Thyroid hormone plus dual-specificity phosphatase-5 siRNA increases the number of cardiac muscle cells and improves left ventricular contractile function in chronic doxorubicin-injured hearts

• Type of Material: Article
• Authors: Lin Tan, Emory University; Nikolay Bogush, Emory University; Emmen Naqvi, Emory University; John Calvert, Emory University; Robert M Graham, Victor Chang Cardiac Research Institute; W. Robert Taylor, Emory University; Nawazish Naqvi, Emory University; Ahsan Husain, Emory University
• Language: English
• Date: 2021-01-01
• Publisher: IVYSPRING INT PUBL
• Publication Version: Final Published Version
• Copyright Statement: © The author(s)
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.7150/thno.57456
• Title of Journal or Parent Work: THERANOSTICS
• Volume: 11
• Issue: 10
• Start Page: 4790
• End Page: 4808
• Grant/Funding Information: This work was supported by grants from the Department of Medicine, Emory University, the Carlyle Fraser Heart Center, Emory University Hospital Midtown, the NIH (HL079040, HL127726, HL098481, T32HL007745, HL092141, HL093579, HL094373 and HL113452), the American Heart Association (13SDG16460006; 17GRNT33670975), the Fondation Leducq Transatlantic Network, National Health and Medical Research Council, Australia (APP1074386), R.T. Hall estate, and the Australian Research Council Stem Cells Australia Special Initiative in Stem Cell Science grant (SR1101002).
• Keywords: cardiomyocytes; Science & Technology; DUSP5; Research & Experimental Medicine; heart failure; thyroid hormone; Life Sciences & Biomedicine; doxorubicin cardiotoxicity; Medicine, Research & Experimental
• Research Categories: Engineering, Biomedical; Health Sciences, Medicine and Surgery

Abstract

Rationale: Doxorubicin is a widely used anticancer drug. However, its major side effect, cardiotoxicity, results from cardiomyocyte loss that causes left ventricle (LV) wall thinning, chronic LV dysfunction and heart failure. Cardiomyocyte number expansion by thyroid hormone (T3) during preadolescence is suppressed by the developmental induction of an ERK1/2-specific dual specificity phosphatase 5 (DUSP5). Here, we sought to determine if a brief course of combined DUSP5 suppression plus T3 therapy replaces cardiomyocytes lost due to preexisting doxorubicin injury and reverses heart failure. Methods: We used in vivo-jetPEI to deliver DUSP5 or scrambled siRNA to ∼5-week-old C57BL6 mice followed by 5 daily injections of T3 (2 ng/μg body weight). Genetic lineage tracing using Myh6-MerCreMer::Rosa26fs-Confetti mice and direct cardiomyocyte number counting, along with cell cycle inhibition (danusertib), was used to test if this treatment leads to de novo cardiomyocyte generation and improves LV contractile function. Three doses of doxorubicin (20 μg/g) given at 2-weekly intervals, starting at 5-weeks of age in C57BL6 mice, caused severe heart failure, as evident by a decrease in LV ejection fraction. Mice with an ∼40 percentage point decrease in LVEF post-doxorubicin injury were randomized to receive either DUSP5 siRNA plus T3, or scrambled siRNA plus vehicle for T3. Age-matched mice without doxorubicin injury served as controls. Results: In uninjured adult mice, transient therapy with DUSP5 siRNA and T3 increases cardiomyocyte numbers, which is required for the associated increase in LV contractile function, since both are blocked by danusertib. In mice with chronic doxorubicin injury, DUSP5 siRNA plus T3 therapy rebuilds LV muscle by increasing cardiomyocyte numbers, which reverses LV dysfunction and prevents progressive chamber dilatation. Conclusion: RNA therapies are showing great potential. Importantly, a GMP compliant in vivo-jetPEI system for delivery of siRNA is already in use in humans, as is T3. Given these considerations, our findings provide a potentially highly translatable strategy for addressing doxorubicin cardiomyopathy, a currently untreatable condition.