Compartmental analysis of proteomic biomarkers during intra-uterine infections

描述（由申请人提供）：本研究和职业发展提案的目的是利用羊膜内和绒毛膜蜕膜接种微小脲原体的非人灵长类动物实验模型来表征生物标志物表达谱之间的机制和时间相互作用（即，IGFBP-1蛋白水解片段，钙颗粒蛋白A和B以及膜联蛋白II）。我们的假设是，宫颈阴道液（CVF）、羊水、母体和胎儿血液中特定生物标志物的空间和时间特征将作为子宫内感染进展阶段的替代物;同样，母体治疗干预（抗生素和抗炎剂）期间生物标志物表达谱的变化将作为预后指标。胎儿对宫内感染的生理适应将在感染的中期和晚期以及对母体治疗的反应中进行评估。补充的体外研究将确定候选生物标志物的组织来源和产生速率，并解决基质金属蛋白酶在IGFBP-1的蛋白水解裂解中的作用。 生理学监测与蛋白质组学和分子学研究将解决以下问题：（1）在可接近的采样部位（CVF或母体血液）中进行特异性生物标志物的免疫检测是否与子宫内感染/炎症的进展或消退相关？(2)胎儿血液和/或羊水中的特定生物标志物谱是否与微生物定位和胎儿血流动力学反应有关？(3)子宫活动是否与绒毛膜蜕膜或羊水中U。与母体或胎儿采样部位生物标志物谱的改变有关？将确定一些特定的机制终点，以建立支原体感染进展与早产机制之间的因果关系。子宫收缩力;羊水中PGE 2、PGF 2 α、细胞因子和MMP的水平将与CVF、羊水和母体和胎儿血液中IGFBP-1蛋白水解片段、钙颗粒蛋白A和B以及膜联蛋白II的表达相关。胎儿对宫内感染的生理适应将通过胎儿呼吸参数（胎儿动脉pH、pO 2、sO2%）和胎儿心血管血流动力学的多普勒超声检查来确定。通过测量皮质醇、DHEAS、雌二醇、孕酮和促炎细胞因子来评价内分泌-免疫相互作用和胎儿HPA轴。所提出的工作是独特的，其诊断和干预策略的结合使用在非人灵长类动物模型的子宫内感染（实验绒毛膜蜕膜和羊膜内阶段）。我们期望这些研究的结果将促进临床管理，并促进早期诊断因宫内感染而有早产风险的妇女。

DESCRIPTION (provided by applicant): The objectives of this research and career development proposal are to utilize a nonhuman primate experimental model of intra-amniotic and choriodecidual inoculation with Ureaplasma parvum to characterize the mechanistic and temporal interactions among biomarker expression profiles (i.e., IGFBP-1 proteolytic fragments, calgranulins A and B and annexin II), in maternal and fetal compartments during defined stages of ascending uterine infection. It is our hypothesis that spatial and temporal characteristics of specific biomarkers in cervical vaginal fluid (CVF), amniotic fluid, maternal and fetal blood, will act as surrogates for the stage of progression of intra-uterine infection; similarly, changes in biomarker expression profiles during maternal therapeutic interventions (antibiotic and anti-inflammatory agents), will serve as prognostic indicators. Fetal physiological adaptations to intra-uterine infection will be assessed at intermediate and advanced stages of infection, and in response to maternal therapy. Complementary in vitro studies will determine the tissue sources and production rates of candidate biomarkers, and address the role of matrix metalloproteinases in the proteolytic cleavage of IGFBP-1. Physiological monitoring together with proteomic and molecular studies will address the following questions: (1) Will the immunodetection of specific biomarkers in accessible sampling sites (CVF or maternal blood), correlate with the progression or resolution of intra-uterine infection/inflammation? (2) Will specific biomarker profiles in fetal blood and/or amniotic fluid be linked to microbial localization and fetal hemodynamic responses? (3) Will uterine activity correlate with the choriodecidual or amniotic presence of U. parvum, and with alterations in biomarker profiles in maternal or fetal sampling sites? A number of specific mechanistic endpoints will be ascertained to establish the causal links among progression of Ureaplasma infection and mechanisms of preterm labor. Uterine contractility; amniotic fluid levels of PGE2, PGF2a, cytokines and MMPs will be correlated with the expression of IGFBP-1 proteolytic fragments, calgranulins A and B and annexin II in CVF, amniotic fluid and maternal and fetal blood. Fetal physiologic adaptations to intra-uterine infection will be determined by fetal respiratory parameters (fetal arterial pH, pO2, sO2%), and by Doppler ultrasonography of fetal cardiovascular hemodynamics. Endocrine-immune interactions and the fetal HPA axis will be evaluated by measurements of cortisol, DHEAS, estradiol, progesterone and pro-inflammatory cytokines. The work proposed is unique in its combined use of diagnostic and interventional strategies in a nonhuman primate model of intra-uterine infection (experimental choriodecidual and intra-amniotic stages). It is our expectation that the results of these studies will advance clinical management and facilitate the early diagnosis of women that are at risk for preterm delivery as a consequence of intra-uterine infection.