Modeling and Therapeutic Approaches for Genetic Vasculopathies

总结 平滑肌功能障碍综合征是一种罕见的疾病，全世界已知病例不到50例。所致 ACTA 2基因中的一种特定基因突变会影响平滑肌细胞。平滑肌细胞 存在于身体的许多不同器官中。这些包括在周围运送血液的大血管。 身体（主动脉），脑血管，肺，瞳孔肌肉，肠道，膀胱，甚至女性的子宫。的 受这种特定ACTA 2突变影响的儿童具有非常复杂的医学问题，涉及许多身体 系统.由于供应大脑的血管形状异常， 缩小了在青少年时期，主动脉可能会变弱并分离，需要进行大手术。有些孩子 需要呼吸支持或家庭吸氧这些孩子患有一种严重的复杂疾病， 进行性神经功能障碍我们的目标是为ACTA 2疾病的儿童开发一种基因疗法 可以治疗所有受影响的器官在本提案中，我们将在体外广泛表征 ACTA 2 R179 H平滑肌细胞功能（AIM 1），评价神经血管和随后的行为 ACTA 2 R179 H突变的后果在一种新的小鼠模型（AIM 2），并最终使用 ACTA 2 R179 H小鼠模型，以研究缺血性中风（AIM 3）。研究治疗方案 在整个提案中，我们将利用一种新的CRISPR-cas9系统与定制的指导RNA来恢复（碱基）。 编辑）或破坏（等位基因靶向）ACTA 2 R179 H等位基因，在体外和体内递送该系统，以及 定量测量基因打靶的表型后果

Summary Smooth Muscle Dysfunction Syndrome is a rare disease with less than 50 known cases worldwide. It is caused by a specific genetic mutation in the ACTA2 gene that affects smooth muscle cells. Smooth muscle cells are found in many different organs in the body. These include the large blood vessels that carry blood around the body (aorta), brain blood vessels, lungs, eye pupil muscles, gut, bladder and even the womb in women. The children affected by this specific ACTA2 mutation have very complex medical problems involving many body systems. Patients experience repeated strokes as blood vessels supplying the brain are abnormal in shape and narrowed. As adolescents, the aorta can weaken and dissect, requiring major surgery. Some children have need respiratory support or home oxygen. These children suffer from a severe complex disease that can result in progressive neurological disability. Our aim is to develop a gene therapy for children with ACTA2 disease that can treat all the different organs affected. In this proposal we will extensively characterize in vitro ACTA2R179H smooth muscle cell function (AIM1), evaluate the neurovascular and subsequent behavioral consequences of the ACTA2R179H mutation in a novel mouse model (AIM2), and finally use the ACTA2R179H mouse model to study the ischemic strokes (AIM3). To investigate therapeutic options throughout the proposal we will utilize a novel CRISPR-cas9 system with custom guide RNAs to revert (base editing) or destroy (allele targeting) the ACTA2R179H allele, delivering the system in vitro and in vivo, and quantitatively measuring the phenotypic consequences of gene targeting