Structure-guided evolution of antigenically distinct adeno-associated virus variants for immune evasion

Tse, Longping V.; Klinc, Kelli A.; Madigan, Victoria J.; Rivera, Ruth M. Castellanos; Wells, Lindsey F; Havlik, L. Patrick; Smith, J. W. G.; Agbandje‐McKenna, Mavis; Asokan, Aravind

doi:10.1073/pnas.1704766114

Cited by 167 publications

(155 citation statements)

References 30 publications

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“…Multiple reports have demonstrated that natural isolates AAV5 (Zabner et al, 2000), AAV6 (Limberis et al, 2009), and AAV9 (Adam et al, 2014) as well as engineered vectors AAV2.5 (Li et al, 2009) and AAV2.5T (Excoffon et al, 2009) are able to transduce human lung epithelial cells, including primary human airway epithelial (HAE) cultures. Although there is a high prevalence of nAbs against AAVs in the human population, new technologies have been developed to engineer AAV to evade humoral immune responses (Tse et al, 2015(Tse et al, , 2017. These developments potentially allow for the delivery of CoV vaccines or immunotherapeutic directly to the mucosal compartments of the lung.…”

Section: Aav As a Vector For Passive Immunization Against Emerging Covmentioning

confidence: 99%

“…Fortunately, multiple strategies have been developed to address these hurdles (Tse et al, 2015). For instance, through vector engineering, a new generation of AAV vectors can evade pre-existing nAbs while retaining a good transduction profile in the respiratory system (Li et al, 2009;Tse et al, 2017). For rare genetic diseases, life-long gene expression is important for therapeutic purposes but could cause toxicity.…”

Section: The Challengesmentioning

confidence: 99%

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The Current and Future State of Vaccines, Antivirals and Gene Therapies Against Emerging Coronaviruses

et al. 2020

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Emerging coronaviruses (CoV) are constant global public health threats to society. Multiple ongoing clinical trials for vaccines and antivirals against CoVs showcase the availability of medical interventions to both prevent and treat the future emergence of highly pathogenic CoVs in human. However, given the diverse nature of CoVs and our close interactions with wild, domestic and companion animals, the next epidemic zoonotic CoV could resist the existing vaccines and antivirals developed, which are primarily focused on Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS CoV). In late 2019, the novel CoV (SARS-CoV-2) emerged in Wuhan, China, causing global public health concern. In this review, we will summarize the key advancements of current vaccines and antivirals against SARS-CoV and MERS-CoV as well as discuss the challenge and opportunity in the current SARS-CoV-2 crisis. At the end, we advocate the development of a "plug-and-play" platform technologies that could allow quick manufacturing and administration of broad-spectrum countermeasures in an outbreak setting. We will discuss the potential of AAV-based gene therapy technology for in vivo therapeutic antibody delivery to combat SARS-CoV-2 outbreak and the future emergence of severe CoVs.

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Section: Aav As a Vector For Passive Immunization Against Emerging Covmentioning

confidence: 99%

Section: The Challengesmentioning

confidence: 99%

The Current and Future State of Vaccines, Antivirals and Gene Therapies Against Emerging Coronaviruses

et al. 2020

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“…Four rAAV1 PTMs within the disordered N-terminal region of the capsid were shared between the human and baculo-rAAV1 vectors: N-terminal acetylation on the initial start methionine, methylation of lysine 61, and phosphorylation on serines 149 and 153. For the remaining residues 217-736 on rAAV1, we detected numerous PTMs within known capsid antigenic motifs that react with known neutralizing antibodies against rAAV1 (456-AQNK-459), (492-TKTDNNNS-499), and (588-STDPATGDVH-597) 27 . One PTM was detected within the 4E4 neutralizing epitope residue (456-459 and 492-498) 28,29 , and several additional PTMs were detected near the 5H7 Here again, HCP impurities were present and different between platforms (Table S6), and Sf9 insect HCP impurities were detected with N-linked glycans ( Table S7).…”

Section: Presence Of Capsid Alterations and Hcp Impurity Levels Are Cmentioning

confidence: 99%

Methods Matter -- Standard Production Platforms for Recombinant AAV Produce Chemically and Functionally Distinct Vectors

Rumachik

Malaker

Poweleit

et al. 2019

Preprint

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Different manufacturing approaches have been used in the production of recombinant adeno-associated virus (rAAV). We sought to characterize differences in rAAV vectors when produced by the two leading manufacturing platforms; live baculovirus infection of Sf9 insect cells and transiently-transfected human HEK293 cells. We used multiple analytical approaches, including proteomic profiling by mass spectrometry, isoelectric focusing, cryo-EM, denaturation assays, next-generation sequencing of packaged genomes, human cytokine profiling in response to vector transduction, and comparative functional transduction assessments in vivo and in vitro. Our data support the following findings: 1) rAAV capsids have post-translational modifications (PTMs); 2) vector lot modifications included glycosylation, acetylation, phosphorylation, methylation and deamidation; 3) capsid PTMs differ when produced in either platform; 4) impurities were different in vectors when produced in either platform; 5) impurities can also have their own PTMs, including N-linked glycans; 6) capsid PTMs and impurities were seen across all rAAV serotypes, manufacturers, and purification types; 7) there was no difference in the packaged rAAV genome sequence in either platform; 8) baculovirus-Sf9 vector lots have insect and baculoviral impurities and can have poorer packaging percentages than human-produced vector; 9) rAAV capsids have no significant structural differences when produced in either platform; 10) when given at the same vector genome dose, human-produced rAAVs can be more potent than baculovirus-Sf9 vectors in vitro and in vivo (P<0.05-0.0001); and lastly, 11) regardless of the manufacturing platform, functional rAAV transduction is sexually dimorphic in the liver when administered IV, but not in skeletal muscle when administered IM. Our results demonstrate that baculovirus-Sf9 and human manufacturing platforms can produce vector lots exhibiting chemical and functional differences. These findings were reproducible across numerous rAAV vendors, including commercial manufacturers, academic core facilities, and individual laboratories. These differences may have implications for receptor binding, trafficking, expression kinetics, stability, and immunogenicity.

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“…Strategies that are currently being evaluated to circumvent pre-existing humoral immunity to AAV vectors are either early in development, ineffective or prone to causing undesirable side effects. These include the engineering of new AAV variants with reduced NAb recognition(18, 19), plasmapheresis or immunoadsorption to reduce the overall levels of circulating antibodies in patient serum prior to AAV administration(20–23), use of capsid decoys(24) or immunosuppression to decrease the B cell population and consequently antibody levels in general(25, 26). While these approaches have demonstrated varying success and efficiency in addressing the problem of circulating antibodies and remain under evaluation, a one-solution-fits-all approach that resolves this challenge is unlikely.…”

Section: Introductionmentioning

confidence: 99%

Rescuing AAV gene transfer from antibody neutralization with an IgG-degrading enzyme

Elmore

Simon

et al. 2020

Preprint

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20Pre-existing humoral immunity to recombinant adeno-associated viral (AAV) vectors 21 restricts the treatable patient population and efficacy of human gene therapies. Approaches to 22 clear neutralizing antibodies (NAbs), such as plasmapheresis and immunosuppression are either 23 ineffective or cause undesirable side effects. Here, we describe a clinically relevant strategy to 24 rapidly and transiently degrade NAbs prior to AAV administration using an IgG degrading 25 enzyme (IdeZ). We demonstrate that recombinant IdeZ efficiently cleaves IgG in dog, monkey 26 and human antisera. Prophylactically administered IdeZ cleaves circulating, human IgG in mice 27 and prevents AAV neutralization in vivo. In macaques, a single intravenous dose of IdeZ rescues 28 AAV transduction by transiently reversing seropositivity. Importantly, IdeZ efficiently cleaves 29 NAbs and rescues AAV transduction in mice passively immunized with individual human donor 30 sera representing a diverse population. Our antibody clearance approach presents a new 31 paradigm for expanding the prospective patient cohort and improving efficacy of AAV gene 32 therapy. 34Human gene therapy using recombinant AAV vectors continues to advance steadily as a 37 treatment paradigm for rare, monogenic disorders. This is highlighted by the recent FDA approval 38 and clinical success of Zolgensma®, an intravenously dosed AAV vector delivering a functional 39 copy of the SMN1 gene in children with Spinal Muscular Atrophy (SMA)(1). Further, the list of 40 systemically dosed AAV-based gene therapies for rare disorders such as Hemophilia A & B, 41 Duchenne Muscular Dystrophy (DMD), X-linked myotubularin myopathy (XLMTM) and Pompe 42 disease amongst others continues to grow(2, 3). These promising clinical examples have 43 concurrently highlighted important challenges that include manufacturing needs, patient 44 recruitment, and the potential for toxicity at high AAV doses. One such challenge that limits the 45 recruitment of patients for gene therapy clinical trials and adversely affects the efficacy of AAV 46 gene therapy is the prevalence of pre-existing neutralizing antibodies (NAbs) to AAV capsids in 47 the human population. Such NAbs arise due to natural infection or cross-reactivity between 48 different AAV serotypes(4-7). NAbs can mitigate AAV infection through multiple mechanisms 49 by (a) binding to AAV capsids and blocking critical steps in transduction such as cell surface 50 attachment and uptake, endosomal escape, productive trafficking to the nucleus or uncoating and 51 (b) promoting AAV opsonization by phagocytic cells, thereby mediating their rapid clearance from 52 the circulation. Multiple preclinical studies in different animal models have demonstrated that pre-53 existing NAbs impede systemic gene transfer by AAV vectors(8-11). 54In humans, serological studies reveal a high prevalence of NAbs in the worldwide 55 population, with about 67% of people having antibodies against AAV1, 72% against AAV2, and 56 ~ 40% against AAV serotypes 5 thr...

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Structure-guided evolution of antigenically distinct adeno-associated virus variants for immune evasion

Cited by 167 publications

References 30 publications

The Current and Future State of Vaccines, Antivirals and Gene Therapies Against Emerging Coronaviruses

The Current and Future State of Vaccines, Antivirals and Gene Therapies Against Emerging Coronaviruses

Methods Matter -- Standard Production Platforms for Recombinant AAV Produce Chemically and Functionally Distinct Vectors

Rescuing AAV gene transfer from antibody neutralization with an IgG-degrading enzyme

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