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Molecular mechanisms of recessive and dominant mutations in the small vessel disease-related high temperature requirement protease HTRA1

小血管疾病相关高温蛋白酶 HTRA1 隐性和显性突变的分子机制

基本信息

批准号：
320697423
负责人：
Dr. Nathalie Beaufort
金额：
--
依托单位：
Institut für Schlaganfall- und Demenzforschung (ISD)
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2017
资助国家：
德国
起止时间：
2016-12-31 至 2020-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/320697423?language=en
关键词：
Molecular mechanisms recessive dominant mutations

项目摘要

Cerebral small vessel diseases (SVDs) are a major cause of both stroke and dementia but therapeutic options are still very limited. Mendelian conditions have been instrumental in defining the molecular, cellular, and pathophysiological basis of SVDs. Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is a recessive early onset familial SVD caused by loss-of-function mutations in the high temperature requirement protein HTRA1 gene. HTRA1, a secreted protease, assembles as trimers and higher order oligomers to form a mature and proteolytically active complex. Our previous results indicate that SVD-related mutations in HTRA1 result in molecularly diverse effects including but not limited to assembly defects. In collaboration with others, we further found that heterozygous mutations in HTRA1 associate with autosomal dominant late onset SVD but the distinguishing features of recessive and dominant HTRA1 mutations remain to be defined. The current application addresses three major aims: i) to determine the effects of pathogenic mutations on key molecular and cellular aspects of HTRA1 function (mRNA stability, protein stability, trimer assembly, ECM integration, substrate recognition); ii) to investigate, whether heterozygous HTRA1 mutations have a dominant negative effect on enzyme function and determine the underlying mechanisms; iii) to explore strategies to restore HTRA1 function as a potential basis for future targeted therapies. This aim is motivated by our preliminary results indicating it is be possible to restore enzymatic activity for individual mutations. To achieve these aims we will employ a variety of genetic tools, biochemical techniques and cell biology approaches, which will be applied to both transfected cells and primary cells from patients and control subjects. Regardless of the specific outcomes this project will provide novel insights into key mechanisms of CARASIL as well as autosomal dominant forms of HTRA1-related SVD. In addition, the project might open a therapeutic perspective for patients carrying mutations with defined molecular properties.

脑部小血管疾病(SVD)是中风和痴呆症的主要原因，但治疗选择仍然非常有限。孟德尔条件有助于确定SVDS的分子、细胞和病理生理学基础。常染色体隐性遗传性脑动脉病伴皮质下梗塞和白质脑病(CARASIL)是一种由高温需求蛋白HTRA1基因功能缺失突变引起的隐性早发性家族性SVD。HTRA1是一种分泌型蛋白水解酶，以三聚体和高阶低聚物的形式聚集在一起，形成成熟的、具有蛋白分解活性的复合体。我们以前的结果表明，HTRA1中与SVD相关的突变会导致分子上的不同影响，包括但不限于组装缺陷。与其他人合作，我们进一步发现HTRA1杂合突变与常染色体显性遗传迟发性SVD有关，但隐性和显性HTRA1突变的特征仍有待确定。目前的应用有三个主要目标：i)确定致病突变对HTRA1功能的关键分子和细胞方面(mRNA稳定性、蛋白质稳定性、三聚体组装、ECM整合、底物识别)的影响；ii)研究杂合子HTRA1突变是否对酶功能有明显的负面影响并确定潜在的机制；iii)探索恢复HTRA1功能的策略，作为未来靶向治疗的潜在基础。这一目标是由我们的初步结果推动的，这些结果表明有可能恢复单个突变的酶活性。为了实现这些目标，我们将使用各种遗传工具、生化技术和细胞生物学方法，这些方法将应用于患者和对照对象的转基因细胞和原代细胞。无论具体结果如何，该项目都将为CARASIL的关键机制以及与HTRA1相关的常染色体显性SVD提供新的见解。此外，该项目可能会为携带具有明确分子特性的突变的患者打开治疗前景。