Regeneration of Anti-Hypoxic Myocardial Cells by Transduction of Mesenchymal Stem Cell-Derived Exosomes Containing Tat-Metallothionein Fusion Proteins

Qurrat Ul Ain; Young Sun Woo; Jee Young Chung; Yong-Hee Kim

Macromolecular Research, Vol.26, No.8, 709-716, August, 2018

DOI10.1007/s13233-018-6101-5 Export Citation

Regeneration of Anti-Hypoxic Myocardial Cells by Transduction of Mesenchymal Stem Cell-Derived Exosomes Containing Tat-Metallothionein Fusion Proteins

Qurrat Ul Ain, Young Sun Woo, Jee Young Chung, and Yong-Hee Kim^†

Department of Bioengineering, Institute for Bioengineering and Biopharmaceutical Research, Room 908, IT/BT building, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul 04763, Korea

E-mail:

Stem cells secrete many extracellular vesicles such as micro vesicles, exosomes and membrane particles. Exosomes represent characteristics similar to their native cells and exosomes secreted from human mesenchymal stem cells (hMSCs) have demonstrated cardio protective effects. In this study, we examined the synergistic effects of exosomes derived from hMSCs expressing metallothionein (MT), a well-known therapeutic protein to treat myocardial infarction, for recovery of cell viability in vitro in hypoxic conditions. Tat-metallothionein (Tat-MT) recombinant fusion proteins were prepared by a recombinant method to increase the transduction of metallothionein into exosomes via Tat’s transduction characteristic. Exosomes from hMSCs were transduced with Tat-MT, and characterized by transmission electron microscopy and immunoblotting. Cellular uptake of exosomes and protein was analyzed by confocal microscopy. The cytoprotective effects of exosomes transfected with Tat-MT (Exo/Tat-MT) on cardiomyocytes were evaluated by accessing cell viability. Exo/Tat-MT significantly upregulated cell viability and downregulated apoptosis in cardiomyocytes. The therapeutic potential of exosome-mediated therapeutic protein delivery was demonstrated by strong cell viability (70-80%) under in vitro hypoxic conditions. This study reveals the dual benefits of exosomes derived from hMSCs and highlights a new method of intercellular stem cells mediation for the stem cell-derived treatment of myocardial infarction.

Keywords:exosomes;human mesenchymal stem cells;tat-metallothionein fusion proteins;hypoxia;myocardial infarction

[References]

Lloyd-Jones D, Adams R, Carnethon M, De Simone G, Ferguson TB, et al., Circulation, 119, e21 (2009)
Pittenger MF, Martin BJ, Circ. Res., 95, 9 (2004)
Hatzistergos KE, Quevedo H, Oskouei BN, Hu Q, Feigenbaum GS, Margitich IS, Mazhari R, Boyle AJ, Zambrano JP, Rodriguez JE, Circ. Res., 107, 913 (2010)
Nagaya N, Kangawa K, Itoh T, Iwase T, Murakami S, Miyahara Y, Fujii T, Uematsu M, Ohgushi H, Yamagishi M, Circulation, 112, 1128 (2005)
Piao H, Youn TJ, Kwon JS, Kim YH, Bae JW, Kim DW, Cho MC, Lee MM, Park YB, Eur. J. Heart Fail., 7, 730 (2005)
Quevedo HC, Hatzistergos KE, Oskouei BN, Feigenbaum GS, Rodriguez JE, Valdes D, Pattany PM, Zambrano JP, Hu Q, McNiece I, Proc. Natl. Acad. Sci., 106, 14022 (2009)
Stamm C, Westphal B, Kleine HD, Petzsch M, Kittner C, Klinge H, Schumichen C, Nienaber CA, Freund M, Steinhoff G, Lancet, 361, 45 (2003)
Meyer GP, Wollert KC, Lotz J, Steffens J, Lippolt P, Fichtner S, Hecker H, Schaefer A, Arseniev L, Hertenstein B, Circulation, 113, 1287 (2006)
Kwak BR. Mach F, Nat. Med., 11, 367 (2005)
Duran JM, Makarewich CA, Sharp TE, Starosta T, Zhu F, Hoffman NE, Chiba Y, Madesh M, Berretta RM, Kubo H, Circ. Res., 113, 539 (2013)
Mureli S, Gans CP, Bare DJ, Geenen DL, Kumar NM, Banach K, Am. J. Physiol. Heart Circ. Physiol., 304, H600 (2013)
Mummery CL, Davis RP, Krieger JE, Sci. Transl. Med., 2, 27ps17 (2010)
Caplan AI, Dennis JE, J. Cell. Biochem., 98, 1076 (2006)
Gnecchi M, Danieli P, Cervio E, Vascul. Pharmacol., 57, 48 (2012)
Chimenti I, Smith RR, Li TS, Gerstenblith G, Messina E, Giacomello A, Marban E, Circ. Res., 106, 971 (2010)
Gnecchi M, He H, Noiseux N, Liang OD, Zhang L, Morello F, Mu H, Melo LG, Pratt RE, Ingwall JS, Faseb J., 20, 661 (2006)
Deuse T, Peter C, Fedak PW, Doyle T, Reichenspurner H, Zimmermann WH, Eschenhagen T, Stein W, Wu JC, Robbins RC, Circulation, 120, S247 (2009)
Gnecchi M, Zhang Z, Ni A, Dzau VJ, Circ. Res., 103, 1204 (2008)
Rogers TB, Pati S, Gaa S, Riley D, Khakoo AY, Patel S, Wardlow RD, Frederick CA, Hall G, He LP, J. Mol. Cell. Cardiol., 50, 346 (2011)
Kota J, Chivukula RR, O'Donnell KA, Wentzel EA, Montgomery CL, Hwang HW, Chang TC, Vivekanandan P, Torbenson M, Clark KR, Cell, 137, 1005 (2009)
Alvarez-Erviti L, Seow Y, Yin H, Betts C, Lakhal S, Wood MJ, Nat. Biotechnol., 29, 341 (2011)
Li H, Zuo S, He Z, Yang Y, Pasha Z, Wang Y, Xu M, Am. J. Physiol. Heart Circ. Physiol., 299, H1772 (2010)
Timmers L, Lim SK, Arslan F, Armstrong JS, Hoefer IE, Doevendans PA, Piek JJ, El Oakley RM, Choo A, Lee CN, Stem Cell Res., 1, 129 (2008)
Lai RC, Arslan F, Lee MM, Sze NSK, Choo A, Chen TS, Salto-Tellez M, Timmers L, Lee CN, El Oakley RM, Stem. Cell. Res., 4, 214 (2010)
Lim KS, Cha MJ, Kim JK, Park EJ, Chae JW, Rhim T, Hwang KC, Kim YH, J. Control. Release, 169, 306 (2013)
Kang YJ, Li Y, Sun X, Sun X, Am. J. Physiol. Heart Circ. Physiol, 163, 1579 (2003)
Kang YJ, Li G, Saari JT, Am. J. Physiol. Heart Circ. Physiol, 276, H993 (1999)
Watson R, Redmond HP, Wang JH, Bouchier-Hayes D, J. Leukoc. Biol., 60, 625 (1996)
Rehman J, Traktuev D, Li J, Merfeld-Clauss S, Temm-Grove CJ, Bovenkerk JE, Pell CL, Johnstone BH, Considine RV, March KL, Circulation, 109, 1292 (2004)
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR, Science, 284, 143 (1999)
Tang YL, Zhao Q, Zhang YC, Cheng L, Liu M, Shi J, Yang YZ, Pan C, Ge J, Phillips MI, Regul. Pept., 117, 3 (2004)
Zhuang X, Xiang X, Grizzle W, Sun D, Zhang S, Axtell RC, Ju S, Mu J, Zhang L, Steinman L, Mol. Ther., 19, 1769 (2011)
Zhang Y, Chopp M, Meng Y, Katakowski M, Xin H, Mahmood A, Xiong Y, J. Neurosurg., 122, 856 (2015)
Jiang XC, Gao JQ, Int. J. Pharm., 521, 167 (2017)
Kumar L, Verma S, Vaidya B, Gupta V, Curr. Pharm. Des., 21, 4556 (2015)
Huang L, Ma W, Ma Y, Feng D, Chen H, Cai B, Int. J. Biol. Sci., 11, 238 (2015)
William R, Watson G, Redmond HP, Wang JH, Bouchier-Hayes D, J. Leukoc. Biol., 60, 625 (1996)

[Related Articles]

[1] Lloyd-Jones D, Adams R, Carnethon M, De Simone G, Ferguson TB, et al., Circulation, 119, e21 (2009)

[2] Pittenger MF, Martin BJ, Circ. Res., 95, 9 (2004)

[3] Hatzistergos KE, Quevedo H, Oskouei BN, Hu Q, Feigenbaum GS, Margitich IS, Mazhari R, Boyle AJ, Zambrano JP, Rodriguez JE, Circ. Res., 107, 913 (2010)

[4] Nagaya N, Kangawa K, Itoh T, Iwase T, Murakami S, Miyahara Y, Fujii T, Uematsu M, Ohgushi H, Yamagishi M, Circulation, 112, 1128 (2005)

[5] Piao H, Youn TJ, Kwon JS, Kim YH, Bae JW, Kim DW, Cho MC, Lee MM, Park YB, Eur. J. Heart Fail., 7, 730 (2005)

[6] Quevedo HC, Hatzistergos KE, Oskouei BN, Feigenbaum GS, Rodriguez JE, Valdes D, Pattany PM, Zambrano JP, Hu Q, McNiece I, Proc. Natl. Acad. Sci., 106, 14022 (2009)

[7] Stamm C, Westphal B, Kleine HD, Petzsch M, Kittner C, Klinge H, Schumichen C, Nienaber CA, Freund M, Steinhoff G, Lancet, 361, 45 (2003)

[8] Meyer GP, Wollert KC, Lotz J, Steffens J, Lippolt P, Fichtner S, Hecker H, Schaefer A, Arseniev L, Hertenstein B, Circulation, 113, 1287 (2006)

[9] Kwak BR. Mach F, Nat. Med., 11, 367 (2005)

[10] Duran JM, Makarewich CA, Sharp TE, Starosta T, Zhu F, Hoffman NE, Chiba Y, Madesh M, Berretta RM, Kubo H, Circ. Res., 113, 539 (2013)

[11] Mureli S, Gans CP, Bare DJ, Geenen DL, Kumar NM, Banach K, Am. J. Physiol. Heart Circ. Physiol., 304, H600 (2013)

[12] Mummery CL, Davis RP, Krieger JE, Sci. Transl. Med., 2, 27ps17 (2010)

[13] Caplan AI, Dennis JE, J. Cell. Biochem., 98, 1076 (2006)

[14] Gnecchi M, Danieli P, Cervio E, Vascul. Pharmacol., 57, 48 (2012)

[15] Chimenti I, Smith RR, Li TS, Gerstenblith G, Messina E, Giacomello A, Marban E, Circ. Res., 106, 971 (2010)

[16] Gnecchi M, He H, Noiseux N, Liang OD, Zhang L, Morello F, Mu H, Melo LG, Pratt RE, Ingwall JS, Faseb J., 20, 661 (2006)

[17] Deuse T, Peter C, Fedak PW, Doyle T, Reichenspurner H, Zimmermann WH, Eschenhagen T, Stein W, Wu JC, Robbins RC, Circulation, 120, S247 (2009)

[18] Gnecchi M, Zhang Z, Ni A, Dzau VJ, Circ. Res., 103, 1204 (2008)

[19] Rogers TB, Pati S, Gaa S, Riley D, Khakoo AY, Patel S, Wardlow RD, Frederick CA, Hall G, He LP, J. Mol. Cell. Cardiol., 50, 346 (2011)

[20] Kota J, Chivukula RR, O'Donnell KA, Wentzel EA, Montgomery CL, Hwang HW, Chang TC, Vivekanandan P, Torbenson M, Clark KR, Cell, 137, 1005 (2009)

[21] Alvarez-Erviti L, Seow Y, Yin H, Betts C, Lakhal S, Wood MJ, Nat. Biotechnol., 29, 341 (2011)

[22] Li H, Zuo S, He Z, Yang Y, Pasha Z, Wang Y, Xu M, Am. J. Physiol. Heart Circ. Physiol., 299, H1772 (2010)

[23] Timmers L, Lim SK, Arslan F, Armstrong JS, Hoefer IE, Doevendans PA, Piek JJ, El Oakley RM, Choo A, Lee CN, Stem Cell Res., 1, 129 (2008)

[24] Lai RC, Arslan F, Lee MM, Sze NSK, Choo A, Chen TS, Salto-Tellez M, Timmers L, Lee CN, El Oakley RM, Stem. Cell. Res., 4, 214 (2010)

[25] Lim KS, Cha MJ, Kim JK, Park EJ, Chae JW, Rhim T, Hwang KC, Kim YH, J. Control. Release, 169, 306 (2013)

[26] Kang YJ, Li Y, Sun X, Sun X, Am. J. Physiol. Heart Circ. Physiol, 163, 1579 (2003)

[27] Kang YJ, Li G, Saari JT, Am. J. Physiol. Heart Circ. Physiol, 276, H993 (1999)

[28] Watson R, Redmond HP, Wang JH, Bouchier-Hayes D, J. Leukoc. Biol., 60, 625 (1996)

[29] Rehman J, Traktuev D, Li J, Merfeld-Clauss S, Temm-Grove CJ, Bovenkerk JE, Pell CL, Johnstone BH, Considine RV, March KL, Circulation, 109, 1292 (2004)

[30] Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR, Science, 284, 143 (1999)

[31] Tang YL, Zhao Q, Zhang YC, Cheng L, Liu M, Shi J, Yang YZ, Pan C, Ge J, Phillips MI, Regul. Pept., 117, 3 (2004)

[32] Zhuang X, Xiang X, Grizzle W, Sun D, Zhang S, Axtell RC, Ju S, Mu J, Zhang L, Steinman L, Mol. Ther., 19, 1769 (2011)

[33] Zhang Y, Chopp M, Meng Y, Katakowski M, Xin H, Mahmood A, Xiong Y, J. Neurosurg., 122, 856 (2015)

[34] Jiang XC, Gao JQ, Int. J. Pharm., 521, 167 (2017)

[35] Kumar L, Verma S, Vaidya B, Gupta V, Curr. Pharm. Des., 21, 4556 (2015)

[36] Huang L, Ma W, Ma Y, Feng D, Chen H, Cai B, Int. J. Biol. Sci., 11, 238 (2015)

[37] William R, Watson G, Redmond HP, Wang JH, Bouchier-Hayes D, J. Leukoc. Biol., 60, 625 (1996)