Validation of Novel Infertility Biomarker

新型不孕不育生物标志物的验证

• 批准号: 8241025
• 负责人: PETER Sutovsky SUTOVSKY
• 金额: $ 18.83万
• 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 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8241025
• 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; public health relevance; 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差距，即缺乏可靠的，客观的方法论无法准确诊断人类男性不育。我们的目标是验证精子特异性硫氧还蛋白SPTRX3作为人类精子质量的新型生物标志物。我们将研究男性不育症患者的精子SPTRX3水平与未能构想，自发流产和不必要的女性伴侣中有多余的分娩相关的女性伴侣（ART）如何相关。人类精液分析和人类男性不育症的诊断的当前方法在很大程度上取决于主观光显微镜评估。这是不准确的，因为肥沃男性和不育男人的精液参数之间存在显着重叠。存在目标，自动化的精子分析方法，但昂贵，并且仅限于专门的参考实验室，而不是廉价的医生办公室生育测试。我们已经确定SPTRX3蛋白仅由缺陷，形态上的人类精子携带。我们的初步数据表明，携带SPTRX3的精子的精液含量增加，与ART后的妊娠率降低相吻合。相比之下，与男性伴侣表现出低精液SPTRX3水平的艺术伴侣产生了更多的多个出生。这个为期两年的项目具有一个具体的目标，即证明精液SPTRX3水平与受精衰竭的发生，自发妊娠丧失和多次怀孕之间的统计关系。我们的假设是，SPTRX3水平较低的男性的女性伴侣更有可能通过艺术构想，并且不太可能自发流产，但他们也更容易出现多个出生。精液SPTRX3含量将通过自动流式细胞仪分析和免疫标记精子的表荧光细胞仪分析以及通过半高素质的蛋白质印刷/致密技术来评估来自不育夫妇的原始和梯度/游泳纯化的精液样品。高精液SPTRX3含量的影响将根据治疗结果的基本度量进行评估，包括受精和首次胚胎裂解率，成功辅助施肥和胚胎转移后自发流产的发生率以及在ART处理的夫妇中多次妊娠的发生率。为了了解SPTRX3结构如何影响人类精子功能的机械理解，我们将与精子染色质结构相关联，这可能会受到含Sptrx3的核液泡的影响，以及流式细胞仪SPTRX3数据。我们还将评估从高/低SPTRX3样品中的精子异源ICSI到仓鼠OVA后的男性前核发育，这是一种通常用于预测人类精子施肥能力和发育潜力的测定。基于我们的临床合作者的反馈，其中之一将参加该项目，我们预计基于SPTRX3的测试将使临床医生能够在一般性不育症临床人群中与IVF/ICSI之间的宫内授精（IUI）做出治疗决定。在IVF/ICSI治疗的夫妻中，将促进应转移多少个胚胎以获得单胎怀孕的决定，而不是没有怀孕或不需要的多重出生。经过拟议的研究验证后，可以以探测器的形式将这种新颖的生物标志物快速转移到临床实践中，以进行参考实验室测试，或用于医生办公室使用的横向流动盒（量子）；也可以开发出柜台家庭生育测试。 公共卫生相关性：该项目将探索并验证人类不孕症的新型生物标志物，即最近发现的精子阳离子蛋白SPTRX3。初步数据表明，该生物标志物可用于更准确地诊断男性不育症，并预测辅助施肥后多生出生和自发怀孕损失的可能性。拟议的探索性研究对于未来的诊断技术开发是必要的，从而导致在美国不孕症诊所引入这种创新的诊断工具。每年，在美国诊所进行了135,000多个不育夫妇的不育治疗，在美国诊所中，治疗决策仍然依赖于过时的，高度主观的光学显微镜方法的精液评估方法。

项目成果

Negative biomarker based male fertility evaluation: Sperm phenotypes associated with molecular-level anomalies.

• DOI: 10.4103/1008-682x.153847
• 发表时间: 2015-07
• 期刊: Asian journal of andrology
• 影响因子: 2.9
• 作者: Sutovsky P;Aarabi M;Miranda-Vizuete A;Oko R
• 通讯作者: Oko R

Semen levels of spermatid-specific thioredoxin-3 correlate with pregnancy rates in ART couples.

• DOI: 10.1371/journal.pone.0061000
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Buckman C;Ozanon C;Qiu J;Sutovsky M;Carafa JA;Rawe VY;Manandhar G;Miranda-Vizuete A;Sutovsky P
• 通讯作者: Sutovsky P