Safe and efficient in vivo hematopoietic stem cell transduction in nonhuman primates using HDAd5/35++ vectors

Chang Li; Hongjie Wang; Sucheol Gil; Audrey Germond; Connie Fountain; Audrey Baldessari; Jiho Kim; Zhinan Liu; Aphrodite Georgakopoulou; Stefan Radtke; Tamás Raskó; Amit Pande; Christina Chiang; Eli Chin; Evangelia Yannaki; Zsuzsanna Izsvák; Thalia Papayannopoulou; Hans-Peter Kiem; André Lieber

doi:10.1016/j.omtm.2021.12.003

Safe and efficient in vivo hematopoietic stem cell transduction in nonhuman primates using HDAd5/35++ vectors

Mol Ther Methods Clin Dev. 2021 Dec 6:24:127-141. doi: 10.1016/j.omtm.2021.12.003. eCollection 2022 Mar 10.

Authors

Chang Li¹, Hongjie Wang¹, Sucheol Gil¹, Audrey Germond², Connie Fountain², Audrey Baldessari², Jiho Kim¹, Zhinan Liu¹, Aphrodite Georgakopoulou^{1

3}, Stefan Radtke⁴, Tamás Raskó⁵, Amit Pande⁴, Christina Chiang¹, Eli Chin¹, Evangelia Yannaki³, Zsuzsanna Izsvák⁵, Thalia Papayannopoulou⁶, Hans-Peter Kiem^{4

7}, André Lieber^{1

8}

Affiliations

¹ University of Washington, Department of Medicine, Division of Medical Genetics, Box 357720, Seattle, WA 98195, USA.
² Washington National Primate Research Center, Seattle, WA 98195, USA.
³ Gene and Cell Therapy Center, Hematology Department, George Papanicolaou Hospital, Thessaloniki 57010, Greece.
⁴ Stem and Gene Therapy Program, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
⁵ AG "Mobile DNA Lab" Max Delbrück Center for Molecular Medicine, 13125 Berlin-Buch, Germany.
⁶ University of Washington, Department of Medicine, Division of Hematology, Box 357720, Seattle, WA 98195, USA.
⁷ University of Washington, Department of Medicine, Division of Medical Oncology, Box 357720, Seattle, WA 98195, USA.
⁸ University of Washington, Department of Pathology, Seattle, WA 98195, USA.

Abstract

We tested a new in vivo hematopoietic stem cell (HSC) transduction/selection approach in rhesus macaques using HSC-tropic, integrating, helper-dependent adenovirus vectors (HDAd5/35++) designed for the expression of human γ-globin in red blood cells (RBCs) to treat hemoglobinopathies. We show that HDAd5/35++ vectors preferentially transduce HSCs in vivo after intravenous injection into granulocyte colony-stimulating factor (G-CSF)/AMD3100-mobilized animals and that transduced cells return to the bone marrow and spleen. The approach was well tolerated, and the activation of proinflammatory cytokines that are usually associated with intravenous adenovirus vector injection was successfully blunted by pre-treatment with dexamethasone in combination with interleukin (IL)-1 and IL-6 receptor blockers. Using our MGMT^P140K-based in vivo selection approach, γ-globin⁺ RBCs increased in all animals with levels up to 90%. After selection, the percentage of γ-globin⁺ RBCs declined, most likely due to an immune response against human transgene products. Our biodistribution data indicate that γ-globin⁺ RBCs in the periphery were mostly derived from mobilized HSCs that homed to the spleen. Integration site analysis revealed a polyclonal pattern and no genotoxicity related to transgene integrations. This is the first proof-of-concept study in nonhuman primates to show that in vivo HSC gene therapy could be feasible in humans without the need for high-dose chemotherapy conditioning and HSC transplantation.

Keywords: adenovirus vector; gamma globin; hematopoietic stem cells; hemoglobinopathies; in vivo; nonhuman primates.

Abstract

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