CROI 2018 Abstract eBook

Abstract eBook

Poster Abstracts

636 IMPACT OF OCCULT HCV INFECTION (OCI) ON SYSTEMIC IMMUNE ACTIVATION AFTER DAA THERAPY

637 POTENTIAL FOR SERUM MICRO-RNAs TO PREDICT FIBROSIS REGRESSION DURING HBV TREATMENT Cody Orr 1 , Rob Myers 2 , David Gong 2 , Biao Li 2 , Zhaoshi Jiang 2 , John Flaherty 2 , Anuj Gaggar 2 , G. Mani Subramanian 2 , Eric G. Meissner 1 1 Medical University of South Carolina, Charleston, SC, USA, 2 Gilead Sciences, Inc, Foster City, CA, USA Background: Long-term anti-viral therapy of patients with chronic hepatitis B virus (HBV) infection results in regression of fibrosis and cirrhosis in many, but not all patients. Fibrosis regression likely reduces the risk of hepatic decompensation and hepatocellular carcinoma. We asked whether serum micro-RNAs (miRNAs) could serve as biomarkers reflective of fibrosis regression. Methods: We used serum collected from subjects with chronic HBV infection and biopsy proven cirrhosis (Ishak score 5 or 6) who were treated for 5 years with tenofovir disoproxil fumarate (TDF) in clinical trials NCT00117676 and NCT00116805. In these trials, 71/96 subjects had biopsy-proven regression of cirrhosis at year 5. Serum samples from 14 subjects with fibrosis regression and 14 patients without fibrosis regression were available at baseline, year 1, and year 5 of treatment. mi-RNAs were isolated from serum and analyzed by quantitative PCR (qPCR) for 179 species using the Exiqon platform. Hemolyzed samples were excluded (n=3). Data were normalized to housekeeping miRNA species and then analyzed using non-parametric assumptions. Results: qPCR detected an average 159 miRNAs per sample, with 45 miRNAs detected in all samples. A comparison of patients with and without cirrhosis regression identified a number of miRNAs that differed at baseline (miR-421, miR-454-3p, miR-15b-5p, miR-141-3p), at year 1 of treatment (miR-199a-5p, miR-223-3p), and at year 5 of treatment (miR-199a-3p, miR-423-3p, miR-142-3p, miR-let-7d-5p). In addition, several species had differential change between baseline and 1 year of treatment (miR-21-5p, mir-29a-3p, miR-22-3p, miR-425- 3p, miR-30a-5p) and between baseline and year 5 of treatment (miR-103a-3p, miR-107, miR-34a, miR-885) in the two groups. Multiple of the identified miRNA species, including mir-21 and miR-199a-5p, have plausible mechanistic relationships to pathways associated with stellate cell activation and known mechanisms of fibrogenesis. Conclusion: In cirrhotic patients with chronic HBV infection treated with TDF, a number of serummi-RNAs differ both before and after treatment based on fibrosis regression. Further validation will determine their potential clinical utility as biomarkers. 638 HEPATIC EXPRESSION OF HSA-MIR-125A-5P AND FIBROSIS IN OVERT AND OCCULT HBV INFECTION Lorenzo Onorato 1 , Aniello Russo 1 , Giorgio de Stefano 2 , Carmine Minichini 1 , Mario Starace 1 , Pietro Filippini 1 , Loredana Alessio 1 , Nunzia Farella 2 , Nicoletta Potenza 1 , Evangelista Sagnelli 1 , Nicola Coppola 1 1 Second University of Naples, Caserta, Italy, 2 AORN dei Colli, Naples, Italy Background: MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression at post-transcriptional level; hsa-miR-125a-5p, a microRNA expressed in human liver, was shown to bind a specific sequence of S gene of HBV, inhibiting the expression of HBsAg in vitro. We aimed to correlate the hepatic expression pattern of hsa-miR-125a-5p with the concentrations of

David M. Asmuth 1 , Annie Y. Chen 2 , Netanya S. Utay 3 , Natalie Torok 1 , Patricia L. Poole 1 , Joy Vongspanich 1 , Kathleen R. Haight 1 , Anoma D. Sumasunderam 3 , Enkhmaa Byambaa 1 , Xiao-Dong Li 1 , Stuart T. Cohen 1 , Lars Berglund 1 , Mark Holodniy 4 , Tomasz I. Michalak 2 1 University of California Davis, Davis, CA, USA, 2 Memorial University of Newfoundland, St. John’s, NL, Canada, 3 University of Texas at Houston, Houston, TX, USA, 4 Veterans Affairs Palo Alto Health Care System, Palo Alto, CA, USA Background: OCI is the presence of low level HCV detection in plasma or cells when commercial assays report undetectable viral loads and was well described in the IFNα-era following sustained virologic response (SVR). The occurrence and frequency of OCI following Directly Acting Agent (DAA) therapy has not been reported. Further, the relationship between OCI and persistent immune activation following SVR is unknown. Methods: HCV/HIV co- and HCV monoinfected patients (pts) with prolonged SVR following DAA Rx were enrolled. 4 mL of plasma underwent 300,000g ultracentrifugation for 22 hrs. Extracted RNA from the plasma pellet and 5x10 6 PBMC underwent 2 rounds of PCR amplification using HCV 5’-UTR primers applying stringent precautions/controls in validated methods. Signal specificity was confirmed by nucleic acid hybridization (NAH) with a 5’-UTR-E2 32 P probe. Sensitivity of the RT-PCR/NAH assay is ∠ 10 viral equivalents/mL or ∠ 2.5 ve/ μg RNA (≈4 IU/mL or 1 IU/μg RNA, respectively). NS5A is sequenced for DAA resistance. Biomarkers of inflammation were measured using ELISA and Luminex kits. Nonparametric statistical methods are reported (median, IQR). Results: 12 HCV/HIV pts on chronic ART and 12 HCV-monoinfected pts with a median of 13.5 (9.5, 17) months since SVR12 provided a single blood sample (ie, 16.5 mo since any DAA). HCV/HIV pts had 663 T-cells (426, 916) and ∠ 20 HIV load. All but 1 were genotype (GN) 1 and all received either 12 or 24 weeks of sofosbuvir with ledipasvir except 1 who received ribavirin (GN 2). 50% in each cohort had documented cirrhosis. 7/12 (58%) co-infected and 4/12 (33%) mono- infected pts had HCV RNA positive-strand detected from plasma and/or PBMCs. Overall, only 3 were discordant with OCI in cells but not plasma (n=2) or plasma but not cells (n=1). In total, 3 had detectable HCV RNA negative-strand. sCD14 and I-FABP were higher than uninfected pts (p<0.0001) and mono-infected OCI positive pts had higher sCD14 than OCI negative pts (P<0.05). Others were not increased. Cirrhosis status did not impact OCI detection or biomarkers. Conclusion: These findings support the hypothesis that persistent OCI following DAA could contribute to systemic immune activation observed in these pts. Since B-cells and monocytes are known to harbor HCV, sorted cells and/or identification of the tissue reservoir may reveal higher OCI frequency. Despite OCI not leading to relapse historically, its impact on HCV-related comorbidities/HCC in cured pts needs further investigation.

Poster Abstracts

CROI 2018 236