Amplification of Replication Competent HIV-1 by Adoptive Transfer of Human Cells From Infected Humanized Mice

Su, Hang; Sravanam, Sruthi; Gorantla, Santhi; Kaminski, Rafal; Khalili, Kamel; Poluektova, Larisa Y.; Gendelman, Howard E.; Dash, Prasanta K.

doi:10.3389/fcimb.2020.00038

Cited by 10 publications

(12 citation statements)

References 59 publications

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“…However, at the same view, HIV-1p24 signal was not detected (data not shown). Reflective of our prior ( Su, et al 2020 ) studies, after 12 weeks of first-generation ART administration, brain HIV-1 infection was restricted where viral RNA and DNA were below the detection limit (10 copies/10 6 human CD45+ cells). This finding was observed in all the donor hu-HSC mice; measured using semi-nested qRT-PCR (data not shown).…”

Section: Resultsmentioning

confidence: 83%

“…ART was initiated at 2 weeks after infection then maintained for 12 weeks before donor hu-HSC mice were sacrificed and adoptive transfer was performed. This treatment scheme induced vigorous HIV-1 suppression in both hu-HSC mouse PBMC and lymphoid and brain tissues ( Dash, et al 2019 ; Su, et al 2020 ). Before animal euthanasia, plasma HIV-1 RNA was confirmed below detection limit (200 copies/ml) in all 4 donors ( Figure 2A ).…”

Section: Resultsmentioning

confidence: 99%

“…Recently, our group extended MVOA using humanized mice as donors and enabled a humanized mouse-to-mouse VOA (hu-MVOA) detection system. By adoptively transferring PBMCs from virally suppressed into naive humanized mice, HIV-1 recovery was made ( Su, et al 2020 ). In addition, these works allowed studies of human tissue cells that carry latent virus present in virally suppressed donor humanized mouse including splenocytes and bone marrow (BM) cells.…”

Section: Introductionmentioning

confidence: 99%

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Recovery of Latent HIV-1 from Brain Tissue by Adoptive Cell Transfer in Virally Suppressed Humanized Mice

Sravanam

Sillman

et al. 2021

J Neuroimmune Pharmacol

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Defining the latent human immunodeficiency virus type 1 (HIV-1) burden in the human brain during progressive infection is limited by sample access. Human hematopoietic stem cells (hu-HSCs)-reconstituted humanized mice provide an opportunity for this study. The model mimics, in measure, HIV-1 pathophysiology, transmission, treatment, and elimination in an infected human host. However, to date, brain HIV-1 latency in hu-HSC mice during suppressive antiretroviral therapy (ART) was not studied. To address this need, hu-HSC mice were administered long acting (LA) ART 14 days after HIV-1 infection was established. Animals were maintained under suppressive ART for 3 months, at which time HIV-1 infection was detected at low levels in brain tissue by droplet digital polymerase chain reaction (ddPCR) test on DNA. Notably, adoptive transfer of cells acquired from the hu-HSC mouse brains and placed into naive hu-HSC mice demonstrated viral recovery. These proof-of-concept results demonstrate replication-competent HIV-1 reservoir can be established in hu-HSC mouse brains that persists during long-term ART treatment. Hu-HSC mice-based mouse viral outgrowth assay (hu-MVOA) serves as a sensitive tool to interrogate latent HIV-1 brain reservoirs.

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Section: Resultsmentioning

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Recovery of Latent HIV-1 from Brain Tissue by Adoptive Cell Transfer in Virally Suppressed Humanized Mice

Sravanam

Sillman

et al. 2021

J Neuroimmune Pharmacol

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“…For example, some studies have established optimal proviral targets for viral deactivation. The LTR is the most common HIV-1 CRISPR/Cas9 target investigated thus far (Ebina et al, 2013 ; Dampier et al, 2014 , 2017 ; Hu et al, 2014 ; Zhu et al, 2015 ; Bialek et al, 2016 ; Ji et al, 2016 ; Kaminski et al, 2016a , b , c ; Limsirichai et al, 2016 ; Saayman et al, 2016 ; Ueda et al, 2016 ; Wang et al, 2016a , b ; Yin et al, 2016 ; Lebbink et al, 2017 ; Zhao et al, 2017 ; Bella et al, 2018 ; Kunze et al, 2018 ; Roychoudhury et al, 2018 ; Campbell et al, 2019 ; Darcis et al, 2019 ; Dash et al, 2019 ; Kaushik et al, 2019 ; Su et al, 2020 ). The ability of CRISPR/Cas9 to deactivate the virus in cell lines, primary cells ex vivo and human primary cells in engrafted in mice has also been established (Ebina et al, 2013 ; Hu et al, 2014 ; Kaminski et al, 2016a , b ; Lebbink et al, 2017 ; Bella et al, 2018 ; Ophinni et al, 2018 ; Campbell et al, 2019 ; Darcis et al, 2019 ).…”

Section: Application To the Hiv Gene Editing Fieldmentioning

confidence: 99%

Off-Target Analysis in Gene Editing and Applications for Clinical Translation of CRISPR/Cas9 in HIV-1 Therapy

et al. 2021

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As genome-editing nucleases move toward broader clinical applications, the need to define the limits of their specificity and efficiency increases. A variety of approaches for nuclease cleavage detection have been developed, allowing a full-genome survey of the targeting landscape and the detection of a variety of repair outcomes for nuclease-induced double-strand breaks. Each approach has advantages and disadvantages relating to the means of target-site capture, target enrichment mechanism, cellular environment, false discovery, and validation of bona fide off-target cleavage sites in cells. This review examines the strengths, limitations, and origins of the different classes of off-target cleavage detection systems including anchored primer enrichment (GUIDE-seq), in situ detection (BLISS), in vitro selection libraries (CIRCLE-seq), chromatin immunoprecipitation (ChIP) (DISCOVER-Seq), translocation sequencing (LAM PCR HTGTS), and in vitro genomic DNA digestion (Digenome-seq and SITE-Seq). Emphasis is placed on the specific modifications that give rise to the enhanced performance of contemporary techniques over their predecessors and the comparative performance of techniques for different applications. The clinical relevance of these techniques is discussed in the context of assessing the safety of novel CRISPR/Cas9 HIV-1 curative strategies. With the recent success of HIV-1 and SIV-1 viral suppression in humanized mice and non-human primates, respectively, using CRISPR/Cas9, rigorous exploration of potential off-target effects is of critical importance. Such analyses would benefit from the application of the techniques discussed in this review.

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“…These mice can also be used to test how best to interrupt viral integration, activation, and replication [ 55 , 56 ]. Recently our group employed humanized mice to examine tissue viral reservoirs and to recapitulate latent HIV-1 in vivo [ 57 , 58 ]. These works demonstrated that mature macrophages are a cell reservoir in antiretroviral therapy (ART)-suppressed HIV-infected humanized mice [ 59 ].…”

Section: Hiv-1 Infection Pathogenesis Prevention and Antiretroviral Testingmentioning

confidence: 99%

Humanized Mice for Infectious and Neurodegenerative disorders

et al. 2021

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Humanized mice model human disease and as such are used commonly for research studies of infectious, degenerative and cancer disorders. Recent models also reflect hematopoiesis, natural immunity, neurobiology, and molecular pathways that influence disease pathobiology. A spectrum of immunodeficient mouse strains permit long-lived human progenitor cell engraftments. The presence of both innate and adaptive immunity enables high levels of human hematolymphoid reconstitution with cell susceptibility to a broad range of microbial infections. These mice also facilitate investigations of human pathobiology, natural disease processes and therapeutic efficacy in a broad spectrum of human disorders. However, a bridge between humans and mice requires a complete understanding of pathogen dose, co-morbidities, disease progression, environment, and genetics which can be mirrored in these mice. These must be considered for understanding of microbial susceptibility, prevention, and disease progression. With known common limitations for access to human tissues, evaluation of metabolic and physiological changes and limitations in large animal numbers, studies in mice prove important in planning human clinical trials. To these ends, this review serves to outline how humanized mice can be used in viral and pharmacologic research emphasizing both current and future studies of viral and neurodegenerative diseases. In all, humanized mouse provides cost-effective, high throughput studies of infection or degeneration in natural pathogen host cells, and the ability to test transmission and eradication of disease.

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Amplification of Replication Competent HIV-1 by Adoptive Transfer of Human Cells From Infected Humanized Mice

Cited by 10 publications

References 59 publications

Recovery of Latent HIV-1 from Brain Tissue by Adoptive Cell Transfer in Virally Suppressed Humanized Mice

Recovery of Latent HIV-1 from Brain Tissue by Adoptive Cell Transfer in Virally Suppressed Humanized Mice

Off-Target Analysis in Gene Editing and Applications for Clinical Translation of CRISPR/Cas9 in HIV-1 Therapy

Humanized Mice for Infectious and Neurodegenerative disorders

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