الفهرس 
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Abstract
Most HCV cases become chronic hepatitis C (CHC), which may progress to liver fibrosis, cirrhosis, and hepatocellular carcinoma.
Rapid disease progression correlates with increasing number of fibrotic polymorphisms, emphasizing the complexity of the role played by genetic variation in the pathogenesis of HCV-related diseases. The present study included 150 HCV infected patients attending the Outpatient Clinic of Minia Health Insurance hospital, a referral center for treatment of HCV-infections in Egypt from January 2019 to July 2019. Fifty healthy volunteers were also included in the study as a control group. The patients were scheduled to receive (sofosbuvir (400 mg) + daclatasvir (60 mg) + ribavirin (600–1,000 mg) in divided doses for 12 weeks. All patients subjected to:
Thorough clinical history, clinical examination and laboratory investigations including: Complete Blood Count (CBC), Liver function tests (bilirubin, AST, ALT, total proteins and albumin) and AFP.
Fibrosis assessment by Fib-4 index was calculated using Sterling formula to assess fibrosis state. The detection of the relative expression levels of serum miR-21, miR-181c, miR-199 and miR-448 was performed using Real-time quantitative reverse transcription polymerase chain reaction (RT- qPCR) after microRNA extraction. SNORD68 gene was used as a house keeping gene for relative quantification of micro RNAs.
Detection of interleukin 28B polymorphisms at areas, rs12980275 and rs8099917 was carried out by PCR-RFLP assay. Out of 150 HCV- patients, 141 (94%) were responders to (SOF/ DCV) + RIB regimen and Only 9 patients were resistant to the treatment (6%).
Regarding fibrosis, out of 141 responders, 112 were belonging to stage 0- 1and 29 were belonging to stage2-3 while all non-responders were belonging to stage 2-3 of fibrosis.
MiR-21, miR-181c, miR-199 and miR-448 were upregulated in HCV- patients compared to controls with significant fold increase (P<0.001, for each). Analyzing serum miRNAs profiles in HCV- patients regarding treatment response revealed that only miR-199 showed significant fold increase in non- responders’ patients (P<0.001). However, the other studied serum miRNAs profiles showed no significant difference between responders and non- responders patients. ROC curve analysis for diagnostic accuracy of serum miRNA-199, showed that miRNA-199 was significantly upregulated in non- responders patients (p value <0.001*). Both miRNAs 181c and 448 showed a significant decrease in late stages than early stages fibrosis with significant p value < 0.05. MiRNA-199 showed a significant rise in late stages fibrosis than in early stages with a highly significant p value of <0.001. However, miRNA-21 showed no significant rise between the 2 groups. ROC curve analysis for diagnostic accuracy of serum miRNAs as diagnostic biomarkers differentiating patients with early-stage fibrosis from patients with advanced fibrosis showed that MiRNA-199 was significantly upregulated (p<0.001*) and miRNA-448 was significantly down regulated (p<0.001*).
Regarding SNPs of IL28B at positions rs12980275and IL28B rs8099917, the results were as follows: GG genotype of IL28B (rs8099917) was associated with an increased susceptibility to infection while TT genotype was the predominant in controls, in responders and in patients with early fibrosis (p values<.0.05). There were no statistically significant differences between the different genotypes of IL28B rs12980275 among the study population.
Conclusion
In conclusion, our results indicate that the new regimen by DCV and SOF with ribavirin had high efficacy among HCV genotype 4 patients. These findings indicated that there is association between therapeutic failure and stage of fibrosis. Our results suggested that circulating miR-199 and IL28Brs8099917 polymorphism may be used as a predictor molecular marker for response to DAA in HCV genotype 4 patient.
Also, miR-199, 448 and IL28Brs8099917 may be used as a noninvasive marker to predict stages of fibrosis as they are easily accessible from only blood sample and has a very good stability and can be detected easily through real time PCR. Also, our results suggested GG genotype of IL28Brs8099917 is associated with an increased susceptibility to infection and TT genotype may be a protective gene and is associated with achieving SVR to combined DCV and SOF therapy in genotype 4 infected HCV patients. Moreover, no statistically significant difference was observed between the different genotypes of IL28B rs12980275 between responders and non-responders.