Selecting sperm with distinct metabolic phenotypes to increase ART efficiency

项目总结/摘要 在美国，每年有超过80，000例出生是由于辅助生殖技术（ART）。的 ART的成功通常需要多个昂贵的周期，这些周期加在一起超过了许多美国家庭的年费用 家庭收入。多个抗逆转录病毒治疗周期的必要性部分源于健康的人数量不足。 植入前胚胎健康的胚胎是最高质量配子的直接产物， 存在鉴定最高质量精子的方法。在女性生殖道内，精子中 受精能力是基于功能参数-运动模式，趋化性， 和顶体反应。最好的受精能力的精子也有最低水平的氧化和 DNA损伤。不幸的是，目前临床上选择精子进行卵胞浆内单精子注射的方法 （ICSI）不利用这些参数。本申请的广泛目标是定义生物化学物质， 精子进行运动转换和受精能力的机制，结果将同时 提高基础知识水平，优化体外精子选择技术。的 优化ART的精子选择反过来将：1）增加健康胚胎的产量; 2）降低平均 成本高昂的周期数量;因此3）减轻低收入家庭的成本负担。在目标1中，我们将利用 互补的综合生物能量表型，计算机辅助运动分析（CASA），以及 新的脱氢酶筛选，以确定代谢物可预测的潜在机制 调节小鼠精子的基本运动模式。在目标2中，我们将测试可预测的假设， 对代谢物的反应中的运动性变化可用于选择最佳小鼠精子-那些具有正常 形态学和低水平的DNA损伤，向趋化信号导航并启动 过度活跃的运动，并完成顶体反应。然后我们将采用体外受精（IVF）， 确定基于这些基于代谢的运动特性选择的小鼠精子是否具有增强的能力， 以产生健康的胚胎。在目标3中，我们将利用我们对小鼠精子的研究经验来定义 人类精子基于代谢的运动反应的根本差异。该项目预计 确定微环境与精子功能的联系机制，从而提供概念验证 用于快速开发优化临床选择具有受精能力的人类精子的方法， 条

PROJECT SUMMARY/ABSTRACT More than 80,000 births in the US occur annually as a result of assisted reproductive technology (ART). The success of ART typically requires multiple expensive cycles that together exceed many US families’ yearly household incomes. The necessity of multiple ART cycles stems in part from an insufficient number of healthy preimplantation embryos. Healthy embryos are a direct product of the highest-quality gametes, yet limited methods exist to identify the highest-quality sperm. Within the female reproductive tract, sperm with the highest fertilization competence are naturally selected based on functional parameters – motility patterns, chemotaxis, and the acrosome reaction. The best fertilization-competent sperm also have the lowest levels of oxidative and DNA damage. Unfortunately, current clinical methods for selecting sperm for intracytoplasmic sperm injection (ICSI) do not leverage these parameters. The broad objective of this application is to define the biochemical mechanisms by which sperm undergo motility switching and fertilization competence, and results will both advance the state of basic knowledge and enable optimization of in vitro sperm selection techniques. The optimization of sperm selection for ART will in turn: 1) increase production of healthy embryos; 2) reduce average numbers of costly cycles; and thus 3) lessen the cost burden on lower-income families. In Aim 1, we will utilize complementary comprehensive bioenergetic phenotyping, computer assisted motility analysis (CASA), and a novel dehydrogenase screen to define the underlying mechanisms through which metabolites predictably modulate the essential motility patterns of mouse sperm. In Aim 2, we will test the hypothesis that predictable motility changes in response to metabolites can be used to select the best mouse sperm – those with normal morphology and low levels of DNA damage, the ability to navigate towards a chemotactic signal and initiate hyperactive motility, and complete the acrosome reaction. We will then employ in vitro fertilization (IVF) to determine whether mouse sperm selected based on these metabolism-based motility traits have enhanced ability to generate healthy embryos. In Aim 3, we will exploit our experience working with mouse sperm to define the fundamental differences in the metabolism-based motility responses of human sperm. This project is expected to identify the mechanisms connecting microenvironment to sperm function and thereby provide proof-of-concept for the rapid development of methods to optimize clinical selection of fertilization-competent human sperm for ART.