Enhanced Transduction of Human Hematopoietic Stem Cells by AAV6 Vectors: Implications in Gene Therapy and Genome Editing

Hua Yang; Keyun Qing; Geoffrey D Keeler; Ling Yin; Mario Mietzsch; Chen Ling; Brad E Hoffman; Mavis Agbandje-McKenna; Mengqun Tan; Wei Wang; Arun Srivastava

doi:10.1016/j.omtn.2020.03.009

Enhanced Transduction of Human Hematopoietic Stem Cells by AAV6 Vectors: Implications in Gene Therapy and Genome Editing

Mol Ther Nucleic Acids. 2020 Jun 5:20:451-458. doi: 10.1016/j.omtn.2020.03.009. Epub 2020 Mar 29.

Authors

Hua Yang¹, Keyun Qing², Geoffrey D Keeler², Ling Yin³, Mario Mietzsch⁴, Chen Ling⁵, Brad E Hoffman⁶, Mavis Agbandje-McKenna⁴, Mengqun Tan⁷, Wei Wang⁸, Arun Srivastava⁹

Affiliations

¹ Department of Radiology, Institute of Cell and Gene Therapy, The Third Xiangya Hospital, Central South University, Changsha, China; Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA; Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA.
² Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA; Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA.
³ Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA; Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA; State Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, China.
⁴ Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA; Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville, FL, USA.
⁵ State Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, China.
⁶ Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA; Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA; Department of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USA.
⁷ Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA; Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA; Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, China.
⁸ Department of Radiology, Institute of Cell and Gene Therapy, The Third Xiangya Hospital, Central South University, Changsha, China. Electronic address: cjr.wangwei@vip.163.com.
⁹ Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA; Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA; Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, FL, USA. Electronic address: aruns@peds.ufl.edu.

Abstract

We have reported that of the 10 most commonly used adeno-associated virus (AAV) serotype vectors, AAV6 is the most efficient in transducing primary human hematopoietic stem cells (HSCs) in vitro, as well as in vivo. More recently, polyvinyl alcohol (PVA), was reported to be a superior replacement for human serum albumin (HSA) for ex vivo expansion of HSCs. Since HSA has been shown to increase the transduction efficiency of AAV serotype vectors, we evaluated whether PVA could also enhance the transduction efficiency of AAV6 vectors in primary human HSCs. We report here that up to 12-fold enhancement in the transduction efficiency of AAV6 vectors can be achieved in primary human HSCs with PVA. We also demonstrate that the improvement in the transduction efficiency is due to PVA-mediated improved entry and intracellular trafficking of AAV6 vectors in human hematopoietic cells in vitro, as well as in murine hepatocytes in vivo. Taken together, our studies suggest that the use of PVA is an attractive strategy to further improve the efficacy of AAV6 vectors. This has important implications in the optimal use of these vectors in the potential gene therapy and genome editing for human hemoglobinopathies such as β-thalassemia and sickle cell disease.

Keywords: AAV vectors; gene transfer; genome editing; hematopoietic stem cells; hemoglobinopathies.

Abstract

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