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miRNA-26b Downregulation in Peripheral Blood Mononuclear Cells of Patients with Hepatitis C Associated Lymphomas is Restored by Successful Interferon-Free Antiviral Therapy

miRNA-26b Downregulation in Peripheral Blood Mononuclear Cells of Patients with Hepatitis C... Antiviral Therapy 2019; 24:437–442 (doi: 10.3851/IMP3322) Original article miRNA-26b downregulation in peripheral blood mononuclear cells of patients with hepatitis C associated lymphomas is restored by successful interferon-free antiviral therapy 1,2,3 1,3 1 4 4 Jan Peveling-Oberhag *, Katrin Bankov , Georg Dultz , Olivier Ballo , Julian Lohmeyer , 4 4 1 1 3 1 Uta Brunnberg , Vasile Marcu , Dirk Walter , Stefan Zeuzem , Martin-Leo Hansmann , Tania M Welzel , Johannes Vermehren Department of Internal Medicine 1, University Hospital Frankfurt, Frankfurt am Main, Germany Department of Internal Medicine 1, Robert-Bosch-Hospital, Stuttgart, Germany Senckenberg Institute for Pathology, University Hospital Frankfurt, Frankfurt am Main, Germany Department of Internal Medicine 2, University Hospital Frankfurt, Frankfurt am Main, Germany *Corresponding author e-mail: jan.peveling-oberhag@rbk.de Background: Patients with chronic HCV infection are at at different time points during antiviral therapy for all increased risk of developing B-cell non-Hodgkin lym- included patients as well as for a total of 10 controls with phoma (B-NHL). Regression of HCV-associated B-NHL (n=5) and without (n=5) chronic HCV infection. (HCV-NHL) can be achieved through HCV eradication Results: All patients had marginal zone lymphoma subtype using interferon (IFN). However, only about two-thirds and received different DAA regimens for 12–24 weeks. of patients with sustained virological response (SVR) also All four patients achieved SVR, but only three patients had a consecutive lymphoma response. miRNA-26b is also had lymphoma response (one complete response, two associated with HCV-NHL response to antiviral therapy. partial responses). One patient showed progression to a Recent data suggest that IFN-free direct-acting antiviral high-grade lymphoma subtype after SVR. miRNA-26b (DAA) regimens also have anti-lymphoma activity in this expression was generally decreased in patients with HCV- patient population. NHL. Moreover, miRNA-26b expression was restored in Methods: We report four patients with HCV-NHL who those HCV-NHL patients with lymphoma response after 6 were treated with different IFN-free DAA regimens as months (P=0.009). oncological monotherapy in our centre between 2015 Conclusions: We have demonstrated that IFN-free DAA and 2016. We analysed the virological and lymphoprolif- treatment of HCV can improve or even cure NHL. miRNA- erative disease response. Moreover, we analysed miRNA- 26b-levels could be a potentially useful biomarker to pre- 26b expression in peripheral blood mononuclear cells dict lymphoma response in HCV-NHL patients. Introduction Worldwide, an estimated 71 million people are causal relationship between HCV and B-NHL is made chronically infected with HCV, a hepatotropic and by interventional studies demonstrating that antiviral potentially lymphotropic virus [1–4]. In the last two therapy using interferon (IFN) or pegylated IFN with decades, evidence from epidemiological studies, bio- and without addition of ribavirin (RBV) frequently logical insights, and especially therapeutic approaches leads to regression of HCV-associated lymphomas provided strong support for an association between (HCV-NHL) [7,8]. HCV and B-cell non-Hodgkin’s lymphomas (NHL) [5]. Piluso et al. [9] showed downregulation of the HCV-related B-cell proliferation represents an impor- miRNA miR-26b in peripheral blood mononuclear tant model of virus-driven autoimmune/neoplastic dis- cells (PBMCs) of patients with HCV-NHL compared order leading to mixed cryoglobulinaemia (MC) and/or with controls. Downregulation of the tumour suppres- NHL [6]. However, the most compelling argument for a sive microRNA-26b (miR-26b) was first connected to ©2019 International Medical Press 1359-6535 (print) 2040-2058 (online) 437 AVT-19-OA-4510_Peveling-Oberhag.indd 437 AVT-19-OA-4510_Peveling-Oberhag.indd 437 22/11/2019 13:11:10 22/11/2019 13:11:10 J Peveling-Oberhag et al. HCV-NHL in microdissected primary tissue of HCV- 12 weeks after completion of therapy. HCV viral load associated splenic marginal zone lymphoma [10]. Con- was measured using the COBAS AmpliPrep/COBAS secutively, the same miRNA was found downregulated TaqMan HCV Test, v2.0 (Roche Diagnostics, Man- in PBMCs of patients with MC and HCV-NHL, also nheim, Germany). HCV genotypes were determined demonstrating restoration of miR-26b expression by the VERSANT HCV Genotype 2.0 assay (Siemens in MC patients who achieved a sustained virological Healthcare Diagnostics, Erlangen, Germany). response (SVR) after treatment with IFN and RBV [11]. These findings indicate that miR-26b could be involved miR-26b expression analysis in the mechanism of HCV-driven lymphomagenesis PBMCs were isolated from fresh anticoagulated blood and could potentially serve as a valuable biomarker to by gradient precipitation on Lymphoprep (Axis-Shield monitor lymphoma response during antiviral therapy. PoC AS, Oslo, Norway) according to the manufactur- Antiviral therapy of HCV underwent a revolution er’s instructions. After the second wash, the cells were within the last several years. After almost 25 years of counted and stored at -80°C. RNA extraction was IFN-based therapies, highly effective direct-acting anti- performed using Trizol reagent (Invitrogen, Carlsbad, viral (DAA) drugs are now standard of care for treat- CA, USA) from 5×106 PBMCs according to the manu- ment of chronic HCV infection. With these DAAs, viral facturer’s instructions. Reverse transcription was done eradication can now be achieved in approximately 95% using the TaqMan MicroRNA RT kit (Applied Biosys- of all patients across different genotypes and fibrosis tems, Foster City, CA, USA) and 100 ng of total RNA. stages [12]. Expression levels of human miR-let-7g and miR-26b Data on IFN-free treatment of HCV-NHL is still were evaluated by real-time PCR using specific TaqMan scarce and mainly based on case reports [13–17]. Now MicroRNA Assays (Applied Biosystems), according to the first multicentre retrospective series of HCV-NHL the manufacturer’s instructions. treated with IFN-free antiviral therapy shows promis- Relative expression levels of the different miRNAs ing results but also reports on patients with SVR who were evaluated with the delta-Ct method, using miR- did not show favourable oncological response [18–20]. let-7d as internal control to normalize miRNA expres- The aim of this study was to investigate the clinical sion levels, as previously described [11]. course of HCV-NHL patients from our centre treated with IFN-free DAA regimen. In addition, we evaluated Statistical analysis miR-26b expression as a biomarker for lymphoma Statistical analyses and illustrations were performed response in PBMC of HCV-NHL patients undergoing using GraphPad Prism 5.0 (GraphPad Software, La Jolla, DAA therapies. CA, USA). Student’s two-sample t-test for either paired or unpaired samples was used to compare miRNA expression values. All P-values were two-tailed and were Methods considered statistically significant for levels ≤ 0.05. Study design and patients The current study is a retrospective analysis, which was Results approved by the local Ethics Committee of the Univer- Cases sity Hospital Frankfurt, Germany (Ethik-Kommission des Fachbereichs Medizin der Goethe-Universität). Writ- Case 1 ten informed consent was obtained from all patients. The 49-year-old male was diagnosed with HCV GT3a Clinical data were abstracted from electronic records. infection in 2005 following intravenous drug use. Viral Patients with HCV-NHL (n=4) and matched individu- load was 541,000 IE/ml. Pretreatment transient elas- als with chronic HCV without lymphoma (n=5) were tography (FibroScan, Echosens, Paris, France) showed a recruited from our outpatient clinic specialized in HCV liver stiffness of 5.9 KPa, corresponding to F0–F1 fibro- treatment. Additionally, PBMCs from healthy volun- sis. In 2015, the patient presented to his primary care teers (n=5) were acquired from blood donors. physician with abdominal pain in the left upper quad- rant. There were no B symptoms present. On physical Clinical and virological end points examination, the treating physician diagnosed spleno- The primary oncological end point was overall tumour megaly, and the blood count showed an increased leuko- 3 3 response. Response evaluation was based on Lugano cyte/lymphocyte cell count (60,000/mm /42,000/mm ). classification criteria for lymphomas [12]. Bone mar - The patient was referred to a specialized centre for hae- row biopsy as well as CT scans were performed before matology/oncology. The high lymphocyte count was start of antiviral therapy and repeated at the end and 6 confirmed showing a monoclonal B-cell immune phe- months after treatment. SVR was defined as undetecta- notype with the morphology of villous lymphocytes. ble HCV viral load with a sensitive real-time PCR assay Cryoglobulinaemia was ruled out. CT scan showed 438 ©2019 International Medical Press AVT-19-OA-4510_Peveling-Oberhag.indd 438 AVT-19-OA-4510_Peveling-Oberhag.indd 438 22/11/2019 13:11:10 22/11/2019 13:11:10 miR-26b expression during DAA therapy for hepatitis C splenomegaly (longitudinal diameter of 27 cm) and an daily) for 12 weeks. HCV RNA was undetectable from enlargement of paraaortic, paracaval and mesenteric as week 2 and remained undetectable at 12 and 24 weeks well as supraclavicular and mediastinal lymph nodes. after the end of treatment. The patient was symptom Bone marrow biopsy showed lymphoid nodular infil- free 8 weeks after initiation of antiviral therapy. The tration of more than 40%. The final diagnosis was stage staging CT scan 6 months after the start of treatment IV leukaemic splenic marginal zone lymphoma. As the showed a minimal decrease in spleen size (21 to 18 cm) patient was asymptomatic at presentation at the hae- but several mediastinal lymph nodes were noted which matology centre, he was then referred to our centre for had grown in size. Biopsies of the mediastinal lymph antiviral therapy. The patient received a combination nodes were taken using endobronchial ultrasound. His- of sofosbuvir 400 mg once daily and daclatasvir 60 mg tology showed a high grade B-NHL with high prolif- once daily for 12 weeks. HCV RNA was undetectable eration (Ki67 index 80%) consistent with diffuse large from week 2 and remained undetectable at 12 and 24 B-cell lymphoma (DLBCL). Therefore, response to weeks after the end of treatment. At week 4 of treat- antiviral treatment was classified as progressive disease ment, the patient presented with abdominal pain due with transformation into a high-grade lymphoma. Con- to splenomegaly and symptom control was achieved secutively the patient received immuno-chemotherapy with use of non-steroidal anti-inflammatory drugs in (R-CHOP) with partial response after three cycles and combination with transdermal fentanyl. At week 8 of is currently planned to receive three additional cycles. treatment, the pain subsided gradually with no need for any pain medication after week 12. The blood lympho- Case 3 cyte count increased initially from baseline to week 2 The 78-year-old female was diagnosed with splenic (44,000 to 54,000/mm ), but steadily decreased (39,000 marginal zone lymphoma in 2015 from an ultrasound at follow-up week 12) to nearly normal values 1 year guided breast biopsy, after the patient noted a lump in after antiviral therapy, with the B-cell clone still detect- the right breast. Subsequent work-up revealed chronic able. Moreover, bone marrow biopsy 3 months after infection with HCV genotype-2, while no typical risk initiation of treatment showed residual but reduced factors for contracting HCV infection were noted. Viral lymphoma infiltration (25%). The CT scan 6 months load was 1,090,000 IE/ml. Transient elastography was after antiviral therapy showed decrease of all lymph not available but conventional ultrasound did not show node diameters by more than 50% and a decrease in signs of cirrhosis. The patient reported weight loss of spleen size to nearly normal size. The response was clas- about 4 kilograms but no fevers or night sweats. Stag- sified as partial response. ing CT scans revealed that the breast and mediastinal lymph nodes were the only localizations of the lym- Case 2 phoma. There was no bone marrow involvement. The A 56-year-old male, who was diagnosed with HCV peripheral blood count showed normal results. The final genotype-1b infection in 2013 following intravenous diagnosis was stage IVA low-grade splenic marginal drug use in the 1990s. Viral load was 943,000 IE/ml. zone lymphoma. The patient was then referred to our Pretreatment transient elastography showed a liver centre for HCV treatment. In line with guideline recom- stiffness of 15.1 KPa, corresponding to F4 fibrosis mendations at the time, the patient received sofosbu- score and liver morphology in the abdominal ultra- vir 400 mg once daily plus weight-based RBV for 12 sound was consistent with compensated cirrhosis. At weeks. HCV RNA was undetectable from week 4 and the time of HCV diagnosis, the patient was also diag- remained undetectable at 12 and 24 weeks after the end nosed with a low-grade B-NHL. The patient suffered of treatment. The staging CT scan 6 months after the from occasional fever episodes and night sweats. CT start of therapy showed complete response with mini- scan showed splenomegaly (longitudinal diameter of mal scarring tissue at the breast biopsy site, which did 20.7 cm) and paraaortic, mesenteric as well as medi- not show residual lymphoma after re-biopsy. astinal lymph node enlargement. Bone marrow biopsy showed monoclonal lymphoid infiltration of more than Case 4 50% with positivity for CD79, CD3, CD20 and CD34. The 60-year-old male patient was diagnosed with Peripheral blood count showed mild anaemia but oth- chronic HCV genotype-1b infection following diagnosis erwise normal results. The final diagnosis was stage IVB of splenic marginal zone lymphoma in 2011. He was ini- low-grade marginal zone lymphoma. The patient was tially treated with splenectomy because of symptomatic followed-up with staging CT scans every 3 to 6 months splenomegaly. After splenectomy, the lymphoma per- and showed a stable course. In June 2015, after inter- sisted in the bone marrow with a tumour cell infiltration disciplinary discussion, the patient received antiviral of 40–50%. There were no other disease manifestations therapy with ombitasvir/paritaprevir (12.5 mg/75 mg and the patient had no B symptoms. The final diag- two tablets once daily) and dasabuvir (250 mg twice nosis was stage IV splenic marginal zone lymphoma. Antiviral Therapy 24.6 439 AVT-19-OA-4510_Peveling-Oberhag.indd 439 AVT-19-OA-4510_Peveling-Oberhag.indd 439 22/11/2019 13:11:10 22/11/2019 13:11:10 J Peveling-Oberhag et al. The patient was then referred to our centre for HCV P=0.01; Figure 2). When comparing miR-26b expression treatment in 2014. Pretreatment elastography showed between HCV-NHL at baseline and end-of-treatment a liver stiffness of 4.8 KPa, corresponding to F0–F1 or 6 months after treatment, respectively, a restoration fibrosis. The patient received a combination of sofosbu- of miR-26b levels could be observed in all patients but vir 400 mg once daily and daclatasvir 60 mg once daily one (patient 2 with no lymphoma response to antiviral plus weight-based RBV for 24 weeks as a participant of therapy). Mean miR-26b expression in this comparison a daclatasvir compassionate use programme [21]. HCV was not statistically significant. Again, however, when RNA was undetectable from week 2 and remained unde- case 2 was removed from the analysis, we observed tectable at 12 and 24 weeks after the end of treatment. a significant increase in miR-26b following success- There was no evidence of lymphoma manifestation in ful HCV eradication over time. When analysing the follow-up CT scans and follow-up bone marrow biopsy individual HCV-NHL cases, we observed an increase 6 months after the start of HCV therapy showed a in miR-26b expression after HCV treatment for all decrease in lymphoma infiltration to 10%. The response patients who also showed oncological response, reach- was classified as partial response. ing levels of the control population. Case 2, whose lym- phoma progressed and transformed into DLBCL had a miR-26b expression in PBMCs higher baseline level of miR-26b and did not show an For all patients, miR-26b expression was analysed from increase in expression after HCV was cured. PBMCs at the start of antiviral therapy, at the end of treatment and 6 months thereafter (clinical patient data Discussion are summarized in Table 1). To evaluate baseline miR-26b expression in HCV- We present a case series of four patients with HCV-NHL NHL cases, values of matched healthy volunteers and who received IFN-free DAA therapy with the intention patients with chronic HCV infection without NHL to treat the underlying malignant disease. We found were added as controls (demographic data are shown in evidence that the expression of the tumour-suppressive Additional file 1). There was no difference in miR-26b miRNA, miR-26b, correlates inversely with successful expression between healthy volunteers and patients lymphoma treatment. with chronic HCV infection without NHL. HCV-NHL So far, few case reports have been published report- patients showed a trend to lower miR-26b expression ing on IFN-free DAA treatment of HCV-NHL patients compared with controls (Figure 1; P=0.06). Moreover, [14–16]. In these publications successful HCV eradi- when the one patient whose lymphoma did not respond cation in patients with marginal zone lymphoma or to HCV therapy was removed from the analysis, the follicular lymphoma led to a complete and lasting onco- baseline miR-26b level was significantly lower in the logical response. Furthermore, Carrier et al. [13] reported HCV-NHL group compared with the HCV-negative on five patients with HCV-NHL treated with DAAs, out and HCV-positive controls, respectively (P=0.001 and of which three received exclusive anti-HCV treatment. Table 1. Clinical data of HCV-NHL patients Patient 1 Patient 2 Patient 3 Patient 4 Gender Male Male Female Male Age at start of HCV therapy 49 56 78 60 Lymphoma subtype SMZL MZL MZL SMZL Ann Arbor Stage IVA IVB IVA IVA Baseline HCV RNA, log IU/ml 5.7 6.0 7.0 7.0 HCV genotype 3a 1b 2 1b Cirrhosis No Yes No No Previous chemotherapy No No No No Previous antiviral therapy No No No No Antiviral therapy SOF/DCV 3D/RBV SOF/RBV SOF/DCV/RBV Treatment duration, weeks 12 12 12 24 Major adverse events associated with antiviral therapy No No No No SVR12/24 Yes Yes Yes Yes NHL response PR PD CR PR Duration of response, months 11 0 15 8 CR, complete response; DCV, daclatasvir; MZL, marginal zone lymphoma; NHL, non-Hodgkin lymphoma; PD, progressive disease; PR, partial response; RBV, ribavirin; SMZL, splenic marginal zone lymphoma; SOF, sofosbuvir; SVR, sustained virological response; 3D, ombitasvir/paritaprevir + dasabuvir. 440 ©2019 International Medical Press AVT-19-OA-4510_Peveling-Oberhag.indd 440 AVT-19-OA-4510_Peveling-Oberhag.indd 440 22/11/2019 13:11:10 22/11/2019 13:11:10 miR-26b expression during DAA therapy for hepatitis C Figure 1. Relative expression of miR-26b for all patients and Figure 2. Relative expression of miR-26b with data from patient controls 2 (with no lymphoma response to antiviral therapy) removed P=0.12 P=0.01 P=0.06 P=0.001 P=0.009 P=0.05 Delta Ct values were calculated with let-7g as housekeeping RNA. Black bar: mean, whiskers: standard deviation, black triangle: patient 2 (with no lymphoma Delta Ct values were calculated with let-7g as housekeeping RNA. Black bar: mean, whiskers: standard deviation. EOT, end of treatment; NHL, non-Hodgkin response to antiviral therapy). EOT, end of treatment; NHL, non-Hodgkin lymphoma; SVR, sustained virological reponse. lymphoma; SVR, sustained virological reponse. All treated patients in these publications reached com- delaying oncological treatment could worsen the prog- plete virological as well as oncological response. nosis of the respective patient and concomitant antivi- Recently, multicentre retrospective series of HCV-NHL ral therapy and chemotherapy has been proposed [23]. treated with IFN-free antiviral therapy showed promis- However, to spare patients from potentially toxic ing results but also reported on patients with SVR who chemotherapy, a biomarker that can predict oncologi- did not show favourable oncological response [18–20]. cal outcomes after HCV eradication at an early time Our recent meta-analysis on HCV-NHL treated with point would be desirable. In the past, miR-26b, meas- IFN-based regimens showed that overall lymphoma ured in lymphoma tissue as well as patient blood, has response rate (either complete or partial response) was been shown to be in a close relationship with HCV- 73% (95% CI, 67, 78%) [22]. Across the different NHL [9,10,24]. Moreover, a restoration of this tumour- included studies there was a strong association between suppressive miRNA to normal levels could be observed SVR and lymphoma response (overall response, 83%; in patients successfully cured of mixed cryoglobulinae- 95% CI, 76, 88%) compared with a failure in achiev- mia by anti-HCV therapy [11]. In the current study, we ing SVR (overall response, 53%; 95% CI, 39, 67%; observed an inverse correlation of HCV viral load as P=0.0002). Thus, almost one-fifth of patients with well as lymphoma burden with miR-26b expression in IFN-induced SVR did not achieve lymphoma response. PBMCs of HCV-NHL patients. More importantly, the Although HCV cure rates now exceed 90% across dif- one patient who did not show lymphoma response fol- ferent patient populations, DAA therapy may not be lowing HCV cure did not have a significantly decreased able to improve the oncological outcome in all HCV- miR-26b level before therapy and did also not show any NHL patients. One possible explanation may be that increase in miR-26b expression after achieving SVR. some patients suffer from lymphomas that are coinci- In summary, this case series demonstrates that suc- dent and primarily independent of HCV or, being pri- cessful HCV eradication following treatment of HCV marily driven by HCV, have become independent of with novel DAAs can lead to lymphoma regression or the viral stimulus over time. Patients with low-grade even cure. However, we also show that one lymphoma HCV-NHL and lymphoma progression or even trans- patient showed progressive disease despite achieving formation into high-malignant lymphoma types can be SVR. miR-26b-levels could be a potential biomarker treated with immune-chemotherapy in succession, as to predict lymphoma response in HCV-NHL patients. was done in patient number 4 in our study. However, Given the relatively low prevalence of HCV-NHL, Antiviral Therapy 24.6 441 AVT-19-OA-4510_Peveling-Oberhag.indd 441 AVT-19-OA-4510_Peveling-Oberhag.indd 441 22/11/2019 13:11:11 22/11/2019 13:11:11 Healthy volunteers HCV+, no NHL HCV-NHL d -1 HCV-NHL SVR 12 HCV-NHL EOT+6 mo Healthy volunteers HCV+, no NHL HCV-NHL d -1 HCV-NHL SVR 12 HCV-NHL EOT+6 mo delta Ct delta Ct J Peveling-Oberhag et al. 7. Arcaini L, Merli M, Volpetti S, Rattotti S, Gotti M, Zaja F. international collaborative studies are required to Indolent B-cell lymphomas associated with HCV infection: clinical and virological features and role of antiviral therapy. address open questions with regard to the potential use Clin Dev Immunol 2012; 2012:638185. of miR-26b and other biomarkers for tumour outcome 8. Hermine O, Lefrere F, Bronowicki JP, et al. Regression of prediction. Larger, and preferably prospective studies splenic lymphoma with villous lymphocytes after treatment of hepatitis C virus infection. N Engl J Med 2002; 347:89–94. could help to evaluate whether the lack of miR-26b 9. Piluso A, Gragnani L, Fognani E, et al. Deregulation of downregulation before the start of antiviral therapy microRNA expression in peripheral blood mononuclear expresses an independence of the lymphoma from the cells from patients with HCV-related malignancies. Hepatol Int 2015; 9:586–593. viral stimulus that could hint to a lack of oncological 10. Peveling-Oberhag J, Crisman G, Schmidt A, et al. efficacy of antiviral therapy. Moreover, it should be fur - Dysregulation of global microRNA expression in splenic ther evaluated whether a lack in restoration or increase marginal zone lymphoma and influence of chronic hepatitis C virus infection. Leukemia 2012; 26:1654–1662. in miR-26b expression during and after antiviral ther- 11. Fognani E, Giannini C, Piluso A, et al. Role of microRNA apy might predict worse oncological outcomes. profile modifications in hepatitis C virus-related mixed cryoglobulinemia. PLoS One 2013; 8:e62965. Acknowledgements 12. Pawlotsky JM, Feld JJ, Zeuzem S, Hoofnagle JH. From non-A, non-B hepatitis to hepatitis C virus cure. J Hepatol 2015; 62:S87–S99. This work was supported by the Deutsche Forschun- 13. Carrier P, Jaccard A, Jacques J, et al. HCV-associated B-cell gsgemeinschaft (PE2152/2-1). non-Hodgkin lymphomas and new direct antiviral agents. Liver Int 2015; 35:2222–2227. 14. Rossotti R, Travi G, Pazzi A, Baiguera C, Morra E, Puoti M. Disclosure statement Rapid clearance of HCV-related splenic marginal zone lymphoma under an interferon-free, NS3/NS4A inhibitor- based treatment. A case report. J Hepatol 2015; 62:234–237. SZ: consultancy for AbbVie, Gilead, Janssen, Merck/ 15. Sultanik P, Klotz C, Brault P, Pol S, Mallet V. Regression of an MSD. TMW: consultancies for AbbVie, Bristol-Myers HCV-associated disseminated marginal zone lymphoma under Squibb, Gilead. JV: lectures/consultancies for AbbVie, IFN-free antiviral treatment. Blood 2015; 125:2446–2447. Gilead, Merck/MSD. The other authors have no conflict 16. Hattori N, Ikeda H, Nakano H, et al. Curative effects for B-cell lymphoma accomplished by direct-acting antiviral of interest to declare in context of this manuscript. agents of hepatitis C. Open Forum Infect Dis 2017; 4:ofx057. 17. Galati G, Rampa L, Vespasiani-Gentilucci U, et al. Hepatitis Additional file C and double-hit B cell lymphoma successfully treated by antiviral therapy. World J Hepatol 2016; 8:1244–1250. 18. Arcaini L, Besson C, Frigeni M, et al. Interferon-free Additional file 1: A table showing clinical data of antiviral treatment in B-cell lymphoproliferative disorders matched control patients and healthy control subjects associated with hepatitis C virus infection. Blood 2016; 128:2527–2532. can be found at https://www.intmedpress.com/uploads/ 19. Alric L, Besson C, Lapidus N, et al. Antiviral treatment of documents/4510_Peveling-Oberhag_Addfile1.pdf HCV-infected patients with B-cell non-Hodgkin lymphoma: ANRS HC-13 Lympho-C Study. PLoS One 2016; 11:e0162965. References 20. Michot JM, Canioni D, Driss H, et al. Antiviral therapy is 1. WHO. Hepatitis C Fact sheet No164. (Updated October associated with a better survival in patients with hepatitis 2017. Accessed 1 December 2018.) Available from www. C virus and B-cell non-Hodgkin lymphomas, ANRS HC-13 who.int/mediacentre/factsheets/fs164/en/ lympho-C study. Am J Hematol 2015; 90:197–203. 2. Durand T, Di Liberto G, Colman H, et al. Occult infection 21. Welzel TM, Petersen J, Herzer K, et al. Daclatasvir plus of peripheral B cells by hepatitis C variants which have low sofosbuvir, with or without ribavirin, achieved high translational efficiency in cultured hepatocytes. Gut 2010; sustained virological response rates in patients with HCV 59:934–942. infection and advanced liver disease in a real-world cohort. Gut 2016; 65:1861–1870. 3. Sung VM, Shimodaira S, Doughty AL, et al. Establishment of B-cell lymphoma cell lines persistently infected with 22. Peveling-Oberhag J, Arcaini L, Bankov K, Zeuzem S, hepatitis C virus in vivo and in vitro: the apoptotic effects of Herrmann E. The anti-lymphoma activity of antiviral virus infection. J Virol 2003; 77:2134–2146. therapy in HCV-associated B-cell non-Hodgkin lymphomas: a meta-analysis. J Viral Hepat 2016; 23:536–544. 4. Polaris Observatory HCV Collaborators. Global prevalence and genotype distribution of hepatitis C virus infection in 23. Persico M, Aglitti A, Caruso R, et al. Efficacy and safety 2015: a modelling study. Lancet Gastroenterol Hepatol of new direct antiviral agents in hepatitis C virus-infected 2017; 2:161–176. patients with diffuse large B-cell non-Hodgkin’s lymphoma. Hepatology 2018; 67:48–55. 5. Peveling-Oberhag J, Arcaini L, Hansmann ML, Zeuzem S. Hepatitis C-associated B-cell non-Hodgkin lymphomas. 24. Gragnani L, Fognani E, Piluso A, Zignego AL. Hepatitis Epidemiology, molecular signature and clinical C-associated B-cell non-Hodgkin lymphomas: the emerging management. J Hepatol 2013; 59:169–177. role of miRNA-26b. J Hepatol 2013; 59:1362–1363. 6. Ferri C, Sebastiani M, Giuggioli D, et al. Hepatitis C virus syndrome: a constellation of organ- and non-organ specific autoimmune disorders, B-cell non-Hodgkin’s lymphoma, and cancer. World J Hepatol 2015; 7:327–343. Accepted 28 May 2019; published online 10 June 2019 442 ©2019 International Medical Press AVT-19-OA-4510_Peveling-Oberhag.indd 442 AVT-19-OA-4510_Peveling-Oberhag.indd 442 22/11/2019 13:11:11 22/11/2019 13:11:11 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Antiviral Therapy SAGE

miRNA-26b Downregulation in Peripheral Blood Mononuclear Cells of Patients with Hepatitis C Associated Lymphomas is Restored by Successful Interferon-Free Antiviral Therapy

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Abstract

Antiviral Therapy 2019; 24:437–442 (doi: 10.3851/IMP3322) Original article miRNA-26b downregulation in peripheral blood mononuclear cells of patients with hepatitis C associated lymphomas is restored by successful interferon-free antiviral therapy 1,2,3 1,3 1 4 4 Jan Peveling-Oberhag *, Katrin Bankov , Georg Dultz , Olivier Ballo , Julian Lohmeyer , 4 4 1 1 3 1 Uta Brunnberg , Vasile Marcu , Dirk Walter , Stefan Zeuzem , Martin-Leo Hansmann , Tania M Welzel , Johannes Vermehren Department of Internal Medicine 1, University Hospital Frankfurt, Frankfurt am Main, Germany Department of Internal Medicine 1, Robert-Bosch-Hospital, Stuttgart, Germany Senckenberg Institute for Pathology, University Hospital Frankfurt, Frankfurt am Main, Germany Department of Internal Medicine 2, University Hospital Frankfurt, Frankfurt am Main, Germany *Corresponding author e-mail: jan.peveling-oberhag@rbk.de Background: Patients with chronic HCV infection are at at different time points during antiviral therapy for all increased risk of developing B-cell non-Hodgkin lym- included patients as well as for a total of 10 controls with phoma (B-NHL). Regression of HCV-associated B-NHL (n=5) and without (n=5) chronic HCV infection. (HCV-NHL) can be achieved through HCV eradication Results: All patients had marginal zone lymphoma subtype using interferon (IFN). However, only about two-thirds and received different DAA regimens for 12–24 weeks. of patients with sustained virological response (SVR) also All four patients achieved SVR, but only three patients had a consecutive lymphoma response. miRNA-26b is also had lymphoma response (one complete response, two associated with HCV-NHL response to antiviral therapy. partial responses). One patient showed progression to a Recent data suggest that IFN-free direct-acting antiviral high-grade lymphoma subtype after SVR. miRNA-26b (DAA) regimens also have anti-lymphoma activity in this expression was generally decreased in patients with HCV- patient population. NHL. Moreover, miRNA-26b expression was restored in Methods: We report four patients with HCV-NHL who those HCV-NHL patients with lymphoma response after 6 were treated with different IFN-free DAA regimens as months (P=0.009). oncological monotherapy in our centre between 2015 Conclusions: We have demonstrated that IFN-free DAA and 2016. We analysed the virological and lymphoprolif- treatment of HCV can improve or even cure NHL. miRNA- erative disease response. Moreover, we analysed miRNA- 26b-levels could be a potentially useful biomarker to pre- 26b expression in peripheral blood mononuclear cells dict lymphoma response in HCV-NHL patients. Introduction Worldwide, an estimated 71 million people are causal relationship between HCV and B-NHL is made chronically infected with HCV, a hepatotropic and by interventional studies demonstrating that antiviral potentially lymphotropic virus [1–4]. In the last two therapy using interferon (IFN) or pegylated IFN with decades, evidence from epidemiological studies, bio- and without addition of ribavirin (RBV) frequently logical insights, and especially therapeutic approaches leads to regression of HCV-associated lymphomas provided strong support for an association between (HCV-NHL) [7,8]. HCV and B-cell non-Hodgkin’s lymphomas (NHL) [5]. Piluso et al. [9] showed downregulation of the HCV-related B-cell proliferation represents an impor- miRNA miR-26b in peripheral blood mononuclear tant model of virus-driven autoimmune/neoplastic dis- cells (PBMCs) of patients with HCV-NHL compared order leading to mixed cryoglobulinaemia (MC) and/or with controls. Downregulation of the tumour suppres- NHL [6]. However, the most compelling argument for a sive microRNA-26b (miR-26b) was first connected to ©2019 International Medical Press 1359-6535 (print) 2040-2058 (online) 437 AVT-19-OA-4510_Peveling-Oberhag.indd 437 AVT-19-OA-4510_Peveling-Oberhag.indd 437 22/11/2019 13:11:10 22/11/2019 13:11:10 J Peveling-Oberhag et al. HCV-NHL in microdissected primary tissue of HCV- 12 weeks after completion of therapy. HCV viral load associated splenic marginal zone lymphoma [10]. Con- was measured using the COBAS AmpliPrep/COBAS secutively, the same miRNA was found downregulated TaqMan HCV Test, v2.0 (Roche Diagnostics, Man- in PBMCs of patients with MC and HCV-NHL, also nheim, Germany). HCV genotypes were determined demonstrating restoration of miR-26b expression by the VERSANT HCV Genotype 2.0 assay (Siemens in MC patients who achieved a sustained virological Healthcare Diagnostics, Erlangen, Germany). response (SVR) after treatment with IFN and RBV [11]. These findings indicate that miR-26b could be involved miR-26b expression analysis in the mechanism of HCV-driven lymphomagenesis PBMCs were isolated from fresh anticoagulated blood and could potentially serve as a valuable biomarker to by gradient precipitation on Lymphoprep (Axis-Shield monitor lymphoma response during antiviral therapy. PoC AS, Oslo, Norway) according to the manufactur- Antiviral therapy of HCV underwent a revolution er’s instructions. After the second wash, the cells were within the last several years. After almost 25 years of counted and stored at -80°C. RNA extraction was IFN-based therapies, highly effective direct-acting anti- performed using Trizol reagent (Invitrogen, Carlsbad, viral (DAA) drugs are now standard of care for treat- CA, USA) from 5×106 PBMCs according to the manu- ment of chronic HCV infection. With these DAAs, viral facturer’s instructions. Reverse transcription was done eradication can now be achieved in approximately 95% using the TaqMan MicroRNA RT kit (Applied Biosys- of all patients across different genotypes and fibrosis tems, Foster City, CA, USA) and 100 ng of total RNA. stages [12]. Expression levels of human miR-let-7g and miR-26b Data on IFN-free treatment of HCV-NHL is still were evaluated by real-time PCR using specific TaqMan scarce and mainly based on case reports [13–17]. Now MicroRNA Assays (Applied Biosystems), according to the first multicentre retrospective series of HCV-NHL the manufacturer’s instructions. treated with IFN-free antiviral therapy shows promis- Relative expression levels of the different miRNAs ing results but also reports on patients with SVR who were evaluated with the delta-Ct method, using miR- did not show favourable oncological response [18–20]. let-7d as internal control to normalize miRNA expres- The aim of this study was to investigate the clinical sion levels, as previously described [11]. course of HCV-NHL patients from our centre treated with IFN-free DAA regimen. In addition, we evaluated Statistical analysis miR-26b expression as a biomarker for lymphoma Statistical analyses and illustrations were performed response in PBMC of HCV-NHL patients undergoing using GraphPad Prism 5.0 (GraphPad Software, La Jolla, DAA therapies. CA, USA). Student’s two-sample t-test for either paired or unpaired samples was used to compare miRNA expression values. All P-values were two-tailed and were Methods considered statistically significant for levels ≤ 0.05. Study design and patients The current study is a retrospective analysis, which was Results approved by the local Ethics Committee of the Univer- Cases sity Hospital Frankfurt, Germany (Ethik-Kommission des Fachbereichs Medizin der Goethe-Universität). Writ- Case 1 ten informed consent was obtained from all patients. The 49-year-old male was diagnosed with HCV GT3a Clinical data were abstracted from electronic records. infection in 2005 following intravenous drug use. Viral Patients with HCV-NHL (n=4) and matched individu- load was 541,000 IE/ml. Pretreatment transient elas- als with chronic HCV without lymphoma (n=5) were tography (FibroScan, Echosens, Paris, France) showed a recruited from our outpatient clinic specialized in HCV liver stiffness of 5.9 KPa, corresponding to F0–F1 fibro- treatment. Additionally, PBMCs from healthy volun- sis. In 2015, the patient presented to his primary care teers (n=5) were acquired from blood donors. physician with abdominal pain in the left upper quad- rant. There were no B symptoms present. On physical Clinical and virological end points examination, the treating physician diagnosed spleno- The primary oncological end point was overall tumour megaly, and the blood count showed an increased leuko- 3 3 response. Response evaluation was based on Lugano cyte/lymphocyte cell count (60,000/mm /42,000/mm ). classification criteria for lymphomas [12]. Bone mar - The patient was referred to a specialized centre for hae- row biopsy as well as CT scans were performed before matology/oncology. The high lymphocyte count was start of antiviral therapy and repeated at the end and 6 confirmed showing a monoclonal B-cell immune phe- months after treatment. SVR was defined as undetecta- notype with the morphology of villous lymphocytes. ble HCV viral load with a sensitive real-time PCR assay Cryoglobulinaemia was ruled out. CT scan showed 438 ©2019 International Medical Press AVT-19-OA-4510_Peveling-Oberhag.indd 438 AVT-19-OA-4510_Peveling-Oberhag.indd 438 22/11/2019 13:11:10 22/11/2019 13:11:10 miR-26b expression during DAA therapy for hepatitis C splenomegaly (longitudinal diameter of 27 cm) and an daily) for 12 weeks. HCV RNA was undetectable from enlargement of paraaortic, paracaval and mesenteric as week 2 and remained undetectable at 12 and 24 weeks well as supraclavicular and mediastinal lymph nodes. after the end of treatment. The patient was symptom Bone marrow biopsy showed lymphoid nodular infil- free 8 weeks after initiation of antiviral therapy. The tration of more than 40%. The final diagnosis was stage staging CT scan 6 months after the start of treatment IV leukaemic splenic marginal zone lymphoma. As the showed a minimal decrease in spleen size (21 to 18 cm) patient was asymptomatic at presentation at the hae- but several mediastinal lymph nodes were noted which matology centre, he was then referred to our centre for had grown in size. Biopsies of the mediastinal lymph antiviral therapy. The patient received a combination nodes were taken using endobronchial ultrasound. His- of sofosbuvir 400 mg once daily and daclatasvir 60 mg tology showed a high grade B-NHL with high prolif- once daily for 12 weeks. HCV RNA was undetectable eration (Ki67 index 80%) consistent with diffuse large from week 2 and remained undetectable at 12 and 24 B-cell lymphoma (DLBCL). Therefore, response to weeks after the end of treatment. At week 4 of treat- antiviral treatment was classified as progressive disease ment, the patient presented with abdominal pain due with transformation into a high-grade lymphoma. Con- to splenomegaly and symptom control was achieved secutively the patient received immuno-chemotherapy with use of non-steroidal anti-inflammatory drugs in (R-CHOP) with partial response after three cycles and combination with transdermal fentanyl. At week 8 of is currently planned to receive three additional cycles. treatment, the pain subsided gradually with no need for any pain medication after week 12. The blood lympho- Case 3 cyte count increased initially from baseline to week 2 The 78-year-old female was diagnosed with splenic (44,000 to 54,000/mm ), but steadily decreased (39,000 marginal zone lymphoma in 2015 from an ultrasound at follow-up week 12) to nearly normal values 1 year guided breast biopsy, after the patient noted a lump in after antiviral therapy, with the B-cell clone still detect- the right breast. Subsequent work-up revealed chronic able. Moreover, bone marrow biopsy 3 months after infection with HCV genotype-2, while no typical risk initiation of treatment showed residual but reduced factors for contracting HCV infection were noted. Viral lymphoma infiltration (25%). The CT scan 6 months load was 1,090,000 IE/ml. Transient elastography was after antiviral therapy showed decrease of all lymph not available but conventional ultrasound did not show node diameters by more than 50% and a decrease in signs of cirrhosis. The patient reported weight loss of spleen size to nearly normal size. The response was clas- about 4 kilograms but no fevers or night sweats. Stag- sified as partial response. ing CT scans revealed that the breast and mediastinal lymph nodes were the only localizations of the lym- Case 2 phoma. There was no bone marrow involvement. The A 56-year-old male, who was diagnosed with HCV peripheral blood count showed normal results. The final genotype-1b infection in 2013 following intravenous diagnosis was stage IVA low-grade splenic marginal drug use in the 1990s. Viral load was 943,000 IE/ml. zone lymphoma. The patient was then referred to our Pretreatment transient elastography showed a liver centre for HCV treatment. In line with guideline recom- stiffness of 15.1 KPa, corresponding to F4 fibrosis mendations at the time, the patient received sofosbu- score and liver morphology in the abdominal ultra- vir 400 mg once daily plus weight-based RBV for 12 sound was consistent with compensated cirrhosis. At weeks. HCV RNA was undetectable from week 4 and the time of HCV diagnosis, the patient was also diag- remained undetectable at 12 and 24 weeks after the end nosed with a low-grade B-NHL. The patient suffered of treatment. The staging CT scan 6 months after the from occasional fever episodes and night sweats. CT start of therapy showed complete response with mini- scan showed splenomegaly (longitudinal diameter of mal scarring tissue at the breast biopsy site, which did 20.7 cm) and paraaortic, mesenteric as well as medi- not show residual lymphoma after re-biopsy. astinal lymph node enlargement. Bone marrow biopsy showed monoclonal lymphoid infiltration of more than Case 4 50% with positivity for CD79, CD3, CD20 and CD34. The 60-year-old male patient was diagnosed with Peripheral blood count showed mild anaemia but oth- chronic HCV genotype-1b infection following diagnosis erwise normal results. The final diagnosis was stage IVB of splenic marginal zone lymphoma in 2011. He was ini- low-grade marginal zone lymphoma. The patient was tially treated with splenectomy because of symptomatic followed-up with staging CT scans every 3 to 6 months splenomegaly. After splenectomy, the lymphoma per- and showed a stable course. In June 2015, after inter- sisted in the bone marrow with a tumour cell infiltration disciplinary discussion, the patient received antiviral of 40–50%. There were no other disease manifestations therapy with ombitasvir/paritaprevir (12.5 mg/75 mg and the patient had no B symptoms. The final diag- two tablets once daily) and dasabuvir (250 mg twice nosis was stage IV splenic marginal zone lymphoma. Antiviral Therapy 24.6 439 AVT-19-OA-4510_Peveling-Oberhag.indd 439 AVT-19-OA-4510_Peveling-Oberhag.indd 439 22/11/2019 13:11:10 22/11/2019 13:11:10 J Peveling-Oberhag et al. The patient was then referred to our centre for HCV P=0.01; Figure 2). When comparing miR-26b expression treatment in 2014. Pretreatment elastography showed between HCV-NHL at baseline and end-of-treatment a liver stiffness of 4.8 KPa, corresponding to F0–F1 or 6 months after treatment, respectively, a restoration fibrosis. The patient received a combination of sofosbu- of miR-26b levels could be observed in all patients but vir 400 mg once daily and daclatasvir 60 mg once daily one (patient 2 with no lymphoma response to antiviral plus weight-based RBV for 24 weeks as a participant of therapy). Mean miR-26b expression in this comparison a daclatasvir compassionate use programme [21]. HCV was not statistically significant. Again, however, when RNA was undetectable from week 2 and remained unde- case 2 was removed from the analysis, we observed tectable at 12 and 24 weeks after the end of treatment. a significant increase in miR-26b following success- There was no evidence of lymphoma manifestation in ful HCV eradication over time. When analysing the follow-up CT scans and follow-up bone marrow biopsy individual HCV-NHL cases, we observed an increase 6 months after the start of HCV therapy showed a in miR-26b expression after HCV treatment for all decrease in lymphoma infiltration to 10%. The response patients who also showed oncological response, reach- was classified as partial response. ing levels of the control population. Case 2, whose lym- phoma progressed and transformed into DLBCL had a miR-26b expression in PBMCs higher baseline level of miR-26b and did not show an For all patients, miR-26b expression was analysed from increase in expression after HCV was cured. PBMCs at the start of antiviral therapy, at the end of treatment and 6 months thereafter (clinical patient data Discussion are summarized in Table 1). To evaluate baseline miR-26b expression in HCV- We present a case series of four patients with HCV-NHL NHL cases, values of matched healthy volunteers and who received IFN-free DAA therapy with the intention patients with chronic HCV infection without NHL to treat the underlying malignant disease. We found were added as controls (demographic data are shown in evidence that the expression of the tumour-suppressive Additional file 1). There was no difference in miR-26b miRNA, miR-26b, correlates inversely with successful expression between healthy volunteers and patients lymphoma treatment. with chronic HCV infection without NHL. HCV-NHL So far, few case reports have been published report- patients showed a trend to lower miR-26b expression ing on IFN-free DAA treatment of HCV-NHL patients compared with controls (Figure 1; P=0.06). Moreover, [14–16]. In these publications successful HCV eradi- when the one patient whose lymphoma did not respond cation in patients with marginal zone lymphoma or to HCV therapy was removed from the analysis, the follicular lymphoma led to a complete and lasting onco- baseline miR-26b level was significantly lower in the logical response. Furthermore, Carrier et al. [13] reported HCV-NHL group compared with the HCV-negative on five patients with HCV-NHL treated with DAAs, out and HCV-positive controls, respectively (P=0.001 and of which three received exclusive anti-HCV treatment. Table 1. Clinical data of HCV-NHL patients Patient 1 Patient 2 Patient 3 Patient 4 Gender Male Male Female Male Age at start of HCV therapy 49 56 78 60 Lymphoma subtype SMZL MZL MZL SMZL Ann Arbor Stage IVA IVB IVA IVA Baseline HCV RNA, log IU/ml 5.7 6.0 7.0 7.0 HCV genotype 3a 1b 2 1b Cirrhosis No Yes No No Previous chemotherapy No No No No Previous antiviral therapy No No No No Antiviral therapy SOF/DCV 3D/RBV SOF/RBV SOF/DCV/RBV Treatment duration, weeks 12 12 12 24 Major adverse events associated with antiviral therapy No No No No SVR12/24 Yes Yes Yes Yes NHL response PR PD CR PR Duration of response, months 11 0 15 8 CR, complete response; DCV, daclatasvir; MZL, marginal zone lymphoma; NHL, non-Hodgkin lymphoma; PD, progressive disease; PR, partial response; RBV, ribavirin; SMZL, splenic marginal zone lymphoma; SOF, sofosbuvir; SVR, sustained virological response; 3D, ombitasvir/paritaprevir + dasabuvir. 440 ©2019 International Medical Press AVT-19-OA-4510_Peveling-Oberhag.indd 440 AVT-19-OA-4510_Peveling-Oberhag.indd 440 22/11/2019 13:11:10 22/11/2019 13:11:10 miR-26b expression during DAA therapy for hepatitis C Figure 1. Relative expression of miR-26b for all patients and Figure 2. Relative expression of miR-26b with data from patient controls 2 (with no lymphoma response to antiviral therapy) removed P=0.12 P=0.01 P=0.06 P=0.001 P=0.009 P=0.05 Delta Ct values were calculated with let-7g as housekeeping RNA. Black bar: mean, whiskers: standard deviation, black triangle: patient 2 (with no lymphoma Delta Ct values were calculated with let-7g as housekeeping RNA. Black bar: mean, whiskers: standard deviation. EOT, end of treatment; NHL, non-Hodgkin response to antiviral therapy). EOT, end of treatment; NHL, non-Hodgkin lymphoma; SVR, sustained virological reponse. lymphoma; SVR, sustained virological reponse. All treated patients in these publications reached com- delaying oncological treatment could worsen the prog- plete virological as well as oncological response. nosis of the respective patient and concomitant antivi- Recently, multicentre retrospective series of HCV-NHL ral therapy and chemotherapy has been proposed [23]. treated with IFN-free antiviral therapy showed promis- However, to spare patients from potentially toxic ing results but also reported on patients with SVR who chemotherapy, a biomarker that can predict oncologi- did not show favourable oncological response [18–20]. cal outcomes after HCV eradication at an early time Our recent meta-analysis on HCV-NHL treated with point would be desirable. In the past, miR-26b, meas- IFN-based regimens showed that overall lymphoma ured in lymphoma tissue as well as patient blood, has response rate (either complete or partial response) was been shown to be in a close relationship with HCV- 73% (95% CI, 67, 78%) [22]. Across the different NHL [9,10,24]. Moreover, a restoration of this tumour- included studies there was a strong association between suppressive miRNA to normal levels could be observed SVR and lymphoma response (overall response, 83%; in patients successfully cured of mixed cryoglobulinae- 95% CI, 76, 88%) compared with a failure in achiev- mia by anti-HCV therapy [11]. In the current study, we ing SVR (overall response, 53%; 95% CI, 39, 67%; observed an inverse correlation of HCV viral load as P=0.0002). Thus, almost one-fifth of patients with well as lymphoma burden with miR-26b expression in IFN-induced SVR did not achieve lymphoma response. PBMCs of HCV-NHL patients. More importantly, the Although HCV cure rates now exceed 90% across dif- one patient who did not show lymphoma response fol- ferent patient populations, DAA therapy may not be lowing HCV cure did not have a significantly decreased able to improve the oncological outcome in all HCV- miR-26b level before therapy and did also not show any NHL patients. One possible explanation may be that increase in miR-26b expression after achieving SVR. some patients suffer from lymphomas that are coinci- In summary, this case series demonstrates that suc- dent and primarily independent of HCV or, being pri- cessful HCV eradication following treatment of HCV marily driven by HCV, have become independent of with novel DAAs can lead to lymphoma regression or the viral stimulus over time. Patients with low-grade even cure. However, we also show that one lymphoma HCV-NHL and lymphoma progression or even trans- patient showed progressive disease despite achieving formation into high-malignant lymphoma types can be SVR. miR-26b-levels could be a potential biomarker treated with immune-chemotherapy in succession, as to predict lymphoma response in HCV-NHL patients. was done in patient number 4 in our study. However, Given the relatively low prevalence of HCV-NHL, Antiviral Therapy 24.6 441 AVT-19-OA-4510_Peveling-Oberhag.indd 441 AVT-19-OA-4510_Peveling-Oberhag.indd 441 22/11/2019 13:11:11 22/11/2019 13:11:11 Healthy volunteers HCV+, no NHL HCV-NHL d -1 HCV-NHL SVR 12 HCV-NHL EOT+6 mo Healthy volunteers HCV+, no NHL HCV-NHL d -1 HCV-NHL SVR 12 HCV-NHL EOT+6 mo delta Ct delta Ct J Peveling-Oberhag et al. 7. Arcaini L, Merli M, Volpetti S, Rattotti S, Gotti M, Zaja F. international collaborative studies are required to Indolent B-cell lymphomas associated with HCV infection: clinical and virological features and role of antiviral therapy. address open questions with regard to the potential use Clin Dev Immunol 2012; 2012:638185. of miR-26b and other biomarkers for tumour outcome 8. Hermine O, Lefrere F, Bronowicki JP, et al. Regression of prediction. Larger, and preferably prospective studies splenic lymphoma with villous lymphocytes after treatment of hepatitis C virus infection. N Engl J Med 2002; 347:89–94. could help to evaluate whether the lack of miR-26b 9. Piluso A, Gragnani L, Fognani E, et al. Deregulation of downregulation before the start of antiviral therapy microRNA expression in peripheral blood mononuclear expresses an independence of the lymphoma from the cells from patients with HCV-related malignancies. Hepatol Int 2015; 9:586–593. viral stimulus that could hint to a lack of oncological 10. Peveling-Oberhag J, Crisman G, Schmidt A, et al. efficacy of antiviral therapy. Moreover, it should be fur - Dysregulation of global microRNA expression in splenic ther evaluated whether a lack in restoration or increase marginal zone lymphoma and influence of chronic hepatitis C virus infection. Leukemia 2012; 26:1654–1662. in miR-26b expression during and after antiviral ther- 11. Fognani E, Giannini C, Piluso A, et al. Role of microRNA apy might predict worse oncological outcomes. profile modifications in hepatitis C virus-related mixed cryoglobulinemia. PLoS One 2013; 8:e62965. Acknowledgements 12. Pawlotsky JM, Feld JJ, Zeuzem S, Hoofnagle JH. From non-A, non-B hepatitis to hepatitis C virus cure. J Hepatol 2015; 62:S87–S99. This work was supported by the Deutsche Forschun- 13. Carrier P, Jaccard A, Jacques J, et al. HCV-associated B-cell gsgemeinschaft (PE2152/2-1). non-Hodgkin lymphomas and new direct antiviral agents. Liver Int 2015; 35:2222–2227. 14. Rossotti R, Travi G, Pazzi A, Baiguera C, Morra E, Puoti M. Disclosure statement Rapid clearance of HCV-related splenic marginal zone lymphoma under an interferon-free, NS3/NS4A inhibitor- based treatment. A case report. J Hepatol 2015; 62:234–237. SZ: consultancy for AbbVie, Gilead, Janssen, Merck/ 15. Sultanik P, Klotz C, Brault P, Pol S, Mallet V. Regression of an MSD. TMW: consultancies for AbbVie, Bristol-Myers HCV-associated disseminated marginal zone lymphoma under Squibb, Gilead. JV: lectures/consultancies for AbbVie, IFN-free antiviral treatment. Blood 2015; 125:2446–2447. Gilead, Merck/MSD. The other authors have no conflict 16. Hattori N, Ikeda H, Nakano H, et al. Curative effects for B-cell lymphoma accomplished by direct-acting antiviral of interest to declare in context of this manuscript. agents of hepatitis C. Open Forum Infect Dis 2017; 4:ofx057. 17. Galati G, Rampa L, Vespasiani-Gentilucci U, et al. Hepatitis Additional file C and double-hit B cell lymphoma successfully treated by antiviral therapy. World J Hepatol 2016; 8:1244–1250. 18. Arcaini L, Besson C, Frigeni M, et al. Interferon-free Additional file 1: A table showing clinical data of antiviral treatment in B-cell lymphoproliferative disorders matched control patients and healthy control subjects associated with hepatitis C virus infection. Blood 2016; 128:2527–2532. can be found at https://www.intmedpress.com/uploads/ 19. Alric L, Besson C, Lapidus N, et al. Antiviral treatment of documents/4510_Peveling-Oberhag_Addfile1.pdf HCV-infected patients with B-cell non-Hodgkin lymphoma: ANRS HC-13 Lympho-C Study. PLoS One 2016; 11:e0162965. References 20. Michot JM, Canioni D, Driss H, et al. Antiviral therapy is 1. WHO. Hepatitis C Fact sheet No164. (Updated October associated with a better survival in patients with hepatitis 2017. Accessed 1 December 2018.) Available from www. C virus and B-cell non-Hodgkin lymphomas, ANRS HC-13 who.int/mediacentre/factsheets/fs164/en/ lympho-C study. Am J Hematol 2015; 90:197–203. 2. Durand T, Di Liberto G, Colman H, et al. Occult infection 21. Welzel TM, Petersen J, Herzer K, et al. Daclatasvir plus of peripheral B cells by hepatitis C variants which have low sofosbuvir, with or without ribavirin, achieved high translational efficiency in cultured hepatocytes. Gut 2010; sustained virological response rates in patients with HCV 59:934–942. infection and advanced liver disease in a real-world cohort. Gut 2016; 65:1861–1870. 3. Sung VM, Shimodaira S, Doughty AL, et al. Establishment of B-cell lymphoma cell lines persistently infected with 22. Peveling-Oberhag J, Arcaini L, Bankov K, Zeuzem S, hepatitis C virus in vivo and in vitro: the apoptotic effects of Herrmann E. The anti-lymphoma activity of antiviral virus infection. J Virol 2003; 77:2134–2146. therapy in HCV-associated B-cell non-Hodgkin lymphomas: a meta-analysis. J Viral Hepat 2016; 23:536–544. 4. Polaris Observatory HCV Collaborators. Global prevalence and genotype distribution of hepatitis C virus infection in 23. Persico M, Aglitti A, Caruso R, et al. Efficacy and safety 2015: a modelling study. Lancet Gastroenterol Hepatol of new direct antiviral agents in hepatitis C virus-infected 2017; 2:161–176. patients with diffuse large B-cell non-Hodgkin’s lymphoma. Hepatology 2018; 67:48–55. 5. Peveling-Oberhag J, Arcaini L, Hansmann ML, Zeuzem S. Hepatitis C-associated B-cell non-Hodgkin lymphomas. 24. Gragnani L, Fognani E, Piluso A, Zignego AL. Hepatitis Epidemiology, molecular signature and clinical C-associated B-cell non-Hodgkin lymphomas: the emerging management. J Hepatol 2013; 59:169–177. role of miRNA-26b. J Hepatol 2013; 59:1362–1363. 6. Ferri C, Sebastiani M, Giuggioli D, et al. Hepatitis C virus syndrome: a constellation of organ- and non-organ specific autoimmune disorders, B-cell non-Hodgkin’s lymphoma, and cancer. World J Hepatol 2015; 7:327–343. Accepted 28 May 2019; published online 10 June 2019 442 ©2019 International Medical Press AVT-19-OA-4510_Peveling-Oberhag.indd 442 AVT-19-OA-4510_Peveling-Oberhag.indd 442 22/11/2019 13:11:11 22/11/2019 13:11:11

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Antiviral Therapy – SAGE

Published: Aug 1, 2019

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