Po-Jui Hsu1,2,3, Horng-Dar Wang2, Yung-Che Tseng4, Shao-Wei Pan1, Bonifasius Putera Sampurna1, Yuh-Jyh Jong5,6,7,8, Chiou-Hwa Yuh9,10,11,12. 1. Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli, Taiwan. 2. Institute of Biotechnology, National Tsing Hua University, Hsinchu, Taiwan. 3. Department of Laboratory Medicine, Mackay Memorial Hospital, Taipei, Taiwan. 4. Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, I-Lan County, Taiwan. 5. Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan. yjjong@gap.kmu.edu.tw. 6. Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. yjjong@gap.kmu.edu.tw. 7. Departments of Pediatrics and Laboratory Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. yjjong@gap.kmu.edu.tw. 8. Translational Research Center of Neuromuscular Diseases, Kaohsiung Medical University, Kaohsiung, Taiwan. yjjong@gap.kmu.edu.tw. 9. Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli, Taiwan. chyuh@nhri.edu.tw. 10. Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan. chyuh@nhri.edu.tw. 11. Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan. chyuh@nhri.edu.tw. 12. Ph.D. Program in Environmental and Occupational Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. chyuh@nhri.edu.tw.
Abstract
BACKGROUND: Congenital myopathy (CM) is a group of clinically and genetically heterogeneous muscle disorders, characterized by muscle weakness and hypotonia from birth. Currently, no definite treatment exists for CM. A de novo mutation in Tropomyosin 3-TPM3(E151G) was identified from a boy diagnosed with CM, previously TPM3(E151A) was reported to cause CM. However, the role of TPM3(E151G) in CM is unknown. METHODS: Histopathological, swimming behavior, and muscle endurance were monitored in TPM3 wild-type and mutant transgenic fish, modelling CM. Gene expression profiling of muscle of the transgenic fish were studied through RNAseq, and mitochondria respiration was investigated. RESULTS: While TPM3(WT) and TPM3(E151A) fish show normal appearance, amazingly a few TPM3(E151G) fish display either no tail, a crooked body in both F0 and F1 adults. Using histochemical staining for the muscle biopsy, we found TPM3(E151G) displays congenital fiber type disproportion and TPM3(E151A) resembles nemaline myopathy. TPM3(E151G) transgenic fish dramatically swimming slower than those in TPM3(WT) and TPM3(E151A) fish measured by DanioVision and T-maze, and exhibit weaker muscle endurance by swimming tunnel instrument. Interestingly, L-carnitine treatment on TPM3(E151G) transgenic larvae significantly improves the muscle endurance by restoring the basal respiration and ATP levels in mitochondria. With RNAseq transcriptomic analysis of the expression profiling from the muscle specimens, it surprisingly discloses large downregulation of genes involved in pathways of sodium, potassium, and calcium channels, which can be rescued by L-carnitine treatment, fatty acid metabolism was differentially dysregulated in TPM3(E151G) fish and rescued by L-carnitine treatment. CONCLUSIONS: These results demonstrate that TPM3(E151G) and TPM3(E151A) exhibit different pathogenicity, also have distinct gene regulatory profiles but the ion channels were downregulated in both mutants, and provides a potential mechanism of action of TPM3 pathophysiology. Our results shed a new light in the future development of potential treatment for TPM3-related CM.
BACKGROUND:Congenital myopathy (CM) is a group of clinically and genetically heterogeneous muscle disorders, characterized by muscle weakness and hypotonia from birth. Currently, no definite treatment exists for CM. A de novo mutation in Tropomyosin 3-TPM3(E151G) was identified from a boy diagnosed with CM, previously TPM3(E151A) was reported to cause CM. However, the role of TPM3(E151G) in CM is unknown. METHODS: Histopathological, swimming behavior, and muscle endurance were monitored in TPM3 wild-type and mutant transgenic fish, modelling CM. Gene expression profiling of muscle of the transgenic fish were studied through RNAseq, and mitochondria respiration was investigated. RESULTS: While TPM3(WT) and TPM3(E151A) fish show normal appearance, amazingly a few TPM3(E151G) fish display either no tail, a crooked body in both F0 and F1 adults. Using histochemical staining for the muscle biopsy, we found TPM3(E151G) displays congenital fiber type disproportion and TPM3(E151A) resembles nemaline myopathy. TPM3(E151G) transgenic fish dramatically swimming slower than those in TPM3(WT) and TPM3(E151A) fish measured by DanioVision and T-maze, and exhibit weaker muscle endurance by swimming tunnel instrument. Interestingly, L-carnitine treatment on TPM3(E151G) transgenic larvae significantly improves the muscle endurance by restoring the basal respiration and ATP levels in mitochondria. With RNAseq transcriptomic analysis of the expression profiling from the muscle specimens, it surprisingly discloses large downregulation of genes involved in pathways of sodium, potassium, and calcium channels, which can be rescued by L-carnitine treatment, fatty acid metabolism was differentially dysregulated in TPM3(E151G) fish and rescued by L-carnitine treatment. CONCLUSIONS: These results demonstrate that TPM3(E151G) and TPM3(E151A) exhibit different pathogenicity, also have distinct gene regulatory profiles but the ion channels were downregulated in both mutants, and provides a potential mechanism of action of TPM3 pathophysiology. Our results shed a new light in the future development of potential treatment for TPM3-related CM.
Authors: Josine M de Winter; Barbara Joureau; Eun-Jeong Lee; Balázs Kiss; Michaela Yuen; Vandana A Gupta; Christopher T Pappas; Carol C Gregorio; Ger J M Stienen; Simon Edvardson; Carina Wallgren-Pettersson; Vilma-Lotta Lehtokari; Katarina Pelin; Edoardo Malfatti; Norma B Romero; Baziel G van Engelen; Nicol C Voermans; Sandra Donkervoort; C G Bönnemann; Nigel F Clarke; Alan H Beggs; Henk Granzier; Coen A C Ottenheijm Journal: Ann Neurol Date: 2016-04-30 Impact factor: 10.422
Authors: A C Gimenes; D M Bravo; L M Nápolis; M T Mello; A S B Oliveira; J A Neder; L E Nery Journal: Braz J Med Biol Res Date: 2015-02-24 Impact factor: 2.590
Authors: Michaela Yuen; Sandra T Cooper; Steve B Marston; Kristen J Nowak; Elyshia McNamara; Nancy Mokbel; Biljana Ilkovski; Gianina Ravenscroft; John Rendu; Josine M de Winter; Lars Klinge; Alan H Beggs; Kathryn N North; Coen A C Ottenheijm; Nigel F Clarke Journal: Hum Mol Genet Date: 2015-08-24 Impact factor: 6.150