In vivo engineered B cells secrete high titers of broadly neutralizing anti-HIV antibodies in mice.
Journal
Nature biotechnology
ISSN: 1546-1696
Titre abrégé: Nat Biotechnol
Pays: United States
ID NLM: 9604648
Informations de publication
Date de publication:
08 2022
08 2022
Historique:
received:
15
02
2021
accepted:
22
04
2022
pubmed:
11
6
2022
medline:
16
8
2022
entrez:
10
6
2022
Statut:
ppublish
Résumé
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
Identifiants
pubmed: 35681059
doi: 10.1038/s41587-022-01328-9
pii: 10.1038/s41587-022-01328-9
pmc: PMC7613293
mid: EMS144534
doi:
Substances chimiques
Antibodies, Neutralizing
0
Broadly Neutralizing Antibodies
0
HIV Antibodies
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1241-1249Subventions
Organisme : NIAID NIH HHS
ID : R01 AI167003
Pays : United States
Organisme : European Research Council
ID : 759296
Pays : International
Organisme : NIAID NIH HHS
ID : R01 AI073148
Pays : United States
Organisme : NIAID NIH HHS
ID : U01 AI157189
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI128836
Pays : United States
Commentaires et corrections
Type : CommentIn
Type : CommentIn
Type : CommentIn
Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.
Références
Mendoza, P. et al. Combination therapy with anti-HIV-1 antibodies maintains viral suppression. Nature 561, 479–484 (2018).
pubmed: 30258136
pmcid: 6166473
doi: 10.1038/s41586-018-0531-2
Bar-On, Y. et al. Safety and antiviral activity of combination HIV-1 broadly neutralizing antibodies in viremic individuals. Nat. Med. 24, 1701–1707 (2018).
pubmed: 30258217
pmcid: 6221973
doi: 10.1038/s41591-018-0186-4
Cohen, Y. Z. et al. Safety, pharmacokinetics, and immunogenicity of the combination of the broadly neutralizing anti-HIV-1 antibodies 3BNC117 and 10-1074 in healthy adults: a randomized, phase 1 study. PLoS ONE 14, e0219142 (2019).
pubmed: 31393868
pmcid: 6687118
doi: 10.1371/journal.pone.0219142
Johnson, P. R. et al. Vector-mediated gene transfer engenders long-lived neutralizing activity and protection against SIV infection in monkeys. Nat. Med. 15, 901–906 (2009).
pubmed: 19448633
pmcid: 2723177
doi: 10.1038/nm.1967
Balazs, A. B. et al. Vectored immunoprophylaxis protects humanized mice from mucosal HIV transmission. Nat. Med. 20, 296–300 (2014).
pubmed: 24509526
pmcid: 3990417
doi: 10.1038/nm.3471
Priddy, F. H. et al. Adeno-associated virus vectored immunoprophylaxis to prevent HIV in healthy adults: a phase 1 randomised controlled trial. Lancet HIV 6, e230–e239 (2019).
pubmed: 30885692
pmcid: 6443625
doi: 10.1016/S2352-3018(19)30003-7
Nahmad, A. D. et al. Engineered B cells expressing an anti-HIV antibody enable memory retention, isotype switching and clonal expansion. Nat. Commun. 11, 5851 (2020).
pubmed: 33203857
pmcid: 7673991
doi: 10.1038/s41467-020-19649-1
Huang, D. et al. Vaccine elicitation of HIV broadly neutralizing antibodies from engineered B cells. Nat. Commun. 11, 5850 (2020).
pubmed: 33203876
pmcid: 7673113
doi: 10.1038/s41467-020-19650-8
Fusil, F. et al. A lentiviral vector allowing physiologically regulated membrane-anchored and secreted antibody expression depending on B-cell maturation status. Mol. Ther. 23, 1734–1747 (2015).
pubmed: 26281898
pmcid: 4817946
doi: 10.1038/mt.2015.148
Greiner, V. et al. CRISPR-mediated editing of the B cell receptor in primary human B cells. Science 12, 369–378 (2019).
Voss, J. E. et al. Reprogramming the antigen specificity of B cells using genome-editing technologies. eLife 8, e42995 (2019).
pubmed: 30648968
pmcid: 6355199
doi: 10.7554/eLife.42995
Hartweger, H. et al. HIV-specific humoral immune responses by CRISPR/Cas9-edited B cells. J. Exp. Med. 216, 1301–1310 (2019).
pubmed: 30975893
pmcid: 6547862
doi: 10.1084/jem.20190287
Moffett, H. F. et al. B cells engineered to express pathogen-specific antibodies protect against infection. Sci. Immunol. 4, eaax0644 (2019).
pubmed: 31101673
pmcid: 6913193
doi: 10.1126/sciimmunol.aax0644
Smith, T. T. et al. In situ programming of leukaemia-specific t cells using synthetic DNA nanocarriers. Nat. Nanotechnol. 12, 813–822 (2017).
pubmed: 28416815
pmcid: 5646367
doi: 10.1038/nnano.2017.57
Agarwal, S. et al. In vivo generation of CAR T cells selectively in human CD4
pubmed: 32485137
pmcid: 7403353
doi: 10.1016/j.ymthe.2020.05.005
Agarwal, S., Weidner, T., Thalheimer, F. B. & Buchholz, C. J. In vivo generated human CAR T cells eradicate tumor cells. Oncoimmunology 8, e167161 (2019).
doi: 10.1080/2162402X.2019.1671761
Frank, A. M. et al. Combining T-cell-specific activation and in vivo gene delivery through CD3-targeted lentiviral vectors. Blood Adv. 4, 5702–5715 (2020).
pubmed: 33216892
pmcid: 7686896
Frank, A. M. & Buchholz, C. J. Surface-engineered lentiviral vectors for selective gene transfer into subtypes of lymphocytes. Mol. Ther. Methods Clin. Dev. 12, 19–31 (2019).
pubmed: 30417026
doi: 10.1016/j.omtm.2018.10.006
Frank, A. M. et al. CD8-specific designed ankyrin repeat proteins improve selective gene delivery into human and primate T lymphocytes. Hum. Gene Ther. 31, 679–691 (2020).
pubmed: 32160795
doi: 10.1089/hum.2019.248
Pfeiffer, A. et al. In vivo generation of human CD19-CAR T cells results in B-cell depletion and signs of cytokine release syndrome. EMBO Mol. Med. 10, e9158 (2018).
pubmed: 30224381
pmcid: 6220327
doi: 10.15252/emmm.201809158
Michels, A. et al. Lentiviral and adeno-associated vectors efficiently transduce mouse T lymphocytes when targeted to murine CD8. Mol. Ther. Methods Clin. Dev. 23, 334–347 (2021).
pubmed: 34729380
pmcid: 8531454
doi: 10.1016/j.omtm.2021.09.014
Münch, R. C. et al. Off-target-free gene delivery by affinity-purified receptor-targeted viral vectors. Nat. Commun. 6, 2–10 (2015).
doi: 10.1038/ncomms7246
Breuer, C. B. et al. In vivo engineering of lymphocytes after systemic exosome-associated AAV delivery. Sci. Rep. 10, 4544 (2020).
pubmed: 32161326
pmcid: 7066196
doi: 10.1038/s41598-020-61518-w
Nawaz, W. et al. AAV-mediated in vivo CAR gene therapy for targeting human T-cell leukemia. Blood Cancer J. 11, 119 (2021).
pubmed: 34162832
pmcid: 8222347
doi: 10.1038/s41408-021-00508-1
Rurik, J. G. et al. CAR T cells produced in vivo to treat cardiac injury. Science 375, 91–96 (2022).
pubmed: 34990237
doi: 10.1126/science.abm0594
Parayath, N. N., Stephan, S. B., Koehne, A. L., Nelson, P. S. & Stephan, M. T. In vitro-transcribed antigen receptor mRNA nanocarriers for transient expression in circulating T cells in vivo. Nat. Commun. 11, 6080 (2020).
pubmed: 33247092
pmcid: 7695830
doi: 10.1038/s41467-020-19486-2
Grimm, D. et al. In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J. Virol. 82, 5887–5911 (2008).
pubmed: 18400866
pmcid: 2395137
doi: 10.1128/JVI.00254-08
Ran, F. A. et al. In vivo genome editing using Staphylococcus aureus Cas9. Nature 520, 186–191 (2015).
pubmed: 25830891
pmcid: 4393360
doi: 10.1038/nature14299
Scheid, J. F. et al. Sequence and structural convergence of broad and potent HIV antibodies that mimic CD4 binding. Science 333, 1633–1637 (2011).
pubmed: 21764753
pmcid: 3351836
doi: 10.1126/science.1207227
Hung, K. L. et al. Engineering protein-secreting plasma cells by homology-directed repair in primary human B cells. Mol. Ther. 26, 456–467 (2018).
pubmed: 29273498
doi: 10.1016/j.ymthe.2017.11.012
Mesin, L., Ersching, J. & Victora, G. D. Germinal center B cell dynamics. Immunity 45, 471–482 (2016).
pubmed: 27653600
pmcid: 5123673
doi: 10.1016/j.immuni.2016.09.001
Sok, D. & Burton, D. R. HIV broadly neutralizing antibodies: taking good care of the 98%. Immunity 45, 958–960 (2016).
pubmed: 27851923
pmcid: 5378060
doi: 10.1016/j.immuni.2016.10.033
Garber, D. A. et al. Durable protection against repeated penile exposures to simian-human immunodeficiency virus by broadly neutralizing antibodies. Nat. Commun. 11, 3195 (2020).
pubmed: 32581216
pmcid: 7314794
doi: 10.1038/s41467-020-16928-9
Taylor, J. J., Pape, K. A., Steach, H. R. & Jenkins, M. K. Apoptosis and antigen affinity limit effector cell differentiation of a single naïve B cell. Science 347, 11214–11218 (2015).
Abbott, R. K. et al. Precursor frequency and affinity determine B cell competitive fitness in germinal centers, tested with germline-targeting HIV vaccine immunogens. Immunity 48, 133–145.e6 (2017).
pubmed: 29287996
pmcid: 5773359
doi: 10.1016/j.immuni.2017.11.023
Dosenovic, P. et al. Anti-HIV-1 B cell responses are dependent on B cell precursor frequency and antigen binding affinity. Proc. Natl Acad. Sci. USA 115, 4743–4748 (2018).
pubmed: 29666227
pmcid: 5939114
doi: 10.1073/pnas.1803457115
Dey, B. et al. Structure-based stabilization of HIV-1 gp120 enhances humoral immune responses to the induced co-receptor binding site. PLoS Pathog. 5, e1000445 (2009).
pubmed: 19478876
pmcid: 2680979
doi: 10.1371/journal.ppat.1000445
Steichen, J. M. et al. HIV vaccine design to target germline precursors of glycan-dependent broadly neutralizing antibodies. Immunity 45, 483–496 (2016).
pubmed: 27617678
pmcid: 5040827
doi: 10.1016/j.immuni.2016.08.016
Lecomte, E. et al. Advanced characterization of DNA molecules in rAAV vector preparations by single-stranded virus next-generation sequencing. Mol. Ther. Nucleic Acids 4, e260 (2015).
pubmed: 26506038
pmcid: 4881760
doi: 10.1038/mtna.2015.32
Lazzarotto, C. R. et al. CHANGE-seq reveals genetic and epigenetic effects on CRISPR–Cas9 genome-wide activity. Nat. Biotechnol. 38, 1317–1327 (2020).
pubmed: 32541958
pmcid: 7652380
doi: 10.1038/s41587-020-0555-7
Busch, S. et al. Circulating monocytes and tumor‐associated macrophages express recombined immunoglobulins in glioblastoma patients. Clin. Transl. Med. 8, 18 (2019).
pubmed: 31155685
pmcid: 6545295
doi: 10.1186/s40169-019-0235-8
Gong, X. et al. Macrophage-derived immunoglobulin M inhibits inflammatory responses via modulating endoplasmic reticulum stress. Cells 10, 2812 (2021).
pubmed: 34831038
pmcid: 8616491
doi: 10.3390/cells10112812
Fuchs, T. et al. Expression of combinatorial immunoglobulins in macrophages in the tumor microenvironment. PLoS ONE 13, e0204108 (2018).
pubmed: 30240437
pmcid: 6150476
doi: 10.1371/journal.pone.0204108
Moreau, T., Bardin, F., Imbert, J., Chabannon, C. & Tonnelle, C. Restriction of transgene expression to the B-lymphoid progeny of human lentivirally transduced CD34
pubmed: 15233941
doi: 10.1016/j.ymthe.2004.04.005
McCarron, M. J., Park, P. W. & Fooksman, D. R. CD138 mediates selection of mature plasma cells by regulating their survival. Blood 129, 2749–2759 (2017).
pubmed: 28381397
pmcid: 5437827
doi: 10.1182/blood-2017-01-761643
Nemazee, D. Mechanisms of central tolerance for B cells. Nat. Rev. Immunol. 17, 281–294 (2017).
pubmed: 28368006
pmcid: 5623591
doi: 10.1038/nri.2017.19
Russell, D. M. et al. Peripheral deletion of self-reactive B cells. Nature 354, 308–311 (1991).
pubmed: 1956380
pmcid: 3787863
doi: 10.1038/354308a0
Edraki, A., Mir, A., Ibraheim, R., Gainetdinov, I. & Sontheimer, E. J. A compact, high-accuracy Cas9 with a dinucleotide PAM for in vivo genome editing. Mol. Cell 73, 714–726 (2019).
pubmed: 30581144
doi: 10.1016/j.molcel.2018.12.003
Pausch, P. et al. CRISPR-CasΦ from huge phages is a hypercompact genome editor. Science 337, 333–337 (2020).
doi: 10.1126/science.abb1400
Hartmann, J. et al. A library-based screening strategy for the identification of DARPins as ligands for receptor-targeted AAV and lentiviral vectors. Mol. Ther. Methods Clin. Dev. 10, 128–143 (2018).
pubmed: 30101151
pmcid: 6077149
doi: 10.1016/j.omtm.2018.07.001
Barzel, A. et al. Promoterless gene targeting without nucleases ameliorates haemophilia B in mice. Nature 517, 360–364 (2015).
pubmed: 25363772
doi: 10.1038/nature13864
Magoč, T. & Salzberg, S. L. FLASH: fast length adjustment of short reads to improve genome assemblies. Bioinformatics 27, 2957–2963 (2011).
pubmed: 21903629
pmcid: 3198573
doi: 10.1093/bioinformatics/btr507
Lee, H., Chang, H. Y., Cho, S. W. & Ji, H. P. CRISPRpic: fast and precise analysis for CRISPR-induced mutations via prefixed index counting. NAR Genom. Bioinform. 2, lqaa012 (2020).
pubmed: 32118203
pmcid: 7034628
doi: 10.1093/nargab/lqaa012
Langmead, B. & Salzberg, S. L. Fast gapped-read alignment with Bowtie 2. Nat. Methods 9, 357–359 (2012).
pubmed: 22388286
pmcid: 3322381
doi: 10.1038/nmeth.1923
Stern, A. et al. Selecton 2007: advanced models for detecting positive and purifying selection using a Bayesian inference approach. Nucleic Acids Res. 35, 506–511 (2007).
doi: 10.1093/nar/gkm382
Benjamini, Y. & Hochberg, Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J. R. Stat. Soc. Ser. B 57, 289–300 (1995).
Wieland, A. et al. Defining HPV-specific B cell responses in patients with head and neck cancer. Nature 597, 274–278 (2020).
pubmed: 33208941
doi: 10.1038/s41586-020-2931-3
Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the [Formula: see text] method. Methods 25, 402–408 (2001).
pubmed: 11846609
doi: 10.1006/meth.2001.1262
Nojima, T. et al. In-vitro derived germinal centre B cells differentially generate memory B or plasma cells in vivo. Nat. Commun. 2, 411–465 (2011).
doi: 10.1038/ncomms1475
Ferrari, S. et al. Efficient gene editing of human long-term hematopoietic stem cells validated by clonal tracking. Nat. Biotechnol. 38, 1298–1308 (2020).
pubmed: 32601433
pmcid: 7610558
doi: 10.1038/s41587-020-0551-y
Santana-Magal, N. et al. Melanoma-secreted lysosomes trigger monocyte-derived dendritic cell apoptosis and limit cancer immunotherapy. Cancer Res. 80, 1942–1956 (2020).
pubmed: 32127354
doi: 10.1158/0008-5472.CAN-19-2944
Brinkman, E. K., Chen, T., Amendola, M. & Van Steensel, B. Easy quantitative assessment of genome editing by sequence trace decomposition. Nucleic Acids Res. 42, e168 (2014).
pubmed: 25300484
pmcid: 4267669
doi: 10.1093/nar/gku936
Bitton, A., Nahary, L. & Benhar, I. in Phage Display: Methods and Protocols (eds. Hust, M. & Lim, T. S.) 349–363 (Springer, 2018).