Validation of Novel Infertility Biomarker

新型不孕不育生物标志物的验证

• 批准号: 8115694
• 负责人: PETER Sutovsky SUTOVSKY
• 金额: $ 21.49万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-11 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8115694
• 关键词: Address; Affect; Appearance; Biological Assay; Biological Markers; Chromatin Structure; Clinic; Clinical; Couples; Couples Therapy; Data; Decision Making; Densitometry; Development; Diagnosis; Diagnostic; Embryo; Embryo Transfer; Enzyme-Linked Immunosorbent Assay; Etiology; Evaluation; Exhibits; Failure; Feedback; Female; Female infertility; Fertility; Fertilization; Fertilization failure; Flow Cytometry; Fluorescent Probes; Future; Goals; Hamsters; Home environment; Human; Immunoassay; Incidence; Infertility; Intracytoplasmic Sperm Injections; Judgment; Laboratories; Lateral; Light; Male Infertility; Measurement; Measures; Methodology; Methods; Microscopic; Morphology; Multiple Birth Offspring; Multiple Pregnancy; Nuclear; Outcome; Ovum; Patients; Pattern; Population; Pregnancy; Pregnancy Outcome; Pregnancy Rate; Pregnancy Tests; Pregnancy loss; Process; Proteins; Reproductive Medicine; Research; Research Personnel; Sampling; Seminal fluid; Sperm Count Procedure; Sperm Motility; Spontaneous abortion; Swimming; Techniques; Testing; Thioredoxin; Training; Treatment outcome; Vacuole; Validation; Western Blotting; base; cell motility; clinical practice; design; deviant; human male; innovation; intrauterine insemination; male; men; novel; patient population; reproductive; sperm analysis; sperm cell; sperm function; sperm morphology; sperm quality; success; technology development; therapy outcome; tool

DESCRIPTION (provided by applicant): This R21 exploratory proposal addresses a major gap in U.S. reproductive medicine, namely the lack of reliable, objective methodology for accurate diagnostics of human male infertility. Our goal is to validate the sperm-specific thioredoxin SPTRX3 as a novel biomarker of human sperm quality. We will examine how sperm SPTRX3 levels in male infertility patients correlate to a failure to conceive, to spontaneous abortion and to unwanted multiple births in their female partners treated by assisted reproductive therapies (ART). Current methods of human semen analysis and diagnostics of human male infertility rely heavily on subjective light microscopic evaluation. This is inaccurate because of a significant overlap between semen parameters of fertile and infertile men. Objective, automated methods for sperm analysis exist but are expensive and confined to specialized reference laboratories, as opposed to an inexpensive doctor's office fertility test. We have already established that SPTRX3 protein is carried exclusively by defective, morphologically deviant human spermatozoa. Our preliminary data demonstrate that the increased semen content of SPTRX3-carrying spermatozoa coincides with reduced pregnancy rates after ART. In contrast, ART couples with male partners exhibiting low semen SPTRX3 levels produced significantly more multiple births. This two year project has ONE SPECIFIC AIM, i.e. to demonstrate a statistical relationship between semen SPTRX3 levels and the occurrence of fertilization failure, spontaneous pregnancy loss and multiple pregnancies in couples from a general infertility clinic population. Our HYPOTHESIS is that the female partners of men with low SPTRX3 levels are more likely to conceive by ART and less likely to suffer a spontaneous abortion, but they are also more prone to multiple births. Semen SPTRX3 content will be evaluated in raw and gradient/swim-up purified semen samples from infertile couples by automated flow cytometric analysis and epifluorescence-light microscopic analysis of immunolabeled spermatozoa, and by semi-quantitative western blotting/densitometry technique. The influence of high semen SPTRX3 content will be evaluated against basic measures of treatment outcome, including fertilization and first embryo cleavage rates, incidence of spontaneous abortion following a successful assisted fertilization and embryo transfer, and the incidence of multiple pregnancies in couples treated by ART. To gain mechanistic understanding of how SPTRX3 carryover affects human sperm function, we will correlate sperm chromatin structure, likely affected by the presence of SPTRX3-containing nuclear vacuoles, with flow cytometric SPTRX3 data. We will also assess male pronuclear development after heterologous ICSI of sperm from high/low SPTRX3 samples into hamster ova, an assay commonly used to predict human sperm fertilizing ability and developmental potential. Based on feedback from our clinical collaborators, one of whom will participate in this project, we anticipate that the SPTRX3 based test will allow clinicians to make a treatment decision between intrauterine insemination (IUI) versus IVF/ICSI in the general infertility clinic-population. In the IVF/ICSI treated couples, a decision will be facilitated for how many embryos should be transferred to obtain a singleton pregnancy, as opposed to no pregnancy or an unwanted multiple birth. Upon validation by the proposed research, this novel biomarker can be quickly transferred to clinical practice in the form of probe for a reference laboratory test, or a lateral flow cassette (dipstick) for doctor's office use; an over the counter home fertility test could also be developed. PUBLIC HEALTH RELEVANCE: This project will explore and validate a novel biomarker of human male infertility, the recently discovered sperm borne protein SPTRX3. Preliminary data suggest that this biomarker can be used to more accurately diagnose male infertility and also to predict the likelihood of multiple births and spontaneous pregnancy losses after assisted fertilization. The proposed exploratory research is necessary for future diagnostic technology development leading to the introduction of this innovative diagnostic tool in US infertility clinics. Annually, more than 135,000 infertile couples are treated for infertility in US clinics, where the treatment-decision making still relies on outdated, highly subjective light microscopic method of semen evaluation.

描述（由申请人提供）：该R21探索性提案解决了美国生殖医学的一个主要空白，即缺乏可靠、客观的方法来准确诊断人类男性不育症。我们的目标是验证精子特异性硫氧还蛋白SPTRX 3作为人类精子质量的新生物标志物。我们将研究男性不育患者的精子SPTRX 3水平如何与通过辅助生殖疗法（ART）治疗的女性伴侣的不孕，自然流产和不想要的多胎生育相关。目前的人类精液分析和人类男性不育症的诊断方法严重依赖于主观的光学显微镜评价。这是不准确的，因为有生育能力和不育男性的精液参数之间有明显的重叠。客观地说，精子分析的自动化方法是存在的，但昂贵且仅限于专业的参考实验室，而不是便宜的医生办公室生育力测试。我们已经确定SPTRX 3蛋白只由有缺陷的、形态异常的人类精子携带。我们的初步数据表明，携带SPTRX 3的精子的精液含量增加与ART后妊娠率降低相一致。相反，ART夫妇与男性伴侣表现出低精液SPTRX 3水平产生显着更多的多胞胎。这个为期两年的项目有一个特定的目标，即证明精液SPTRX 3水平与一般不孕症诊所人群中夫妇受精失败，自发流产和多胎妊娠发生率之间的统计关系。我们的假设是，SPTRX 3水平低的男性的女性伴侣更有可能通过ART怀孕，不太可能遭受自然流产，但他们也更容易多胎生育。将通过免疫标记精子的自动流式细胞仪分析和落射荧光显微镜分析以及半定量蛋白质印迹/密度测定技术，评估不孕夫妇的原始和梯度/上游纯化精液样本中的精液SPTRX 3含量。高精液SPTRX 3含量的影响将根据治疗结果的基本测量进行评估，包括受精率和首次胚胎卵裂率，成功辅助受精和胚胎移植后自然流产的发生率，以及接受ART治疗的夫妇中多胎妊娠的发生率。为了获得SPTRX 3携带如何影响人类精子功能的机制理解，我们将精子染色质结构，可能受到含有SPTRX 3的核空泡的存在的影响，具有流式细胞术SPTRX 3数据。我们还将评估高/低SPTRX 3样本中精子异源ICSI进入仓鼠卵后的雄性原核发育，这是一种常用于预测人类精子可重复性和发育潜力的试验。根据我们的临床合作者的反馈，其中一人将参与该项目，我们预计基于SPTRX 3的测试将允许临床医生在一般不孕症临床人群中做出宫腔内人工授精（IUI）与IVF/ICSI之间的治疗决定。在IVF/ICSI治疗的夫妇中，将有助于决定应该转移多少胚胎以获得单胎妊娠，而不是没有妊娠或不想要的多胎妊娠。在拟议的研究验证后，这种新的生物标志物可以快速转移到临床实践中，以探针的形式用于参考实验室测试，或用于医生办公室使用的侧流盒（试纸）;也可以开发非处方家庭生育力测试。 公共卫生关系：该项目将探索和验证人类男性不育的一种新型生物标志物，即最近发现的精子携带蛋白SPTRX 3。初步数据表明，这种生物标志物可用于更准确地诊断男性不育症，并预测辅助受精后多胞胎和自发妊娠丢失的可能性。拟议的探索性研究是必要的，未来的诊断技术的发展，导致这种创新的诊断工具在美国不孕症诊所的介绍。每年有超过135，000对不孕夫妇在美国诊所接受不孕症治疗，其中治疗决策仍然依赖于过时的，高度主观的精液评估光学显微镜方法。