In vivo engineered B cells secrete high titers of broadly neutralizing anti-HIV antibodies in mice

Alessio D. Nahmad, Cicera R. Lazzarotto, Natalie Zelikson, Talia Kustin, Mary Tenuta, Deli Huang, Inbal Reuveni, Daniel Nataf, Yuval Raviv, Miriam Horovitz-Fried, Iris Dotan, Yaron Carmi, Rina Rosin-Arbesfeld, David Nemazee, James E. Voss, Adi Stern, Shengdar Q. Tsai, Adi Barzel*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

Transplantation of B cells engineered ex vivo to secrete broadly neutralizing antibodies (bNAbs) has shown efficacy in disease models. However, clinical translation of this approach would require specialized medical centers, technically demanding protocols and major histocompatibility complex compatibility of donor cells and recipients. Here we report in vivo B cell engineering using two adeno-associated viral vectors, with one coding for Staphylococcus aureus Cas9 (saCas9) and the other for 3BNC117, an anti-HIV bNAb. After intravenously injecting the vectors into mice, we observe successful editing of B cells leading to memory retention and bNAb secretion at neutralizing titers of up to 6.8 µg ml−1. We observed minimal clustered regularly interspaced palindromic repeats (CRISPR)–Cas9 off-target cleavage as detected by unbiased CHANGE-sequencing analysis, whereas on-target cleavage in undesired tissues is reduced by expressing saCas9 from a B cell-specific promoter. In vivo B cell engineering to express therapeutic antibodies is a safe, potent and scalable method, which may be applicable not only to infectious diseases but also in the treatment of noncommunicable conditions, such as cancer and autoimmune disease.

Original languageEnglish
Pages (from-to)1241-1249
Number of pages9
JournalNature Biotechnology
Volume40
Issue number8
DOIs
StatePublished - Aug 2022

Funding

FundersFunder number
Edmond J. Safra Center for Bioinformatics
SAIA Foundation
Varda and Boaz Dotan donation
Veterinary Service Center, Tel Aviv University
Yoran Institute
National Institutes of HealthR01 AI073148, R01 AI167003-01
National Institute of Allergy and Infectious DiseasesR01AI128836
Bill and Melinda Gates FoundationOPP1183956
St. Jude Children's Research Hospital
H2020 European Research Council759296 570
American Lebanese Syrian Associated CharitiesU01AI157189
Israel Science Foundation2157/16, 1632/16, 2876/21
Tel Aviv University

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