Regulation of CAP Protein S-Nitrosation in Preterm Labor

早产中 CAP 蛋白 S-亚硝化的调节

基本信息

批准号：
10221011
负责人：
IAIN L BUXTON
金额：
$ 48.23万
依托单位：
UNIVERSITY OF NEVADA RENO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-15 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10221011
关键词：
Accounting Actins Acute Affect Asthma Biological Assay Biophysics Cavia Contractile Proteins Cyclic GMP Development F-Actin Failure Fingerprint Functional disorder Generations Goals Human In Vitro Infection Length Light MYLK gene Maintenance Measures Mediating Modeling Molecular Morbidity - disease rate Myosin Light Chain Kinase Nature Nitric Oxide Nitric Oxide Donors Nitrosation Oxidoreductase Oxytocin Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Post-Translational Protein Processing Pregnancy Pregnant Women Premature Birth Premature Labor Prevention Protein Dephosphorylation Protein S Proteins Regulation Relaxation Research Resistance Role S-Nitrosoglutathione Signal Transduction Site Smooth Muscle Steroids Testing Therapeutic Tissues Tocolysis Tocolytic Agents Uterine Contraction Uterus Woman antenatal cell motility design experimental study fetal inhibitor/antagonist innovation mortality myometrium myosin phosphatase new therapeutic target novel novel strategies porcine model pregnant prevent profilin 1 protein function telokin treatment strategy

项目摘要

Discovery of the molecular mechanisms subserving human uterine quiescence during pregnancy and their dysregulation in spontaneous preterm labor is the objective of this proposal. We will test the hypothesis that failure of preterm human myometrium to relax to nitric oxide-(NO) is the result of dysregulated S-nitrosation of specific smooth muscle contractile proteins. Our long-term goal is to find new effective tocolytics to treat women who enter labor too soon. Preterm labor leads to preterm delivery, a global problem accounting for 75% of fetal morbidity and mortality. No drugs reliably prevent labor in patients who enter labor preterm, thereby allowing their pregnancies to go to term. Therapeutic approaches to manage spontaneous preterm labor (SPTL) are employed without clear evidence of benefit for acute or maintenance tocolysis. NO-mediated relaxation of myometrium is cGMP-independent. Preterm myometrium fails to relax to NO. Discovering the mechanism of action of S-nitrosated contractile proteins can suggest new therapeutic targets to manage SPTL. We propose that gestational quiescence until term results from regulated post-translational S- nitrosation of myosin light chain kinase (MLCK), the regulatory light chain (MYL9) and profilin-1 (PFN1). Addition of NO relaxes term, but not preterm laboring tissues as a result of S-nitrosation differences that alter the function of these CAPs in SPTL. Discovering the effect of regulated S-nitrosations on the mechanism of contractile protein action in term tissues, term tissues from patients in labor and in SPTL (with controls for gestational timing, tocolytic and antenatal steroid use, infection and gestational length) will establish whether or not NO is an endogenous relaxation signal. Comparison of this S-NO fingerprint with that measured following relaxation of the tissue by NO addition in each pregnancy state is novel because SPTL is not simply early labor, will likely be influenced by infection and/or gestational length and because NO- induced relaxation of spontaneous and oxytocin-induced contractions of preterm myometrium is blunted. S-nitrosation differences between labor and SPTL point to altered quiescence mechanisms. Gestational length comparisons in the guinea pig will establish a model in which to investigate S-nitroso regulation of CAP proteins. We describe innovative experiments employing pregnant guinea pigs and tissues from pregnant women and in vitro functional assays designed to reveal the mechanisms underlying the failure of preterm tissues to relax to NO. Completion of this research will suggest therapeutic strategies for the treatment of SPTL such as the S-nitrosoglutathione reductase that regulates S- nitroso protein levels and is known to provide therapeutic benefit in asthma and for which an inhibitor is in development.

人类妊娠期间子宫静止的分子机制的发现及其影响该提案的目标是解决自发性早产的失调问题。我们将检验失败的假设早产儿子宫肌层松弛为一氧化氮（NO）是特定物质 S-亚硝化失调的结果平滑肌收缩蛋白。我们的长期目标是找到新的有效的宫缩药物来治疗过早临产的妇女。早产线索早产是一个全球性问题，占胎儿发病率和死亡率的 75%。没有可靠的药物预防早产患者的分娩，从而使她们能够足月妊娠。治疗性采用管理自发性早产（SPTL）的方法，但没有明确的证据表明对急性早产有益。或维持安胎。 NO 介导的子宫肌层松弛不依赖于 cGMP。早产子宫肌层未能松弛至NO。发现 S-亚硝化收缩蛋白的作用机制可以提出新的治疗靶点管理 SPTL。我们认为，直到足月为止的妊娠静止是由调节的翻译后 S-引起的。肌球蛋白轻链激酶 (MLCK)、调节轻链 (MYL9) 和 profilin-1 (PFN1) 的亚硝化。添加 NO 可以放松足月组织，但不能放松早产组织，因为 S-亚硝化差异会改变这些组织的功能 SPTL 中的 CAP。发现受调节的 S-亚硝化对足月组织收缩蛋白作用机制的影响，足月来自临产和 SPTL 患者的组织（对妊娠时间、宫缩抑制剂和产前类固醇使用进行控制，感染和妊娠长度）将确定 NO 是否是内源性松弛信号。对比一下这个 S-NO 指纹图谱是在每次怀孕状态下添加 NO 使组织松弛后测量的 S-NO 指纹图谱是新颖的因为 SPTL 不仅仅是早产，可能会受到感染和/或妊娠长度的影响，并且因为 NO- 早产子宫肌层自发收缩和催产素诱导的收缩减弱。 S-亚硝化分娩和 SPTL 之间的差异表明静止机制发生了变化。孕期长度比较豚鼠将建立一个模型来研究 CAP 蛋白的 S-亚硝基调节。我们描述创新使用怀孕豚鼠和孕妇组织进行的实验以及设计的体外功能测定揭示早产组织未能松弛至 NO 的潜在机制。完成这项研究将建议治疗 SPTL 的治疗策略，例如调节 S-亚硝基谷胱甘肽还原酶亚硝基蛋白水平，已知可提供哮喘治疗益处，并且有抑制剂可用于治疗哮喘发展。