Effect of heart ischemia and administration route on biodistribution and transduction efficiency of AAV9 vectors

Abstract

Adeno-associated viruses (AAV) have become one of the most promising tools for gene transfer in clinics. Among all the serotypes, AAV9 has been described as the most efficient for cardiac transduction. In order to achieve optimal therapeutic delivery in heart disease, we have explored AAV9 transduction efficiency in an infarcted heart using different routes of administration and promoters, including a cardiacspecific one. AAV9 vectors carrying luciferase or green fluorescence protein under the control of the ubiquitous elongation-factor-1-alpha or the cardiac-specific troponin-T (TnT) promoters were administered by intramyocardial or intravenous injection, either in healthy or myocardial-infarcted mice. The transduction efficacy and specificity, the time-course expression, and the safety of each vector were tested. High transgene expression levels were found in the heart, but not in the liver, of mice receiving AAV-TnT, which was significantly higher after intramyocardial injection regardless of ischemia-induction. On the contrary, high hepatic transgene expression levels were detected with the elongation-factor-1-alpha-promoter, independently of the administration route and heart damage. Moreover, tissue-specific green fluorescence protein expression was found in cardiomyocytes with the TnT vector, whereas minimal cardiac expression was detected with the ubiquitous one.Interestingly, we found that myocardial infarction greatly increased the transcriptional activity of AAV genomes. Our findings show that the use of cardiac promoters allows for specific and stable cardiac gene expression, which is optimal and robust when intramyocardially injected. Furthermore, our data indicate that the pathological status of the tissue can alter the transcriptional activity of AAV genomes, an aspect that should be carefully evaluated for clinical applications.

iTRAQ‑based quantitative proteomics analysis of the potential application of secretoneurin gene therapy for cardiac hypertrophy induced by DL‑isoproterenol hydrochloride in mice

“…After loss of the righting reflex, the mice were ventilated by intratracheal intubation with a rodent ventilator (Harvard Apparatus). The chest of each mouse was opened to expose the heart and 2x10 10 particles of the Ad-con or Ad-SN (injection volume, 50 µl) were injected into the myocardium at five different sites, as previously described (26,27). Subsequently, the mice received subcutaneous injection of either ISO (5 mg/kg/day) or saline into their back for seven days (28).…”

Development and Application of a Liquid Chromatography-Mass Spectrometry Method for Residual Iodixanol Quantification in AAV-Based Gene Therapy Product Development

“…There have been 13 AAV serotypes reported. 4 Cesium chloride-and iodixanol-based density gradients are the core step among the most common purification methods for serotype-independent AAV. [5][6][7] Cesium chloride approach contains multiple steps and is typically considered to be time-consuming, compared to the iodixanol method.…”

“…Development and application of a liquid chromatography-mass spectrometry method for residual iodixanol quantification in AAVbased gene therapy product development Yi Pu, 1 Russell Katz, 2 Yunqiu Chen, 1 Vic Kostrubsky, 3 Pete Clarner, 4 Shih-Ching Lo, 5 Zoran Sosic, 1 Bernice Yeung 1…”

Overexpression of IκBα in cardiomyocytes alleviates hydrogen peroxide-induced apoptosis and autophagy by inhibiting NF-κB activation

“…Phosphorylation of IκBα, the key inhibitor of the canonical NF-κB pathway, at Ser 32 and Ser 36 is necessary for its degradation, and any mutation of these two serine residues blocks IκBα degradation [6]. Recently, adeno-associated virus serotype 9 (AAV9) was demonstrated to be the best gene carrier due to its high efficiency in the heart [17]. H 2 O 2 , a common reactive oxygen species (ROS), is generally utilized to mimic I/R injury in vitro [12].…”