CROI 2017 Abstract e-Book

Abstract eBook

Poster and Themed Discussion Abstracts

Conclusion: Our data suggest that HIV integrated into specific genes involved with cell cycle regulation, cancer, and potentially immune signaling, may lead to enhanced proliferation and thus persistence during ART. The observed proviral orientation biases suggest that transcription of the provirus may potentially enhance or interfere with host gene expression and impact cell proliferation.

305 ROMIDEPSIN-INDUCED HIV-1 VIREMIA IS OLIGOCLONAL WITH LIMITED DELETERIOUS MUTATIONS

Anni Winckelmann 1 , Kirston Barton 2 , Bonnie Hiener 2 , Timothy E. Schlub 3 , Wei Shao 4 , Thomas A. Rasmussen 1 , Lars Østergaard 1 , Ole S. Søgaard 1 , Martin Tolstrup 1 , Sarah Palmer 2 1 Aarhus Univ Hosp, Aarhus, Denmark, 2 Univ of Sydney, Westmead, Australia, 3 Univ of Sydney, Camperdown, New South Wales, Australia, 4 Leidos Biomed Rsr, Inc, Frederick, MD, USA Background: The administration of the latency-reversing agent romidepsin once weekly for three consecutive weeks to individuals on suppressive antiretroviral therapy (ART) revealed quantifiable increases of cell-associated (CA) and plasma HIV-1 RNA in 5 of 6 participants, which coincided with the romidepsin infusions. However, the origin of the romidepsin-induced plasma HIV-1 RNA is unknown. To address this, we compared HIV-1 DNA and CA RNA sequences from peripheral blood CD4+ T cells to HIV-1 RNA sequences obtained from the plasma during romidepsin treatment. Methods: CD4+ T-cells were obtained at baseline, following the second and third romidepsin infusion, and 10 weeks after the final romidepsin treatment. From three participants we obtained plasma collected 24 and 72 hours following each romidepsin infusion. Using the single-copy assay, we confirmed these plasma samples contained HIV-1 RNA from 3-70 copies/ml and no HIV-1 DNA. Single-genome sequencing of the env region was used to genetically characterize the virus from proviral DNA, the transcribed CA RNA as well as the plasma RNA pool. Results: In all three participants with available plasma samples we identified plasma HIV-1 RNA sequences that were identical to DNA and/or CA RNA sequences from peripheral blood CD4+ T cells. Plasma HIV-1 RNA, DNA and CA RNA sequences intermingled throughout the phylogenetic trees. In two participants, we identified several expansions of identical plasma HIV-1 RNA sequences, corresponding to 62% and 100% of the total plasma RNA sequences, respectively. Proportions of defective viruses, defined as containing hypermutation or stop codons in the regions sequenced, differed significantly between HIV-1 DNA, CA RNA and plasma. Plasma HIV-1 RNA had very low amounts of defective viruses compared to CA RNA (odds ratio 20.85, p<0.001) and to DNA (odds ratio 7.07, p=0.011) during romidepsin therapy. Conclusion: Our findings demonstrate that romidepsin induced transcription from proviruses in peripheral blood cells, which contributed to viremia in patients on suppressive ART. The intermingling of CA HIV-1 RNA with DNA sequences indicates transcription from a diverse range of proviruses. However, the oligoclonal pattern of viremia and low amounts of defective plasma HIV-1 RNA sequences indicate that the romidepsin-induced viremia arises from intact proviruses with highly similar or identical genetic backgrounds. These findings will inform future eradication strategies employing latency reversing agents. 306 MURINE MODEL TO PREDICT VIRAL REBOUND IN HIV+ ALLOTRANSPLANTED SUBJECTS Maria Salgado 1 , Mi Kwon 2 , Cristina Galvez Celada 1 , Monique Nijhuis 3 , Cristina Vilaplana 4 , Alessandra Bandera 5 , Annemarie Wensing 3 , Jose Luis Diez 2 , Javier Martinez-Picado 1 , for the IciStem Consortium 1 IrsiCaixa AIDS Rsr Inst, Badalona, Spain, 2 Gregorio Maranon Hosp, Madrid, Spain, 3 Univ Med Cntr Utrecht, Utrecht, Netherlands, 4 Inst for Hlth Sci Rsr Germans Trias i Pujol, Badalona, Spain, 5 Univ of Milano–Bicocca, Milan, Italy Background: Allogeneic stem cell transplantation (SCT) in HIV-infected subjects with hematological malignancies considerably reduces the viral reservoir. However, because antiretroviral treatment (cART) is maintained in these patients, it is difficult to infer the dynamics of viral rebound if cART is withdrawn. Herein, we explore the mice viral outgrowth approach (mVOA) as a potential tool to predict early viral rebound after cART discontinuation needed to evaluate the success of any eradication strategy. Methods: Blood leukapheresis was collected from 4 subjects that had undergone SCT within the IciStem cohort, and 2 “control” HIV-infected subjects who did not receive allotransplants, all of them on cART. Purified CD4 + T cells were transferred to 5 immunosupressed NSG mice per patient. Mouse health condition as well as human cell engraftment, T-cell subpopulation and T-cell activation were monitored. We quantified HIV RNA in mouse plasma, as well as total HIV-DNA on peripheral blood cells during the follow up and in spleen upon animal euthanasia. All procedures performed were reviewed and approved by the correspondent Ethical Committees and according to the national and European legislations. Results: We transferred 10-50 million CD4 + T cells per mouse. A total of five mice were used per study subject, 76% of them completing the follow up. Just one week post-infusion, we detected up to 10 4 HIV RNA copies/mL in the plasma of mice to which cells from control subjects had been transferred. Total HIV DNA was also detected in peripheral blood cells between weeks 1 and 3 (10 5 copies/million cells). Conversely, after 13 weeks of follow up, no HIV RNA or DNA was detected in either plasma or peripheral blood cells of mice to which CD4 + T cells from transplanted subjects had been transferred. These mice achieved a human lymphocyte engraftment over 40%, with human CD4 + T cells activation levels over 50% in all cases. Finally, total HIV DNA was detected in all spleens of mice from «control» cases (10 6 copies per million cells), but not in the ones infused with cells from allotransplanted cases. Conclusion: Mice qVOA might become a feasible tool to anticipate in vivo viral rebound dynamics after a substantial reduction of the HIV latent reservoir in eradication strategies, such as allogeneic stem cell transplantation. Compared with other assays, mVOA might reflect more physiological conditions of viral reactivation. 307 ROLE OF HIGH-MOBILITY GROUP A1 (HMGA1) GENE EXPRESSION IN REACTIVATION OF LATENT HIV Hosiana Abewe 1 , Amey Mukim 2 , Savitha Deshmukh 2 , Celsa A. Spina 2 , Douglas D. Richman 1 , Nadejda Beliakova-Bethell 2 1 Univ of California San Diego, La Jolla, CA, USA, 2 VA San Diego Hlthcare System, San Diego, CA, USA Background: Histone deacetylase inhibitors (HDACi) induced HIV RNA, but did not reduce the latent HIV reservoir in clinical trials. Previously, we demonstrated that HDACi suberoylanilide hydroxamic acid (SAHA) had mixed effects on host gene and protein expression, some of which would be predicted to promote, and some inhibit HIV reactivation.

Poster and Themed Discussion Abstracts

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