Generation of a Human Induced Pluripotent Stem Cell Line Expressing a Magnetic Resonance Imaging Reporter Gene

Overview

Journal Small Methods

Specialty Biotechnology

Date 2024 May 6

PMID 38708688

Authors

Yura Son

Pengsheng Li

Dakota Ortega

Huiliang Qiu

Hannah Prachyl

Ming Yang

Wuqiang Zhu

Affiliations

Soon will be listed here.

Abstract

The objective of the current study is to develop a new method for tracking transplanted human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs) using magnetic resonance imaging (MRI). The CRISPR/dCas9 activation system is employed to overexpress ferritin heavy chain (FHC) in hiPSC-CMs. The mRNA and protein expression of FHC in hiPSC and hiPSC-CMs significantly increased after transfection. Iron chloride does not affect the cell viability in a concentration range from 0 to 2000 µm. hiPSCs overexpressing FHC (hiPSC- FHC) and hiPSC-CMs overexpressing FHC (hiPSC-CM-FHC) significantly enhanced cellular uptake of iron chloride but with no changes in electrophysiological properties compared to hiPSC-CM-Control. Furthermore, hiPSC-CM-FHC presented robust contrast and lower T* values, signifying their potential as highly effective candidates for cardiac MRI. Next, hiPSC-CM-FHC is injected into mouse hearts and after 3 days of transplantation, MR images are obtained. hiPSC-CM-FHC cells exhibited clear signals in the hearts with lower T* and rapid signal decay. Collectively, data from this proof-of-concept study demonstrated that endogenous labeling with FHC in hiPSC-CMs can be a potent strategy for enhancing the accuracy of cardiac MRI. This technology represents a significant step forward in tracking the transplanted hiPSC-CMs in the hearts of live animals.

Citing Articles

Sequential delivery of cardioactive drugs via microcapped microneedle patches for improved heart function in post myocardial infarction rats.

He F, Andrabi S, Shi H, Son Y, Qiu H, Xie J Acta Biomater. 2024; 192():235-247.

PMID: 39643223 PMC: 11735313. DOI: 10.1016/j.actbio.2024.12.009.

References

Son Y, Zhu W . Gene Therapy for Cardiomyocyte Renewal: Cell Cycle, a Potential Therapeutic Target. Mol Diagn Ther. 2022; 27(2):129-140. PMC: 10123801. DOI: 10.1007/s40291-022-00625-y. View

Ruggiero A, Thorek D, Guenoun J, Krestin G, Bernsen M . Cell tracking in cardiac repair: what to image and how to image. Eur Radiol. 2011; 22(1):189-204. PMC: 3229694. DOI: 10.1007/s00330-011-2190-7. View

Cromer Berman S, Walczak P, Bulte J . Tracking stem cells using magnetic nanoparticles. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2011; 3(4):343-55. PMC: 3193153. DOI: 10.1002/wnan.140. View

Cunningham C, Arai T, Yang P, McConnell M, Pauly J, Conolly S . Positive contrast magnetic resonance imaging of cells labeled with magnetic nanoparticles. Magn Reson Med. 2005; 53(5):999-1005. DOI: 10.1002/mrm.20477. View

Stepniewski J, Tomczyk M, Andrysiak K, Kraszewska I, Martyniak A, Langrzyk A . Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes, in Contrast to Adipose Tissue-Derived Stromal Cells, Efficiently Improve Heart Function in Murine Model of Myocardial Infarction. Biomedicines. 2020; 8(12). PMC: 7762393. DOI: 10.3390/biomedicines8120578. View

Prasad A, Inesi G . Analysis of calcium transients in cardiac myocytes and assessment of the sarcoplasmic reticulum Ca2+-ATPase contribution. Methods Mol Biol. 2011; 798:411-21. DOI: 10.1007/978-1-61779-343-1_24. View

Garbern J, Lee R . Cardiac stem cell therapy and the promise of heart regeneration. Cell Stem Cell. 2013; 12(6):689-98. PMC: 3756309. DOI: 10.1016/j.stem.2013.05.008. View

Smith M, Harris P, Sayre L, Perry G . Iron accumulation in Alzheimer disease is a source of redox-generated free radicals. Proc Natl Acad Sci U S A. 1997; 94(18):9866-8. PMC: 23283. DOI: 10.1073/pnas.94.18.9866. View

Naumova A, Reinecke H, Yarnykh V, Deem J, Yuan C, Murry C . Ferritin overexpression for noninvasive magnetic resonance imaging-based tracking of stem cells transplanted into the heart. Mol Imaging. 2010; 9(4):201-10. PMC: 4082401. View

10.

Fernandez-Aviles F, San Roman J, Garcia-Frade J, Fernandez M, Penarrubia M, de La Fuente L . Experimental and clinical regenerative capability of human bone marrow cells after myocardial infarction. Circ Res. 2004; 95(7):742-8. DOI: 10.1161/01.RES.0000144798.54040.ed. View

11.

Lunde K, Solheim S, Aakhus S, Arnesen H, Abdelnoor M, Egeland T . Intracoronary injection of mononuclear bone marrow cells in acute myocardial infarction. N Engl J Med. 2006; 355(12):1199-209. DOI: 10.1056/NEJMoa055706. View

12.

Rheault-Henry M, White I, Grover D, Atoui R . Stem cell therapy for heart failure: Medical breakthrough, or dead end?. World J Stem Cells. 2021; 13(4):236-259. PMC: 8080540. DOI: 10.4252/wjsc.v13.i4.236. View

13.

Naumova A, Balu N, Yarnykh V, Reinecke H, Murry C, Yuan C . Magnetic Resonance Imaging Tracking of Graft Survival in the Infarcted Heart: Iron Oxide Particles Versus Ferritin Overexpression Approach. J Cardiovasc Pharmacol Ther. 2014; 19(4):358-367. PMC: 4185000. DOI: 10.1177/1074248414525999. View

14.

Cohen B, Dafni H, Meir G, Harmelin A, Neeman M . Ferritin as an endogenous MRI reporter for noninvasive imaging of gene expression in C6 glioma tumors. Neoplasia. 2005; 7(2):109-17. PMC: 1501126. DOI: 10.1593/neo.04436. View

15.

Burridge P, Keller G, Gold J, Wu J . Production of de novo cardiomyocytes: human pluripotent stem cell differentiation and direct reprogramming. Cell Stem Cell. 2012; 10(1):16-28. PMC: 3255078. DOI: 10.1016/j.stem.2011.12.013. View

16.

Acimovic I, Vilotic A, Pesl M, Lacampagne A, Dvorak P, Rotrekl V . Human pluripotent stem cell-derived cardiomyocytes as research and therapeutic tools. Biomed Res Int. 2014; 2014:512831. PMC: 3996932. DOI: 10.1155/2014/512831. View

17.

Joshi J, Xu B, Rubart M, Chang Y, Bao X, Chaliki H . Optogenetic Control of Engrafted Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Live Mice: A Proof-of-Concept Study. Cells. 2022; 11(6). PMC: 8946017. DOI: 10.3390/cells11060951. View

18.

Meyer G, Wollert K, Lotz J, Steffens J, Lippolt P, Fichtner S . Intracoronary bone marrow cell transfer after myocardial infarction: eighteen months' follow-up data from the randomized, controlled BOOST (BOne marrOw transfer to enhance ST-elevation infarct regeneration) trial. Circulation. 2006; 113(10):1287-94. DOI: 10.1161/CIRCULATIONAHA.105.575118. View

19.

Bernstein H, Srivastava D . Stem cell therapy for cardiac disease. Pediatr Res. 2012; 71(4 Pt 2):491-9. DOI: 10.1038/pr.2011.61. View

20.

Tejo-Otero A, Fenollosa-Artes F, Achaerandio I, Rey-Vinolas S, Buj-Corral I, Mateos-Timoneda M . Soft-Tissue-Mimicking Using Hydrogels for the Development of Phantoms. Gels. 2022; 8(1). PMC: 8774477. DOI: 10.3390/gels8010040. View