Abstract
Cellular reprogramming of somatic cells from cardiac patients to induced pluripotent stem cells (iPSCs) enables in vitro modeling of human genetic disorders for pathogenic investigations and therapeutic screens. However, using iPSC-derived cardiomyocytes (iPSC-CMs) to model an adult-onset heart disease remains challenging due to the uncertainty regarding the ability of relatively immature iPSC-CMs to fully recapitulate adult disease phenotypes. Arrhythmogenic right ventricular dysplasia (ARVD) is an inherited cardiomyopathy characterized by pathological fibrofatty infiltration and cardiomyocyte (CM) loss predominantly in the right ventricle (RV), leading to life-threatening ventricular arrhythmias. Over 50 % of affected individuals have desmosome gene mutations, most commonly in PKP2 encoding plakophilin-2. The median age at presentation of ARVD is 26–30 years. We used Yamanaka’s pluripotent factors to generate iPSC lines from two ARVD patients with PKP2 mutations. We first developed a method to induce metabolic maturation of iPSC-CMs and showed that induction of adult-like/fatty acid dominant energetics from an embryonic/glycolytic state is essential to model an adult-onset cardiac disease using patient-specific iPSC-CMs. Furthermore, we demonstrate that coactivation of normal peroxisome proliferator-activated receptor-alpha (PPARα) and abnormal PPARγ pathways led to aggressive lipogenesis, elevated apoptosis, and defective intracellular calcium handling in ARVD iPSC-CMs, recapitulating the pathological signatures of ARVD. PPARγ antagonists rescued all ARVD pathological phenotypes and reactive oxygen species (ROS) scavengers curtailed CM apoptosis in our ARVD in vitro model. Thus, using this model, we revealed novel pathogenic insights that metabolic derangement in an adult-like metabolic milieu underlies ARVD pathologies, enabling us to test novel disease-modifying therapeutic strategies.
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References
Asimaki A, Tandri H, Huang H, Halushka MK, Gautam S, Basso C, Thiene G, Tsatsopoulou A, Protonotarios N, McKenna WJ, Calkins H, Saffitz JE (2009) A new diagnostic test for arrhythmogenic right ventricular cardiomyopathy. N Engl J Med 360(11):1075–1084
Awad MM, Dalal D, Tichnell C, James C, Tucker A, Abraham T, Spevak PJ, Calkins H, Judge DP (2006) Recessive arrhythmogenic right ventricular dysplasia due to novel cryptic splice mutation in PKP2. Hum Mutat 27(11):1157
Awad MM, Calkins H, Judge DP (2008) Mechanisms of disease: molecular genetics of arrhythmogenic right ventricular dysplasia/cardiomyopathy. Nat Clin Pract Cardiovasc Med 5(5):258–267
Basso C, Bauce B, Corrado D, Thiene G (2012) Pathophysiology of arrhythmogenic cardiomyopathy. Nat Rev Cardiol 9(4):223–233
Basso C, Pilichou K, Thiene G (2013) Is it time for plakoglobin immune-histochemical diagnostic test for arrhythmogenic cardiomyopathy in the routine pathology practice? Cardiovasc Pathol 22(5):312–313
Brasaemle DL (2007) Thematic review series: adipocyte biology. The perilipin family of structural lipid droplet proteins: stabilization of lipid droplets and control of lipolysis. J Lipid Res 48(12):2547–2559
Cai CL, Liang X, Shi Y, Chu PH, Pfaff SL, Chen J, Evans S (2003) Isl1 identifies a cardiac progenitor population that proliferates prior to differentiation and contributes a majority of cells to the heart. Dev Cell 5(6):877–889
Calkins H (2015) Arrhythmogenic right ventricular dysplasia/cardiomyopathy- three decades of progress. Circ J 79(5):901–913
Carvajal-Vergara X, Sevilla A, D’Souza SL, Ang YS, Schaniel C, Lee DF, Yang L, Kaplan AD, Adler ED, Rozov R, Ge Y, Cohen N, Edelmann LJ, Chang B, Waghray A, Su J, Pardo S, Lichtenbelt KD, Tartaglia M, Gelb BD, Lemischka IR (2010) Patient-specific induced pluripotent stem-cell-derived models of LEOPARD syndrome. Nature 465(7299):808–812
Cerrone M, Lin X, Zhang M, Agullo-Pascual E, Pfenniger A, Chkourko Gusky H, Novelli V, Kim C, Tirasawadichai T, Judge DP, Rothenberg E, Chen HS, Napolitano C, Priori SG, Delmar M (2014) Missense mutations in plakophilin-2 cause sodium current deficit and associate with a Brugada syndrome phenotype. Circulation 129(10):1092–1103
Chakravarthy MV, Lodhi IJ, Yin L, Malapaka RR, Xu HE, Turk J, Semenkovich CF (2009) Identification of a physiologically relevant endogenous ligand for PPARalpha in liver. Cell 138(3):476–488
Chen HS, Kim C, Mercola M (2009) Electrophysiological challenges of cell-based myocardial repair. Circulation 120(24):2496–2508
Dalal D, Molin LH, Piccini J, Tichnell C, James C, Bomma C, Prakasa K, Towbin JA, Marcus FI, Spevak PJ, Bluemke DA, Abraham T, Russell SD, Calkins H, Judge DP (2006) Clinical features of arrhythmogenic right ventricular dysplasia/cardiomyopathy associated with mutations in plakophilin-2. Circulation 113(13):1641–1649
den Haan AD, Tan BY, Zikusoka MN, Llado LI, Jain R, Daly A, Tichnell C, James C, Amat-Alarcon N, Abraham T, Russell SD, Bluemke DA, Calkins H, Dalal D, Judge DP (2009) Comprehensive desmosome mutation analysis in north Americans with arrhythmogenic right ventricular dysplasia/cardiomyopathy. Circ Cardiovasc Genet 2(5):428–435
Djouadi F, Lecarpentier Y, Hebert JL, Charron P, Bastin J, Coirault C (2009) A potential link between peroxisome proliferator-activated receptor signalling and the pathogenesis of arrhythmogenic right ventricular cardiomyopathy. Cardiovasc Res 84(1):83–90
Ferrick DA, Neilson A, Beeson C (2008) Advances in measuring cellular bioenergetics using extracellular flux. Drug Discov Today 13(5–6):268–274
Garcia-Gras E, Lombardi R, Giocondo MJ, Willerson JT, Schneider MD, Khoury DS, Marian AJ (2006) Suppression of canonical Wnt/beta-catenin signaling by nuclear plakoglobin recapitulates phenotype of arrhythmogenic right ventricular cardiomyopathy. J Clin Invest 116(7):2012–2021
Go AS, Mozaffarian D, Roger VL, Benjamin EJ, Berry JD, Blaha MJ, Dai S, Ford ES, Fox CS, Franco S, Fullerton HJ, Gillespie C, Hailpern SM, Heit JA, Howard VJ, Huffman MD, Judd SE, Kissela BM, Kittner SJ, Lackland DT, Lichtman JH, Lisabeth LD, Mackey RH, Magid DJ, Marcus GM, Marelli A, Matchar DB, McGuire DK, Mohler ER 3rd, Moy CS, Mussolino ME, Neumar RW, Nichol G, Pandey DK, Paynter NP, Reeves MJ, Sorlie PD, Stein J, Towfighi A, Turan TN, Virani SS, Wong ND, Woo D, Turner MB (2014) Heart disease and stroke statistics- 2014 update: a report from the American Heart Association. Circulation 129(3):e28–e292
Gregoire FM, Smas CM, Sul HS (1998) Understanding adipocyte differentiation. Physiol Rev 78(3):783–809
Itzhaki I, Maizels L, Huber I, Zwi-Dantsis L, Caspi O, Winterstern A, Feldman O, Gepstein A, Arbel G, Hammerman H, Boulos M, Gepstein L (2011) Modelling the long QT syndrome with induced pluripotent stem cells. Nature 471(7337):225–229
Kim C, Majdi M, Xia P, Wei KA, Talantova M, Spiering S, Nelson B, Mercola M, Chen HS (2010) Non-cardiomyocytes influence the electrophysiological maturation of human embryonic stem cell-derived cardiomyocytes during differentiation. Stem Cells Dev 19(6):783–795
Kim C, Wong J, Wen J, Wang S, Wang C, Spiering S, Kan NG, Forcales S, Puri PL, Leone TC, Marine JE, Calkins H, Kelly DP, Judge DP, Chen HS (2013) Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs. Nature 494(7435):105–110
Knollmann BC (2013) Induced pluripotent stem cell-derived cardiomyocytes: boutique science or valuable arrhythmia model? Circ Res 112(6):969–976; discussion 976
Li D, Liu Y, Maruyama M, Zhu W, Chen H, Zhang W, Reuter S, Lin SF, Haneline LS, Field LJ, Chen PS, Shou W (2011) Restrictive loss of plakoglobin in cardiomyocytes leads to arrhythmogenic cardiomyopathy. Hum Mol Genet 20(23):4582–4596
Lombardi R, Dong J, Rodriguez G, Bell A, Leung TK, Schwartz RJ, Willerson JT, Brugada R, Marian AJ (2009) Genetic fate mapping identifies second heart field progenitor cells as a source of adipocytes in arrhythmogenic right ventricular cardiomyopathy. Circ Res 104(9):1076–1084
Lopaschuk GD, Ussher JR, Folmes CD, Jaswal JS, Stanley WC (2010) Myocardial fatty acid metabolism in health and disease. Physiol Rev 90(1):207–258
Macfarlane DP, Forbes S, Walker BR (2008) Glucocorticoids and fatty acid metabolism in humans: fuelling fat redistribution in the metabolic syndrome. J Endocrinol 197(2):189–204
Marcus FI, Fontaine GH, Guiraudon G, Frank R, Laurenceau JL, Malergue C, Grosgogeat Y (1982) Right ventricular dysplasia: a report of 24 adult cases. Circulation 65(2):384–398
Marcus FI, McKenna WJ, Sherrill D, Basso C, Bauce B, Bluemke DA, Calkins H, Corrado D, Cox MG, Daubert JP, Fontaine G, Gear K, Hauer R, Nava A, Picard MH, Protonotarios N, Saffitz JE, Sanborn DM, Steinberg JS, Tandri H, Thiene G, Towbin JA, Tsatsopoulou A, Wichter T, Zareba W (2010) Diagnosis of arrhythmogenic right ventricular cardiomyopathy/dysplasia: proposed modification of the Task Force Criteria. Eur Heart J 31(7):806–814
Marfella R, Portoghese M, Ferraraccio F, Siniscalchi M, Babieri M, Di Filippo C, D’Amico M, Rossi F, Paolisso G (2009) Thiazolidinediones may contribute to the intramyocardial lipid accumulation in diabetic myocardium: effects on cardiac function. Heart (Br Card Soc) 95(12):1020–1022
Mercola M, Colas A, Willems E (2013) Induced pluripotent stem cells in cardiovascular drug discovery. Circ Res 112(3):534–548
Moretti A, Bellin M, Welling A, Jung CB, Lam JT, Bott-Flugel L, Dorn T, Goedel A, Hohnke C, Hofmann F, Seyfarth M, Sinnecker D, Schomig A, Laugwitz KL (2010) Patient-specific induced pluripotent stem-cell models for long-QT syndrome. N Engl J Med 363(15):1397–1409
Moretti A, Laugwitz KL, Dorn T, Sinnecker D, Mummery C (2013) Pluripotent stem cell models of human heart disease. Cold Spring Harb Perspect Med 3(11):1–20
Murry CE, Reinecke H, Pabon LM (2006) Regeneration gaps: observations on stem cells and cardiac repair. J Am Coll Cardiol 47(9):1777–1785
Neubauer S (2007) The failing heart – an engine out of fuel. N Engl J Med 356(11):1140–1151
Okano H, Nakamura M, Yoshida K, Okada Y, Tsuji O, Nori S, Ikeda E, Yamanaka S, Miura K (2013) Steps toward safe cell therapy using induced pluripotent stem cells. Circ Res 112(3):523–533
Onay-Besikci A (2006) Regulation of cardiac energy metabolism in newborn. Mol Cell Biochem 287(1–2):1–11
Park IH, Arora N, Huo H, Maherali N, Ahfeldt T, Shimamura A, Lensch MW, Cowan C, Hochedlinger K, Daley GQ (2008) Disease-specific induced pluripotent stem cells. Cell 134(5):877–886
Pettinelli P, Videla LA (2011) Up-regulation of PPAR-gamma mRNA expression in the liver of obese patients: an additional reinforcing lipogenic mechanism to SREBP-1c induction. J Clin Endocrinol Metab 96(5):1424–1430
Qyang Y, Martin-Puig S, Chiravuri M, Chen S, Xu H, Bu L, Jiang X, Lin L, Granger A, Moretti A, Caron L, Wu X, Clarke J, Taketo MM, Laugwitz KL, Moon RT, Gruber P, Evans SM, Ding S, Chien KR (2007) The renewal and differentiation of Isl1+ cardiovascular progenitors are controlled by a Wnt/beta-catenin pathway. Cell Stem Cell 1(2):165–179
Razani B, Zhang H, Schulze PC, Schilling JD, Verbsky J, Lodhi IJ, Topkara VK, Feng C, Coleman T, Kovacs A, Kelly DP, Saffitz JE, Dorn GW 2nd, Nichols CG, Semenkovich CF (2011) Fatty acid synthase modulates homeostatic responses to myocardial stress. J Biol Chem 286(35):30949–30961
Santostefano KE, Hamazaki T, Biel NM, Jin S, Umezawa A, Terada N (2015) A practical guide to induced pluripotent stem cell research using patient samples. Lab Invest 95(1):4–13
Sawant AC, Calkins H (2015) Sports in patients with arrhythmogenic right ventricular dysplasia/cardiomyopathy and desmosomal mutations. Herz 40(3):402–409
Shiba Y, Fernandes S, Zhu WZ, Filice D, Muskheli V, Kim J, Palpant NJ, Gantz J, Moyes KW, Reinecke H, Van Biber B, Dardas T, Mignone JL, Izawa A, Hanna R, Viswanathan M, Gold JD, Kotlikoff MI, Sarvazyan N, Kay MW, Murry CE, Laflamme MA (2012) Human ES-cell-derived cardiomyocytes electrically couple and suppress arrhythmias in injured hearts. Nature 489(7415):322–325
Shinnawi R, Gepstein L (2014) iPCS cell modeling of inherited cardiac arrhythmias. Curr Treat Options Cardiovasc Med 16(9):331
Son NH, Park TS, Yamashita H, Yokoyama M, Huggins LA, Okajima K, Homma S, Szabolcs MJ, Huang LS, Goldberg IJ (2007) Cardiomyocyte expression of PPARgamma leads to cardiac dysfunction in mice. J Clin Invest 117(10):2791–2801
Souders CA, Bowers SL, Baudino TA (2009) Cardiac fibroblast: the renaissance cell. Circ Res 105(12):1164–1176
Swope D, Cheng L, Gao E, Li J, Radice GL (2012) Loss of cadherin-binding proteins beta-catenin and plakoglobin in the heart leads to gap junction remodeling and arrhythmogenesis. Mol Cell Biol 32(6):1056–1067
Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131(5):861–872
Veerman CC, Kosmidis G, Mummery CL, Casini S, Verkerk AO, Bellin M (2015) Immaturity of human stem-cell-derived cardiomyocytes in culture: fatal flaw or soluble problem? Stem Cells Dev 24(9):1035–1052
Waku T, Shiraki T, Oyama T, Maebara K, Nakamori R, Morikawa K (2010) The nuclear receptor PPARgamma individually responds to serotonin- and fatty acid-metabolites. EMBO J 29(19):3395–3407
Willson TM, Lambert MH, Kliewer SA (2001) Peroxisome proliferator-activated receptor gamma and metabolic disease. Annu Rev Biochem 70:341–367
Yang X, Pabon L, Murry CE (2014) Engineering adolescence: maturation of human pluripotent stem cell-derived cardiomyocytes. Circ Res 114(3):511–523
Yazawa M, Hsueh B, Jia X, Pasca AM, Bernstein JA, Hallmayer J, Dolmetsch RE (2011) Using induced pluripotent stem cells to investigate cardiac phenotypes in Timothy syndrome. Nature 471(7337):230–234
Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, Thomson JA (2007) Induced pluripotent stem cell lines derived from human somatic cells. Science (New York, N Y) 318(5858):1917–1920
Zhang Z, Stroud MJ, Zhang J, Fang X, Ouyang K, Kimura K, Mu Y, Dalton ND, Gu Y, Bradford WH, Peterson KL, Cheng H, Zhou X, Chen J (2015) Normalization of Naxos plakoglobin levels restores cardiac function in mice. J Clin Invest 125(4):1708–1712
Acknowledgments
We thank the patients for their participation, microarray core facilities at SBMRI for their support, the Johns-Hopkins ARVD registry for their valuable support, and George W. Rogers from Seahorse Bioscience for assistance in metabolic assays. C-Y. W. is supported by a CIRM training grant (TG2-01162). H-S. V. C. is supported by grants from NIH (RO1 HL105194) and California Institute of Regenerative Medicine (CIRM RB2-01512 & RB4-06276).
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Shah, K. et al. (2016). Modeling Arrhythmogenic Right Ventricular Dysplasia/Cardiomyopathy with Patient-Specific iPSCs. In: Fukuda, K. (eds) Human iPS Cells in Disease Modelling. Springer, Tokyo. https://doi.org/10.1007/978-4-431-55966-5_3
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DOI: https://doi.org/10.1007/978-4-431-55966-5_3
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