Leona Dold

Circulating MicroRNAs as a marker for liver injury in human immunodeficiency virus patients

Human immunodeficiency virus (HIV) and hepatitis virus coinfection amplify and accelerate hepatic injury. MicroRNAs (miRNAs) are small regulatory RNAs suggested as biomarkers for liver injury. We analyzed the circulating levels of miRNAs in HIV patients with regard to the extent and etiology of liver injury. Total RNA was extracted from 335 serum samples of HIV patients and 22 healthy control participants using Qiazol. Comprehensive polymerase chain reaction (PCR) array analyses (768 miRNA) were performed in serum samples of eight HIV, eight HIV/HCV (hepatitis C virus), six HCV patients, and three healthy controls. Reverse transcription (RT)‐PCR measured levels of miRNA‐122, miRNA‐22, and miRNA‐34a in serum samples of 335 patients and 19 healthy control participants. Liver injury and fibrosis in these patients were defined using aspartate aminotransferase (AST) levels, fibrosis‐4 (FIB‐4) index and AST‐to‐platelet ratio index (APRI) score. The miRNA pattern of HIV/HCV samples showed altered expression of 57 and 33 miRNA compared to HCV and HIV infection, respectively. miRNA‐122, miRNA‐22, and miRNA‐34a were highly up‐regulated in HIV/HCV patients. Analyzing the entire cohort, these miRNAs were correlated with liver function tests and were independent predictors of liver injury (AST >2 × ULN). miRNA‐122 and miRNA‐22 were associated with relevant fibrosis (FIB‐4 >1.45; APRI >1). Circulating levels of miRNA‐122 were independent predictors for relevant fibrosis in HIV patients. Interestingly, miRNA‐122 and miRNA‐34a levels were higher in HIV/HCV patients, miRNA‐22 levels were highest in HIV/HBV patients, and circulating levels of miRNA‐34a correlated positively with illicit drug use and ethanol consumption. Conclusion: Circulating miRNA‐122, miRNA‐22, and miRNA‐34a correlates with the etiology of liver injury in HIV patients. These biomarkers not only mirror different mechanisms of hepatic injury, but also are independent predictors of liver injury in HIV patients. (Hepatology 2015;61:46–55)

Genetic polymorphisms associated with fatty liver disease and fibrosis in HIV positive patients receiving combined antiretroviral therapy (cART)

Hepatic steatosis can occur with any antiretroviral therapy (cART). Although single nucleotide polymorphisms (SNPs) have been identified to predispose to alcoholic and non-alcoholic fatty liver disease, their role for treatment-associated steatosis in HIV-positive patients remains unclear. We determined the frequency of PNPLA3 (rs738409), CSPG3/NCAN (rs2228603), GCKR (rs780094), PPP1R3B (rs4240624), TM6SF (rs8542926), LYPLAL1 (rs12137855) and MBOAT7 (rs626283) by RT-PCR in 117 HIV-positive patients on cART and stratified participants based on their “controlled attenuation parameter” (CAP) into probable (CAP: 215–300 dB/m) and definite (CAP >300 dB/m) hepatic steatosis. We analyzed CAP values and routine metabolic parameters according to the allele frequencies. Sixty-five (55.6%) and 13 (11.1%) patients were allocated to probable and definite steatosis. CAP values (p = 0.012) and serum triglycerides (p = 0.043) were increased in carriers of the GCKR (rs780094) A allele. Cox logistic regression identified triglycerides (p = 0.006), bilirubin (p = 0.021) and BMI (p = 0.068), but not the genetic parameters as risk factors for the occurrence of hepatic steatosis. Taken together, according to the limited sample size, this exploratory study generates the hypothesis that genetic polymorphisms seem to exert minor effects on the risk for fatty liver disease in HIV-positive patients on cART. Nevertheless, SNPs may modify metabolic complications once metabolic abnormalities have developed. Hence, subsequent analysis of a larger cohort is needed.

Survival and HLA-B*57 in HIV/HCV Co-Infected Patients on Highly Active Antiretroviral Therapy (HAART)

Background and aims HLA class I alleles, in particular HLA-B*57, constitute the most consistent host factor determining outcomes in untreated HCV- and HIV-infection. In this prospective cohort study, we analysed the impact of HLA class I alleles on all-cause mortality in patients with HIV-, HCV- and HIV/HCV- co-infection receiving HAART. Methods In 2003 HLA-A and B alleles were determined and patients were prospectively followed in 3-month intervals until 2013 or death. HLA-A and B alleles were determined by strand-specific oligonucleotide hybridisation and PCR in 468 Caucasian patients with HCV- (n=120), HIV- (n=186) and HIV/HCV-infection (n=162). All patients with HIV-infection were on HAART. In each patient group, HLA class I-associated survival was analysed by Kaplan-Meier method and Cox regression analysis. Results At recruitment the proportion of patients carrying a HLA-B*57 allele differed between HIV- (12.9%) and HCV-infection (4.2%). Kaplan Meier analysis revealed significantly increased mortality in HLA-B*57-positive patients with HIV-infection (p=0.032) and HIV/HCV-co-infection (p=0.004), which was apparently linked to non-viral infections. Cox logistic regression analysis confirmed HLA-B*57 (p=0.001), serum gamma-glutamyltranspeptidase (p=0.003), serum bilirubin (p=0.022) and CD4 counts (p=0.041) as independent predictors of death in HIV-infected patients. Conclusion Differences in the prevalence of HLA-B*57 at study entry between HIV- and HCV- infected patients may reflect immune selection in the absence of antiviral therapy. When patients were treated with HAART, however, HLA-B*57 was associated with increased mortality and risk to die from bacterial infections and sepsis, suggesting an ambiguous role of HLA-B*57 for survival in HIV/HCV infection depending on the circumstances.

Percutaneous Transgastral biliodigestive diversion as treatment option for BRIC

A 51-year-old woman was hospitalized with jaundice, pruritus and nausea. Four weeks before a common bile duct stone had been removed by endoscopic retrograde cholangiopancreatography (ERCP) with papillotomy. Blood tests showed isolated bilirubinaemia (15 mg/dl), while all liver enzymes were in the normal range. In a second ERCP we saw a wide opened major papilla with unimpaired bile flow and without any stones or strictures. Liver histology showed portal inflammation and cholestasis and genetic testing revealed homozygous mutation in the ABCB11 gene, which encodes a bile salt exporter pump. Therefore, we hypothesized that the jaundice was resulting from benign recurrent intrahepatic cholestasis (BRIC 2). This article is protected by copyright. All rights reserved.

359 HLA FOOTPRINTS IN HIV/HCV CO-INFECTED PATIENTS

Background and Aims: HLA-diversity is a major host genetic risk factor in infectious diseases. In particular, several studies have described associations between distinct HLA alleles and outcomes of HCV and HIV infection. The objective of this study was to analyse the distribution of HLA Class I alleles in HIV/HCV co-infected individuals in order to identify HLA-disease associations that reflect the impact of this dual infection. Methods: Clinical data from 1110 Caucasian patients (HCV: n =477, HIV: n =181, HIV and HCV: n =346, healthy controls: n = 106) were retrieved by retrospective chart review. HLA-A and B allele distribution were determined by molecular methods (SSO and SSP) with HLA-resolution reduced to the 2-digit level to enable uniform comparisons. Fisher’s exact test and Chi-square tests with Yates correction for multiple testing were applied for statistical analysis. Results: Patients with HCV mono-infection were significantly older than patients in each other group (p < 0.001). MSM was the predominant risk factor for HIV infection (52%), while most HIV/HCV and HCV-infected patients had been infected via blood exposure (83%). The frequency of HLA alleles HLA-A*30 (p =0.023), HLA-B*08 (p =0.003), HLA-B*39 (p =0.003), and HLAB*49 (p =0.032) did not differ between the patient groups but was significantly reduced in infected patients as compared to healthy controls irrespective from the underlying type of infection. In contrast, HLA-B*57, which is assumed to increase chances of spontaneous HCV resolution and delay HIV progression, was selectively increased in patients with HIV mono-infection (13%) as compared to each other group (healthy controls: 9%, HIV/HCV and HCV mono-infection: 6% each; p =0.022). Conclusion: Reduced detection rates in HIVor HCV-monoand HIV/HCV co-infected patients indicate that alleles HLA-A*30, HLAB*08, HLA-B*39, and HLA-B*49 carry a poor prognosis but do not reflect interactions between HIVand HCV-infections. Patients with blood-borne infections usually acquire HCV first and HIV later on. Thus, finding a high HLA-B57 carrier rate in HIV+ patients but in none of the other groups indicates a protective role of this allele for both HIV and HCV infection. Taken together long-term exposure of Caucasian patients to HIV and HCV infection seems to induce subtle footprints in their HLA class I repertoire.