Compartmental analysis of proteomic biomarkers during intra-uterine infections

描述（由申请人提供）：本研究和职业发展计划的目标是利用一个非人灵长类动物的羊膜内和绒毛膜内接种细小脲原体的实验模型来表征生物标志物表达谱（即IGFBP-1蛋白水解片段，钙粒蛋白a和B和膜联蛋白II）在母体和胎儿室中在上升子宫感染的特定阶段的机制和时间相互作用。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时空特征将作为子宫内感染进展阶段的替代指标；同样，在母体治疗干预（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。胎儿对子宫内感染的生理适应将在感染的中期和晚期进行评估，并对母体治疗作出反应。补充的体外研究将确定候选生物标志物的组织来源和生产速度，并解决基质金属蛋白酶在IGFBP-1蛋白水解裂解中的作用。

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.