CROI 2018 Abstract eBook

Abstract eBook

Poster Abstracts

HBV-DNA in liver tissue and the progression of fibrosis in 74 consecutive patients with overt or occult HBV infection. Methods: We enrolled all the consecutive HBsAg positive treatment naive patients who underwent a diagnostic liver biopsy (overt HBV group) between April 2007 and April 2015 in one of the three liver units participating in the study. Moreover, of the 68 HBsAg negative patients with hepatocellular carcinoma who underwent a liver biopsy in the same period, those with a positive HBV-DNA in liver tissue were enrolled (occult HBV group). HIV coinfected patients were excluded from the analysis. For each patient we collected a plasma sample and two fragments from the extremities of liver biopsy. Tissue concentrations of HBV-DNA and hsa-miR-125a-5p were analysed by real-time quantitative PCR. Necroinflammatory activity and fibrosis were evaluated according to the Ishak score. Results: 64 HBsAg positive and 10 HBsAg negative patients were included in the study. In the overt HBV group, 54.7% showed a mild fibrosis, 26.6% a moderate fibrosis, while the remaining had a cirrhosis. All patients in the occult HBV group were cirrhotic. Patients with more advanced fibrosis showed a higher mean age as compared to those with mild (p<0.00001) or moderate fibrosis (p<0.00001). Similarly, patients with occult B infection were older than HBsAg positive patients. Liver concentrations of miR-125a-5p were significantly higher in patients with cirrhosis as compared to patients with mild (p=0.0002) or moderate fibrosis (p=0.0006). Moreover we found an inverse correlation, although not statistically significant, between the tissue HBV-DNA levels and the staging of fibrosis. Eventually, patients with higher liver HBV-DNA concentrations showed a slightly lower microRNA expression (p=0.19). Conclusion: This study demonstrates a strong correlation between the tissue expression of hsa-miR-125a-5p and the progression of liver damage in a group of patients with occult or overt HBV infection. However further studies are needed to investigate the role of this miRNA in the pathogenesis of HBV infection.

R-193b-5p, miR-483-5p, miR-1224-5p, miR-125b-1-3p, miR-885-5p, miR-100-5p, miR-192-5p, miR-592, miR-193b-3p, miR-125b-2-3p, miR-629-5p, miR-99a-5p and miR-203a-3p) were upregulated in HIV-1 monoinfected patients. These 13 miRs were also upregulated in HIV-1-HCV coinfected patients. Importantly, these 13 miRs significantly and positively correlated (p< 0.05) with ALT and AST levels with most of the study samples including healthy donors. Two of the former miRs, miR-99a-5p and miR-100-5p, were significantly upregulated in HIV-1-HCV coinfected patients who progressed to liver cirrhosis, although at the time of sampling all compared coinfected patients had liver fibrosis-0. Finally, the comparison of miR profiles of HIV-1 monoinfected patients with elevated ALT or focal nodular hyperplasia with HIV-1 monoinfected patients displaying normal levels of ALT showed significantly altered expression of 25 and 7 miRs, respectively. The two more highly overexpressed miRs in these two cohorts were miR-122-3p and miR-193b-5p. The levels of these 2 miRs significantly correlated with liver fibrosis progression in our cohort of HCV monoinfected patients (p< 0.05). Conclusion: These results reveal that HIV-1 infection affects liver miR metabolism, even in the absence of co-infection with hepatotropic viruses, and highlights the potential of miRs as biomarkers in the progression of liver injury in HIV-1 infected patients. 640 MIRNA PROFILE OF HCV SPONTANEOUS CLEARANCE INDIVIDUALS SHOW PREVIOUS HCV INFECTION Óscar Brochado Kith 1 , Alicia Gómez Sanz 1 , Luis M. Real 2 , Javier Crespo 3 , Pablo Ryan 4 , Juan Macías 2 , Joaquín Cabezas 3 , Juan A. Pineda 2 , María T. Arias-Loste 3 , Isabel Cuesta 1 , Sara Monzón-Fernández 1 , Luz M. Medrano 1 , María Á. Jimenez- Sousa 1 , Salvador Resino 1 , Amanda Fernández-Rodríguez 1 1 Institute de Salud Carlos III, Majadahonda, Spain, 2 Hospital Universitario de Valme, Seville, Spain, 3 Hospital Universitario Marqués de Valdecilla, Santander, Spain, 4 Hospital Universitario Infanta Leonor, Madrid, Spain Background: Hepatitis C virus (HCV) usually progress to chronic infection, but a small percentage of patients clarify the virus spontaneously. Factors involved in HCV clarification seem to be related to interaction between HCV and host innate/adaptive immune system. However, little is known about the cost for the immune system after HCV infection. Genomic background of patients plays also a key role in the antiviral immune response against HCV, being the microRNAs (miRNA) essential. These small RNAs are post-transcriptional regulators of gene expression, which play a key role in the innate and adaptive immune response and they are actively involved in the HCV cycle. In this study, we analyzed the miRNA profile of peripheral blood mononuclear cells (PBMCs), from individuals that spontaneously clarified HCV, chronic patients and healthy donors, to evaluate the effects of HCV infection on immune system. Methods: We have sequenced the miRNA profiles from PBMCs of 96 individuals: 32 HCV chronic patients, 32 individuals that spontaneously clarified HCV, and 32 healthy donors. Library was performed with TruSeq smallRNA (Illumina), and sequenced in an Illumina HiSeq 2500 with 1x50 bp SE reads. Quality control of the reads was evaluated with FastQC. Adapters were removed by Cutadpat. Different methods were used for miRNA quantification, normalization, and differential expression analysis (mirDeep2, edgeR, DESeq and NOIseq). Principal Component Analysis (PCA) was performed. A fold change higher than 2 and false discover rate (FDR) of 5%, were considered. Results: We did not find any differentially expressed miRNAs between HCV spontaneous clarifiers and chronic patients. However, both groups showed similar expression differences with healthy donor individuals (9 and 8 miRNAs respectively). PCA also confirmed that spontaneous clarifier’s patients were more similar to HCV chronic patients than healthy donors (Figure). The potentially altered molecular pathways by these miRNAs (miRPath) belongs manly to fatty acids pathways, which seem to be repressed in donors. MIRNAs related to chronic myeloid leukemia were also altered, showing higher expression within chronic patients. Finally, some miRNAs involved in the strogen signaling pathways were impaired in chronic patients. Conclusion: It seems that the HCV infection leaves a fingerprint in the immune system that do not vanish after HCV clarification. PBMC´s microRNA expression profile of spontaneously clarified individuals and HCV chronic patients were similar

Poster Abstracts

639 CIRCULATING MICRORNAs IN HIV PATIENTS REVEAL SPECIFIC SIGNATURES FOR LIVER DAMAGE

Miguel Angel Martinez 1 , Sandra Franco 1 , Daniela Buccione 2 , Beatriz Mothe 1 , Lidia Ruiz 1 , Maria Nevot 1 , Ana Jordan-Paiz 1 , Raquel Pluvinet 2 , Susanna Aussó 2 , Rosa M. Morillas 2 , Lauro Sumoy 2 , Cristina Tural 2 1 IrsiCaixa Institute for AIDS Research, Badalona, Spain, 2 Hospital Germans Trias i Pujol, Barcelona, Spain Background: HIV-1-induced inflammation and/or long-term antiretroviral drug toxicity may contribute to liver disease evolution. MicroRNAs (miRs) are potential disease biomarkers and therapeutic targets. Here, we investigated circulating plasma miRs among HIV-1 infected patients and their association with liver injury. Methods: Large-scale deep sequencing analysis of small RNA expression was performed on plasma samples from 101 HIV-1 patients with elevated ALT, focal nodular hyperplasia or HCV coinfection. Twenty one healthy uninfected donors and 22 HCV monoinfected patients were also analyzed. Results: A total of 1425 different miRs were identified. When compared with healthy donors, monoinfected HIV-1 or HCV patients showed significant (fold change> 2 and adjusted p< 0.05) altered expression of 25 and 70 miRs, respectively. Of the 25 altered miRs found in HIV-1 patients, 19 were also found in the HCV mono-infected patients. Indeed, 13 of the 14 miRs more highly upregulated (ranging a 9.3-3.4 fold increase) in HCV monoinfected patients (mi-

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