The Low Tumorigenic Risk and Subtypes of Cardiomyocytes Derived from Human-induced Pluripotent Stem Cells

被引:0
作者
Lu, Jizhen [1 ]
Zhang, Lu [1 ]
Cao, Hongxia [1 ]
Ma, Xiaoxue [1 ]
Bai, Zhihui [1 ]
Zhu, Hanyu [1 ]
Qi, Yiyao [1 ]
Zhang, Shoumei [1 ]
Zhang, Peng [2 ,3 ,4 ,5 ]
He, Zhiying [1 ]
Yang, Huangtian [2 ,3 ,4 ,5 ]
Liu, Zhongmin [1 ]
Jia, Wenwen [1 ]
机构
[1] Tongji Univ, Shanghai East Hosp, East Hosp, Natl Stem Cell Translat Resource Ctr,GMP Lab Stem, Shanghai, Peoples R China
[2] Tongji Univ, Shanghai East Hosp, Translat Med Ctr Stem Cell Therapy, Sch Med, Shanghai, Peoples R China
[3] Tongji Univ, Shanghai East Hosp, Inst Heart Failure & Regenerat Med, Sch Med, Shanghai, Peoples R China
[4] Shanghai Inst Stem Cell Res & Clin Translat, Shanghai, Peoples R China
[5] Univ Chinese Acad Sci CAS, Shanghai Inst Nutr & Hlth, Lab Mol Cardiol, Shanghai, Peoples R China
关键词
hiPSC; cardiomyocytes; single-cell RNA sequence; cardiac subtypes; whole exome sequence; tumorigenicity; CARDIAC REGENERATIVE THERAPY; MYOCARDIAL-INFARCTION; HEART; GENE; MUSCLE; CYCLE; DIFFERENTIATION; TRANSPLANTATION; PROLIFERATION; PURIFICATION;
D O I
10.2174/011574888X318139240621051224
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Background Clinical application of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is a promising approach for the treatment of heart diseases. However, the tumorigenicity of hiPSC-CMs remains a concern for their clinical applications and the composition of the hiPSC-CM subtypes need to be clearly identified.Methods In the present study, hiPSC-CMs were induced from hiPSCs via modulation of Wnt signaling followed by glucose deprivation purification. The structure, function, subpopulation composition, and tumorigenic risk of hiPSC-CMs were evaluated by single-cell RNA sequencing (scRNAseq), whole exome sequencing (WES), and integrated molecular biology, cell biology, electrophysiology, and/or animal experiments.Results The high purity of hiPSC-CMs, determined by flow cytometry analysis, was generated. ScRNAseq analysis of differentiation day (D) 25 hiPSC-CMs did not identify the transcripts representative of undifferentiated hiPSCs. WES analysis showed a few newly acquired confidently identified mutations and no mutations in tumor susceptibility genes. Further, no tumor formation was observed after transplanting hiPSC-CMs into NOD-SCID mice for 3 months. Moreover, D25 hiPSC-CMs were composed of subtypes of ventricular-like cells (23.19%) and atrial-like cells (66.45%) in different cell cycle stages or mature levels, based on the scRNAseq analysis. Furthermore, a subpopulation of more mature ventricular cells (3.21%) was identified, which displayed significantly up-regulated signaling pathways related to myocardial contraction and action potentials. Additionally, a subpopulation of cardiomyocytes in an early differentiation stage (3.44%) experiencing nutrient stress-induced injury and heading toward apoptosis was observed.Conclusions This study confirmed the biological safety of hiPSC-CMs and described the composition and expression profile of cardiac subtypes in hiPSC-CMs which provide standards for quality control and theoretical supports for the translational applications of hiPSC-CMs.
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页码:317 / 335
页数:19
相关论文
共 75 条
[21]   Interaction of the Joining Region in Junctophilin-2 With the L-Type Ca2+ Channel Is Pivotal for Cardiac Dyad Assembly and Intracellular Ca2+ Dynamics [J].
Gross, Polina ;
Johnson, Jaslyn ;
Romero, Carlos M. ;
Eaton, Deborah M. ;
Poulet, Claire ;
Sanchez-Alonso, Jose ;
Lucarelli, Carla ;
Ross, Jean ;
Gibb, Andrew A. ;
Garbincius, Joanne F. ;
Lambert, Jonathan ;
Varol, Erdem ;
Yang, Yijun ;
Wallner, Markus ;
Feldsott, Eric A. ;
Kubo, Hajime ;
Berretta, Remus M. ;
Yu, Daohai ;
Rizzo, Victor ;
Elrod, John ;
Sabri, Abdelkarim ;
Gorelik, Julia ;
Chen, Xiongwen ;
Houser, Steven R. .
CIRCULATION RESEARCH, 2021, 128 (01) :92-114
[22]   Ryanodine receptor 2 (RYR2) dysfunction activates the unfolded protein response and perturbs cardiomyocyte maturation [J].
Guo, Yuxuan ;
Cao, Yangpo ;
Jardin, Blake D. ;
Zhang, Xiaoran ;
Zhou, Pingzhu ;
Guatimosim, Silvia ;
Lin, Junsen ;
Chen, Zhan ;
Zhang, Yueyang ;
Mazumdar, Neil ;
Lu, Fujian ;
Ma, Qing ;
Lu, Yao-Wei ;
Zhao, Mingming ;
Wang, Da-Zhi ;
Dong, Erdan ;
Pu, William T. .
CARDIOVASCULAR RESEARCH, 2023, 119 (01) :221-235
[23]   Distinctive Clinical and Pathologic Features of Immature Teratomas Arising from Induced Pluripotent Stem Cell-Derived Beta Cell Injection in a Diabetes Patient [J].
Han, Lei ;
He, Hao ;
Yang, Yihao ;
Meng, Qingyin ;
Ye, Fan ;
Chen, Gong ;
Zhang, Jing .
STEM CELLS AND DEVELOPMENT, 2022, 31 (5-6) :97-101
[24]   CACNA1C (CaV1.2) and other L-type calcium channels in the pathophysiology and treatment of psychiatric disorders: Advances from functional genomics and pharmacoepidemiology [J].
Harrison, Paul J. ;
Husain, Syed M. ;
Lee, Hami ;
De Los Angeles, Alejandro ;
Colbourne, Lucy ;
Mould, Arne ;
Hall, Nicola A. L. ;
Haerty, Wilfried ;
Tunbridge, Elizabeth M. .
NEUROPHARMACOLOGY, 2022, 220
[25]   Delineation of target expression profiles in CD34+/CD38- and CD34+/CD38+ stem and progenitor cells in AML and CML [J].
Herrmann, Harald ;
Sadovnik, Irina ;
Eisenwort, Gregor ;
Ruelicke, Thomas ;
Blatt, Katharina ;
Herndlhofer, Susanne ;
Willmann, Michael ;
Stefanzl, Gabriele ;
Baumgartner, Sigrid ;
Greiner, Georg ;
Schulenburg, Axel ;
Mueller, Niklas ;
Rabitsch, Werner ;
Bilban, Martin ;
Hoermann, Gregor ;
Streube, Berthold ;
Vallera, Daniel A. ;
Sperr, Wolfgang R. ;
Valent, Peter .
BLOOD ADVANCES, 2020, 4 (20) :5118-5132
[26]   Robustness and applicability of transcription factor and pathway analysis tools on single-cell RNA-seq data [J].
Holland, Christian H. ;
Tanevski, Jovan ;
Perales-Paton, Javier ;
Gleixner, Jan ;
Kumar, Manu P. ;
Mereu, Elisabetta ;
Joughin, Brian A. ;
Stegle, Oliver ;
Lauffenburger, Douglas A. ;
Heyn, Holger ;
Szalai, Bence ;
Saez-Rodriguez, Julio .
GENOME BIOLOGY, 2020, 21 (01)
[27]   Establishment of a protocol to administer immunosuppressive drugs for iPS cell-derived cardiomyocyte patch transplantation in a rat myocardial infarction model [J].
Ito, Emiko ;
Kawamura, Ai ;
Kawamura, Takuji ;
Takeda, Maki ;
Harada, Akima ;
Mochizuki-Oda, Noriko ;
Sawa, Yoshiki ;
Miyagawa, Shigeru .
SCIENTIFIC REPORTS, 2023, 13 (01)
[28]   Tumorigenicity assay essential for facilitating safety studies of hiPSC-derived cardiomyocytes for clinical application [J].
Ito, Emiko ;
Miyagawa, Shigeru ;
Takeda, Maki ;
Kawamura, Ai ;
Harada, Akima ;
Iseoka, Hiroko ;
Yajima, Shin ;
Sougawa, Nagako ;
Mochizuki-Oda, Noriko ;
Yasuda, Satoshi ;
Sato, Yoji ;
Sawa, Yoshiki .
SCIENTIFIC REPORTS, 2019, 9 (1)
[29]   Transmural myocardial repair with engineered heart muscle in a rat model of heterotopic heart transplantation - A proof-of-concept study [J].
Jebran, Ahmad-Fawad ;
Tiburcy, Malte ;
Biermann, Daniel ;
Balfanz, Paul ;
Didie, Michael ;
Karikkineth, Bijoy Chandapillai ;
Schoendube, Friedrich ;
Kutschka, Ingo ;
Zimmermann, Wolfram-Hubertus .
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY, 2022, 168 :3-12
[30]   An induced pluripotent stem cell line (EHTJUi003-A) generated from a neonate with c.1377delC mutation in the gene MYBPC3 causing hypertrophic cardiomyopathy [J].
Jia, Wen-Wen ;
Lu, Ji-Zhen ;
Zhang, Lu ;
Cao, Hong-Xia ;
Qi, Yi-Yao ;
Zhu, Han-Yu ;
Bai, Zhi-Hui ;
Zhang, Shou-Mei ;
Qiao, Zhi-Bin ;
Bao, Yan ;
Liu, Zhong-Min .
STEM CELL RESEARCH, 2021, 53