Search for new genes involved in male infertility through novel approaches

通过新方法寻找与男性不育相关的新基因

基本信息

批准号：
10577839
负责人：
Thomas Garcia
金额：
$ 55万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-22 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10577839
关键词：
Affect Alleles Animal Model Area Bioinformatics Biological Process Biological Specimen Banks Biology Blood CRISPR/Cas technology Candidate Disease Gene Cells Child Chromosomes Classification Clinic Clinical Management Code Complex Contraceptive Agents Contraceptive methods Copy Number Polymorphism Couples Development Diagnosis Diagnostic Etiology Evaluation FDA approved Family Fertility Functional disorder Future Genes Genetic Genetic Counseling Genome Genomic Segment Goals Haploid Cells Haploidy Haplotypes Heterozygote Hormonal Human Impairment Individual Infertility Intervention Investigation Knowledge Laboratories Link Male Contraceptive Agents Male Infertility Mediating Meiosis Methods Modeling Molecular Molecular Abnormality Molecular Diagnostic Testing Mus Mutant Strains Mice Mutation Pathway interactions Patients Pharmaceutical Preparations Phenotype Population Research Resolution Role Sampling Spermiogenesis Therapeutic Third Generation Sequencing Time Translating United States National Institutes of Health Untranslated RNA Validation Variant Vertebrates Whole Blood Woman Work analysis pipeline candidate identification clinical infrastructure cohort contraceptive target empowerment gene product gene synthesis gene therapy genetic testing genetic variant genome sequencing improved large datasets loss of function male male fertility men mouse model next generation novel novel strategies reproductive small molecule inhibitor sperm cell standard of care subfertility technology validation transmission process whole genome

项目摘要

SUMMARY To date, the etiology of idiopathic male infertility is not fully understood, and hormonal male contraceptives have not been effective. Through novel approaches to sequencing and sequencing analysis of male infertile patients and controls, this proposal seeks to identify new coding and non-coding genes and genomic regions required for male fertility and to validate the functional requirement of newly identified genes through mouse models. Past limited sequencing-based approaches have so far only been able to provide diagnostic variants to 1.5% of infertile men who present to the clinic, a yield that will improve with identification of more genes underlying infertility in men. One limitation to past progress has been the inability of conventional short-read sequencing to resolve haplotype-specific information: whether two or more mutations in each gene are on the same or separate alleles. Since gene loss-of-function typically requires damage to both alleles, it is of paramount importance to be able to resolve this information. Through long-read sequencing (a newly established and validated technology) we will identify novel damaging mutations at haplotype-specific resolution, correlating multiple novel damaging mutations with potential loss-of-function in our newly sequenced patients. Additionally, since sperm are haploid and previous studies have demonstrated a requirement for newly synthesized genes after meiosis, we will identify mutations lethal to sperm development through sequencing of both whole blood and sperm. Mutations present in blood (representing both maternal and paternal genomes) that are absent in sperm [with haploid genomes representing 8 million (2^23) chromosome combinations] demonstrate a requirement for those genes in the later stages of sperm development. Thus, through our proposed work we will identify for the first-time genes required for male fertility in men who may in fact be fertile. Lastly, we will apply CRISPR/Cas9 in mice to validate the functional requirement of candidate genes that we identify through our sequencing studies. Novel male infertility associated genes that are found to be expressed in sperm or epididymal cells, are evolutionarily conserved in vertebrates, including mice and humans, and rank among the highest in our patients will be prioritized for validation. We will assess the fertility phenotype of mutant mice and further explore molecular pathophysiology, which for some genes may determine candidacy as a contraceptive target.

概括迄今为止，特发性男性不育症的病因尚不完全清楚，激素男性避孕药已没有效果。通过新方法对男性不育患者进行测序和测序分析和控制，该提案旨在确定新的编码和非编码基因以及基因组区域所需的男性生育能力并通过小鼠模型验证新发现的基因的功能需求。过去的迄今为止，有限的基于测序的方法只能为 1.5% 的患者提供诊断变异。到诊所就诊的不育男性，通过鉴定更多潜在基因，产量将会提高男性不育症。过去进展的一个限制是传统的短读长测序无法解析单倍型特异性信息：每个基因中的两个或多个突变是否位于相同或不同的位置等位基因。由于基因功能丧失通常需要同时损害两个等位基因，因此至关重要能够解析该信息。通过长读长测序（一项新建立并经过验证的技术）我们将以单倍型特异性分辨率识别新的破坏性突变，将多个新的破坏性突变关联起来在我们新测序的患者中存在潜在功能丧失的突变。此外，由于精子是单倍体之前的研究已经证明减数分裂后需要新合成的基因，我们将通过对全血和精子进行测序来识别对精子发育致命的突变。突变存在于血液中（代表母本和父本基因组），而精子中不存在[单倍体代表 800 万 (2^23) 个染色体组合的基因组]证明了对这些基因的需求精子发育的后期阶段。因此，通过我们提出的工作，我们将首次识别基因对于实际上可能具有生育能力的男性来说，这是男性生育能力所必需的。最后，我们将在小鼠身上应用CRISPR/Cas9来验证我们通过测序研究确定的候选基因的功能要求。小说男发现在精子或附睾细胞中表达的不孕相关基因在进化上在包括小鼠和人类在内的脊椎动物中保守，并且在我们的患者中排名最高的是优先进行验证。我们将评估突变小鼠的生育表型并进一步探索分子机制病理生理学，对于某些基因来说可能决定作为避孕目标的候选资格。