Mitochondrial DNA mutation m.3243A>G is associated with altered mitochondrial function in peripheral blood mononuclear cells, with heteroplasmy levels and with clinical phenotypes
X. Geng
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
#X.G., Y.Z. and J.Y. contributed equally to this work.Search for more papers by this authorY. Zhang
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Centre for Translational Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
#X.G., Y.Z. and J.Y. contributed equally to this work.Search for more papers by this authorJ. Yan
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Key Laboratory of Embryo Molecular Biology, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Ministry of Health of China and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
#X.G., Y.Z. and J.Y. contributed equally to this work.Search for more papers by this authorC. Chu
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorF. Gao
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorZ. Jiang
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorX. Zhang
Department of Neurology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorY. Chen
Department of Ophthalmology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorX. Wei
Department of Diagnostic Radiology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorY. Feng
Department of Otolaryngology Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorH. Lu
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorCorresponding Author
C. Wang
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Correspondence to: Congrong Wang, e-mail: [email protected]Search for more papers by this authorCorresponding Author
F. Zeng
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Key Laboratory of Embryo Molecular Biology, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Ministry of Health of China and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Correspondence to: Congrong Wang, e-mail: [email protected]Search for more papers by this authorCorresponding Author
W. Jia
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Correspondence to: Congrong Wang, e-mail: [email protected]Search for more papers by this authorX. Geng
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
#X.G., Y.Z. and J.Y. contributed equally to this work.Search for more papers by this authorY. Zhang
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Centre for Translational Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
#X.G., Y.Z. and J.Y. contributed equally to this work.Search for more papers by this authorJ. Yan
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Key Laboratory of Embryo Molecular Biology, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Ministry of Health of China and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
#X.G., Y.Z. and J.Y. contributed equally to this work.Search for more papers by this authorC. Chu
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorF. Gao
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorZ. Jiang
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorX. Zhang
Department of Neurology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorY. Chen
Department of Ophthalmology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorX. Wei
Department of Diagnostic Radiology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorY. Feng
Department of Otolaryngology Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorH. Lu
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Search for more papers by this authorCorresponding Author
C. Wang
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Correspondence to: Congrong Wang, e-mail: [email protected]Search for more papers by this authorCorresponding Author
F. Zeng
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Key Laboratory of Embryo Molecular Biology, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Ministry of Health of China and Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai Children's Hospital, Shanghai Jiao Tong University, Shanghai, China
Correspondence to: Congrong Wang, e-mail: [email protected]Search for more papers by this authorCorresponding Author
W. Jia
Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai Key Laboratory of Diabetes, Shanghai, China
Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Metabolic Diseases Biobank, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China
Correspondence to: Congrong Wang, e-mail: [email protected]Search for more papers by this authorAbstract
Aims
To investigate the associations among heteroplasmy levels (i.e. the proportions of mutant and wild-type mitochondrial DNA in the same cell), mitochondrial function and clinical severity of the m.3243A>G mutation.
Methods
A total of 17 participants carrying the m.3243A>G mutation and 17 sex- and age-matched healthy controls were included in this study. Heteroplasmy levels of the m.3243A>G mutation in leukocytes, saliva and urine sediment were determined by pyrosequencing. The clinical evaluation included endocrinological, audiological and ophthalmological examinations. Mitochondrial function was determined in peripheral blood mononuclear cells isolated from participants.
Results
Heteroplasmy levels in urine sediment were higher than those in leukocytes and saliva. Reduced levels of adenosine triphosphate and mitochondrial membrane potential, and increased reactive oxygen species production were observed in mutant peripheral blood mononuclear cells (all P < 0.05). Linear regression analysis indicated that higher heteroplasmy levels in peripheral blood leukocytes were associated with increased levels of glycated albumin and HbA1c, and decreased total hip bone mineral density T-score after adjustment for age and sex (all P < 0.05). Furthermore, mitochondrial membrane potential was independently associated with bone mineral density T-score at the femoral neck (P < 0.05).
Conclusions
Heteroplasmy levels in peripheral blood leukocytes and mitochondrial membrane potential in peripheral blood mononuclear cells were closely associated with clinical manifestations and were valuable for evaluation of the clinical severity of the m.3243A>G mutation.
Supporting Information
| Filename | Description |
|---|---|
| dme13874-sup-0001-FigS1A-S1H.docxWord document, 497.3 KB | |
| dme13874-sup-0002-TableS1.docxWord document, 16.5 KB | Table S1 Comparision of the clinical features between the diabetic subjects carrying the m.3243A>G mutation and individuals diagnosed with Type 2 diabetes mellitus. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
References
- 1Elliott HR, Samuels DC, Eden JA, Relton CL, Chinnery PF. Pathogenic mitochondrial DNA mutations are common in the general population. Am J Hum Genet 2008; 83: 254–260.
- 2Langdahl JH, Frederiksen AL, Hansen SJ, Andersen PH, Yderstraede KB, Duno M et al. Mitochondrial Point Mutation m.3243A>G Associates With Lower Bone Mineral Density, Thinner Cortices, and Reduced Bone Strength: A Case-Control Study. J Bone Miner Res 2017; 32: 2041–2048.
- 3de Laat P, Koene S, van den Heuvel LP, Rodenburg RJ, Janssen MC, Smeitink JA. Clinical features and heteroplasmy in blood, urine and saliva in 34 Dutch families carrying the m.3243A > G mutation. J Inherit Metab Dis 2012; 35: 1059–1069.
- 4Grady JP, Pickett SJ, Ng YS, Alston CL, Blakely EL, Hardy SA et al. mtDNA heteroplasmy level and copy number indicate disease burden in m.3243A>G mitochondrial disease. EMBO Mol Med 2018; 10: pii: e8262.
- 5Whittaker RG, Blackwood JK, Alston CL, Blakely EL, Elson JL, McFarland R et al. Urine heteroplasmy is the best predictor of clinical outcome in the m.3243A>G mtDNA mutation. Neurology 2009; 72: 568–569.
- 6Fayssoil A, Laforet P, Bougouin W, Jardel C, Lombes A, Becane HM et al. Prediction of long-term prognosis by heteroplasmy levels of the m.3243A>G mutation in patients with the mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes syndrome. Eur J Neurol 2017; 24: 255–261.
- 7Qi Y, Zhang Y, Wang Z, Yang Y, Yuan Y, Niu S et al. Screening of common mitochondrial mutations in Chinese patients with mitochondrial encephalomyopathies. Mitochondrion 2007; 7: 147–150.
- 8Mehrazin M, Shanske S, Kaufmann P, Wei Y, Coku J, Engelstad K et al. Longitudinal changes of mtDNA A3243G mutation load and level of functioning in MELAS. Am J Med Genet A 2009; 149A: 584–587.
- 9Tranah GJ, Katzman SM, Lauterjung K, Yaffe K, Manini TM, Kritchevsky S et al. Mitochondrial DNA m.3243A > G heteroplasmy affects multiple aging phenotypes and risk of mortality. Sci Rep 2018; 8: 11887.
- 10Ohkubo K, Yamano A, Nagashima M, Mori Y, Anzai K, Akehi Y et al. Mitochondrial gene mutations in the tRNA(Leu(UUR)) region and diabetes: prevalence and clinical phenotypes in Japan. Clin Chem 2001; 47: 1641–1648.
- 11Ma Y, Fang F, Yang Y, Zou L, Zhang Y, Wang S et al. The study of mitochondrial A3243G mutation in different samples. Mitochondrion 2009; 9: 139–143.
- 12Janssen GM, Hensbergen PJ, van Bussel FJ, Balog CI, Maassen JA, Deelder AM et al. The A3243G tRNALeu(UUR) mutation induces mitochondrial dysfunction and variable disease expression without dominant negative acting translational defects in complex IV subunits at UUR codons. Hum Mol Genet 2007; 16: 2472–2481.
- 13de Andrade PB, Rubi B, Frigerio F, van den Ouweland JM, Maassen JA, Maechler P. Diabetes-associated mitochondrial DNA mutation A3243G impairs cellular metabolic pathways necessary for beta cell function. Diabetologia 2006; 49: 1816–1826.
- 14Picard M, Zhang J, Hancock S, Derbeneva O, Golhar R, Golik P et al. Progressive increase in mtDNA 3243A>G heteroplasmy causes abrupt transcriptional reprogramming. Proc Natl Acad Sci U S A 2014; 111: E4033–4042.
- 15Mende S, Royer L, Herr A, Schmiedel J, Deschauer M, Klopstock T et al. Whole blood genome-wide expression profiling and network analysis suggest MELAS master regulators. Neurol Res 2011; 33: 638–655.
- 16Hamalainen RH, Manninen T, Koivumaki H, Kislin M, Otonkoski T, Suomalainen A. Tissue- and cell-type-specific manifestations of heteroplasmic mtDNA 3243A>G mutation in human induced pluripotent stem cell-derived disease model. Proc Natl Acad Sci U S A 2013; 110: E3622–3630.
- 17Rudkowska I, Raymond C, Ponton A, Jacques H, Lavigne C, Holub BJ et al. Validation of the use of peripheral blood mononuclear cells as surrogate model for skeletal muscle tissue in nutrigenomic studies. OMICS 2011; 15: 1–7.
- 18Czajka A, Ajaz S, Gnudi L, Parsade CK, Jones P, Reid F et al. Altered Mitochondrial Function, Mitochondrial DNA and Reduced Metabolic Flexibility in Patients With Diabetic Nephropathy. EBioMedicine 2015; 2: 499–512.
- 19Weiss SL, Selak MA, Tuluc F, Perales Villarroel J, Nadkarni VM, Deutschman CS et al. Mitochondrial dysfunction in peripheral blood mononuclear cells in pediatric septic shock. Pediatr Crit Care Med 2015; 16: e4–e12.
- 20Yan JB, Zhang R, Xiong C, Hu C, Lv Y, Wang CR et al. Pyrosequencing is an accurate and reliable method for the analysis of heteroplasmy of the A3243G mutation in patients with mitochondrial diabetes. J Mol Diagn 2014; 16: 431–439.
- 21Obesity: preventing and managing the global epidemic. Report of a WHO consultation. World Health Organ Tech Rep Ser 2000; 894: i–xii, 1–253.
- 22Pascolini D, Smith A. Hearing Impairment in 2008: a compilation of available epidemiological studies. Int J Audiol 2009; 48: 473–485.
- 23Paternostro-Sluga T, Grim-Stieger M, Posch M, Schuhfried O, Vacariu G, Mittermaier C et al. Reliability and validity of the Medical Research Council (MRC) scale and a modified scale for testing muscle strength in patients with radial palsy. J Rehabil Med 2008; 40: 665–671.
- 24Trifunovic A, Wredenberg A, Falkenberg M, Spelbrink JN, Rovio AT, Bruder CE et al. Premature ageing in mice expressing defective mitochondrial DNA polymerase. Nature 2004; 429: 417–423.
- 25Varanasi SS, Francis RM, Berger CE, Papiha SS, Datta HK. Mitochondrial DNA deletion associated oxidative stress and severe male osteoporosis. Osteoporos Int 1999; 10: 143–149.
- 26Kanis JA. Assessment of fracture risk and its application to screening for postmenopausal osteoporosis: synopsis of a WHO report. WHO Study Group. Osteoporos Int 1994; 4: 368–381.
- 27Lang RM, Badano LP, Mor-Avi V, Afilalo J, Armstrong A, Ernande L et al. Recommendations for cardiac chamber quantification by echocardiography in adults: an update from the American Society of Echocardiography and the European Association of Cardiovascular Imaging. J Am Soc Echocardiogr 2015; 28(1–39): e14.
- 28Widlansky ME, Wang J, Shenouda SM, Hagen TM, Smith AR, Kizhakekuttu TJ et al. Altered mitochondrial membrane potential, mass, and morphology in the mononuclear cells of humans with type 2 diabetes. Transl Res 2010; 156: 15–25.
- 29Langdahl JH, Larsen M, Frost M, Andersen PH, Yderstraede KB, Vissing J et al. Lecocytes mutation load declines with age in carriers of the m.3243A>G mutation: A 10-year Prospective Cohort. Clin Genet 2018; 93: 925–928.
- 30Pickett SJ, Grady JP, Ng YS, Gorman GS, Schaefer AM, Wilson IJ et al. Phenotypic heterogeneity in m.3243A>G mitochondrial disease: The role of nuclear factors. Ann Clin Transl Neurol 2018; 5: 333–345.
- 31Mancuso M, Orsucci D, Angelini C, Bertini E, Carelli V, Comi GP et al. The m.3243A>G mitochondrial DNA mutation and related phenotypes. A matter of gender? J Neurol 2014; 261: 504–510.
- 32Laloi-Michelin M, Meas T, Ambonville C, Bellanne-Chantelot C, Beaufils S, Massin P et al. The clinical variability of maternally inherited diabetes and deafness is associated with the degree of heteroplasmy in blood leukocytes. J Clin Endocrinol Metab 2009; 94: 3025–3030.
- 33Lee KM, Chung CY, Kwon SS, Lee SY, Kim TG, Choi Y et al. Factors associated with bone mineral density and risk of fall in Korean adults with type 2 diabetes mellitus aged 50 years and older. J Clin Endocrinol Metab 2014; 99: 4206–4213.
- 34Bartell SM, Kim HN, Ambrogini E, Han L, Iyer S, Serra Ucer S et al. FoxO proteins restrain osteoclastogenesis and bone resorption by attenuating H2O2 accumulation. Nat Commun 2014; 5: 3773.
- 35Miyazaki T, Iwasawa M, Nakashima T, Mori S, Shigemoto K, Nakamura H et al. Intracellular and extracellular ATP coordinately regulate the inverse correlation between osteoclast survival and bone resorption. J Biol Chem 2012; 287: 37808–37823.
- 36Regu GM, Kim H, Kim YJ, Paek JE, Lee G, Chang N et al. Association between Dietary Carotenoid Intake and Bone Mineral Density in Korean Adults Aged 30-75 Years Using Data from the Fourth and Fifth Korean National Health and Nutrition Examination Surveys (2008-2011). Nutrients 2017; 9: pii: E1025.
- 37Ji L, Hou X, Han X. Prevalence and clinical characteristics of mitochondrial tRNAleu(UUR) nt 3243 A–>G and nt 3316 G–>A mutations in Chinese patients with type 2 diabetes. Diabetes Res Clin Pract 2001; 54(Suppl. 2): S35–38.
- 38Guillausseau PJ, Massin P, Dubois-LaForgue D, Timsit J, Virally M, Gin H et al. Maternally inherited diabetes and deafness: a multicenter study. Ann Intern Med 2001; 134: 721–728.
- 39Wang C, Lu J, Lu W, Yu H, Jiang L, Li M et al. Evaluating peripheral nerve function in asymptomatic patients with type 2 diabetes or latent autoimmune diabetes of adults (LADA): results from nerve conduction studies. J Diabetes Complications 2015; 29: 265–269.
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