Genetic Basis of Oligozoospermia in Infertile Males

描述（由申请人提供）：申请人是临床分子遗传学家，符合abmg认可的临床分子遗传学项目认证资格。他的长期目标是成为一名独立的医生科学家，专注于研究影响男性生殖健康的男性不育的遗传基础。该计划将为申请人提供生殖生物学和人类和小鼠遗传学新技术方面的培训。该项目的总体科学目标是鉴定和表征导致少精子症（OS）的男性不育相关基因的突变。在美国，男性不育影响了近400万男性，其中至少25%被诊断为特发性。男性不育的病因通常与精液特征异常（即精子浓度、活力、形态等）有关。少精症是不育男性最常见的精液异常。本研究的具体目的是：1)建立少精子精子样本的cDNA和DNA收集；2)通过直接cDNA测序和体外功能检测，研究os相关突变候选基因；3)利用寡核苷酸微阵列比较基因组杂交（CGH）技术鉴定和定位与严重OS相关的基因组畸变；4)利用小鼠敲除或敲入模型对cgh检测到的与OS相关的候选基因进行表征。os相关候选基因的选择将根据其蛋白质功能证据和基因在改变的染色体区域的位置进行优先排序。申请人将使用cDNA测序方法和全基因组CGH微阵列研究少精子症患者cDNA和DNA样本的突变和基因组异常。一个突变小鼠模型将试图模仿最常见的形式的精液异常，少精子症在人类开发。候选人将通过确定基因消融对精子发生和受精的时空影响来表征突变小鼠。鉴定和表征与少精症相关的特定基因突变将导致新的诊断程序的发展，并将有助于辅助生殖医学的预后。最终，它将深入了解男性不育的分子基础，并最终可能促进非激素避孕药的发明。拟议的研究将在一个以人类和小鼠遗传学合作研究而闻名的环境中进行。申请人的长期职业目标是成为一名独立的内科科学家，专注于研究影响男性生殖系统的遗传疾病。

DESCRIPTION (provided by applicant): The applicant is a clinical molecular geneticist who is eligible for ABMG-accredited certification in the Clinical Molecular Genetics program. His long-term objective is to become an independent physician- scientist focused on studying the genetic bases of male infertility that affects the reproductive health of men. This proposal will provide the applicant with training in reproductive biology and new technologies in human and mouse genetics. The overall scientific goal of this project is to identify and characterize mutations in male infertility- associated genes that result in oligozoospermia (OS). Male infertility affects nearly 4,000,000 men in the USA, of which at least 25% are diagnosed as idiopathic. An etiology of male infertility is often associated with abnormal semen characteristics (i.e., sperm concentration, motility, morphology, etc.). Oligozoospermia is the most common semen abnormality observed among infertile males. The Specific Aims of the proposed study are: 1) Develop cDNA and DNA collections of oligozoospermic semen samples; 2) Study candidate genes for OS-associated mutations by direct cDNA sequencing and in vitro functional assays; 3) Identify and map genomic aberrations associated with severe OS, using oligo microarray comparative genome hybridization (CGH) technology; and 4) Characterize the CGH-detected gene-candidate that is associated with OS, using mouse knockout or knock-in models. The selection of OS-associated candidate genes will be prioritized according to their protein functional evidence and the gene's position in altered chromosome region. The applicant will study mutations and genomic abnormalities in cDNA and DNA samples from oligozoospermic patients, using cDNA sequencing approach and comprehensive genome-wide CGH microarray. A mutant mouse model will be developed in attempt to mimic the most common form of semen abnormality, oligozoospermia in human. The candidate will characterize the mutant mice by determining the spatiotemporal effects of gene ablation on spermatogenesis and fertilization. Identification and characterization specific gene mutations that associate with oligozoospermia will lead to the development of novel diagnostic procedures, and will help with prognosis in assisted reproductive medicine. Ultimately, it will provide insight into the molecular basis of male infertility, and eventually may facilitate the creation of non-hormonal contraceptives. Proposed study will be performed in an environment known for its collaborative studies in human and mouse genetics. The long-term career goals of the applicant is to become an independent physician-scientist focused on studying genetic disorders that affect male reproductive system. PUBLIC HEALTH RELEVANCE: Male infertility associated with low sperm count (oligozoospermia) affects nearly 2% of men worldwide. The genetic defects that cause this type of male infertility are largely unknown, preventing effective prognosis and treatment of these infertile men. The present research proposal aims to determine the genetic defects that cause male infertility, using the novel approach pioneered in our laboratory.