. AI big data modeling enables genetic tumor risk assessment, marking an important progress in the prevention and control of cancer at the source in China. Embryos that do not carry breast cancer susceptibility gene mutations/breast cancer low risk are implanted in the uterus to prevent the birth of offspring susceptible to tumors, reducing the intergenerational transmission risk of hereditary tumor effectively. While the technical route of preventing chronic diseases from the early source of life has been started with preimplantation Genetic Testing for Polygenic Risk Scores (PGT-PRS), it still has a long way to go. The lack of the genetic database and chronic disease database for Chinese people is a major limiting factor for the widespread application. It is urgent to establish a nation-wide personal health record, to include genetic, environmental, and lifestyle information.

Genome Stability Maintenance and Ovarian Aging

1Yingying Qin, Zi-Jiang Chen*

1State Key Laboratory of Reproductive Medicine and Offspring Health, Center for Reproductive Medicine, Institute of Women, Children and Reproductive Health, Shandong University, Jinan, China.

The delay in childbearing age and the rising prevalence of infertility lead to a significant decline in fecundity (Seifer et al., 2024). Abnormal development of germ cells and reproductive aging are the leading causes of decreased fertility. Germ cells are responsible for the generational transmission of genetic material, and their faithful transmission is prerequisite for survival and propagation for species. Germ cells require a stringent DNA damage surveillance mechanism and high-fidelity repair capabilities to ensure genome stability. Genomic instability not only leads to oogenesis deficiency but also cause decreased oocyte quality, which is a key reason for both physiological and pathological ovarian aging (Stankovic et al., 2024, Ruth et al., 2021, Wu et al., 2025). Developing female germ cells face various endogenous and exogenous DNA damage threats, such as replication stress, programmed DNA double-strand breaks, and DNA damage accumulation. During these processes, female germ cells rely on multiple DNA damage repair pathways, including the Fanconi anemia and homologous repair pathways, to ensure the establishment and maintenance of the reproductive reserve (Ke et al., 2023, Yang et al., 2022, Huang et al., 2021). Furthermore, genomic instability increases in oocytes during physiological ovarian aging, manifesting as DNA damage accumulation, increased chromosomal aneuploidy, and elevation of de novo mutation rates, which are significant causes of early pregnancy loss and birth defects (Goldmann et al., 2016, Stankovic et al., 2024, Gruhn et al., 2019, Titus et al., 2013). Studies have shown that blocking the apoptosis signaling pathway or enhancing DNA damage repair capabilities can delay ovarian aging in mice (Kehrloesser et al., 2018, Lena et al., 2021, Strubel et al., 2022), but the long-term safety and clinical efficacy remain unclear. Further exploration is needed to understand the unique genomic threats and stability maintenance mechanisms during female germ cell development, the impact of ovarian aging on offspring health, and develop new strategies to enhance oocyte genomic stability and thereby improve female fertility.

References:

Goldmann, J. M., Wong, W. S., Pinelli, M., Farrah, T., Bodian, D., et al.(2016). Parent-of-origin-specific signatures of de novo mutations (p.935-9). Nat Genet.

Gruhn, J. R., Zielinska, A. P., Shukla, V., Blanshard, R., Capalbo, A., et al.(2019). Chromosome errors in human eggs shape natural fertility over reproductive life span (p.1466-1469). Science.

Huang, C., Guo, T. & Qin, Y.(2021). Meiotic Recombination Defects and Premature Ovarian Insufficiency (p.652407). Front Cell Dev Biol.

Ke, H., Tang, S., Guo, T., Hou, D., Jiao, X., et al.(2023). Landscape of pathogenic mutations in premature ovarian insufficiency (p.483–492). Nat Med.

Kehrloesser, S., Tuppi, M. & Dötsch, V.(2018). CHK2 sets the stage for CK1 in oocyte quality control (p.1007-1009). Cell Death Differ.

Lena, A. M., Rossi, V., Osterburg, S., Smirnov, A., Osterburg, C., et al.(2021). The p63 C-terminus is essential for murine oocyte integrity (p.383). Nat Commun.

Ruth, K. S., Day, F. R., Hussain, J., Martínez-Marchal, A., Aiken, C. E., et al.(2021). Genetic insights into biological mechanisms governing human ovarian ageing (p.393–397). Nature.

Seifer, D. B., Feinberg, E. C. & Hsu, A. L.(2024). Ovarian Aging and Fertility (p.1750-1751). Jama.

Stankovic, S., Shekari, S., Huang, Q. Q., Gardner, E. J., Ivarsdottir, E. V., et al.(2024). Genetic links between ovarian ageing, cancer risk and de novo mutation rates (p.608–614). Nature.

Strubel, A., Münick, P., Chaikuad, A., Dreier, B., Schaefer, J., et al.(2022). Designed Ankyrin Repeat Proteins as a tool box for analyzing p63 (p.2445-2458). Cell Death Differ.

Titus, S., Li, F., Stobezki, R., Akula, K., Unsal, E., et al.(2013). Impairment of BRCA1-related DNA double-strand break repair leads to ovarian aging in mice and humans (p.172ra21). Sci Transl Med.

Wu, C., Chen, D., Stout, M. B., Wu, M. & Wang, S.(2025). Hallmarks of ovarian aging (p.418–439). Trends Endocrinol Metab.

Yang, Y., Xu, W., Gao, F., Wen, C., Zhao, S., et al.(2022). Transcription-replication conflicts in primordial germ cells necessitate the Fanconi anemia pathway to safeguard genome stability (p.e2203208119). Proc Natl Acad Sci U S A.

Somatic cell determinants of oocyte developmental competence: the balance between genetics and environment

1David F. Albertini

1Bedford Research Foundation

Mammalian oocytes acquire developmental competence during the protracted and sequential stages of follicle differentiation that take place prior to ovulation. Integration of oogenesis with folliculogenesis draws upon reciprocal gene expression and signaling modalities between the somatic compartment of the follicle and the oocyte, the latter being mediated by several oocyte-secreted factors (OSFs) like GDF-9 and BMP-15. OSFs serve to regulate granulosa cell proliferation and differentiation and prevent premature luteinization because of TBGb signaling as well as integration of cell responses via an extensive network of gap junctions. In parallel, cell contact relationships between granulosa cells and oocytes via transzonal projections (TZPs) assure maintenance of meiotic cell arrest prior to ovulation and progression to oocyte maturity in response to the LH surge. The microenvironment of the follicle is a byproduct of the secretory activities of both granulosa and thecal somatic cells conferring yet another layer of paracrine and autocrine signaling on the cell intrinsic patterns of gene expression that dictate the respective fates of somatic and germ cell lineages. Evidence from mouse transgenic lines and recently characterized genetic variants affecting signaling ligands like GDF-9 and their downstream receptor responses will be reviewed in the context of managing infertile patients whose pathology can be traced to genetic or environmental factors intrinsic to the ovarian follicle.

P001. Integrating Multi-Omics to Reveal the Dynamic Mechanisms of a Dnaaf6-Core DNAAFs Module Synergistically Regulating Dynein Subunits during Spermatogenesis

1Chang-Long Xu*, Xi-Ling Huang, Ting-Jun Liu

1Nanning Second People's Hospital, Nanning, China

The assembly of sperm flagellar axonemes is a prerequisite for sperm motility and male fertility, a process precisely regulated by Dynein Axonemal Assembly Factors (DNAAFs) that guide the cytoplasmic pre-assembly of dynein arms. While individual DNAAF functions have been studied, their dynamic expression patterns and synergistic regulatory mechanisms during the complex process of spermatogenesis remain largely unelucidated. This study aimed to systematically analyze the temporal expression of 19 known DNAAFs to unveil their potential synergistic networks in mice. For this, we utilized public single-cell RNA sequencing (scRNA-seq) data (GSE112393) from mouse testes. By performing detailed cell annotation and pseudotime trajectory analysis, we reconstructed the continuous cellular progression of spermatogenesis. This revealed that most DNAAFs are significantly upregulated commencing at the spermatocyte (SPC) stage, a critical period for axoneme synthesis. We identified several temporal co-expression modules, with a key module (containing Dnaaf4, Dnaaf6, Dnaaf7, Dnaaf9, Dnaaf10, and Dnaaf17) showing ordered high expression in early SPCs. Members of this module displayed significant co-expression with essential dynein arm heavy chain genes (e.g., Dnah2, Dnah7, Dnah17). To functionally validate this network, we integrated the transcriptomic findings with testicular proteomic data from Dnaaf6-knockout (KO) mice. This proteomic analysis confirmed a significant downregulation of the corresponding dynein arm heavy chain subunits, thereby corroborating the regulatory role of the Dnaaf6-core module. Finally, to explore the structural basis of their synergy, AlphaFold3 (AF3) was employed. The structural predictions indicated direct protein-protein interactions among core module members (e.g., DNAAF4-DNAAF10, DNAAF10-DNAAF6) and a critical interaction between DNAAF10 and the R2TP complex member RPAP3, highlighting the involvement of chaperones. In conclusion, our integrative multi-omics approach demonstrates that DNAAFs act synergistically through temporally specific modules to regulate dynein subunit assembly, offering a deeper molecular understanding of flagellar formation and new perspectives on related male infertility and ciliopathies.

Funding: This work was supported by the Nanning Jiangnan District Program (Grant Nos. 20230715-07 and 20240826-09); the Key R&D Program for Scientific Research and Technology of Liangqing District, Nanning (Grant No. 202311); and the Nanning Scientific Research and Technology Development Plan (Grant No. 20253037).

Disclosures: The authors declare no competing interests.

Figure 1 Model for the synergistic action of PIH1D3 and early DNAAFs in cytoplasmic preassembly of dynein arms

The model shows the formation of a core complex, processing of DA subunits, and association with the R2TP/HSP90 system and an extended network. Dashed lines represent potential interactions (ipTM 0.6–0.8.6.8).

P002. DIA-Based Proteomic Analysis Reveals the Specific Role of PIH1D3 in Sperm Flagellar Assembly

1Chang-Long Xu*, Ting-Jun Liu, Xi-Ling Huang, et al.

1Nanning Second People's Hospital, Nanning, China

Primary ciliary dyskinesia (PCD) is a hereditary disorder caused by defects in ciliary function, frequently leading to male infertility. The gene Pih1d3 is a known pathogenic factor for this condition; however, its specific molecular role in sperm flagellum formation and ciliary motility remains unclear. To address this knowledge gap, this study aimed to investigate the specific mechanism of PIH1D3 in sperm flagellar preassembly using Data-Independent Acquisition (DIA)-based proteomics and a Pih1d3 knockout mouse model. For the analysis, testes from Pih1d3 knockout (KO) mice were used as the experimental group, while those from wild-type (WT) mice served as the control group, with three biological replicates per group. A comprehensive DIA-based proteomic analysis was performed, and key selected proteins were subsequently validated by Western blotting. The results identified a total of 398 differentially expressed proteins, consisting of 261 upregulated and 137 downregulated proteins. Among these, 23 proteins were found to be cilia-related, including five dynein heavy chains (DNAH2, DNAH6, DNAH8, DNAH10, DNAH17) and DNAAF9, which were newly identified as potential key factors associated with PIH1D3. Notably, no previous studies have reported this relationship. Gene Ontology (GO) enrichment analysis revealed significant enrichment in terms related to cilium assembly, dynein complex function, and the extracellular microenvironment. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that PI3K-Akt signaling, ECM-receptor interaction, and pathways associated with neurological diseases were significantly enriched. In conclusion, our proteomic profiling reveals that Pih1d3 knockout leads to significant alterations in multiple pathways crucial for sperm flagellar preassembly.........

For details, please click the link

https://link.springer.com/article/10.1007/s1

Or click to download the PDF

https://link.springer.com/content/pdf/10.1007/s10815-025-0373