PMID- 34454586 OWN - NLM STAT- MEDLINE DCOM- 20220216 LR - 20240507 IS - 1756-994X (Electronic) IS - 1756-994X (Linking) VI - 13 IP - 1 DP - 2021 Aug 28 TI - Functional significance of gain-of-function H19 lncRNA in skeletal muscle differentiation and anti-obesity effects. PG - 137 LID - 10.1186/s13073-021-00937-4 [doi] LID - 137 AB - BACKGROUND: Exercise training is well established as the most effective way to enhance muscle performance and muscle building. The composition of skeletal muscle fiber type affects systemic energy expenditures, and perturbations in metabolic homeostasis contribute to the onset of obesity and other metabolic dysfunctions. Long noncoding RNAs (lncRNAs) have been demonstrated to play critical roles in diverse cellular processes and diseases, including human cancers; however, the functional importance of lncRNAs in muscle performance, energy balance, and obesity remains elusive. We previously reported that the lncRNA H19 regulates the poly-ubiquitination and protein stability of dystrophin (DMD) in muscular dystrophy. METHODS: Here, we identified mouse/human H19-interacting proteins using mouse/human skeletal muscle tissues and liquid chromatography-mass spectrometry (LC-MS). Human induced pluripotent stem-derived skeletal muscle cells (iPSC-SkMC) from a healthy donor and Becker Muscular Dystrophy (BMD) patients were utilized to study DMD post-translational modifications and associated proteins. We identified a gain-of-function (GOF) mutant of H19 and characterized the effects on myoblast differentiation and fusion to myotubes using iPSCs. We then conjugated H19 RNA gain-of-function oligonucleotides (Rgof) with the skeletal muscle enrichment peptide agrin (referred to as AGR-H19-Rgof) and evaluated AGR-H19-Rgof's effects on skeletal muscle performance using wild-type (WT) C57BL/6 J mice and its anti-obesity effects using high-fat diet (HFD)- and leptin deficiency-induced obese mouse models. RESULTS: We demonstrated that both human and mouse H19 associated with DMD and that the H19 GOF exhibited enhanced interaction with DMD compared to WT H19. DMD was found to associate with serine/threonine-protein kinase MRCK alpha (MRCKalpha) and alpha-synuclein (SNCA) in iPSC-SkMC derived from BMD patients. Inhibition of MRCKalpha and SNCA-mediated phosphorylation of DMD antagonized the interaction between H19 and DMD. These signaling events led to improved skeletal muscle cell differentiation and myotube fusion. The administration of AGR-H19-Rgof improved the muscle mass, muscle performance, and base metabolic rate of WT mice. Furthermore, mice treated with AGR-H19-Rgof exhibited resistance to HFD- or leptin deficiency-induced obesity. CONCLUSIONS: Our study suggested the functional importance of the H19 GOF mutant in enhancing muscle performance and anti-obesity effects. CI - (c) 2021. The Author(s). FAU - Li, Yajuan AU - Li Y AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. FAU - Zhang, Yaohua AU - Zhang Y AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. FAU - Hu, Qingsong AU - Hu Q AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. FAU - Egranov, Sergey D AU - Egranov SD AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. FAU - Xing, Zhen AU - Xing Z AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. AD - Current address: Sanofi U.S., Boston, MA, 02139, USA. FAU - Zhang, Zhao AU - Zhang Z AD - Department of Biochemistry and Molecular Biology, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, 77030, USA. FAU - Liang, Ke AU - Liang K AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. FAU - Ye, Youqiong AU - Ye Y AD - Department of Biochemistry and Molecular Biology, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, 77030, USA. FAU - Pan, Yinghong AU - Pan Y AD - Department of Biochemistry and Biology, University of Houston, Houston, TX, 77204, USA. AD - Current address: UPMC Genome Center, Pittsburgh, PA, 15232, USA. FAU - Chatterjee, Sujash S AU - Chatterjee SS AD - Department of Biochemistry and Biology, University of Houston, Houston, TX, 77204, USA. FAU - Mistretta, Brandon AU - Mistretta B AD - Department of Biochemistry and Biology, University of Houston, Houston, TX, 77204, USA. FAU - Nguyen, Tina K AU - Nguyen TK AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. FAU - Hawke, David H AU - Hawke DH AD - Department of Systems Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. FAU - Gunaratne, Preethi H AU - Gunaratne PH AD - Department of Biochemistry and Biology, University of Houston, Houston, TX, 77204, USA. FAU - Hung, Mien-Chie AU - Hung MC AD - Graduate Institute of Biomedical Sciences, Research Center for Cancer Biology, and Center for Molecular Medicine, China Medical University, Taichung, 404, Taiwan. AD - Department of Biotechnology, Asia University, Taichung, 413, Taiwan. FAU - Han, Leng AU - Han L AD - Department of Biochemistry and Molecular Biology, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, 77030, USA. AD - Center for Epigenetics and Disease Prevention, Institute of Biosciences and Technology, Texas A&M University, Houston, TX, 77030, USA. FAU - Yang, Liuqing AU - Yang L AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. lyang7@mdanderson.org. AD - Center for RNA Interference and Non-Coding RNAs, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. lyang7@mdanderson.org. AD - The Graduate School of Biomedical Sciences, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. lyang7@mdanderson.org. FAU - Lin, Chunru AU - Lin C AUID- ORCID: 0000-0002-6473-8229 AD - Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. clin2@mdanderson.org. AD - Center for RNA Interference and Non-Coding RNAs, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA. clin2@mdanderson.org. LA - eng GR - R01 CA218036/CA/NCI NIH HHS/United States GR - S10 OD012304/OD/NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20210828 PL - England TA - Genome Med JT - Genome medicine JID - 101475844 RN - 0 (Biomarkers) RN - 0 (Carrier Proteins) RN - 0 (DMD protein, human) RN - 0 (Dystrophin) RN - 0 (H19 long non-coding RNA) RN - 0 (RNA, Long Noncoding) SB - IM MH - Animals MH - Biomarkers MH - Carrier Proteins MH - Cell Differentiation/*genetics MH - Cells, Cultured MH - Disease Management MH - Disease Models, Animal MH - Disease Susceptibility MH - Dystrophin/genetics/metabolism MH - Fluorescent Antibody Technique/methods MH - *Gain of Function Mutation MH - Genetic Therapy MH - Humans MH - Immunohistochemistry MH - Induced Pluripotent Stem Cells/metabolism MH - Mice MH - Mice, Knockout MH - Muscle Development/*genetics MH - Muscle, Skeletal/*metabolism MH - Muscular Dystrophy, Duchenne/genetics/metabolism/therapy MH - Obesity/diagnosis/etiology/metabolism/*therapy MH - Phosphorylation MH - Protein Binding MH - RNA, Long Noncoding/*genetics PMC - PMC8403366 OTO - NOTNLM OT - Dystrophin OT - H19 OT - Long noncoding RNA OT - Obesity OT - RNA therapy OT - Skeletal muscle COIS- Zhen Xing is now an employee of Sanofi U.S. The remaining authors declare that they have no competing interests. EDAT- 2021/08/30 06:00 MHDA- 2022/02/17 06:00 PMCR- 2021/08/28 CRDT- 2021/08/29 20:31 PHST- 2020/12/30 00:00 [received] PHST- 2021/07/09 00:00 [accepted] PHST- 2021/08/29 20:31 [entrez] PHST- 2021/08/30 06:00 [pubmed] PHST- 2022/02/17 06:00 [medline] PHST- 2021/08/28 00:00 [pmc-release] AID - 10.1186/s13073-021-00937-4 [pii] AID - 937 [pii] AID - 10.1186/s13073-021-00937-4 [doi] PST - epublish SO - Genome Med. 2021 Aug 28;13(1):137. doi: 10.1186/s13073-021-00937-4.