Optimizing Adeno-Associated Virus Serotype 9 for Studies of Cardiac Chamber–Specific Gene Regulation

Bilal, Alina S.; Blackwood, Erik A; Thuerauf, Donna J.; Glembotski, Christopher C

doi:10.1161/circulationaha.120.052437

Cited by 6 publications

(2 citation statements)

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“…Accordingly, we adapted this technology to measure the rate of ERAD in cardiac myocytes from mice treated in various ways using adeno-associated virus serovar 9 (AAV9) encoding hemagglutinin (HA)–TCRα under the control of the MLC2v promoter, which restricts expression to cardiac myocytes. 21 Mice treated with AAV9-3XHA-TCRα were subjected to TAC-HF. Then adult mouse ventricular myocytes were prepared, and the rate of HA-TCRα degradation was assessed by Western blot (Figure 1I).…”

Section: Resultsmentioning

confidence: 99%

Noncanonical Form of ERAD Regulates Cardiac Hypertrophy

et al. 2023

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BACKGROUND: Cardiac hypertrophy increases demands on protein folding, which causes an accumulation of misfolded proteins in the endoplasmic reticulum (ER). These misfolded proteins can be removed by the adaptive retrotranslocation, polyubiquitylation, and a proteasome-mediated degradation process, ER-associated degradation (ERAD), which, as a biological process and rate, has not been studied in vivo. To investigate a role for ERAD in a pathophysiological model, we examined the function of the functional initiator of ERAD, valosin-containing protein–interacting membrane protein (VIMP), positing that VIMP would be adaptive in pathological cardiac hypertrophy in mice. METHODS: We developed a new method involving cardiac myocyte-specific adeno-associated virus serovar 9–mediated expression of the canonical ERAD substrate, TCRα, to measure the rate of ERAD, ie, ERAD flux, in the heart in vivo. Adeno-associated virus serovar 9 was also used to either knock down or overexpress VIMP in the heart. Then mice were subjected to transverse aortic constriction to induce pressure overload–induced cardiac hypertrophy. RESULTS: ERAD flux was slowed in both human heart failure and mice after transverse aortic constriction. Surprisingly, although VIMP adaptively contributes to ERAD in model cell lines, in the heart, VIMP knockdown increased ERAD and ameliorated transverse aortic constriction–induced cardiac hypertrophy. Coordinately, VIMP overexpression exacerbated cardiac hypertrophy, which was dependent on VIMP engaging in ERAD. Mechanistically, we found that the cytosolic protein kinase SGK1 (serum/glucocorticoid regulated kinase 1) is a major driver of pathological cardiac hypertrophy in mice subjected to transverse aortic constriction, and that VIMP knockdown decreased the levels of SGK1, which subsequently decreased cardiac pathology. We went on to show that although it is not an ER protein, and resides outside of the ER, SGK1 is degraded by ERAD in a noncanonical process we call ERAD-Out. Despite never having been in the ER, SGK1 is recognized as an ERAD substrate by the ERAD component DERLIN1, and uniquely in cardiac myocytes, VIMP displaces DERLIN1 from initiating ERAD, which decreased SGK1 degradation and promoted cardiac hypertrophy. CONCLUSIONS: ERAD-Out is a new preferentially favored noncanonical form of ERAD that mediates the degradation of SGK1 in cardiac myocytes, and in so doing is therefore an important determinant of how the heart responds to pathological stimuli, such as pressure overload.

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Section: Resultsmentioning

confidence: 99%

Noncanonical Form of ERAD Regulates Cardiac Hypertrophy

et al. 2023

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“…The factors controlling the heart-specificity in viral vehicles remain unknown. However, replacing the capsid components and employing cardiac-specific promoters may increase the heart-specificity of AAV vehicles (Williams et al, 2010;Bilal et al, 2021). For non-viral vehicles, surface modification is capable of improving heart-specificity.…”

Section: Route Of Administrationmentioning

confidence: 99%

mRNA therapy for myocardial infarction: A review of targets and delivery vehicles

Wang

Senyo

2022

Front. Bioeng. Biotechnol.

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Cardiovascular diseases are the leading cause of death in the world. This is partly due to the low regenerative capacity of adult hearts. mRNA therapy is a promising approach under development for cardiac diseases. In mRNA therapy, expression of the target protein is modulated by delivering synthetic mRNA. mRNA therapy benefits cardiac regeneration by increasing cardiomyocyte proliferation, reducing fibrosis, and promoting angiogenesis. Because mRNA is translated in the cytoplasm, the delivery efficiency of mRNA into the cytoplasm and nucleus significantly affects its therapeutic efficacy. To improve delivery efficiency, non-viral vehicles such as lipid nanoparticles have been developed. Non-viral vehicles can protect mRNA from enzymatic degradation and facilitate the cellular internalization of mRNA. In addition to non-viral vehicles, viral vectors have been designed to deliver mRNA templates into cardiac cells. This article reviews lipid nanoparticles, polymer nanoparticles, and viral vectors that have been utilized to deliver mRNA into the heart. Because of the growing interest in lipid nanoparticles, recent advances in lipid nanoparticles designed for cardiac mRNA delivery are discussed. Besides, potential targets of mRNA therapy for myocardial infarction are discussed. Gene therapies that have been investigated in patients with cardiac diseases are analyzed. Reviewing mRNA therapy from a clinically relevant perspective can reveal needs for future investigations.

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