Bmpr2 R491W点突变致大鼠自发肺动脉高压  

Bmpr2 R419W Point Mutation Causes Spontaneous Pulmonary Arterial Hypertension in Rats

作　　者：李天骐 潘慧 纪爽 叶梦婷 董伟 马铭婕 张学佳 张连峰 杨晓敏[2] 吴艳 王晓建 LI Tianqi;PAN Hui;JI Shuang;YE Mengting;DONG Wei;MA Mingjie;ZHANG Xuejia;ZHANG Lianfeng;YANG Xiaomin;WU Yan;WANG Xiaojian(Key Laboratory of Pulmonary Vascular Medicine,Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100037,China;Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014040,Inner Mongolia,China;Kecheng District People's Hospital,Quzhou 324000,Zhejiang,China;Institute of Medical Laboratory Animals,Chinese Academy of Medical Sciences,Center for Comparative Medicine,Peking Union Medical College,Key Laboratory of Comparative Medicine for Human Diseases,National Health Commission,Beijing 100037,China;Hypertension Division,Cardiovascular Institute&Fuwei Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100037,China)

机构地区：[1]中国医学科学院阜外医院肺血管医学重点实验室、心血管疾病国家重点实验室、国家心血管病中心、北京协和医学院,北京市100037 [2]内蒙古科技大学包头医学院,包头市014040 [3]衢州市柯城区人民医院,衢州市324000 [4]中国医学科学院医学实验动物研究所、北京协和医学院比较医学中心、国家卫生健康委员会人类疾病比较医学重点实验室、北京协和医学院,北京市100037 [5]中国医学科学院阜外医院、国家心血管病中心、呼吸与肺血管病中心、北京协和医学院,北京市100037

出　　处：《中国分子心脏病学杂志》2023年第2期5326-5332,共7页Molecular Cardiology of China

基　　金：国家自然科学基金(82170408,81870050);北京市自然科学基金(7222142)。

摘　　要：目的Bmpr2是最主要的肺动脉高压致病基因。既往研究多用基因敲除或截断型突变构建Bmpr2动物模型,缺少Bmpr2错义突变的在体动物研究。方法本研究利用CRISPR/Cas9技术单碱基编辑大鼠Bmpr2基因组区域,定点敲入肺动脉高压患者热点突变c.C1471T,构建Bmpr2 p.R491W突变型大鼠。在突变型大鼠2、3、4月龄时通过超声、右心导管以及病理实验检测大鼠表型。结果经过12代培育,15%的Bmpr2+/R491W突变型大鼠子代在3月龄时自发产生肺动脉高压。与同窝未发生肺动脉高压突变型大鼠相比,发病大鼠的肺动脉平均压显著升高[(17.8±1.2)mmHg vs.(37.7±8.5)mmHg,P<0.001],右心前壁厚度增加1倍[(0.8±0.1)mm vs.(1.6±0.3)mm,P<0.001],右室流出道增宽1.26倍[(3.0±0.4)mm vs.(6.8±0.1)mm,P<0.001]。病理检测显示,发病大鼠的肺血管明显重构,肺小血管肌化程度更高。结论15%的Bmpr2+/R491W突变型大鼠在3月龄时自发产生肺动脉高压,突变外显率和病理表型与患者接近,较好地模拟了遗传性肺动脉高压患者发病特征,为BMPR2突变致病的机制研究提供了重要动物模型。Objective Bmpr2 is the most important disease-causing gene for pulmonary arterial hypertension(PAH).In previous studies,gene knockout or truncated mutations have mostly used to construct Bmpr2 animal models,and there is no in vivo animal study of Bmpr2 missense mutations.Methods In this study,we used CRISPR/Cas9 technology to introduce one single-base mutation in the rat Bmpr2 genome region.This mutation,Bmpr2 c.C1471T(p.R491W),is the hot-spot mutant site in PAH patients.The phenotype of mutant rats was assessed by ultrasound,right heart catheterization and histological experiments at 2 months,3 months and 4 months of age.Results After 12 generations of breeding,15%Bmpr2+/R491W mutant rats spontaneously developed PAH at 3 months of age.Compared with mutant littermate without PAH,the affected rats had significantly higher mean pulmonary artery pressure([17.8±1.2]mmHg vs.[37.7±8.5]mmHg,P<0.001),thicker anterior right heart wall([0.8±0.1]mm vs.[1.6±0.3]mm,P<0.001)and widerright ventricular outflow tract(RVOT)([3.0±0.4]mm vs.[6.8±0.1]mm,P<0.001).Pathological detection showed that the pulmonary blood vessels of the affected rats were significantly remodeled,and the degree of muscularization of pulmonary small blood vessels was higher.Conclusion 15%of Bmpr2+/R491W mutant rats spontaneously developed PAH at 3 months of age.The mutation penetrance is close to those of patients.The pathological phenotype mimic the clinical characteristics of heritable PAH patients,providing an novel animal model to study the mechanism of mutation pathogenesis and drug development.

关 键 词：遗传性肺动脉高压 骨形成蛋白受体2 基因突变 基因编辑 动物模型 

分 类 号：R544.1[医药卫生—心血管疾病]