Reprogramming of the heavy-chain CDR3 regions of a human antibody repertoire
Copyright © 2021 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved..
B cells have been engineered ex vivo to express an HIV-1 broadly neutralizing antibody (bNAb). B cell reprograming may be scientifically and therapeutically useful, but current approaches limit B cell repertoire diversity and disrupt the organization of the heavy-chain locus. A more diverse and physiologic B cell repertoire targeting a key HIV-1 epitope could facilitate evaluation of vaccines designed to elicit bNAbs, help identify more potent and bioavailable bNAb variants, or directly enhance viral control in vivo. Here we address the challenges of generating such a repertoire by replacing the heavy-chain CDR3 (HCDR3) regions of primary human B cells. To do so, we identified and utilized an uncharacterized Cas12a ortholog that recognizes PAM motifs present in human JH genes. We also optimized the design of 200 nucleotide homology-directed repair templates (HDRT) by minimizing the required 3'-5' deletion of the HDRT-complementary strand. Using these techniques, we edited primary human B cells to express a hemagglutinin epitope tag and the HCDR3 regions of the bNAbs PG9 and PG16. Those edited with bNAb HCDR3 efficiently bound trimeric HIV-1 antigens, implying they could affinity mature in vivo in response to the same antigens. This approach generates diverse B cell repertoires recognizing a key HIV-1 neutralizing epitope.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:30 |
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| Enthalten in: |
Molecular therapy : the journal of the American Society of Gene Therapy - 30(2022), 1 vom: 05. Jan., Seite 184-197 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Ou, Tianling [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 07.04.2022 Date Revised 17.09.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.ymthe.2021.10.027 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM33278861X |
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| 500 | |a Date Revised 17.09.2023 | ||
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| 520 | |a Copyright © 2021 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a B cells have been engineered ex vivo to express an HIV-1 broadly neutralizing antibody (bNAb). B cell reprograming may be scientifically and therapeutically useful, but current approaches limit B cell repertoire diversity and disrupt the organization of the heavy-chain locus. A more diverse and physiologic B cell repertoire targeting a key HIV-1 epitope could facilitate evaluation of vaccines designed to elicit bNAbs, help identify more potent and bioavailable bNAb variants, or directly enhance viral control in vivo. Here we address the challenges of generating such a repertoire by replacing the heavy-chain CDR3 (HCDR3) regions of primary human B cells. To do so, we identified and utilized an uncharacterized Cas12a ortholog that recognizes PAM motifs present in human JH genes. We also optimized the design of 200 nucleotide homology-directed repair templates (HDRT) by minimizing the required 3'-5' deletion of the HDRT-complementary strand. Using these techniques, we edited primary human B cells to express a hemagglutinin epitope tag and the HCDR3 regions of the bNAbs PG9 and PG16. Those edited with bNAb HCDR3 efficiently bound trimeric HIV-1 antigens, implying they could affinity mature in vivo in response to the same antigens. This approach generates diverse B cell repertoires recognizing a key HIV-1 neutralizing epitope | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a B cell receptor | |
| 650 | 4 | |a Cas12a | |
| 650 | 4 | |a HCDR3 | |
| 650 | 4 | |a HDR | |
| 650 | 4 | |a HIV-1 V2-glycan antibody | |
| 650 | 4 | |a Mb2Cas12a | |
| 650 | 4 | |a PG16 | |
| 650 | 4 | |a PG9 | |
| 650 | 4 | |a SOSIP | |
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| 650 | 7 | |a Broadly Neutralizing Antibodies |2 NLM | |
| 650 | 7 | |a Epitopes |2 NLM | |
| 650 | 7 | |a HIV Antibodies |2 NLM | |
| 700 | 1 | |a He, Wenhui |e verfasserin |4 aut | |
| 700 | 1 | |a Quinlan, Brian D |e verfasserin |4 aut | |
| 700 | 1 | |a Guo, Yan |e verfasserin |4 aut | |
| 700 | 1 | |a Tran, Mai H |e verfasserin |4 aut | |
| 700 | 1 | |a Karunadharma, Pabalu |e verfasserin |4 aut | |
| 700 | 1 | |a Park, Hajeung |e verfasserin |4 aut | |
| 700 | 1 | |a Davis-Gardner, Meredith E |e verfasserin |4 aut | |
| 700 | 1 | |a Yin, Yiming |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Xia |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Haimin |e verfasserin |4 aut | |
| 700 | 1 | |a Zhong, Guocai |e verfasserin |4 aut | |
| 700 | 1 | |a Farzan, Michael |e verfasserin |4 aut | |
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