Rapid and highly efficient inducible cardiac gene knockout in adult mice using AAV-mediated expression of Cre recombinase

Werfel, Stanislas; Jungmann, Andreas; Lehmann, Lorenz; Ksienzyk, Jan; Bekeredjian, Raffi; Kaya, Ziya; Leuchs, Barbara; Nordheim, Alfred; Backs, Johannes; Engelhardt, Stefan; Katus, Hugo A.; Müller, Oliver

doi:10.1093/cvr/cvu174

Cited by 77 publications

(62 citation statements)

References 43 publications

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“…Previous data indicated that combining AAV9 and a chicken minimal truncated TNNT2 promoter gave 4640-fold higher expression in the heart than in other organs in mice 37 . Here, we used a human minimal truncated TNNT2 promoter that showed restriction of FLAG-cMyBP-C protein expression to the heart 34 wks following systemic administration in vivo, supporting recent data using the same combination of AAV9 and TNNT2 promoter 13 wks after injection 38 . Serial echocardiographic analyses were done by a highly experienced investigator blinded to the genotype or treatment group.…”

Section: Discussionsupporting

confidence: 83%

Mybpc3 gene therapy for neonatal cardiomyopathy enables long-term disease prevention in mice

et al. 2014

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Homozygous or compound heterozygous frameshift mutations in MYBPC3 encoding cardiac myosin-binding protein C (cMyBP-C) cause neonatal hypertrophic cardiomyopathy (HCM), which rapidly evolves into systolic heart failure and death within the first year of life. Here we show successful long-term Mybpc3 gene therapy in homozygous Mybpc3-targeted knock-in (KI) mice, which genetically mimic these human neonatal cardiomyopathies. A single systemic administration of adeno-associated virus (AAV9)-Mybpc3 in 1-day-old KI mice prevents the development of cardiac hypertrophy and dysfunction for the observation period of 34 weeks and increases Mybpc3 messenger RNA (mRNA) and cMyBP-C protein levels in a dose-dependent manner. Importantly, Mybpc3 gene therapy unexpectedly also suppresses accumulation of mutant mRNAs. This study reports the first successful long-term gene therapy of HCM with correction of both haploinsufficiency and production of poison peptides. In the absence of alternative treatment options except heart transplantation, gene therapy could become a realistic treatment option for severe neonatal HCM.

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Section: Discussionsupporting

confidence: 83%

Mybpc3 gene therapy for neonatal cardiomyopathy enables long-term disease prevention in mice

et al. 2014

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“…Our AAV9-based approach (in which CTRP9 expression is driven by the cytomegalovirus-enhanced myosin light chain promoter) will lead to expression of CTRP9 in the whole heart, mainly in cardiomyocytes and in the liver. 31 This is different from the physiological situation where CTRP9 in the heart is mainly derived from endothelial cells (discussed in detail below) and where no expression in the liver is observed. However, given the technical difficulty to target endothelial cells by an AAV9 approach and in the light of the following facts that (1) CTRP9 is also expressed in cardiomyocytes (although at a much lower level), (2) it can act in these cells in an autocrine manner, and (3) in general, for a secreted factor, its net functional effects rather than its source might be more important, our approach still reveals important information at least as proof of concept.…”

Section: Discussionmentioning

confidence: 80%

C1q-TNF-Related Protein-9 Promotes Cardiac Hypertrophy and Failure

Appari

Breitbart

Brandes

et al. 2017

Circulation Research

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Rationale: Myocardial endothelial cells promote cardiomyocyte hypertrophy, possibly through the release of growth factors. The identity of these factors, however, remains largely unknown, and we hypothesized here that the secreted CTRP9 (C1q-tumor necrosis factor–related protein-9) might act as endothelial-derived protein to modulate heart remodeling in response to pressure overload. Objective: To examine the source of cardiac CTRP9 and its function during pressure overload. Methods and Results: CTRP9 was mainly derived from myocardial capillary endothelial cells. CTRP9 mRNA expression was enhanced in hypertrophic human hearts and in mouse hearts after transverse aortic constriction (TAC). CTRP9 protein was more abundant in the serum of patients with severe aortic stenosis and in murine hearts after TAC. Interestingly, heterozygous and especially homozygous knock-out C1qtnf9 (CTRP9) gene-deleted mice were protected from the development of cardiac hypertrophy, left ventricular dilatation, and dysfunction during TAC. CTRP9 overexpression, in turn, promoted hypertrophic cardiac remodeling and dysfunction after TAC in mice and induced hypertrophy in isolated adult cardiomyocytes. Mechanistically, CTRP9 knock-out mice showed strongly reduced levels of activated prohypertrophic ERK5 (extracellular signal-regulated kinase 5) during TAC compared with wild-type mice, while CTRP9 overexpression entailed increased ERK5 activation in response to pressure overload. Inhibition of ERK5 by a dominant negative MEK5 mutant or by the ERK5/MEK5 inhibitor BIX02189 blunted CTRP9 triggered hypertrophy in isolated adult cardiomyocytes in vitro and attenuated mouse cardiomyocyte hypertrophy and cardiac dysfunction in vivo, respectively. Downstream of ERK5, we identified the prohypertrophic transcription factor GATA4, which was directly activated through ERK5-dependent phosphorylation. Conclusions: The upregulation of CTRP9 during hypertrophic heart disease facilitates maladaptive cardiac remodeling and left ventricular dysfunction and might constitute a therapeutic target in the future.

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“…Importantly, we were able to demonstrate effective pacing and defibrillation of mouse hearts by epicardial illumination even more than 1 year after the gene transfer. The great advantage of optogenetic defibrillation is that this would be, in contrast to electrical shocks, pain-free, because ChR2 expression would be restricted to cardiomyocytes by cardiomyocyte-specific AAV vector capsids (30) or promoters (31). Notably, AAV-mediated transduction will result in spatial heterogeneity of ChR2 expression, which is not expected to be pro-arrhythmic per se since ChR2 has neither a leak current in the absence of illumination nor does it affect the electrophysiological properties of cardiomyocytes (5).…”

Section: Discussionmentioning

confidence: 99%

Optogenetic defibrillation terminates ventricular arrhythmia in mouse hearts and human simulations

Bruegmann¹,

Boyle²,

Vogt³

et al. 2016

Journal of Clinical Investigation

158

182

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Rapid and highly efficient inducible cardiac gene knockout in adult mice using AAV-mediated expression of Cre recombinase

Abstract: AAV9-mediated expression of Cre is a promising approach for rapid and efficient conditional cardiac gene knockout in adult mice.

Cited by 77 publications

References 43 publications

Mybpc3 gene therapy for neonatal cardiomyopathy enables long-term disease prevention in mice

Mybpc3 gene therapy for neonatal cardiomyopathy enables long-term disease prevention in mice

C1q-TNF-Related Protein-9 Promotes Cardiac Hypertrophy and Failure

Optogenetic defibrillation terminates ventricular arrhythmia in mouse hearts and human simulations

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