The role of RNA binding proteins in heart development and congenital heart defects

摘要 左心发育不全综合征(HLHS)是一种死亡率较高的先天性心脏病。婴儿患有 这种综合征是由于心脏左半部发育不良而导致的先天循环问题，包括 主动脉、左心室、二尖瓣和主动脉瓣。婴儿可能需要接受一系列的心内直视手术 以恢复体循环。即使在手术后，这些患者也会发展为心力衰竭和其他 作为单心室的右心室负荷过重引起的并发症。我们知道的很少 关于HLHS的病因学和发病机制。最近的人类遗传学研究确认了从头开始 与HLHS表型相关的RNA结合蛋白RBFOX2功能突变的丧失。它是 目前尚不清楚RBFOX2如何导致HLHS发育缺陷。这主要是因为我们的 缺乏关于RBFOX2在心血管发育中的作用及其在心血管疾病中的靶RNA的知识 胚胎心脏。使用我们的RBFox2条件性基因敲除小鼠模型，该模型概括了几个表型， HLHS的细胞和分子特征，我们将解决这些紧迫和临床相关的问题 基因、分子/细胞和最先进的定制设计的测序和计算方法。 我们的主要目标是确定RBFOX2调节的RNA网络有助于 心血管疾病的发展，并调查他们对HLHS的贡献。我们的工作将决定 RBFOX2基因缺失导致HLHS发育缺陷的潜在机制最终，我们的发现 将通过恢复异常的RBFOX2调节来推动创新的HLHS治疗策略的发展 未来应用改良反义寡核苷酸治疗HLHS患者的基因表达。

ABSTRACT Hypoplastic left heart syndrome (HLHS) is a congenital heart disease with a high mortality rate. Infants with this syndrome are born with circulation problems due to hypoplasia of the left part of the heart that includes aorta, left ventricle, mitral valve and aortic valve. Infants may need to undergo a series of open-heart surgeries to restore systemic circulation. Even after the surgeries, these patients can develop heart failure and other complications due to the overload on the right ventricle that serves as the single ventricle. Very little is known about the etiology and pathogenic mechanisms of HLHS. Recent human genetic studies identified de novo loss of function mutations in the RNA binding protein RBFOX2 that are linked to the HLHS phenotype. It is currently unknown how RBFOX2 contributes to HLHS developmental defects. This is mainly because of our lack of knowledge regarding the role of RBFOX2 in cardiovascular development and its target RNAs in the embryonic heart. Using our Rbfox2 conditional knockout mouse model, which recapitulates several phenotypic, cellular, and molecular features of HLHS, we will address these urgent and clinically relevant questions utilizing genetic, molecular/cellular and state-of-the art custom designed sequencing and computational approaches. Our main goal is to determine the mechanisms by which RBFOX2-regulated RNA networks contribute to cardiovascular development and investigate their contributions to HLHS. Our work will determine the underlying mechanisms of HLHS developmental defects caused by loss of RBFOX2. Ultimately, our findings will advance development of innovative HLHS therapeutic strategies by restoring aberrant RBFOX2-regulated gene expression in HLHS patients using modified antisense oligonucleotides in the future.