Analysis of hepatic lentiviral vector transduction; implications on preclinical studies and clinical gene therapy protocols. DOI Creative Commons

Peirong Hu,

Yajing Hao, Weihua Tang

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 21, 2024

Lentiviral vector-transduced T-cells were approved by the FDA as gene therapy anti-cancer medications. Little is known about host genetic variation effects on safety and efficacy of lentiviral vector delivery system. To narrow this knowledge-gap, we characterized hepatic vectors across Collaborative Cross (CC) mouse reference population. For 24 weeks, periodically measured luciferase expression from in 41 CC strains. Hepatic splenic copy numbers determined. We report that strains showed highly diverse outcomes following delivery. first time, moderate correlation between strain-specific sleeping patterns transduction efficiency was observed. associated two quantitative trait loci (QTLs) with intra-strain variations phenotypes, which mechanistically relates to phenomenon metastable epialleles. An additional QTL kinetics transgene expression. Genes comprised above QTLs are potential targets personalize protocols. Importantly, identified open new directions characterizing continuous viral silencing HIV latency. Our findings suggest wide-range patient-specific vector-based should be expected. Thus, novel escalating dose-based clinical protocols considered.

Язык: Английский

Cell-free assays reveal that the HIV-1 capsid protects reverse transcripts from cGAS immune sensing DOI Creative Commons
Tiana M. Scott,

Lydia M. Arnold,

Jordan A. Powers

и другие.

PLoS Pathogens, Год журнала: 2025, Номер 21(1), С. e1012206 - e1012206

Опубликована: Янв. 28, 2025

Retroviruses can be detected by the innate immune sensor cyclic GMP-AMP synthase (cGAS), which recognizes reverse-transcribed DNA and activates an antiviral response. However, extent to HIV-1 shields its genome from cGAS recognition remains unclear. To study this process in mechanistic detail, we reconstituted reverse transcription, release, sensing of a cell-free system. We found that wild-type capsids protect viral genomes even after completing transcription. Viral could “deprotected” thermal stress, capsid mutations, or reduced concentrations inositol hexakisphosphate (IP6) destabilize capsid. Strikingly, inhibitor lenacapavir also disrupted cores dramatically potentiated activity, both vitro cellular infections. Our results provide biochemical evidence lattice conceals chemical physical disruption core expose activate signaling.

Язык: Английский

Процитировано

1
Lenacapavir disrupts HIV-1 core integrity while stabilizing the capsid lattice DOI Creative Commons
Chenglei Li, Ryan C. Burdick,

Rokeya Siddiqui

и другие.

Proceedings of the National Academy of Sciences, Год журнала: 2025, Номер 122(14)

Опубликована: Апрель 1, 2025

Lenacapavir (GS-6207; LEN) is a potent HIV-1 capsid inhibitor approved for treating multidrug-resistant infection. LEN binds to hydrophobic pocket between neighboring (CA) proteins in hexamers and stabilizes the lattice, but its effect on capsids not fully understood. Here, we labeled with green fluorescent protein fused CA (GFP-CA) or fluid-phase GFP content marker (cmGFP) assess LEN’s impact capsids. cores GFP-CA, cmGFP, could be immunostained an anti-GFP antibody were less sensitive capsid-binding host restriction factor MX2, demonstrating that GFP-CA incorporated into lattice stability, whereas cmGFP indicator of core integrity. treatment isolated resulted dose-dependent loss signal while preserving signal, indicating disrupts integrity lattice. In contrast, PF-3450074 (PF74) induced Electron microscopy LEN- PF74-treated viral revealed frequent breakage at narrow end other morphological changes. Our results suggest does prevent nuclear envelope docking inhibits import without PF74 blocks by inhibiting cores, highlighting their different mechanisms inhibition.

Язык: Английский

Процитировано

1
In vivo HIV-1 nuclear condensates safeguard against cGAS and license reverse transcription DOI Creative Commons

Selen Ay,

Julien Burlaud‐Gaillard, Anastasia D. Gazi

и другие.

The EMBO Journal, Год журнала: 2024, Номер 44(1), С. 166 - 199

Опубликована: Дек. 2, 2024

Язык: Английский

Процитировано

4
HIV capsids: orchestrators of innate immune evasion, pathogenesis and pandemicity DOI
Kate L. Morling, Mohamed ElGhazaly,

R Milne

и другие.

Journal of General Virology, Год журнала: 2025, Номер 106(1)

Опубликована: Янв. 13, 2025

Human immunodeficiency virus (HIV) is an exemplar virus, still the most studied and best understood a model for mechanisms of viral replication, immune evasion pathogenesis. In this review, we consider earliest stages HIV infection from transport virion contents through cytoplasm to integration genome into host chromatin. We present holistic virus-host interaction during pivotal stage infection. Central process capsid. The last 10 years have seen transformation in way understand capsid structure function. review key discoveries our latest thoughts on as dynamic regulator innate chromatin targeting. also accessory proteins Vpr Vpx because they are incorporated particles where collaborate with capsids manipulate defensive cellular responses argue that effective regulation uncoating immunity define pandemic potential pathogenesis, how comparison different lineages can reveal what makes lentiviruses special.

Язык: Английский

Процитировано

0
Analysis of Hepatic Lentiviral Vector Transduction: Implications for Preclinical Studies and Clinical Gene Therapy Protocols DOI Creative Commons

Peirong Hu,

Yajing Hao, Weihua Tang

и другие.

Viruses, Год журнала: 2025, Номер 17(2), С. 276 - 276

Опубликована: Фев. 17, 2025

Lentiviral vector-transduced T cells were approved by the FDA as gene therapy anti-cancer medications. Little is known about effects of host genetic variation on safety and efficacy lentiviral vector delivery system. To narrow this knowledge gap, we characterized hepatic vectors across Collaborative Cross (CC) mouse reference population. For 24 weeks, periodically measured luciferase expression from in 41 CC strains. Hepatic splenic copy numbers determined. We report that strains showed highly diverse outcomes following delivery. first time, a moderate correlation between mouse-strain-specific sleeping patterns transduction efficiency was observed. associated two quantitative trait loci (QTLs) with intrastrain variations phenotypes, which mechanistically relates to phenomenon metastable epialleles. An additional QTL kinetics transgene expression. Genes found above QTLs are potential targets for personalized protocols. Importantly, identified open new directions characterizing continuous viral silencing HIV latency. Our findings suggest wide-range patient-specific vector-based should be expected. Thus, novel clinical protocols considered non-fatal diseases.

Язык: Английский

Процитировано

0
Considerations for capsid-targeting antiretrovirals in pre-exposure prophylaxis DOI
William M. McFadden,

Mia Færch,

Karen A. Kirby

и другие.

Trends in Molecular Medicine, Год журнала: 2025, Номер unknown

Опубликована: Фев. 1, 2025

Язык: Английский

Процитировано

0
The Complex Interactions Between HIV-1 and Human Host Cell Genome: From Molecular Mechanisms to Clinical Practice DOI Open Access
Manlio Tolomeo,

Francesco Tolomeo,

Antonio Cascio

и другие.

International Journal of Molecular Sciences, Год журнала: 2025, Номер 26(7), С. 3184 - 3184

Опубликована: Март 29, 2025

Antiretroviral therapy (ART) has significantly improved the prognosis of human immunodeficiency virus type 1 (HIV-1) infection. Although ART can suppress plasma viremia below detectable levels, it cannot eradicate HIV-1 DNA (provirus) integrated into host cell genome. This integration often results in unrepaired damage due to HIV-1-induced inhibition repair pathways. Furthermore, infection causes telomere attrition chromosomes, a critical factor contributing CD4+ T senescence and apoptosis. proteins induce damage, block replication, activate responses across various organs. In this review, we explore multiple aspects intricate interactions between genome involved depletion, inflammaging, clonal expansion infected cells long-term-treated patients, viral latency. We discuss molecular mechanisms that contribute comorbidities HIV-1-infected individuals highlight emerging therapeutic strategies targeting provirus.

Язык: Английский

Процитировано

0
Lenacapavir-induced Lattice Hyperstabilization is Central to HIV-1 Capsid Failure at the Nuclear Pore Complex and in the Cytoplasm DOI Creative Commons
Arpa Hudait, Ryan C. Burdick,

Ellie K. Bare

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Апрель 19, 2025

Abstract Lenacapavir (LEN) is the first HIV-1 capsid inhibitor approved for clinical use. It inhibits multiple steps of viral life cycle; however, molecular details effect LEN on structure and mechanistic inhibition are not understood. Recent studies show that intact cone-shaped capsids with LEN-induced breaks can dock at nuclear pore complexes (NPC), but only enter nucleus. In this work, we combined large-scale coarse-grained dynamics simulations live-cell imaging to investigate stepwise mechanism docking LEN-treated into NPC. Capsids bound substoichiometric concentrations reach NPC central channel. As advances end, lattice defects formed pentamer-hexamer interface – primarily narrower end leading pentamer dissociation. Dissociation pentamers detrimental integrity, both rupture narrow destabilization hexamer-hexamer interface. Structural analysis LEN-capsid in our demonstrate heterogeneous hyperstabilization loss essential pliability protein lattice. Live-cell cores labeled two different fluorescent markers showed ruptured were docked imported We conclude contributes elasticity inhibiting entry replication. Significance For replicate, core must nucleus, undergo reverse transcription, uncoat near integration sites. Here, using a computational experimental approach, Lenacapavir-treated stably bind during docking; broken associated defective entry. modulates structures by generating hyperstabilized domains, elasticity, adaptation crowded environment channel import. Our findings altering material properties be an effective strategy designing antiviral drugs disrupt

Язык: Английский

Процитировано

0
Covalent anti-HIV compound that induces HIV-1 capsid multimerization and degradation decomposes the viral core DOI Creative Commons
Tomofumi Nakamura, Nobutoki Takamune,

Mayu Okumura

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Апрель 19, 2025

Abstract Drugs that covalently bind to their target molecules form strong and irreversible interactions with each other. Covalent inhibitors have recently attracted attention because of enhanced structural selectivity within the cavity, which results in prolonged interactions, even at low doses. Here, we report ACAi-001 is a human immunodeficiency virus (HIV)-1 capsid (CA)- targeting compound covalent binding CA, estimating long-lasting effects for HIV inhibition. Using library millions compounds, was identified through silico screening. This designed hydrophobic cavity N-terminal domain CA and, cell-based assay, found inhibit HIV-1 replication. bound induced aberrant multimerization degradation, as assessed by western blotting, size-exclusion chromatography, thermal stability enzyme-linked immunosorbent liquid chromatography-mass spectrometry. ACAi-001, has two putative its chemical structure, binds directly serine 16 domain-targeting cysteine 198 or 218 C-terminal CA. unique profile induces multiple dysfunctions such resulting disruption core, thereby inhibiting infection degrades an manner, may reveal new aspects core assembly disassembly promising candidate next-generation anti-HIV-1 inhibitor. Significance Statement Lenacapavir (LEN) first-in-class inhibitor used clinically LEN long-acting convenient dosing regimen effective both treating multidrug-resistant pre-exposure prophylaxis. targets specific cavities distinct manner from LEN. forms leading degradation cause dysfunctions. Therefore, more potent derivatives serve inhibitors. shows potential against can be details on life cycle. illustration

Язык: Английский

Процитировано

0
How HIV-1 Uses the Metabolite Inositol Hexakisphosphate to Build Its Capsid DOI Creative Commons
Leo C. James

Viruses, Год журнала: 2025, Номер 17(5), С. 689 - 689

Опубликована: Май 9, 2025

The HIV-1 capsid is one of virology's most iconic structures, yet how it assembles has long remained elusive. Remarkably, the made from just a single protein, CA, which forms lattice ~250 hexamers and exactly 12 pentamers. Conical capsids form inside budded virions during maturation, but early efforts to reproduce this in vitro resulted instead open-ended tubes with purely hexameric lattice. missing component assembly was finally identified as metabolite inositol hexakisphosphate (IP6). Simply mixing soluble CA protein IP6 sufficient drive spontaneous conical similar size shape those infectious virions. Equally important, stabilises once formed, increasing their stability minutes hours. Indeed, such dependence on that virus actively packages into production. These discoveries have stimulated work multiple labs role importance replication, subject review.

Язык: Английский

Процитировано

0