[논문]Dysfunctional pancreatic cells differentiated from induced pluripotent stem cells with mitochondrial DNA mutations

So, Seongjun; Lee, Song; Lee, Yeonmi; Han, Jongsuk; Kang, Soonsuk; Choi, Jiwan; Kim, Bitnara; Kim, Deokhoon; Yoo, Hyun-Ju; Shim, In-Kyong; Oh, Ju-Yun; Lee, Yu-Na; Kim, Song-Cheol; Kang, Eunju

doi:10.5483/bmbrep.2022.55.9.023

Dysfunctional pancreatic cells differentiated from induced pluripotent stem cells with mitochondrial DNA mutations 원문보기

BMB reports, v.55 no.9, 2022년, pp.453 - 458

So, Seongjun (Department of Convergence Medicine & Stem Cell Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine) , Lee, Song (Division of Hepato-Biliary and Pancreatic Surgery, Department of Surgery, Asan Medical Center, University of Ulsan College of Medicine) , Lee, Yeonmi (Department of Biomedical Science, College of Life Science and Center for Embryo & Stem Cell Research, CHA Advanced Research Institute, CHA University) , Han, Jongsuk (Department of Biomedical Science, College of Life Science and Center for Embryo & Stem Cell Research, CHA Advanced Research Institute, CHA University) , Kang, Soonsuk (Department of Biomedical Science, College of Life Science and Center for Embryo & Stem Cell Research, CHA Advanced Research Institute, CHA University) , Choi, Jiwan (Department of Biomedical Science, College of Life Science and Center for Embryo & Stem Cell Research, CHA Advanced Research Institute, CHA University) , Kim, Bitnara (Department of Biomedical Science, College of Life Science and Center for Embryo & Stem Cell Research, C) , Kim, Deokhoon , Yoo, Hyun-Ju , Shim, In-Kyong , Oh, Ju-Yun , Lee, Yu-Na , Kim, Song-Cheol , Kang, Eunju

Abstract ▼ AI-Helper

Diabetes mellitus (DM) is a serious disease in which blood sugar levels rise abnormally because of failed insulin production or decreased insulin sensitivity. Although many studies are being conducted for the treatment or early diagnosis of DM, it is not fully understood how mitochondrial genome (mtDNA) abnormalities appear in patients with DM. Here, we induced iPSCs from fibroblasts, PBMCs, or pancreatic cells of three patients with type 2 DM (T2D) and three patients with non-diabetes counterpart. The mtDNA mutations were detected randomly without any tendency among tissues or patients. In T2D patients, 62% (21/34) of iPSC clones harbored multiple mtDNA mutations, of which 37% were homoplasmy at the 100% mutation level compared to only 8% in non-diabetes. We next selected iPSC clones that were a wild type or carried mutations and differentiated into pancreatic cells. Oxygen consumption rates were significantly lower in cells carrying mutant mtDNA. Additionally, the mutant cells exhibited decreased production of insulin and reduced secretion of insulin in response to glucose. Overall, the results suggest that screening mtDNA mutations in iPSCs from patients with T2D is an essential step before pancreatic cell differentiation for disease modeling or autologous cell therapy.

주제어

표/그림 (4)

표 Table 1. mtDNA mutations in iPSCs derived from patients with T2D
표 Table 2. mtDNA mutations in iPSCs derived from patients with ND
그림 Fig. 1. mtDNA mutations in patients with ND- and T2D-iPSCs. (A) The number of mtDNA mutations in iPSC clones derived from patients with ND and T2D. There were no significant differences between ND and T2D. (B) The proportion of iPSCs with and without mtDNA mutations in ND and T2D. n = the number of iPSC clones. (C) Heteroplasmy proportion of whole mtDNA mutations in iPSCs. The proportion of homoplasmic mutations was only 8% in ND compared to 37% in T2D. N = the number of mtDNA mutations (D) Heteroplasmy distribution of mtDNA mutations found in ND and T2D. The heteroplasmy of mutations found in T2D was higher than that of ND (*P < 0.05). (E) Probability of obtaining iPSCs with homoplasmic mtDNA mutations. iPSC clones with homoplasmy were found with a probability of 7% in ND and 24% in T2D. (F) Distribution of mtDNA mutations in ND and T2D. The ratio of mutations in the RNA region was higher in T2D than in ND (23% vs. 8%). iPSC: induced pluripotent stem cell; mtDNA: mitochondrial DNA; ND: non-diabetes; T2D: type 2 diabetes.
그림 Fig. 2. mtDNA mutations affect the dysfunction of differentiated pancreatic cells. (A) The gene expression of PPC, PDX1, NGN3, and NKX6.1, relative to GAPDH in T2D-2 BiPSCs. All gene expressions in clone 1 were significantly lower compared to other clones (*P G and 13% mt7970G>A mutations in the non-syn protein-coding region and clone 2 had only 5% mt874G>A mutations in the rRNA region. Both clones showed similar spheroid formation. Scale bar = 200 μm. (D) Immunocytochemistry examining the INSULIN (green) expression of pancreatic spheroids. Nuclei were stained with DAPI (blue). Scale bar = 20 μm. (E) Analysis of OCR levels in differentiated pancreatic cells. In clone 1, all OCRs were significantly lower compared with clone 2 (*P < 0.05, 2 biological replicates, and 3 technical replicates). (F) Insulin production in pancreatic spheroids derived from iPSC clones. The insulin in clone 2 was significantly higher than that of clone 1 (*P < 0.05). (G) GSIS analysis of pancreatic spheroids. Clone 2- derived pancreatic spheroids increased the amount of insulin secretion when glucose concentration increased, whereas clone 1 did not (*P < 0.05, 2 biological replicates, 3 technical replicates). PPC: pancreatic progenitor cell; DAPI: 4',6-diamidino-2-phenylindole; OCR: oxygen consumption rate; GSIS: glucose-stimulated insulin secretion.

참고문헌 (29)

1 Mather H Nisbet JA Burton G 1979 Hypomagnesaemia in diabetes Arch Med Res 95 235 242 10.1016/0009-8981(79)90364-4 527222

상세보기
2 Caballero AEJAomr 2005 Metabolic and vascular abnormalities in subjects at risk for type 2 diabetes: the early start of a dangerous situation Arch Med Res 36 241 249 10.1016/j.arcmed.2005.03.013 15925014

상세보기
3 Mellitus D 2005 Diagnosis and classification of diabetes mellitus Diabetes Care 28 S5 S10 10.2337/diacare.27.2007.S5 14693921

상세보기
4 Lebovitz HE 1999 Type 2 diabetes: an overview Clin Chem 45 1339 1345 10.1093/clinchem/45.8.1339 10430816

상세보기
5 Leahy JL 2005 Pathogenesis of type 2 diabetes mellitus Arch Med Res 36 197 209 10.1016/j.arcmed.2005.01.003 15925010

상세보기
6 Robertson RP Zhou H Zhang T Harmon JS 2007 Chronic oxidative stress as a mechanism for glucose toxicity of the beta cell in type 2 diabetes Cell Biochem Biophys 48 139 146 10.1007/s12013-007-0026-5 17709883

상세보기
7 Kieffer TJ Woltjen K Osafune K Yabe D Inagaki N 2018 Beta‐cell replacement strategies for diabetes J Diabetes Investig 9 457 463 10.1111/jdi.12758 28984038

상세보기
8 Suchy F Yamaguchi T Nakauchi H 2018 iPSC-derived organs in vivo: challenges and promise Cell Stem Cell 22 21 24 10.1016/j.stem.2017.12.003 29304339

상세보기
9 Nakagawa M Koyanagi M Tanabe K 2008 Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts Nat Biotechnol 26 101 106 10.1038/nbt1374 18059259

상세보기
10 Okano H Nakamura M Yoshida K 2013 Steps toward safe cell therapy using induced pluripotent stem cells Circ Res 112 523 533 10.1161/CIRCRESAHA.111.256149 23371901

상세보기
11 Park J Lee Y Shin J 2019 Mitochondrial genome mutations in mesenchymal stem cells derived from human dental induced pluripotent stem cells BMB Rep 52 689 10.5483/BMBRep.2019.52.12.045 31234953

원문보기 상세보기
12 Reardon W Pembrey M Trembath R 1992 Diabetes mellitus associated with a pathogenic point mutation in mitochondrial DNA Lancet 340 1376 1379 10.1016/0140-6736(92)92560-3 1360090

상세보기
13 Park CB Larsson N-GJJocb 2011 Mitochondrial DNA mutations in disease and aging J Cell Biol 193 809 818 10.1083/jcb.201010024 21606204

상세보기
14 Wallace DC 1994 Mitochondrial DNA sequence variation in human evolution and disease Proc Natl Acad Sci U S A 91 8739 8746 10.1073/pnas.91.19.8739 8090716

상세보기
15 Maechler P Wollheim CBJN 2001 Mitochondrial function in normal and diabetic β-cells Nature 414 807 812 10.1038/414807a 11742413

상세보기
16 Ryan CMJNoa 2005 Diabetes, aging, and cognitive decline Neurobiol Aging 26 21 25 10.1016/j.neurobiolaging.2005.09.006 16213627

상세보기
17 Kang E Wang X Tippner-Hedges R 2016 Age-related accumulation of somatic mitochondrial DNA mutations in adult-derived human iPSCs Cell Stem Cell 18 625 636 10.1016/j.stem.2016.02.005 27151456

상세보기
18 Ziada AS Ignas‐Menzies J Lu MY 2019 Mitochondrial DNA somatic mutation burden and heteroplasmy are associated with chronological age, smoking, and HIV infection Aging Cell 18 e13018 10.1111/acel.13018 31407474

상세보기
19 Plecitá-Hlavatá L Jabůrek M Holendová B 2020 Glucose-stimulated insulin secretion fundamentally requires H2O2 signaling by NADPH oxidase 4 Diabetes 69 1341 1354 10.2337/db19-1130 32245800

상세보기
20 MacDonald PE El-kholy W Riedel MJ Salapatek AMF Light PE Wheeler MB 2002 The multiple actions of GLP-1 on the process of glucose-stimulated insulin secretion Diabetes 51 S434 S442 10.2337/diabetes.51.2007.S434 12475787

상세보기
21 Ma H Folmes CD Wu J 2015 Metabolic rescue in pluripotent cells from patients with mtDNA disease Nature 524 234 238 10.1038/nature14546 26176921

상세보기
22 Li S Pan H Tan C 2018 Mitochondrial dysfunctions contribute to hypertrophic cardiomyopathy in patient iPSC-derived cardiomyocytes with MT-RNR2 mutation Stem Cell Rep 10 808 821 10.1016/j.stemcr.2018.01.013 29456182

상세보기
23 Hatakeyama H Goto Y 2016 Concise review: heteroplasmic mitochondrial DNA mutations and mitochondrial diseases: toward iPSC-based disease modeling, drug discovery, and regenerative therapeutics Stem Cells 34 801 808 10.1002/stem.2292 26850516

상세보기
24 Suzuki Y Muramatsu T Taniyama M 1996 Mitochondrial aldehyde dehydrogenase in diabetes associated with mitochondrial tRNALeu (UUR) mutation at position 3243 Diabetes Care 19 1423 1425 10.2337/diacare.19.12.1423 8941476

상세보기
25 Rorsman P Braun MJArop 2013 Regulation of insulin secretion in human pancreatic islets Annu Rev Physiol 75 155 179 10.1146/annurev-physiol-030212-183754 22974438

상세보기
26 Kristinsson H Smith DM Bergsten P Sargsyan EJE 2013 FFAR1 is involved in both the acute and chronic effects of palmitate on insulin secretion Endocrinology 154 4078 4088 10.1210/en.2013-1352 24035997

상세보기
27 Choi JY Jung JH Shin S Kim YH Han DJJPO 2017 Association between the pancreas transplantation and survival of patients with diabetes: a single center experience PLoS One 12 e0186827 10.1371/journal.pone.0186827 29095903

상세보기
28 Shepherd RK Checcarelli N Naini A De Vivo DC DiMauro S Sue CM 2006 Measurement of ATP production in mitochondrial disorders J Inherit Metab Dis 29 86 91 10.1007/s10545-006-0148-8 16601873

상세보기
29 Kennedy ED Maechler P Wollheim CB 1998 Effects of depletion of mitochondrial DNA in metabolism secretion coupling in INS-1 cells Diabetes 47 374 380 10.2337/diabetes.47.3.374 9519742

상세보기

LOADING...

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증