Stromal-epithelial interactions regulate cervical function during pregnancy

基质-上皮相互作用调节妊娠期间的宫颈功能

• 批准号: 7897213
• 负责人: ALF STEFAN ANDERSSON
• 金额: $ 18.75万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7897213
• 关键词: Biological; Biological Assay; Cell Culture System; Cell Culture Techniques; Cell Extracts; Cells; Cervical; Cervical Ripening; Cervix Uteri; Child; Competence; Conditioned Culture Media; Culture Media; DNA Microarray Chip; Data; Diagnosis; Endocervix; Enzymes; Epithelial; Epithelial Cells; Epithelial-Stromal Communication; Epithelium; Equilibrium; Estradiol; Estrogens; Event; Gene Expression; Genes; Goals; Growth Factor; Growth Factor Gene; High Pressure Liquid Chromatography; Human; Hydroxysteroid Dehydrogenases; Incidence; Laboratories; Maintenance; Mediating; Messenger RNA; Microarray Analysis; Molecular; Peptides; Pregnancy; Premature Birth; Premature Infant; Process; Progesterone; Progestins; Property; Proteins; Proteomics; RNA; Regulation; Research; Role; Series; Stromal Cells; Testing; Time; Tissues; Validation; Woman; abstracting; cell stroma; interest; paracrine; polypeptide; premature; prevent; public health relevance; research study; response; tandem mass spectrometry; therapeutic development; tool; two-dimensional

DESCRIPTION (provided by applicant): ABSTRACT 17b-hydroxysteroid dehydrogenase type 2 (17b-HSD type 2) is the central enzyme in maintaining progesterone and estrogen balance in the human endocervix during pregnancy. Data from our laboratory suggest that cervical competence during pregnancy is maintained by expression of 17b-HSD type 2 in the cervical epithelial cells resulting in inactivation of estradiol and maintenance of progesterone by to its oxidative 17b- and 20-HSD activities. During labor, however, the enzyme is down-regulated thereby increasing intracellular estradiol levels and decreasing progesterone levels, which in turn increases cervical ripening. In recent experiments, we have demonstrated that conditioned media from progestin treated cervical stroma cells dramatically increases 17b-HSD type 2 mRNA in cultured cervical epithelial cells. This observation suggests that the regulation of 17b2-HSD type 2 in the cervical epithelium is not a direct effect of progesterone, but an indirect effect, via the cervical stroma. Hence, we hypothesize that this factor, in a paracrine fashion, is involved in the regulation of 17b-HSD type 2, and perhaps other genes important for the process of cervical ripening. The long range goal of the research proposed, therefore, is to define the stromal-derived factor and to elucidate the molecular mechanism by which it regulates 17b-HSD type 2 gene expression. We speculate that dysregulation and expression of this putative paracrine factor may be a crucial component in a series of events leading to premature cervical ripening and preterm birth. The intermediate goals of this research are presented in 3 specific aims. (i) identification of progesterone- induced gene expression using DNA microarray analysis, (ii) identification of bioactive polypeptides present in conditioned media from progestin-treated stromal cells by proteomics, and (iii) assessing expression of candidate peptides/mRNAs in cervical stromal tissues and validation of efficacy of selected polypeptides using cultured cervical epithelial cells. PUBLIC HEALTH RELEVANCE: In this application, we will determine the factors produced by human cervical stromal cells in response to progesterone that mediate 17b-HSD type 2 expression in endocervical epithelial cells. The proposed studies will define an important role for stromal-epithelial interactions in mediating cervical competency during pregnancy and the mechanisms by which progesterone leads to prolongation of pregnancy in women with premature shortening of the cervix during gestation. The results will not only answer important questions regarding mechanisms in the initiation of labor, but will also provide new tools for its diagnosis thereby leading to the development of therapeutic strategies to prevent or abrogate delivery of preterm infants.

描述（申请人提供）：摘要17 b-羟类固醇脱氢酶2型（17 b-HSD 2型）是妊娠期间维持人子宫颈内孕酮和雌激素平衡的中心酶。来自我们实验室的数据表明，妊娠期间的子宫颈能力是通过子宫颈上皮细胞中17 b-HSD 2型的表达来维持的，其导致雌二醇的失活和孕酮的维持，这是通过其氧化17 b-和20-HSD活性来实现的。然而，在分娩过程中，这种酶被下调，从而增加细胞内雌二醇水平和降低孕酮水平，这反过来又增加了宫颈成熟。在最近的实验中，我们已经证明，条件培养基中的resistin处理的宫颈基质细胞显着增加17 b-HSD 2型mRNA在培养的宫颈上皮细胞。这一观察结果表明，子宫颈上皮中17 b2-HSD 2型的调节不是孕酮的直接作用，而是通过子宫颈基质的间接作用。因此，我们假设，这个因素，在旁分泌的方式，参与了17 b-HSD 2型的调节，也许是其他基因的宫颈成熟的过程中很重要。因此，提出的研究的长期目标是确定基质衍生因子，并阐明其调节17 b-HSD 2型基因表达的分子机制。我们推测，这种假定的旁分泌因子的失调和表达可能是导致宫颈早熟和早产的一系列事件的关键组成部分。本研究的中期目标分为三个具体目标。(i)使用DNA微阵列分析鉴定孕酮诱导的基因表达，（ii）通过蛋白质组学鉴定孕激素处理的基质细胞的条件培养基中存在的生物活性多肽，和（iii）评估候选肽/mRNA在宫颈基质组织中的表达，并使用培养的宫颈上皮细胞验证所选多肽的功效。 公共卫生关系：在本申请中，我们将确定由人宫颈基质细胞响应于孕酮介导的宫颈内膜上皮细胞中17 b-HSD 2型表达所产生的因子。拟议的研究将确定一个重要的作用，基质-上皮细胞的相互作用，在怀孕期间介导的子宫颈的能力和孕激素导致妊娠期妇女的子宫颈过早缩短在怀孕期间延长的机制。这些结果不仅将回答有关分娩启动机制的重要问题，而且还将为其诊断提供新的工具，从而导致预防或废除早产儿分娩的治疗策略的发展。