Efficient gene transduction in pigs and macaques with the engineered AAV vector AAV.GT5 for hemophilia B gene therapy

Yuji Kashiwakura; Kazuhiro Endo; Atsushi Ugajin; Tomohiro Kikuchi; Shuji Hishikawa; Hitoyasu Nakamura; Yuko Katakai; Nemekhbayar Baatartsogt; Takafumi Hiramoto; Morisada Hayakawa; Nobuhiko Kamoshita; Shoji Yamazaki; Akihiro Kume; Harushi Mori; Naohiro Sata; Yoichi Sakata; Shin-Ichi Muramatsu; Tsukasa Ohmori

doi:10.1016/j.omtm.2023.08.016

Efficient gene transduction in pigs and macaques with the engineered AAV vector AAV.GT5 for hemophilia B gene therapy

Mol Ther Methods Clin Dev. 2023 Aug 22:30:502-514. doi: 10.1016/j.omtm.2023.08.016. eCollection 2023 Sep 14.

Authors

Yuji Kashiwakura¹, Kazuhiro Endo^{2

3}, Atsushi Ugajin⁴, Tomohiro Kikuchi⁴, Shuji Hishikawa^{2

3}, Hitoyasu Nakamura⁴, Yuko Katakai⁵, Nemekhbayar Baatartsogt¹, Takafumi Hiramoto¹, Morisada Hayakawa^{1

6}, Nobuhiko Kamoshita^{1

6}, Shoji Yamazaki⁷, Akihiro Kume^{6

7}, Harushi Mori⁴, Naohiro Sata^{2

3}, Yoichi Sakata¹, Shin-Ichi Muramatsu^{6

8

9}, Tsukasa Ohmori^{1

6}

Affiliations

¹ Department of Biochemistry, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.
² Department of Surgery, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.
³ Center for Development of Advanced Medical Technology, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.
⁴ Department of Radiology, Jichi Medical University School of Medicine, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.
⁵ The Corporation for Production and Research of Laboratory Primates, 1-16-2 Sakura, Tsukuba, Ibaraki 305-0003, Japan.
⁶ Center for Gene Therapy Research, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.
⁷ Clinical Research Center, Jichi Medical University Hospital, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.
⁸ Division of Neurological Gene Therapy, Center for Open Innovation, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.
⁹ Center for Gene and Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan.

Abstract

Gene therapy using adeno-associated virus (AAV)-based vectors has become a realistic therapeutic option for hemophilia. We examined the potential of a novel engineered liver-tropic AAV3B-based vector, AAV.GT5, for hemophilia B gene therapy. In vitro transduction with AAV.GT5 in human hepatocytes was more than 100 times higher than with AAV-Spark100, another bioengineered vector used in a clinical trial. However, liver transduction following intravenous injection of these vectors was similar in mice with a humanized liver and in macaques. This discrepancy was due to the low recovery and short half-life of AAV.GT5 in blood, depending on the positive charge of the heparin-binding site in the capsid. Bypassing systemic clearance with the intra-hepatic vascular administration of AAV.GT5, but not AAV-Spark100, enhanced liver transduction in pigs and macaques. AAV.GT5 did not develop neutralizing antibodies (NAbs) in two of four animals, while AAV-Spark100 induced serotype-specific NAbs in all macaques tested (4 of 4). The NAbs produced after AAV-Spark100 administration were relatively serotype specific, and challenge with AAV.GT5 through the hepatic artery successfully boosted liver transduction in one animal previously administered AAV-Spark100. In summary, AAV.GT5 showed different vector kinetics and NAb induction compared with AAV-Spark100, and intra-hepatic vascular administration may minimize the vector dose required and vector dissemination.

Keywords: adeno-associated virus vectors; gene therapy; hemophilia B; intra-hepatic vascular administration; vector recoverability.