Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein granules within processing bodies.
Gain-of-function cardiomyopathic mutations in RBM20 rewire splicing regulation and re-distribute ribonucleoprotein granules within processing bodies.
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DOI：
10.1038/s41467-021-26623-y
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发表时间：
2021-11-03
影响因子：
16.6
通讯作者：
Salomonis N
中科院分区：
文献类型：
--
作者：
Fenix AM;Miyaoka Y;Bertero A;Blue SM;Spindler MJ;Tan KKB;Perez-Bermejo JA;Chan AH;Mayerl SJ;Nguyen TD;Russell CR;Lizarraga PP;Truong A;So PL;Kulkarni A;Chetal K;Sathe S;Sniadecki NJ;Yeo GW;Murry CE;Conklin BR;Salomonis N
心脏剪接因子RBM20的突变可导致恶性扩张型心肌病（DCM）。为了了解RBM20相关DCM的机制，我们在RBM20和RBM20敲除（KO） iPSCs中设计了具有DCM相关错义突变的等基因iPSCs。由这些细胞系制成的ipsc衍生的工程心脏组织再现了rbm20相关DCM的收缩功能障碍，并显示出比KO更大的错义突变功能障碍。通过eCLIP对RBM20 RNA结合的分析显示，突变体RBM20对与肌萎缩性侧索硬化症（ALS）和加工体相关RNA结合蛋白共享的3 ' UTR序列具有功能获得偏好（FUS, DDX6）。深度RNA测序结果显示，RBM20 R636S突变体具有不同于RBM20 KO的独特基因、剪接、聚腺苷化和环状RNA缺陷。超分辨率显微镜证实突变体RBM20保持了非常有限的核定位潜力；相反，突变蛋白在基本条件下与细胞质加工体（DDX6）相关，在急性应激后与应激颗粒（G3BP1）相关。综上所述，我们的研究结果通过剪接依赖性和非依赖性途径强调了心脏病的致病机制。剪接因子RBM20的突变导致侵袭性扩张型心肌病。在这里，作者在人类ipsc衍生的心肌细胞中产生了RBM20 R636S突变体并敲除。突变体RBM20表现出不同的靶RNA结合、剪接和细胞质加工体定位的改变。
Mutations in the cardiac splicing factor RBM20 lead to malignant dilated cardiomyopathy (DCM). To understand the mechanism of RBM20-associated DCM, we engineered isogenic iPSCs with DCM-associated missense mutations in RBM20 as well as RBM20 knockout (KO) iPSCs. iPSC-derived engineered heart tissues made from these cell lines recapitulate contractile dysfunction of RBM20-associated DCM and reveal greater dysfunction with missense mutations than KO. Analysis of RBM20 RNA binding by eCLIP reveals a gain-of-function preference of mutant RBM20 for 3′ UTR sequences that are shared with amyotrophic lateral sclerosis (ALS) and processing-body associated RNA binding proteins (FUS, DDX6). Deep RNA sequencing reveals that the RBM20 R636S mutant has unique gene, splicing, polyadenylation and circular RNA defects that differ from RBM20 KO. Super-resolution microscopy verifies that mutant RBM20 maintains very limited nuclear localization potential; rather, the mutant protein associates with cytoplasmic processing bodies (DDX6) under basal conditions, and with stress granules (G3BP1) following acute stress. Taken together, our results highlight a pathogenic mechanism in cardiac disease through splicing-dependent and -independent pathways. Mutations in the splicing factor RBM20 cause aggressive Dilated Cardiomyopathy. Here the authors generated RBM20 R636S mutants and knockout in human iPSC-derived cardiomyocytes. Mutant RBM20 showed different target RNA binding, altered splicing and localization to cytoplasmic processing bodies.
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影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
影响因子：
4.6
作者：
Ihara K;Sasano T;Hiraoka Y;Togo-Ohno M;Soejima Y;Sawabe M;Tsuchiya M;Ogawa H;Furukawa T;Kuroyanagi H
通讯作者：
Kuroyanagi H
影响因子：
20.1
作者：
Campostrini G;Windt LM;van Meer BJ;Bellin M;Mummery CL
通讯作者：
Mummery CL
影响因子：
64.8
作者：
Hondele, Maria;Sachdev, Ruchika;Weis, Karsten
通讯作者：
Weis, Karsten
影响因子：
10.5
作者：
Conlon EG;Manley JL
通讯作者：
Manley JL