PMID- 32518806 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20200928 IS - 2329-0501 (Print) IS - 2329-0501 (Electronic) IS - 2329-0501 (Linking) VI - 17 DP - 2020 Jun 12 TI - Variability in Cardiac miRNA-122 Level Determines Therapeutic Potential of miRNA-Regulated AAV Vectors. PG - 1190-1201 LID - 10.1016/j.omtm.2020.05.006 [doi] AB - Systemically delivered adeno-associated viral vector serotype 9 (AAV9) effectively transduces murine heart, but provides transgene expression also in liver and skeletal muscles. Improvement of the selectivity of transgene expression can be achieved through incorporation of target sites (TSs) for miRNA-122 and miRNA-206 into the 3' untranslated region (3' UTR) of the expression cassette. Here, we aimed to generate such miRNA-122- and miRNA-206-regulated AAV9 vector for a therapeutic, heart-specific overexpression of heme oxygenase-1 (HO-1). We successfully validated the vector functionality in murine cell lines corresponding to tissues targeted by AAV9. Next, we evaluated biodistribution of transgene expression following systemic vector delivery to HO-1-deficient mice of mixed C57BL/6J x FVB genetic background. Although AAV genomes were present in the hearts of these animals, HO-1 protein expression was either absent or significantly impaired. We found that miRNA-122, earlier described as liver specific, was present also in the hearts of C57BL/6J x FVB mice. Various levels of miRNA-122 expression were observed in the hearts of other mouse strains, in heart tissues of patients with cardiomyopathy, and in human induced pluripotent stem cell-derived cardiomyocytes in which we also confirmed such posttranscriptional regulation of transgene expression. Our data clearly indicate that therapeutic utilization of miRNA-based regulation strategy needs to consider inter-individual variability. CI - (c) 2020 The Author(s). FAU - Kraszewska, Izabela AU - Kraszewska I AD - Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland. FAU - Tomczyk, Mateusz AU - Tomczyk M AD - Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland. FAU - Andrysiak, Kalina AU - Andrysiak K AD - Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland. FAU - Biniecka, Monika AU - Biniecka M AD - Kardio-Med Silesia, 41-800 Zabrze, Poland. FAU - Geisler, Anja AU - Geisler A AD - Department of Applied Biochemistry, Institute of Biotechnology, Technische Universitat Berlin, 13355 Berlin, Germany. FAU - Fechner, Henry AU - Fechner H AD - Department of Applied Biochemistry, Institute of Biotechnology, Technische Universitat Berlin, 13355 Berlin, Germany. FAU - Zembala, Michal AU - Zembala M AD - Department of Cardiac Surgery, Heart and Lung Transplantation and Mechanical Circulatory Support, Silesian Center for Heart Diseases, 41-800 Zabrze, Poland. FAU - Stepniewski, Jacek AU - Stepniewski J AD - Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland. FAU - Dulak, Jozef AU - Dulak J AD - Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland. AD - Kardio-Med Silesia, 41-800 Zabrze, Poland. FAU - Jazwa-Kusior, Agnieszka AU - Jazwa-Kusior A AD - Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland. LA - eng PT - Journal Article DEP - 20200512 PL - United States TA - Mol Ther Methods Clin Dev JT - Molecular therapy. Methods & clinical development JID - 101624857 PMC - PMC7270145 EDAT- 2020/06/11 06:00 MHDA- 2020/06/11 06:01 PMCR- 2020/05/12 CRDT- 2020/06/11 06:00 PHST- 2020/02/18 00:00 [received] PHST- 2020/05/07 00:00 [accepted] PHST- 2020/06/11 06:00 [entrez] PHST- 2020/06/11 06:00 [pubmed] PHST- 2020/06/11 06:01 [medline] PHST- 2020/05/12 00:00 [pmc-release] AID - S2329-0501(20)30093-0 [pii] AID - 10.1016/j.omtm.2020.05.006 [doi] PST - epublish SO - Mol Ther Methods Clin Dev. 2020 May 12;17:1190-1201. doi: 10.1016/j.omtm.2020.05.006. eCollection 2020 Jun 12.