Functional evaluation of kinesin gene variants associated with female subfertility and egg aneuploidy.

不孕症是影响美国11%育龄妇女的一种常见病。女性的主要原因 不孕症是卵子或胚胎的非整倍体。尽管非整倍体率与母亲的年龄有关，但非整倍体 在特定的产妇年龄组内，发病率各不相同。因此，对于一些患者来说，母亲的年龄不能作为预测因素。 最近的证据表明，母亲的遗传变异使非整倍体受孕的易感性增加。 然而，目前还没有对女性非整倍体受孕风险的基因测试。因此，这个项目的 目的是确定易患非整倍体受孕的母体变异。vbl.使用 非整倍体受孕率相对较高或较低的妇女的全外显子组测序数据 孕妇年龄，我们发现编码运动蛋白的基因变异在高血压病患者中过度表达。 非整倍体受孕率。运动蛋白，微管相关的马达蛋白，是很有希望的候选蛋白 非整倍体风险的基因，因为它们是染色体分离和细胞分裂的关键调节因素。我 假设编码运动蛋白的基因的变异是导致易感性的因果变异 非整倍体受孕。我将在目标1中通过评估这些候选基因变体来解决这一假设 它们在小鼠卵子中引起非整倍体的能力，对于那些筛查呈阳性的小鼠，决定卵子 中断的开发事件。然后，我将评估硅胶工具中的VIA变种。在《目标2》中，我将建造 根据我的初步数据，显示候选基因KIF18A的变异会导致灾难性的 小鼠卵子中的纺锤体缺陷，我将剖析KIF18A的运动域是如何对 通过小鼠模型和蛋白质结构分析进行非整倍体研究。这些数据将有助于建立 非整倍体中的原因母体遗传变异和对KIF18A特定机制的洞察 非整倍体中的运动区变异。这些研究是研究性训练中的一项综合训练 这项计划将为成为女性生殖健康领域的内科科学家奠定坚实的基础。通过 拟议的研究，我将在鸡蛋生物学的严谨和创造性的研究方法方面获得经验， 女性不孕的分子基础，小鼠模型中的遗传病研究，以及计算基因组学。 这项工作将在新泽西州人类遗传学研究所进行，使用最先进的显微镜 设施、现代化的小鼠卵母细胞研究设施、强大的计算遗传学资源和丰富的师资队伍 专业知识。我的研究经验将通过有针对性的生殖高级培训课程来增强， 高分辨率显微镜和基因组学。我的整个训练将与一个纵向的临床 培训方案包括生殖内分泌学和不孕症、围产期遗传学和母婴 罗格斯大学是该州最大的学术医疗保健中心。受指导的研究和 建议的全面培训计划经过精心设计，以增强员工的独立性 富有成效的翻译生殖健康事业。

Infertility is a common disease affecting 11% of reproductive-age women in the US. A leading cause of female infertility is egg or embryo aneuploidy. Although aneuploidy rates are coupled to maternal age, aneuploidy rates vary within a given maternal age group. Thus, for some patients, maternal age is an insufficient predictor. Recent evidence suggests that maternal genetic variation confers predisposition to aneuploid conception. However, no genetic test for a woman’s risk of aneuploid conception exists. Therefore, this project’s objective is to identify causal maternal variants for predisposition to aneuploid conception. Using whole-exome sequencing data from women with either high or low rates of aneuploid conception relative to maternal age, we found that variants in genes encoding kinesins are overrepresented in patients with high rates of aneuploid conception. Kinesins, microtubule-associated motor proteins, are promising candidate genes for aneuploidy risk because they are key regulators of chromosome segregation and cell division. I hypothesize that variants in genes encoding kinesins are causal variants in conferring predisposition to aneuploid conception. I will address this hypothesis in Aim 1 by assessing these candidate genetic variants for their ability to cause aneuploidy in mouse eggs and, for those that screen positive, determine the egg development event that is disrupted. I will then evaluate the variants via in silico tools. In Aim 2, I will build upon my preliminary data, which reveal that variants in one candidate gene, KIF18A, cause catastrophic spindle defects in mouse eggs, and I will dissect how the kinesin motor domain of KIF18A contributes to aneuploidy via mouse models and in silico protein structure analysis. These data will facilitate establishment of causal maternal genetic variants in aneuploidy and provide insight into the specific mechanisms of KIF18A motor domain variation in aneuploidy. These studies are the research training arm of a comprehensive training plan that will build a strong foundation for a career as a physician-scientist in women’s reproductive health. Via the proposed research, I will gain experience in rigorous and creative research approaches to egg biology, the molecular basis of female infertility, genetic disease research in mouse models, and computational genomics. This work will be conducted at the Human Genetics Institute of New Jersey, with state-of-the-art microscopy facilities, modern mouse oocyte research facilities, powerful computational genetics resources, and rich faculty expertise. My research experience will be enhanced by targeted advanced training courses in reproduction, high-resolution microscopy, and genomics. The entirety of my training will be coupled to a longitudinal clinical training program including reproductive endocrinology and infertility, perinatal genetics, and maternal-fetal medicine at the state’s largest academic healthcare center, Rutgers University. The mentored research and comprehensive training plan proposed has been carefully designed to build increasing independence for a productive translational reproductive health career.