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

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

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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.

发现妊娠期间人子宫静止的分子机制及自发早产的失调是该提议的目的。我们将测试失败的假设早产的人肌矩阵放松至一氧化氮 - （NO）是特异性S-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溶症剂来治疗过早进入劳动的妇女。早产劳动为了早产，一个全球问题占胎儿发病率和死亡率的75％。没有可靠的药物防止进入劳动早产的患者的劳动，从而允许怀孕。治疗性管理自发早产劳动（SPTL）的方法，没有明确的急性利益证据或维持淋巴结。无介导的肌层松弛无关CGMP。早产子宫骨肌无法放松至否。发现S-硝化收缩蛋白的作用机理可以提出新的治疗靶标管理SPTL。我们提出妊娠静止，直到被调节后的后s-产生期限肌球蛋白轻链激酶（MLCK），调节轻链（MYL9）和Profilin-1（PFN1）的硝化化。加法没有放松期限，而不是由于S-硝化差异而改变的，而不是早产。 sptl中的上限。发现调节的S-硝化作用对学期组织中收缩蛋白作用机理的影响，术语劳动和SPTL患者的组织（具有妊娠时机，淋利性和产前类固醇的对照，感染和妊娠长度）将确定内源性弛豫信号是否是内源性信号。比较 S-NO指纹与在每种妊娠状态下不加添加后在组织放松后测量的指纹是新颖的由于SPTL不仅是早期劳动，因此可能会受到感染和/或妊娠长度的影响，并且没有 - 诱导的自发性和催产素诱导的早产收缩的松弛被钝化。 S-硝化劳动力和SPTL之间的差异指向改变静止机制。妊娠长度比较豚鼠将建立一种研究帽蛋白的S-硝基调节的模型。我们描述了创新使用孕妇的孕妇猪和组织以及设计的体外功能测定的实验揭示了早产组织失败的基础机制，无法放松。这项研究的完成将建议治疗SPTL的治疗策略，例如调节S- 硝基蛋白水平，已知可以在哮喘中提供治疗益处发展。