Vaccine elicitation of HIV broadly neutralizing antibodies from engineered B cells

Huang, Deli; Tran, Jenny Tuyet; Olson, Alex; Vollbrecht, Thomas; Tenuta, Mary; Guryleva, Mariia V.; Fuller, Roberta P.; Schiffner, Torben; Abadejos, Justin R.; Couvrette, Lauren; Blane, Tanya R.; Saye, Karen; Li, Wenjuan; Landais, Elise; González-Martín, Alicia; Schief, William R.; Murrell, Ben; Burton, Dennis R.; Nemazee, David; Voss, James E.

doi:10.1038/s41467-020-19650-8

Cited by 63 publications

(82 citation statements)

References 39 publications

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“…One personalized approach is to collect memory B cells from an individual living with HIV and replace the BCR with a bNAb of choice through CRISPR/Cas9 editing, then infuse the engineered cells back into the individual, mirroring the approach used for CAR-T cell therapy approved for some lymphomas ( 159 , 160 ). This approach has shown success in mouse models performed by different groups introducing bNAb VRC01 ( 161 ) or 3BNC117 ( 162 ) as the BCR, with in vivo transfer resulting in a memory B cell population that retains the ability to class switch, undergo somatic hypermutation, and clonally expand and differentiate into plasmablasts following immunization with HIV Env.…”

Section: Cultivating Bnab Development For Treatment and Preventionmentioning

confidence: 99%

Advancing HIV Broadly Neutralizing Antibodies: From Discovery to the Clinic

Spencer

Shapiro

Haigwood

et al. 2021

Front. Public Health

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Despite substantial progress in confronting the global HIV-1 epidemic since its inception in the 1980s, better approaches for both treatment and prevention will be necessary to end the epidemic and remain a top public health priority. Antiretroviral therapy (ART) has been effective in extending lives, but at a cost of lifelong adherence to treatment. Broadly neutralizing antibodies (bNAbs) are directed to conserved regions of the HIV-1 envelope glycoprotein trimer (Env) and can block infection if present at the time of viral exposure. The therapeutic application of bNAbs holds great promise, and progress is being made toward their development for widespread clinical use. Compared to the current standard of care of small molecule-based ART, bNAbs offer: (1) reduced toxicity; (2) the advantages of extended half-lives that would bypass daily dosing requirements; and (3) the potential to incorporate a wider immune response through Fc signaling. Recent advances in discovery technology can enable system-wide mining of the immunoglobulin repertoire and will continue to accelerate isolation of next generation potent bNAbs. Passive transfer studies in pre-clinical models and clinical trials have demonstrated the utility of bNAbs in blocking or limiting transmission and achieving viral suppression. These studies have helped to define the window of opportunity for optimal intervention to achieve viral clearance, either using bNAbs alone or in combination with ART. None of these advances with bNAbs would be possible without technological advancements and expanding the cohorts of donor participation. Together these elements fueled the remarkable growth in bNAb development. Here, we review the development of bNAbs as therapies for HIV-1, exploring advances in discovery, insights from animal models and early clinical trials, and innovations to optimize their clinical potential through efforts to extend half-life, maximize the contribution of Fc effector functions, preclude escape through multiepitope targeting, and the potential for sustained delivery.

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Section: Cultivating Bnab Development For Treatment and Preventionmentioning

confidence: 99%

Advancing HIV Broadly Neutralizing Antibodies: From Discovery to the Clinic

Spencer

Shapiro

Haigwood

et al. 2021

Front. Public Health

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“…1,7,8 A number of groups have begun to explore an alternative to conventional vaccines in which B cells themselves are reprogrammed. [9][10][11][12][13] This approach employs CRISPR-mediated editing of the BCR loci so that the edited B-cell expresses a mature HIV-1 bNAb. In addition to its long-term potential for reprograming human immune responses, BCR editing can be applied more immediately to generate animal models useful for assessing vaccination strategies, and for developing more potent and bioavailable bNAb variants.…”

Section: Introductionmentioning

confidence: 99%

“…To date, investigators have bypassed this challenge by targeting an unvarying intron between the recombined variable region and the IgM constant region (Cµ). 9,[11][12][13] This strategy introduces a single cassette encoding an exogenous promoter and bNAb heavy-and light-chain sequences into this heavy-chain intron. By design, these constructs halt expression of the native variable heavy chain.…”

Section: Introductionmentioning

confidence: 99%

Efficient reprogramming of the heavy-chain CDR3 regions of a human antibody repertoire

Wang

et al. 2021

Preprint

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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 identity 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 and murine JH genes. We also optimized the design of 200 nucleotide homology-directed repair templates (HDRT) by minimizing the required 3'-5' resection 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 CH01. 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.

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“…5–7 These procedures can result in high frequency modification of hematopoietic cells, with editing efficiencies ranging from 20-80% across cell types and genomic loci. 2, 8–20…”

Section: Introductionmentioning

confidence: 99%

Optimization of AAV6 transduction enhances site-specific genome editing of primary human lymphocytes

Rogers

Huang

Clark

et al. 2021

Preprint

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Adeno-associated virus serotype 6 (AAV6) is a valuable reagent for genome editing of hematopoietic cells due to its ability to serve as a homology donor template. However, a comprehensive study of AAV6 transduction of hematopoietic cells in culture, with the goal of maximizing ex vivo genome editing, has not been reported. Here, we evaluated how the presence of serum, culture volume, transduction time, and electroporation parameters could influence AAV6 transduction. Based on these results, we identified an optimized protocol for genome editing of human lymphocytes based on a short, highly concentrated AAV6 transduction in the absence of serum, followed by electroporation with a targeted nuclease. In human CD4+ T cells and B cells, this protocol improved editing rates up to 7-fold and 21-fold respectively, when compared to standard AAV6 transduction protocols described in the literature. As a result, editing frequencies could be maintained using 50-100-fold less AAV6, which also reduced cellular toxicity. Our results highlight the important contribution of cell culture conditions for ex vivo genome editing with AAV6 vectors and provide a blueprint for improving AAV6-mediated homology-directed editing of human T and B cells.

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Vaccine elicitation of HIV broadly neutralizing antibodies from engineered B cells

Cited by 63 publications

References 39 publications

Advancing HIV Broadly Neutralizing Antibodies: From Discovery to the Clinic

Advancing HIV Broadly Neutralizing Antibodies: From Discovery to the Clinic

Efficient reprogramming of the heavy-chain CDR3 regions of a human antibody repertoire

Optimization of AAV6 transduction enhances site-specific genome editing of primary human lymphocytes

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