Dysfunctional pancreatic cells differentiated from induced pluripotent stem cells with mitochondrial DNA mutations
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Diabetes mellitus (DM) is a serious disease in which blood sugarlevels rise abnormally because of failed insulin production ordecreased insulin sensitivity. Although many studies are beingconducted for the treatment or early diagnosis of DM, it is notfully understood how mitochondrial genome (mtDNA) abnormalitiesappear in patients with DM. Here, we induced iPSCs fromfibroblasts, PBMCs, or pancreatic cells of three patients with type2 DM (T2D) and three patients with non-diabetes counterpart. The mtDNA mutations were detected randomly without anytendency among tissues or patients. In T2D patients, 62% (21/34)of iPSC clones harbored multiple mtDNA mutations, of which37% were homoplasmy at the 100% mutation level comparedto only 8% in non-diabetes. We next selected iPSC clones thatwere a wild type or carried mutations and differentiated into pancreaticcells. Oxygen consumption rates were significantly lowerin cells carrying mutant mtDNA. Additionally, the mutant cellsexhibited decreased production of insulin and reduced secretionof insulin in response to glucose. Overall, the results suggest thatscreening mtDNA mutations in iPSCs from patients with T2Dis an essential step before pancreatic cell differentiation for diseasemodeling or autologous cell therapy.
Abstract
Diabetes mellitus (DM) is a serious disease in which blood sugarlevels rise abnormally because of failed insulin production ordecreased insulin sensitivity. Although many studies are beingconducted for the treatment or early diagnosis of DM, it is notfully understood how mitochondrial genome (mtDNA) abnormalitiesappear in patients with DM. Here, we induced iPSCs fromfibroblasts, PBMCs, or pancreatic cells of three patients with type2 DM (T2D) and three patients with non-diabetes counterpart. The mtDNA mutations were detected randomly without anytendency among tissues or patients. In T2D patients, 62% (21/34)of iPSC clones harbored multiple mtDNA mutations, of which37% were homoplasmy at the 100% mutation level comparedto only 8% in non-diabetes. We next selected iPSC clones thatwere a wild type or carried mutations and differentiated into pancreaticcells. Oxygen consumption rates were significantly lowerin cells carrying mutant mtDNA. Additionally, the mutant cellsexhibited decreased production of insulin and reduced secretionof insulin in response to glucose. Overall, the results suggest thatscreening mtDNA mutations in iPSCs from patients with T2Dis an essential step before pancreatic cell differentiation for diseasemodeling or autologous cell therapy.
참고문헌 (29)
- Mather H, "Hypomagnesaemia in diabetes", Arch Med Res, 95, pp.235~242 (1979)
- Caballero AEJAomr, "Metabolic and vascular abnormalities in subjects at risk for type 2 diabetes : the early start of a dangerous situation", Arch Med Res, 36, pp.241~249 (2005)
- Mellitus D, "Diagnosis and classification of diabetes mellitus", Diabetes Care, 28, pp.S5~ (2005)
- Lebovitz HE, "Type 2 diabetes : an overview", Clin Chem, 45, pp.1339~1345 (1999)
- Leahy JL, "Pathogenesis of type 2 diabetes mellitus", Arch Med Res, 36, pp.197~209 (2005)
- Robertson RP, "Chronic oxidative stress as a mechanism for glucose toxicity of the beta cell in type 2 diabetes", Cell Biochem Biophys, 48, pp.139~146 (2007)
- Kieffer TJ, "Beta-cell replacement strategies for diabetes", J Diabetes Investig, 9, pp.457~463 (2018)
- Suchy F, "iPSC-derived organs in vivo : challenges and promise", Cell Stem Cell, 22, pp.21~24 (2018)
- Nakagawa M, "Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts", Nat Biotechnol, 26, pp.101~106 (2008)
- Okano H, "Steps toward safe cell therapy using induced pluripotent stem cells", Circ Res, 112, pp.523~533 (2013)
- 박주미, "Mitochondrial genome mutations in mesenchymal stem cells derived from human dental induced pluripotent stem cells", BMB Reports, 52(12), pp.689~694 (2019), 생화학분자생물학회[DOI]
- Reardon W, "Diabetes mellitus associated with a pathogenic point mutation in mitochondrial DNA", Lancet, 340, pp.1376~1379 (1992)
- Park CB, "Mitochondrial DNA mutations in disease and aging", J Cell Biol, 193, pp.809~818 (2011)
- Wallace DC, "Mitochondrial DNA sequence variation in human evolution and disease", Proc Natl Acad Sci U S A, 91, pp.8739~8746 (1994)
- Maechler P, "Mitochondrial function in normal and diabetic β-cells", Nature, 414, pp.807~812 (2001)
- Ryan CMJNoa, "Diabetes, aging, and cognitive decline", Neurobiol Aging, 26, pp.21~25 (2005)
- Kang E, "Age-related accumulation of somatic mitochondrial DNA mutations in adult-derived human iPSCs", Cell Stem Cell, 18, pp.625~636 (2016)
- Ziada AS, "Mitochondrial DNA somatic mutation burden and heteroplasmy are associated with chronological age, smoking, and HIV infection", Aging Cell, 18, pp.e13018~ (2019)
- Plecitá-Hlavatá L, "Glucose-stimulated insulin secretion fundamentally requires H2O2 signaling by NADPH oxidase 4", Diabetes, 69, pp.1341~1354 (2020)
- MacDonald PE, "The multiple actions of GLP-1 on the process of glucose-stimulated insulin secretion", Diabetes, 51, pp.S434~ (2002)
- Ma H, "Metabolic rescue in pluripotent cells from patients with mtDNA disease", Nature, 524, pp.234~238 (2015)
- Li S, "Mitochondrial dysfunctions contribute to hypertrophic cardiomyopathy in patient iPSC-derived cardiomyocytes with MT-RNR2 mutation", Stem Cell Rep, 10, pp.808~821 (2018)
- Hatakeyama H, "Concise review : heteroplasmic mitochondrial DNA mutations and mitochondrial diseases : toward iPSC-based disease modeling, drug discovery, and regenerative therapeutics", Stem Cells, 34, pp.801~808 (2016)
- Suzuki Y, "Mitochondrial aldehyde dehydrogenase in diabetes associated with mitochondrial tRNALeu(UUR)mutation at position 3243", Diabetes Care, 19, pp.1423~1425 (1996)
- Rorsman P, "Regulation of insulin secretion in human pancreatic islets", Annu Rev Physiol, 75, pp.155~179 (2013)
- Kristinsson H, "FFAR1 is involved in both the acute and chronic effects of palmitate on insulin secretion", Endocrinology, 154, pp.4078~4088 (2013)
- Choi JY, "Association between the pancreas transplantation and survival of patients with diabetes : a single center experience", PLoS One, 12, pp.e0186827~ (2017)
- Shepherd RK, "Measurement of ATP production in mitochondrial disorders", J Inherit Metab Dis, 29, pp.86~91 (2006)
- Kennedy ED, "Effects of depletion of mitochondrial DNA in metabolism secretion coupling in INS-1 cells", Diabetes, 47, pp.374~380 (1998)