SPIA5抑制滋养细胞侵袭参与子痫前期发生的分子机制及其临床诊断价值研究

Eclampsia threatens the life of the patient，so the early diagnosis and monitoring of preeclampsia (PE) is the key to prevent the occurrence of eclampsia. It is known that the invasion of EVTs is the pathogenesis of PE, while urokinase-type plasminogen activator (uPA) can promote EVTs invasion. Our preliminary findings revealed that as an inhibitor of uPA, SPIA5 correlated positively with the severity of PE, and the expression of circR_TPTE2(CT2) were up-regulated in the blood samples of PE patients. The circRNA can up-regulate the expression of SPIA5 by miR-330-5p. Therefore, we hypothesized that CT2-miR-330-5p-SPIA5 contributed to the pathogenesis of PE, can be used as molecular markers for the early diagnosis and monitoring of PE. This project is to 1. reveal the temporal change of SPIA5 expression in PE patients; 2. Explore the effects on invasion of EVTs by overexpression of CT2, SPIA5, and knockdown of miR-330-5p and uPA; 3. Study the effect of SPIA5 on PE phenotype with PE mouse model. This project aims to explore the mechanistic role of SPIA5 in the pathogenesis of PE, and to assess the clinical value of SPIA5 as a novel bio-marker of PE, so as to provide a new approach and scientific basis for PE prediction and disease monitoring.

子痫危及病人生命，子痫前期(PE)的早期诊断和病情监测是阻止子痫发生的关键。有证据显示绒毛外滋养细胞侵袭不足是PE发生的病理基础，尿激酶型纤溶酶原激活剂(uPA)可促进滋养细胞侵袭。我们前期工作发现，确诊PE前患者血中uPA的抑制因子SPIA5的浓度与PE发展呈正相关，同时伴circR_TPTE2（CT2）高表达；CT2可通过miR-330-5p上调SPIA5，据此提出CT2-miR-330-5p-SPIA5构成PE致病的上游分子网络，可作为PE早期诊断和监测的分子标志。本项目拟1.揭示SPIA5分子的临床变化规律；2.在滋养细胞或胎盘组织中过表达CT2和SPIA5、敲弱其MiRNA和uPA探明其对滋养细胞侵袭的作用；3.用PE鼠模型探索SPIA5对PE表型的影响。以明确SPIA5参与PE发生的分子机制，确认其作为PE新型标志物的临床价值，以期为PE的预测和病情监测提供新的途径和科学依据。

子痫前期（pre-eclampsia，PE）是孕20周后出现血压升高伴蛋白尿的一种综合征，是造成孕产妇多器官系统损害、产前出血和胎儿早产的重要原因。PE尚无有效的治疗方法。对高患病风险的妊娠进行早期干预有助于预防PE发生。PE的发病机制尚不明确，滋养细胞侵袭不足是公认的PE病理生理表现之一。滋养细胞侵袭不足的病理生理学机制需要进一步深入探究，以寻找到一些新早期筛查标志物，建立更为精准的早期预测方法。本课题组通过前瞻性队列结合蛋白组学检测发现血清SERPINA5水平在PE患病前明显升高，且在PE患者胎盘中高表达。我们假设高表达SERPINA5可通过影响孕早期滋养细胞的功能参与子痫前期的发病过程。本研究发现：（1）前瞻性研究和横断面分析发现SERPINA5具有作为PE新的早期预测标志物的潜力，以SERPINA5为中心开发并验证了一个早期预测模型（AUC=0.85）。（2）体外实验证明高表达SERPINA5通过抑制绒毛外滋养细胞（EVT）侵袭参与PE的发生。在体内也可诱导孕大鼠出现高血压和蛋白尿等子痫前期样症状。（3）RAP及双荧光素酶报告实验证明circPOX1通过吸附miR-23a-3p解除对SERPINA5表达的抑制。（4）酶活性实验表明高表达SERPINA5阻断uPA/uPAR信号转导，抑制滋养细胞的侵袭。以上结果从临床、体内及体外水平证实了SERPINA5参与子痫前期的发病，提出circPOX1/miR-23a-3p/SERPINA5/uPA/uPAR分子网络调控机制参与了子痫前期的发生。本研究开发了一个有价值的PE早期预测模型，有助于临床上精准预测诊断，以指导临床针对性的预防和治疗。