Pre-mRNA alternative splicing is a common means by which cells generate multiple, functionally distinct isoforms from a single gene. Developmentally regulated splicing has been described individually for many genes, yet little is known about how programs of alternative splicing regulation contribute to embryonic development. Members of the CUG-BP and ETR-3-like factor (CELF) and muscleblind-like (MBNL) protein families have been shown to antagonistically regulate pre-mRNA alternative splicing in the heart. We hypothesize that CELF/MBNL-mediated alternative splicing programs play independent as well as overlapping roles in embryonic heart development. We have shown that changes in CELF and MBNL protein expression during cardiac morphogenesis are accompanied by transitions in alternative splicing. Additional preliminary data suggest that the CELF and/or MBNL proteins are critical for normal cardiac morphogenesis and function. The goal of this proposal is to elucidate the roles that CELF/MBNL-mediated alternative splicing programs play during embryonic heart development. We will use chicken and mouse model systems to investigate the effects of disrupting CELF/MBNL-mediated alternative splicing on cardiac morphogenesis and function, and identify the subset of pre-mRNAs subject to CELF/MBNL regulation in the heart. Understanding the roles that CELF/MBNL-mediated splicing programs play in normal embryonic heart development will provide insight into an understudied yet important mechanism of developmentally regulated gene expression. Furthermore, these studies will shed light on how disruption of normal alternative splicing contributes to disease states resulting from developmental perturbation

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL089376-05
Application #
8605210
Study Section
Development - 2 Study Section (DEV2)
Program Officer
Schramm, Charlene A
Project Start
2009-07-01
Project End
2015-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
5
Fiscal Year
2014
Total Cost
$346,185
Indirect Cost
$125,685
Name
Cleveland Clinic Lerner
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
135781701
City
Cleveland
State
OH
Country
United States
Zip Code
44195
Dasgupta, Twishasri; Stillwagon, Samantha J; Ladd, Andrea N (2013) Gene expression analyses implicate an alternative splicing program in regulating contractile gene expression and serum response factor activity in mice. PLoS One 8:e56590
Blech-Hermoni, Y; Stillwagon, S J; Ladd, A N (2013) Diversity and conservation of CELF1 and CELF2 RNA and protein expression patterns during embryonic development. Dev Dyn 242:767-77
Blech-Hermoni, Yotam; Ladd, Andrea N (2013) RNA binding proteins in the regulation of heart development. Int J Biochem Cell Biol 45:2467-78
Dasgupta, Twishasri; Ladd, Andrea N (2012) The importance of CELF control: molecular and biological roles of the CUG-BP, Elav-like family of RNA-binding proteins. Wiley Interdiscip Rev RNA 3:104-21
Vajda, Natalie A; Brimacombe, Kyle R; LeMasters, Kathryn E et al. (2009) Muscleblind-like 1 is a negative regulator of TGF-beta-dependent epithelial-mesenchymal transition of atrioventricular canal endocardial cells. Dev Dyn 238:3266-72