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Publication Abstract Display
Type: Published Abstract
Title: Characterizing the HIV DNA Reservoirs in Whole-Body Tissues in the Last Gift Cohort.
Authors: Oliveira M, Murrell B, Vollbrecht T, Concha-Garcia S, Kumar V, Porrachia M, Scott B, Laymman L, Ignacio K, Smith D, Gianella S
Year: 2019
Publication: CROI
Volume: Issue: Pages:
Abstract:HIV persistence in cellular reservoirs is the main barrier to a cure. The size and composition of HIV DNA populations in solid tissues during suppressive antiretroviral therapy (ART) is not well characterized. We examined the distribution and genotypic composition of the HIV DNA populations across paired post-mortem tissues from one virally suppressed person living with HIV (PLWH) from the Last Gift (LG) Cohort. The LG cohort enrolls altruistic, terminally-ill PLWH, who are closely followed until the time of death and donate their whole-body for HIV research. Blood and tissues are collected by rapid-autopsy and cryopreserved within 6h from death. From each sample, we extracted total DNA and quantified levels of HIV DNA (gag) by droplet-digital PCR. The genotypic composition of the HIV DNA in tissues was evaluated using a high-throughput single genome amplification and the PacBio platform to deeply sequence full-length HIV envelope (FL HIV-env). The participant was a 72-year-old man with chronic HIV infection and metastatic pancreatic cancer. He enrolled in the LG study 5 months before death. He was on ART and had undetectable HIV RNA in blood plasma, up to 7 hours before death (<20 copies/ml). From 26 paired post-mortem tissues, HIV DNA was detected in 24 samples, including brain (3-11 cps HIV/106 cells), gastrointestinal (45-211), urogenital tract (46-377), lymphoid (22-243) and adipose (13-874) tissues. HIV DNA was undetectable in parietal and motor cortex. We obtained 107 individual FL HIV-env sequences across 10 tissues (median 10.5 sequences/tissue), of which 60 were unique. The maximum likelihood phylogeny showed a deep divergence, segregating the tree into two lineages, which differed by co-receptor tropism, based on in silico tropism prediction (geno2pheno). Interestingly, 100 FL HIV-env sequences were genetically intact, while 7 sequences were non-functional, with major deletions, frame shifts, and stop codon mutations. HIV-env migration appeared to be extensive, with many identical sequences sampled in multiple tissue compartments. HIV DNA was detected in most anatomic tissues despite long-term ART and confirmed undetectable HV RNA at the time of death. Based on the FL HIV-env sequencing, most HIV reservoirs appeared to be intact provirus and may present different viral tropisms. The LG cohort poses a unique opportunity to characterize the HIV reservoirs in anatomic compartments, which is crucial to provide insights for future HIV cure strategies.

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