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Efficient cardiac gene transfer and early-onset expression of a synthetic adeno-associated viral vector, Anc80L65, after intramyocardial administration

      Abstract

      Objective

      Gene therapy is a promising approach in the treatment of cardiovascular diseases. Preclinical and clinical studies have demonstrated that adeno-associated viral vectors are the most attractive vehicles for gene transfer. However, preexisting immunity, delayed gene expression, and postinfection immune response limit the success of this technology. The aim of this study was to investigate the efficacy of the first synthetic adeno-associated viral lineage clone, Anc80L65, for cardiac gene therapy.

      Methods

      By combining 2 different reporter approaches by fluorescence with green fluorescent protein and bioluminescence (Firefly luciferase), we compared transduction efficiency of Anc80L65 and adeno-associated virus, serotype 9 in neonatal rat cardiomyocytes ex vivo and rat hearts in vivo after intramyocardial and intracoronary administration.

      Results

      In cardiomyocytes, Anc80L65 provided a green fluorescent protein expression of 28.9% (36.4 ± 3.34 cells/field) at 24 hours and approximately 100% on day 7. In contrast, adeno-associated virus, serotype 9 green fluorescent protein provided minimal green fluorescent protein expression of 5.64% at 24 hours and 11.8% on day 7. After intramyocardial injection, vector expression peaked on day 7 with Anc80L65; however, with adeno-associated virus, serotype 9 the peak expression was during week 6. Administration of Anc80L65 demonstrated significantly more efficient expression of reporter gene than after adeno-associated virus, serotype 9 at 6 weeks (6.81 ± 0.64 log10 gc/100 ng DNA vs 6.49 ± 0.28 log10 gc/100 ng DNA, P < .05). These results were consistent with the amount of genome copy per cell observed in the heart.

      Conclusions

      Anc80L65 vector allows fast and robust gene transduction compared with adeno-associated virus, serotype 9 vector in cardiac gene therapy. Anc80L65 did not adversely affect cardiac function and caused no inflammatory response or toxicity.

      Graphical abstract

      Key Words

      Abbreviations and Acronyms:

      AAV (adeno-associated virus), AAV9 (adeno-associated virus, serotype 9), Anc80L65 (synthetic adeno-associated virus lineage clone), CM (cardiomyocyte), CR (complete revascularization), GFP (green fluorescent protein), IR (incomplete revascularization), Luc (Luciferase), LV (left ventricle), PBS (phosphate-buffered saline), qPCR (quantitative polymerase chain reaction)
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      References

        • Benjamin E.J.
        • Muntner P.
        • Alonso A.
        • Bittencourt M.S.
        • Callaway C.W.
        • Carson A.P.
        • et al.
        Heart disease and stroke statistics—2019 update: a report from the American Heart Association.
        Circulation. 2019; 139: e56-e528
        • Torabi A.
        • Cleland J.G.
        • Rigby A.S.
        • Sherwi N.
        Development and course of heart failure after a myocardial infarction in younger and older people.
        J Geriatr Cardiol. 2014; 11: 1-12
        • Torabi A.
        • Cleland J.G.
        • Khan N.K.
        • Loh P.H.
        • Clark A.L.
        • Alamgir F.
        • et al.
        The timing of development and subsequent clinical course of heart failure after a myocardial infarction.
        Eur Heart J. 2008; 29: 859-870
        • Garcia S.
        • Sandoval Y.
        • Roukoz H.
        • Adabag S.
        • Canoniero M.
        • Yannopoulos D.
        • et al.
        Outcomes after complete versus incomplete revascularization of patients with multivessel coronary artery disease.
        J Am Coll Cardiol. 2013; 62: 1421-1431
        • Leviner D.B.
        • Torregrosssa G.
        • Puskas J.D.
        Incomplete revascularization: what the surgeon needs to know.
        Ann Cardiothorac Surg. 2018; 7: 463-469
        • Serruys P.W.
        • Morice M.C.
        • Kappetein A.P.
        • Colombo A.
        • Holmes D.R.
        • Mack M.J.
        • et al.
        Percutaneous coronary intervention versus coronary-artery bypass grafting for severe coronary artery disease.
        N Engl J Med. 2009; 360: 961-972
        • Katz M.G.
        • Gubara S.M.
        • Hadas Y.
        • Weber T.
        • Kumar A.
        • Eliyahu E.
        • et al.
        Effects of genetic transfection on calcium cycling pathways mediated by double-stranded adeno-associated virus in postinfarction remodeling.
        J Thorac Cardiovasc Surg. 2020; 159: 1809-1819
        • Mathison M.
        • Singh V.P.
        • Chiuchiolo M.J.
        • Sanagasetti D.
        • Mao Y.
        • Patel V.P.
        • et al.
        In situ reprogramming to transdifferentiate fibroblasts into cardiomyocytes using adenoviral vectors: implications for clinical myocardial regeneration.
        J Thorac Cardiovasc Surg. 2017; 153: 329-339
        • Katz M.G.
        • Fargnoli A.S.
        • Williams R.D.
        • Bridges C.R.
        Cell and gene therapies for cardiovascular disease.
        in: Templeton N.S. Gene and Cell Therapy. CRC Press, New York2015: 861-901
        • Katz M.G.
        • Fargnoli A.S.
        • Williams R.D.
        • Bridges C.R.
        Gene therapy delivery systems for enhancing viral and nonviral vectors for cardiac diseases: current concepts and future applications.
        Hum Gene Ther. 2013; 24: 914-927
        • Duan D.
        Systemic delivery of adeno-associated viral vectors.
        Curr Opin Virol. 2016; 21: 16-25
        • Rangarajan S.
        • Walsh L.
        • Lester W.
        • Perry D.
        • Madan B.
        • Laffan M.
        • et al.
        AAV5-factor VIII gene transfer in severe hemophilia A.
        N Engl J Med. 2017; 377: 2519-2530
        • Keeler A.M.
        • Flotte T.R.
        Recombinant adeno-associated virus gene therapy in light of luxturna (and zolgensma and glybera): where are we, and how did we get here?.
        Annu Rev Virol. 2019; 6: 601-621
        • Gao G.
        • Vandenberghe L.H.
        • Alvira M.R.
        • Lu Y.
        • Calcedo R.
        • Zhou X.
        • et al.
        Clades of adeno-associated viruses are widely disseminated in human tissues.
        J Virol. 2004; 78: 6381-6388
        • Pacak C.A.
        • Mah C.S.
        • Thattaliyath B.D.
        • Conlon T.J.
        • Lewis M.A.
        • Cloutier D.E.
        • et al.
        Recombinant adeno-associated virus serotype 9 leads to preferential cardiac transduction in vivo.
        Circ Res. 2006; 99: e3-e9
        • Inagaki K.
        • Fuess S.
        • Storm T.A.
        • Gibson G.A.
        • Mctiernan C.F.
        • Kay M.A.
        • et al.
        Robust systemic transduction with AAV9 vectors in mice: efficient global cardiac gene transfer superior to that of AAV8.
        Mol Ther. 2006; 14: 45-53
        • Bish L.T.
        • Morine K.
        • Sleeper M.M.
        • Sanmiguel J.
        • Wu D.
        • Gao G.
        • et al.
        Adeno-associated virus (AAV) serotype 9 provides global cardiac gene transfer superior to AAV1, AAV6, AAV7, and AAV8 in the mouse and rat.
        Hum Gene Ther. 2008; 19: 1359-1368
        • Orlowski A.
        • Katz M.G.
        • Gubara S.M.
        • Fargnoli A.S.
        • Fish K.M.
        • Weber T.
        Successful transduction with AAV Vectors after selective depletion of anti-AAV antibodies by immunoadsorption.
        Mol Ther Methods Clin Dev. 2020; 16: 192-203
        • Asokan A.
        • Samulski R.J.
        An emerging adeno-associated viral vector pipeline for cardiac gene therapy.
        Hum Gene Ther. 2013; 24: 906-913
        • Zinn E.
        • Pacouret S.
        • Khaychuk V.
        • Turunen H.T.
        • Carvalho L.S.
        • Andres-Mateos E.
        • et al.
        In silico reconstruction of the viral evolutionary lineage yields a potent gene therapy vector.
        Cell Rep. 2015; 12: 1056-1068
        • Hudry E.
        • Andres-Mateos E.
        • Lerner E.P.
        • Volak A.
        • Cohen O.
        • Hyman B.T.
        • et al.
        Efficient gene transfer to the central nervous system by single-stranded Anc80L65.
        Mol Ther Methods Clin Dev. 2018; 10: 197-209
        • Carvalho L.S.
        • Xiao R.
        • Wassmer S.J.
        • Langsdorf A.
        • Zinn E.
        • Pacouret S.
        • et al.
        Synthetic adeno-associated viral vector efficiently targets mouse and nonhuman primate retina in vivo.
        Hum Gene Ther. 2018; 29: 771-784
        • Katz M.G.
        • Fargnoli A.S.
        • Gubara S.M.
        • Chepurko E.
        • Bridges C.R.
        • Hajjar R.J.
        Surgical and physiological challenges in the development of left and right heart failure in rat models.
        Heart Fail Rev. 2019; 24: 759-777
        • Sellke F.W.
        Gene therapy in cardiac surgery: is there a role?.
        J Thorac Cardiovasc Surg. 2003; 125: 994-997
        • Katz M.G.
        • Fargnoli A.S.
        • Kendle A.P.
        • Hajjar R.J.
        • Bridges C.R.
        Gene therapy in cardiac surgery: clinical trials, challenges, and perspectives.
        Ann Thorac Surg. 2016; 101: 2407-2416
        • Scimia M.C.
        • Gumpert A.M.
        • Koch W.J.
        Cardiovascular gene therapy for myocardial infarction.
        Expert Opin Biol Ther. 2014; 14: 183-195
        • Lähteenvuo J.
        • Ylä-Herttuala S.
        Advances and challenges in cardiovascular gene therapy.
        Hum Gene Ther. 2017; 28: 1024-1032
        • Ylä-Herttuala S.
        • Baker A.H.
        Cardiovascular gene therapy: past, present, and future.
        Mol Ther. 2017; 25: 1095-1106

      E-References

        • Zinn E.
        • Pacouret S.
        • Khaychuk V.
        • Turunen H.T.
        • Carvalho L.S.
        • Andres-Mateos E.
        • et al.
        In silico reconstruction of the viral evolutionary lineage yields a potent gene therapy vector.
        Cell Rep. 2015; 12: 1056-1068
        • Lock M.
        • Alvira M.
        • Vandenberghe L.H.
        • Samanta A.
        • Toelen J.
        • Debyser Z.
        • et al.
        Rapid, simple, and versatile manufacturing of recombinant adeno-associated viral vectors at scale.
        Hum Gene Ther. 2010; 21: 1259-1271

      Linked Article

      • Commentary: Can the new synthetic adeno-associated virus vector deliver the promise of cardiac gene therapy?
        The Journal of Thoracic and Cardiovascular SurgeryVol. 164Issue 6
        • Preview
          Katz and colleagues1 have reported the use of a synthetic adeno-associated virus (AAV) lineage clone, Anc80L65, and compared it with AAV9. They showed that transfer of reporter genes with Anc80L65 in rat cardiomyocytes and rat hearts was more efficient and robust than AAV9.1 This was not associated with off-target transfection in other organs, lymphocyte or neutrophil activation, alteration in inflammatory cytokines, or disturbance of cardiac function. These findings are consistent with favorable reports of Anc80 in gene therapy in the retina and the central nervous system.
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      • Commentary: A synthetic adeno-associated viral vector as a means of future precision treatment
        The Journal of Thoracic and Cardiovascular SurgeryVol. 164Issue 6
        • Preview
          Treating acute myocardial infarction should not be like shooting flies with a cannon; the target is the ischemic heart only, and a specifically targeted local treatment decreases risks of systematic side effects. The key is to bring the treatment to the specific tissue area intended to heal without influencing unnecessarily the whole of the body.
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