Seamless and Non-destructive Monitoring of Extracellular MicroRNAs During Cardiac Differentiation from Human Pluripotent Stem Cells

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2023 Sep 22

PMID 37738969

Authors

Otoya Sekine

Sayaka Kanaami

Kanako Masumoto

Yuki Aihara

Yuika Morita-Umei

Hidenori Tani

Yusuke Soma

Tomohiko C Umei

Kotaro Haga

Taijun Moriwaki

Yujiro Kawai

Masatoshi Ohno

Yoshikazu Kishino

Hideaki Kanazawa

Keiichi Fukuda

Masaki Ieda

Shugo Tohyama

Affiliations

Soon will be listed here.

Abstract

Monitoring cardiac differentiation and maturation from human pluripotent stem cells (hPSCs) and detecting residual undifferentiated hPSCs are indispensable for the development of cardiac regenerative therapy. MicroRNA (miRNA) is secreted from cells into the extracellular space, and its role as a biomarker is attracting attention. Here, we performed an miRNA array analysis of supernatants during the process of cardiac differentiation and maturation from hPSCs. We demonstrated that the quantification of extracellular miR-489-3p and miR-1/133a-3p levels enabled the monitoring of mesoderm and cardiac differentiation, respectively, even in clinical-grade mass culture systems. Moreover, extracellular let-7c-5p levels showed the greatest increase with cardiac maturation during long-term culture. We also verified that residual undifferentiated hPSCs in hPSC-derived cardiomyocytes (hPSC-CMs) were detectable by measuring miR-302b-3p expression, with a detection sensitivity of 0.01%. Collectively, we demonstrate that our method of seamlessly monitoring specific miRNAs secreted into the supernatant is non-destructive and effective for the quality evaluation of hPSC-CMs.

References

Miura K, Okada Y, Aoi T, Okada A, Takahashi K, Okita K . Variation in the safety of induced pluripotent stem cell lines. Nat Biotechnol. 2009; 27(8):743-5. DOI: 10.1038/nbt.1554. View

Zhou M, Hara H, Dai Y, Mou L, Cooper D, Wu C . Circulating Organ-Specific MicroRNAs Serve as Biomarkers in Organ-Specific Diseases: Implications for Organ Allo- and Xeno-Transplantation. Int J Mol Sci. 2016; 17(8). PMC: 5000630. DOI: 10.3390/ijms17081232. View

Kuppusamy K, Jones D, Sperber H, Madan A, Fischer K, Rodriguez M . Let-7 family of microRNA is required for maturation and adult-like metabolism in stem cell-derived cardiomyocytes. Proc Natl Acad Sci U S A. 2015; 112(21):E2785-94. PMC: 4450404. DOI: 10.1073/pnas.1424042112. View

Skelton R, Brady B, Khoja S, Sahoo D, Engel J, Arasaratnam D . CD13 and ROR2 Permit Isolation of Highly Enriched Cardiac Mesoderm from Differentiating Human Embryonic Stem Cells. Stem Cell Reports. 2016; 6(1):95-108. PMC: 4720015. DOI: 10.1016/j.stemcr.2015.11.006. View

Lakshmipathy U, Davila J, Hart R . miRNA in pluripotent stem cells. Regen Med. 2010; 5(4):545-55. PMC: 3077715. DOI: 10.2217/rme.10.34. View

Nakanishi H, Miki K, Komatsu K, Umeda M, Mochizuki M, Inagaki A . Monitoring and visualizing microRNA dynamics during live cell differentiation using microRNA-responsive non-viral reporter vectors. Biomaterials. 2017; 128:121-135. DOI: 10.1016/j.biomaterials.2017.02.033. View

Khush K, Cherikh W, Chambers D, Goldfarb S, Hayes Jr D, Kucheryavaya A . The International Thoracic Organ Transplant Registry of the International Society for Heart and Lung Transplantation: Thirty-fifth Adult Heart Transplantation Report-2018; Focus Theme: Multiorgan Transplantation. J Heart Lung Transplant. 2018; 37(10):1155-1168. DOI: 10.1016/j.healun.2018.07.022. View

Bock C, Kiskinis E, Verstappen G, Gu H, Boulting G, Smith Z . Reference Maps of human ES and iPS cell variation enable high-throughput characterization of pluripotent cell lines. Cell. 2011; 144(3):439-52. PMC: 3063454. DOI: 10.1016/j.cell.2010.12.032. View

Zhang B, Ji S, Ma F, Ma Q, Lu X, Chen X . miR-489 acts as a tumor suppressor in human gastric cancer by targeting PROX1. Am J Cancer Res. 2016; 6(9):2021-2030. PMC: 5043111. View

10.

Cordes K, Srivastava D . MicroRNA regulation of cardiovascular development. Circ Res. 2009; 104(6):724-32. PMC: 2664538. DOI: 10.1161/CIRCRESAHA.108.192872. View

11.

Pigati L, Yaddanapudi S, Iyengar R, Kim D, Hearn S, Danforth D . Selective release of microRNA species from normal and malignant mammary epithelial cells. PLoS One. 2010; 5(10):e13515. PMC: 2958125. DOI: 10.1371/journal.pone.0013515. View

12.

Kawaguchi S, Soma Y, Nakajima K, Kanazawa H, Tohyama S, Tabei R . Intramyocardial Transplantation of Human iPS Cell-Derived Cardiac Spheroids Improves Cardiac Function in Heart Failure Animals. JACC Basic Transl Sci. 2021; 6(3):239-254. PMC: 7987543. DOI: 10.1016/j.jacbts.2020.11.017. View

13.

Funakoshi S, Miki K, Takaki T, Okubo C, Hatani T, Chonabayashi K . Enhanced engraftment, proliferation, and therapeutic potential in heart using optimized human iPSC-derived cardiomyocytes. Sci Rep. 2016; 6:19111. PMC: 4705488. DOI: 10.1038/srep19111. View

14.

Bustin S, Benes V, Garson J, Hellemans J, Huggett J, Kubista M . The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009; 55(4):611-22. DOI: 10.1373/clinchem.2008.112797. View

15.

Karbassi E, Fenix A, Marchiano S, Muraoka N, Nakamura K, Yang X . Cardiomyocyte maturation: advances in knowledge and implications for regenerative medicine. Nat Rev Cardiol. 2020; 17(6):341-359. PMC: 7239749. DOI: 10.1038/s41569-019-0331-x. View

16.

Ma X, Li C, Sun L, Huang D, Li T, He X . Lin28/let-7 axis regulates aerobic glycolysis and cancer progression via PDK1. Nat Commun. 2014; 5:5212. DOI: 10.1038/ncomms6212. View

17.

Tohyama S, Fujita J, Hishiki T, Matsuura T, Hattori F, Ohno R . Glutamine Oxidation Is Indispensable for Survival of Human Pluripotent Stem Cells. Cell Metab. 2016; 23(4):663-74. DOI: 10.1016/j.cmet.2016.03.001. View

18.

Lagos-Quintana M, Rauhut R, Yalcin A, Meyer J, Lendeckel W, Tuschl T . Identification of tissue-specific microRNAs from mouse. Curr Biol. 2002; 12(9):735-9. DOI: 10.1016/s0960-9822(02)00809-6. View

19.

Schwarzenbach H, Hoon D, Pantel K . Cell-free nucleic acids as biomarkers in cancer patients. Nat Rev Cancer. 2011; 11(6):426-37. DOI: 10.1038/nrc3066. View

20.

Wang L, Jia Q, Xinnong C, Xie Y, Yang Y, Zhang A . Role of cardiac progenitor cell-derived exosome-mediated microRNA-210 in cardiovascular disease. J Cell Mol Med. 2019; 23(11):7124-7131. PMC: 6815838. DOI: 10.1111/jcmm.14562. View