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 language | English |
|---|---|
| Pages (from-to) | 1241-1249 |
| Number of pages | 9 |
| Journal | Nature Biotechnology |
| Volume | 40 |
| Issue number | 8 |
| DOIs | |
| State | Published - Aug 2022 |
| Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.
Funding
We thank the Veterinary Service Center, Tel Aviv University for animal husbandry. The IDRFU, Genomic Research Unit and SICF units, Tel Aviv University for logistic support and council. We also thank L. Vardi, M. Gelbart, H. Kobo, D. Burstein, I. Benhar, N. Freund, M. Kay, T. Akriv and N. Gritsenko for reagents and feedback. This research was funded by the Varda and Boaz Dotan donation (A.B.), the H2020 European Research Council grant no. 759296 570 (A.B.) and the Israel Science Foundation grant nos. 1632/16, 2157/16 and 2876/21 (A.B.), The Bill and Melinda Gates Foundation grant no. OPP1183956 (J.E.V.), National Institutes of Health grant nos. R01 AI167003-01 (A.B.) AI128836 and R01 AI073148 (D.N.), Edmond J. Safra Center for Bioinformatics fellowship (T.K. and A.S.), St. Jude Children’s Research Hospital and ALSAC, National Institutes of Health Office Of The Director, Somatic Cell Genome Editing initiative grant no. U01AI157189 (S.Q.T.), the Gertner Institute Scholarship, the Yoran Institute Scholarship and the SAIA Foundation (A.D.N.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. We thank the Veterinary Service Center, Tel Aviv University for animal husbandry. The IDRFU, Genomic Research Unit and SICF units, Tel Aviv University for logistic support and council. We also thank L. Vardi, M. Gelbart, H. Kobo, D. Burstein, I. Benhar, N. Freund, M. Kay, T. Akriv and N. Gritsenko for reagents and feedback. This research was funded by the Varda and Boaz Dotan donation (A.B.), the H2020 European Research Council grant no. 759296 570 (A.B.) and the Israel Science Foundation grant nos. 1632/16, 2157/16 and 2876/21 (A.B.), The Bill and Melinda Gates Foundation grant no. OPP1183956 (J.E.V.), National Institutes of Health grant nos. R01 AI167003-01 (A.B.) AI128836 and R01 AI073148 (D.N.), Edmond J. Safra Center for Bioinformatics fellowship (T.K. and A.S.), St. Jude Children’s Research Hospital and ALSAC, National Institutes of Health Office Of The Director, Somatic Cell Genome Editing initiative grant no. U01AI157189 (S.Q.T.), the Gertner Institute Scholarship, the Yoran Institute Scholarship and the SAIA Foundation (A.D.N.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
| Funders | Funder 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 Health | R01 AI073148, R01 AI167003-01 |
| National Institute of Allergy and Infectious Diseases | R01AI128836 |
| Bill and Melinda Gates Foundation | OPP1183956 |
| St. Jude Children's Research Hospital | |
| H2020 European Research Council | 759296 570 |
| American Lebanese Syrian Associated Charities | U01AI157189 |
| Israel Science Foundation | 2157/16, 1632/16, 2876/21 |
| Tel Aviv University |
