MicroRNA-based therapies that target cardiomyocyte proliferation have great potential for the treatment of myocardial infarction. In previous work, we showed that the miR-302/367 cluster regulates cardiomyocyte proliferation in the prenatal and postnatal heart. Here, we describe the development and application of an injectable hyaluronic acid hydrogel for the local and sustained delivery of miR-302 mimics to the heart. We show that the miR-302 mimics released in vitro promoted cardiomyocyte proliferation over one week, and that a single injection of the hydrogel in the mouse heart led to local and sustained cardiomyocyte proliferation for two weeks. After myocardial infarction, gel–miR-302 injection caused local clonal proliferation and increased cardiomyocyte numbers in the border zone of a Confetti mouse model. Gel–miR-302 further decreased cardiac end-diastolic (39%) and end-systolic (50%) volumes, and improved ejection fraction (32%) and fractional shortening (64%) four weeks after myocardial infarction and injection, compared with controls. Our findings suggest that biomaterial-based miRNA delivery systems can lead to improved outcomes via cardiac regeneration after myocardial infarction.
This is a preview of subscription content
Subscribe to Nature+
Get immediate online access to the entire Nature family of 50+ journals
Subscribe to Journal
Get full journal access for 1 year
only $4.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Townsend, N. et al. Cardiovascular disease in Europe: epidemiological update 2016. Eur. Heart J.37, 3232–3245 (2016).
Mozaffarian, D. et al. Heart disease and stroke statistics—2015 update: a report from the American Heart Association. Circulation131, e29–322 (2014).
Pasumarthi, K. B. S. Cardiomyocyte cell cycle regulation. Circ. Res.90, 1044–1054 (2002).
Jameel, M. N. & Zhang, J. Stem cell therapy for ischaemic heart disease. Antioxid. Redox Signal.13, 1879–1897 (2010).
Leone, M., Magadum, A. & Engel, F. B. Cardiomyocyte proliferation in cardiac development and regeneration: a guide to methodologies and interpretations. Am. J. Physiol. Heart Circ. Physiol.309, H1237–H1250 (2015).
Laflamme, M. A. & Murry, C. E. Heart regeneration. Nature473, 326–335 (2011).
Li, Y. et al. Acute myocardial infarction induced functional cardiomyocytes to re-enter the cell cycle. Am. J. Transl. Res.5, 327–335 (2013).
Orlic, D. et al. Bone marrow cells regenerate infarcted myocardium. Nature410, 701–705 (2001).
Elnakish, M. T. et al. Mesenchymal stem cells for cardiac regeneration: translation to bedside reality. Stem Cells Int.2012, 1–14 (2012).
Gavira, J. J. et al. Autologous skeletal myoblast transplantation in patients with nonacute myocardial infarction: 1-year follow-up. J. Thorac. Cardiovasc. Surg.131, 799–804 (2006).
Hodgson, D. M. et al. Stable benefit of embryonic stem cell therapy in myocardial infarction. Am. J. Physiol. Heart Circ. Physiol.287, H471–H479 (2004).
Lalit, P. A., Hei, D. J., Raval, A. N. & Kamp, T. J. Induced pluripotent stem cells for post-myocardial infarction repair: remarkable opportunities and challenges. Circ. Res.114, 1328–1345 (2014).
Korf-Klingebiel, M. et al. Myeloid-derived growth factor (C19orf10) mediates cardiac repair following myocardial infarction. Nat. Med.21, 140–149 (2015).
Ni, T. T. et al. Discovering small molecules that promote cardiomyocyte generation by modulating Wnt signaling. Chem. Biol.18, 1658–68 (2011).
Cheng, Y.-Y. et al. Reprogramming-derived gene cocktail increases cardiomyocyte proliferation for heart regeneration. EMBO Mol. Med.9, 251–264 (2017).
Liang, D. et al. miRNA-204 drives cardiomyocyte proliferation via targeting Jarid2. Int. J. Cardiol.201, 38–48 (2015).
Eulalio, A. et al. Functional screening identifies miRNAs inducing cardiac regeneration. Nature492, 376–381 (2012).
Tian, Y. et al. A microRNA-Hippo pathway that promotes cardiomyocyte proliferation and cardiac regeneration in mice. Sci. Transl. Med.7, 279ra38 (2015).
Panda, N. C. et al. Improved conduction and increased cell retention in healed MI using mesenchymal stem cells suspended in alginate hydrogel. J. Interv. Card. Electrophysiol.41, 117–127 (2014).
Teng, C. J., Luo, J., Chiu, R. C. J. & Shum-Tim, D. Massive mechanical loss of microspheres with direct intramyocardial injection in the beating heart: implications for cellular cardiomyoplasty. J. Thorac. Cardiovasc. Surg.132, 628–632 (2006).
Tous, E., Purcell, B., Ifkovits, J. L. & Burdick, J. A. Injectable acellular hydrogels for cardiac repair. J. Cardiovasc. Transl. Res.4, 528–542 (2011).
Gaffey, A. C. et al. Injectable shear-thinning hydrogels used to deliver endothelial progenitor cells, enhance cell engraftment, and improve ischaemic myocardium. J. Thorac. Cardiovasc. Surg.150, 1268–1277 (2015).
Rodell, C. B. et al. Shear-thinning supramolecular hydrogels with secondary autonomous covalent crosslinking to modulate viscoelastic properties in vivo. Adv. Funct. Mater.25, 636–644 (2014).
Rodell, C. B., Kaminski, A. L. & Burdick, J. A. Rational design of network properties in guest–host assembled and shear-thinning hyaluronic acid hydrogels. Biomacromolecules14, 4125–4134 (2013).
Rodell, C. B. et al. Injectable shear-thinning hydrogels for minimally invasive delivery to infarcted myocardium to limit left ventricular remodeling. Circ. Cardiovasc. Interv.9, e004058 (2016).
Wang, L. L. et al. Injectable, guest–host assembled polyethylenimine hydrogel for siRNA delivery. Biomacromolecules18, 77–86 (2016).
Seif-Naraghi, S. B. et al. Safety and efficacy of an injectable extracellular matrix hydrogel for treating myocardial infarction. Sci. Transl. Med.5, 173ra25 (2013).
Guvendiren, M., Lu, H. D. & Burdick, J. A. Shear-thinning hydrogels for biomedical applications. Soft Matter8, 260–272 (2012).
Mealy, J. E., Rodell, C. B. & Burdick, J. A. Sustained small molecule delivery from injectable hyaluronic acid hydrogels through host–guest mediated retention. J. Mater. Chem. B3, 8010–8019 (2015).
Liu, Y. M. et al. Cholesterol-conjugated let-7a mimics: antitumor efficacy on hepatocellular carcinoma in vitro and in a preclinical orthotopic xenograft model of systemic therapy. BMC Cancer14, 889 (2014).
Shim, M. S. & Kwon, Y. J. Efficient and targeted delivery of siRNA in vivo. FEBS J.277, 4814–27 (2010).
Wolfrum, C. et al. Mechanisms and optimization of in vivo delivery of lipophilic siRNAs. Nat. Biotechnol.25, 1149–1157 (2007).
Wang, L. L. & Burdick, J. A. Engineered hydrogels for local and sustained delivery of RNA-interference therapies. Adv. Healthc. Mater.6, 1601041 (2016).
van de Manakker, F., van der Pot, M., Vermonden, T., van Nostrum, C. F. & Hennink, W. E. Self-assembling hydrogels based on β-cyclodextrin/cholesterol inclusion complexes. Macromolecules41, 1766–1773 (2008).
López, C. A. et al. Molecular mechanism of cyclodextrin mediated cholesterol extraction. PLoS Comput. Biol.7, e1002020 (2011).
Politzer, I. R. et al. Effect of β-cyclodextrin on the fluorescence, absorption and lasing of rhodamine 6G, rhodamine B and fluorescein disodium salt in aqueous solutions. Chem. Phys. Lett.159, 258–262 (1989).
Mondal, A. & Jana, N. R. Fluorescent detection of cholesterol using β-cyclodextrin functionalized graphene. Chem. Commun.48, 7316 (2012).
López, C. A., de Vries, A. H. & Marrink, S. J. Molecular mechanism of cyclodextrin mediated cholesterol extraction. PLoS Comput. Biol.7, e1002020 (2011).
Senyo, S. E. et al. Mammalian heart renewal by pre-existing cardiomyocytes. Nature493, 433–436 (2012).
Ali, S. R. et al. Existing cardiomyocytes generate cardiomyocytes at a low rate after birth in mice. Proc. Natl Acad. Sci. USA111, 8850–8855 (2014).
Snippert, H. J. et al. Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell143, 134–144 (2010).
Lescroart, F. et al. Early lineage restriction in temporally distinct populations of Mesp1 progenitors during mammalian heart development. Nat. Cell Biol.16, 829–840 (2014).
Rios, A. C., Fu, N. Y., Lindeman, G. J. & Visvader, J. E. In situ identification of bipotent stem cells in the mammary gland. Nature506, 322–327 (2014).
Sohal, D. S. et al. Temporally regulated and tissue-specific gene manipulations in the adult and embryonic heart using a tamoxifen-inducible Cre protein. Circ. Res.89, 20–25 (2001).
Frank, D. B. et al. Emergence of a wave of Wnt signaling that regulates lung alveologenesis by controlling epithelial self-renewal and differentiation. Cell Rep.17, 2312–2325 (2016).
Farin, H. F. et al. Visualization of a short-range Wnt gradient in the intestinal stem-cell niche. Nature530, 340–343 (2016).
Farin, H. F., Van Es, J. H. & Clevers, H. Redundant sources of Wnt regulate intestinal stem cells and promote formation of Paneth cells. Gastroenterology143, 1518–1529.e7 (2012).
Burdick, J. A. & Prestwich, G. D. Hyaluronic acid hydrogels for biomedical applications. Adv. Mater.23, H41–H56 (2011).
Tian, Y. & Morrisey, E. E. Importance of myocyte-nonmyocyte interactions in cardiac development and disease. Circ. Res.110, 1023–1034 (2012).
Yang, Y. et al. MicroRNA-34a plays a key role in cardiac repair and regeneration following myocardial infarction. Circ. Res.117, 450–459 (2015).
Lesizza, P. et al. Single-dose intracardiac injection of pro-regenerative microRNAs improves cardiac function after myocardial infarction. Circ. Res.120, 1298–1304 (2017).
Monaghan, M. G. et al. Exogenous miR-29B delivery through a hyaluronan-based injectable system yields functional maintenance of the infarcted myocardium. Tissue Eng. Part A https://doi.org/10.1089/ten.TEA.2016.0527 (2017).
Pandey, R. et al. MicroRNA-1825 induces proliferation of adult cardiomyocytes and promotes cardiac regeneration post ischaemic injury. Am. J. Transl. Res.9, 3120–3137 (2017).
Patel, R. S. et al. High resolution of microRNA signatures in human whole saliva. Arch. Oral Biol.56, 1506–1513 (2011).
Shcherbakova, D. M. & Verkhusha, V. V. Near-infrared fluorescent proteins for multicolor in vivo imaging. Nat. Methods10, 751–754 (2013).
The authors thank C. Loebel for assistance with manuscript revisions, C. Chen, C. Venkataraman, A. Trubelja, S. Zaman, J. Gordon and F. Arisi for assistance with mouse surgeries and histology, J. Galarraga and C. Rodell for material contribution and helpful discussion, S. Schultz of the Penn Small Animal Imaging Facility for assistance with echocardiography, and the Penn Histology and Gene Expression Core. This work was made possible by financial support from the American Heart Association through an established investigator award (J.A.B.) and predoctoral fellowship (L.L.W.), and the National Institutes of Health (F30 HL134255, UO1 HL100405, U01 HL134745).
Provisional patents concerning the technology described in this work have been filed.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Wang, L.L., Liu, Y., Chung, J.J. et al. Sustained miRNA delivery from an injectable hydrogel promotes cardiomyocyte proliferation and functional regeneration after ischaemic injury. Nat Biomed Eng 1, 983–992 (2017). https://doi.org/10.1038/s41551-017-0157-y
NPG Asia Materials (2022)
Matrix Metalloproteinase-Targeted SPECT/CT Imaging for Evaluation of Therapeutic Hydrogels for the Early Modulation of Post-Infarct Myocardial Remodeling
Journal of Cardiovascular Translational Research (2022)
Cell Regeneration (2021)
Cell Regeneration (2021)
Minimally invasive delivery of therapeutic agents by hydrogel injection into the pericardial cavity for cardiac repair
Nature Communications (2021)