Engineered AAVs for non-invasive gene delivery to rodent and non-human primate nervous systems

Creators: Chen, Xinhong; Ravindra Kumar, Sripriya; Adams, Cameron D.; Yang, Daping; Wang, Tongtong; Wolfe, Damien A.; Arokiaraj, Cynthia M.; Ngo, Victoria; Campos, Lillian J.; Griffiths, Jessica A.; Ichiki, Takako; Mazmanian, Sarkis K.; Osborne, Peregrine B.; Keast, Janet R.; Miller, Cory T.; Fox, Andrew S.; Chiu, Isaac M.; Gradinaru, Viviana¹

1. California Institute of Technology

Abstract

Gene therapy offers great promise in addressing neuropathologies associated with the central and peripheral nervous systems (CNS and PNS). However, genetic access remains difficult, reflecting the critical need for the development of effective and non-invasive gene delivery vectors across species. To that end, we evolved adeno-associated virus serotype 9 (AAV9) capsid in mice and validated two capsids, AAV-MaCPNS1 and AAV-MaCPNS2, across rodent species (mice and rats) and non-human primate (NHP) species (marmosets and rhesus macaques). Intravenous administration of either AAV efficiently transduced the PNS in rodents and both the PNS and CNS in NHPs. Furthermore, we used AAV-MaCPNS1 in mice to systemically deliver the following: (1) the neuronal sensor jGCaMP8s to record calcium signal dynamics in nodose ganglia and (2) the neuronal actuator DREADD to dorsal root ganglia to mediate pain. This conclusively demonstrates the translatability of these two systemic AAVs across four species and their functional utility through proof-of-concept studies in mice.

Additional Information

© 2022 Published by Elsevier. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Received 9 November 2021, Revised 18 February 2022, Accepted 2 May 2022, Available online 27 May 2022. We thank Yaping Lei for help with virus production; Elisha Mackey for mouse colony management; Dr. Zhe Qu and Erin Sullivan for lab management; Miguel Chuapoco for discussions on the NHP experiments; Patricia Anguiano for administrative assistance; the entire Gradinaru group and Dr. Tim Miles for discussions. We also thank Prof. Yuki Oka at Caltech for discussions on nodose calcium imaging experiments; I. Antoshechkin and the Millard and Muriel Jacobs Genetics and Genomics Laboratory at Caltech for providing sequencing service; members of the Miller lab at UCSD for performing marmoset injections and tissue collection; the staff at the California National Primate Research Center for their support with macaque experiments; and Catherine Oikonomou for help with manuscript editing. The diagrams in the figures were created with BioRender.com. This work was primarily supported by grants from the National Institutes of Health (NIH) to V.G.: NIH BRAIN R01MH117069, NIH Pioneer DP1OD025535, and SPARC 1OT2OD024899. Additional funding includes the following: (to V.G.) the Vallee Foundation, the Moore Foundation, the CZI Neurodegeneration Challenge Network, and the NSF NeuroNex Technology Hub grant 1707316; a grant from the Pew Charitable Trusts, the Heritage Medical Research Institute, and the Beckman Institute for CLARITY, optogenetics and vector engineering research (CLOVER) for technology development and dissemination; NIH R01 DK127257 (to I.M.C.); CZI Neurodegeneration Challenge Network (to I.M.C.); NIH R01 DC012087 (to C.T.M.); NIH P51 OD011107 (to A.S.F.); the BRAIN Initiative Armamentarium UF1MH128336 (to V.G., A.S.F., and C.T.M.); the Department of Defense grant W81XWH-17-1-0588 (to S.K.M. and V.G.); and NIH SPARC 3OT2OD023872 (to J.R.K.). Author contributions. X.C., S.R.K., and V.G. designed the experiments; X.C., S.R.K., C.D.A., J.R.K., D.Y., T.W., D.A.W., V.N., and L.J.C. performed the experiments; C.D.A., J.R.K., and P.B.O. assisted with the characterization of variants in rats; D.Y. and I.M.C. assisted with the characterization of the virus in pups and performed the pain experiment; T.W. and T.I. assisted with the calcium imaging experiment; D.A.W. assisted with the library selection and characterization of the virus in mice; C.M.A. assisted with the characterization of the DRG and spinal cord of NHPs; V.N. and C.T.M. assisted with the characterization of the virus in marmoset with the support of vet staff at UCSD; L.J.C. and A.S.F. assisted with the characterization of the virus in rhesus macaque with the support of staff at the California National Primate Research Center; J.A.G. and S.K.M. assisted with the characterization of the virus in mice ENS; X.C. and S.R.K. prepared the figures with input from all authors; X.C., S.R.K., and V.G. wrote the manuscript with input from all authors; V.G. supervised all aspects of the work. Declaration of interests. The California Institute of Technology has filed and licensed patent applications for some of the work described in this manuscript, with X.C., S.R.K., and V.G. listed as inventors. V.G. is a member of the Neuron advisory board and a co-founder and board member of Capsida Biotherapeutics, a fully integrated AAV engineering and gene therapy company. Data and code availability. This paper did not report original code. All data are available upon request.

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