DECLINE OF PREVALENCE OF RESISTANCE ASSOCIATED SUBSTITUTIONS TO NS3 AND NS5A INHIBITORS AT DAA- FAILURE IN HEPATITIS C VIRUS IN ITALY OVER THE YEARS 2015 TO 2018

996A AASLD ABSTRACTS (Poster) HEPATOLOGY, October, 2019

 Denotes AASLD Presidential Poster of Distinction  Denotes AASLD Foundation Abstract Award Recipient

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PERITUMORAL TISSUE IN PATIENTS DIAGNOSED WITH

HEPATOCELLULAR CANCER AFTER HCV CURE SHOWS

STUBBORN B CELLS AND MACROPHAGE ENRICHMENT

Zhibin Zhu1_{, Andrea D. Branch}2_{, Erin Doyle}2_{, Alyx Vogle}1_,

Lucas Fass1_{, Youngmin Anna Lee}2_{, James Carter}2_{, Frank}

Eng2_{, Wei Lam}1_{, Miran Kim}1 _{and Costica Aloman}1_{, (1)}

Department of Hepatology, Rush University Medical Center, (2)Division of Liver Diseases, Icahn School of Medicine at Mount Sinai

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DQWLYLUDOGUXJVDFKLHYHKLJKVXVWDLQHGYLUDOUHVSRQVH695 UDWHVLQFLUUKRWLFSDWLHQWV'HVSLWHFOLQLFDO+&9FXUHOLYHULQMXU\ DQGLQÀDPPDWLRQLQFOXGLQJ%FHOODEQRUPDOLWLHVSHUVLVWLQ 66% of these patients and HCC risk remains elevated. It is LPSRUWDQWWRGH¿QHWKHPHFKDQLVPVRISHUVLVWHQWLQÀDPPDWLRQ DQGWRGHWHUPLQHWKHLUUHODWLRQVKLSWRWXPRULJHQHVLVDIWHU+&9 cure. CXCL13 (also called B cell-attracting chemokine 1) is LQYROYHG LQ KHSDWLF %O\PSKRF\WH WUDI¿FNLQJ DQG O\PSKRLG follicle development. 0HWKRGVLiver samples were obtained from 29 HCC patients who underwent liver transplant in Rush University Medical Center from 2015 to 2017. Three groups ZHUH GH¿QHG +&9 DQWLERG\ $%++&951$- _{(N=7, cured),}

+&9$%++&951$+ 1  FKURQLF DQG +&9$%- _(N=11,

controls) (Table 1). Immunostaining of non-tumor areas of the OLYHU GHOLQHDWHG WKH LQÀDPPDWRU\ LQ¿OWUDWH DQWL&' &' CD38, CD68); anti-CXCL13 measured B cell recruitment and DQWL0'$ LQGLFDWHG UHVSRQVHV WR LQWHUIHURQĮ VWLPXODWLRQ 6WDLQLQJZDVREMHFWLYHO\TXDQWL¿HGE\LQWHJUDWHGGHQVLW\ZLWK Image J. The Tabula Muris database, which was used to identify liver cells that express CXCL13 ÀDJJHG WKH PDFURSKDJH population. 5HVXOWV The most striking result was the high H[SUHVVLRQRI&;&/LQSDWLHQWVEHIRUHDQGDIWHU+&9FXUH Expression was similar in patients with chronic infection and LQSDWLHQWVFXUHGRI+&9WKHOHYHOLQERWKZDVVLJQL¿FDQWO\ KLJKHUWKDQLQFRQWUROVS”([SUHVVLRQRI&'%FHOO PDUNHU&'PDFURSKDJH.XSIIHUFHOOPDUNHUDQG0'$ ZHUHVLJQL¿FDQWO\LQFUHDVHGLQERWKJURXSVRI+&9H[SRVHG patients compared to controls. There was a statistically VLJQL¿FDQWFRUUHODWLRQU S EHWZHHQ&'VWDLQLQJ and CXCL13 staining, indicating that hepatic macrophages FRQWULEXWH WR % FHOO UHFUXLWPHQW EHIRUH DQG DIWHU +&9 FXUH 7KH WZR JURXSV RI +&9H[SRVHG SDWLHQWV DQG WKH FRQWUROV KDGVLPLODUVWDLQLQJRI&'7DQG1.7FHOOVDQG&'D marker of plasma cells). &RQFOXVLRQPatients who develop +&&LQWKH¿UVW\HDUDIWHU+&9FXUHKDYHLPPXQRKLVWRORJLFDO HYLGHQFHRISHUVLVWHQW,QWHUIHURQĮVWLPXODWHGJHQHDFWLYDWLRQ DQGHQULFKHG%FHOOLQÀDPPDWLRQSRWHQWLDOO\WKHFRQVHTXHQFH of CXCL13 production by resident long lived self-renewable hepatic macrophages. 'LVFORVXUHV 7KHIROORZLQJSHRSOHKDYHQRWKLQJWRGLVFORVH=KLELQ=KX$QGUHD'%UDQFK (ULQ'R\OH<RXQJPLQ$QQD/HH-DPHV&DUWHU0LUDQ.LP&RVWLFD$ORPDQ 'LVFORVXUHLQIRUPDWLRQQRWDYDLODEOHDWWKHWLPHRISXEOLFDWLRQ$O\[9RJOH/XFDV )DVV)UDQN(QJ:HL/DP

1 6 5 6

DECLINE OF PREVALENCE OF RESISTANCE ASSOCIATED

SUBSTITUTIONS TO NS3 AND NS5A INHIBITORS AT

DAA-FAILURE IN HEPATITIS C VIRUS IN ITALY OVER THE YEARS

2015 TO 2018

Barbara Rossetti1_{, David Redi}1,2_{, Velia Chiara Di Maio}3_,

Marianna Aragri3_{, Stefania Paolucci}4_{, Lorenzo Paglicci}1,2_,

Chiara Masetti5_{, Bianca Bruzzone}6_{, Carmine Minichini}7_,

Francesca Montagnani1,2_{, Valeria Micheli}8_{, Simona Landonio}9_,

Giacomo Zanelli1,2_{, Elisabetta Degasperi}10_{, Renato Maserati}11_,

Ivana Maida12_{, Annapaola Callegaro}13_{, Silvia Barbaliscia}3_,

Ada Bertoli3_{, Claudio Paternoster}14_{, Simona Marenco}15_,

Filomena Morisco16_{, Vincenza Calvaruso}17_{, Gloria Taliani}18_,

Massimo Puoti19_{, Giovanni Cenderello}20_{, Adriano De Santis}21_,

Miriam Lichtner22_{, Nicola Coppola}7_{, Roberto Gulminetti}11_,

Valeria Cento23_{, Maria Rendina}24_{, Elisabetta Teti}25_{, Giustino}

Parruti26_{, Tina Ruggiero}27_{, Valeria Ghisetti}27_{, Caterina}

Pasquazzi28_{, Laura Ambra Nicolini}29_{, Vincenzo Vullo}30_,

Adriano Pellicelli31_{, Tullio Prestileo}32_{, Raffaele Cozzolongo}33_,

Vincenzo Sangiovanni34_{, Marco Biolato}35_{, Ilaria Lenci}5_{, Anna}

Licata36_{, Antonio Ciaccio}37_{, Valeria Pace Palitti}38_{, Alessia}

Giorgini39_{, Giuseppe Cariti}40_{, Alessia Ciancio}41_{, Alessio}

Aghemo42_{, Vanni Borghi}43_{, Pietro Andreone}44_{, Maurizia R.}

Brunetto45_{, Teresa Pollicino}46_{, Teresa Antonia Santantonio}47_,

Nunzia Cuomo48_{, Cinzia Caudai}49_{, Sergio Babudieri}12_,

Pietro Lampertico10_{, Giovanni Battista Gaeta}7_{, Giovanni}

Raimondo46_{, Massimo Andreoni}25_{, Giuliano Rizzardini}9_,

Mario Angelico5_{, Carlo Federico Perno}50_{, Antonio Craxì}17_,

Maurizio Zazzi2_{, Francesca Ceccherini Silberstein}3 _{and HCV}

Virology Italian Resistance Network (Vironet C), (1)Infectious Diseases Unit, Aou Senese, Siena, Italy, (2)Department of Medical Biotechnology, University of Siena, Siena, Italy, (3) Department of Experimental Medicine, University of Rome Tor Vergata, Rome, Italy, (4)Molecular Virology Unit, Microbiology and Virology Department, Irccs Policlinic Foundation San Matteo, Pavia, Italy, (5)Hepatology Unit, University Hospital of Rome Tor Vergata, Rome, Italy, (6)Hygiene Unit, Irccs Aou San Martino-IST, Genoa, Italy, (7)Infectious Diseases Unit, University of Campania L. Vanvitelli, Naples, Italy, (8)Clinical Microbiology, Virology and Bioemergencies Diagnosis, L. Sacco University Hospital, Milan, Italy, (9)Division of Infectious Diseases, Asst Fatebenefratelli Sacco, Milan, Italy, (10)CRC “a.M. e a. Migliavacca” Center for Liver Diseases, Division of Gastroenterology and Hepatology, Fondazione Irccs Ca’ Granda Ospedale Maggiore Policlinico, University of Milan, Milan, Italy, (11)Institute of Infectious Diseases, University of Pavia, Pavia, Italy, (12)Medical, Surgical and Experimental Sciences, University of Sassari, Sassari, Italy, (13)Department of Laboratory Medicine, Asst Papa Giovanni XXIII, Bergamo, Italy, (14)Infectious Disease Unit, Ospedale Di Trento, Trento, Italy, (15)Division of Hepatology, University of Genoa-Aou Irccs San Martino-IST, Genoa, Italy, (16)Department of Clinical Medicine and Surgery, University “Federico II” of Naples, Naples, Italy, (17)Gastroenterology, “P. Giaccone” University Hospital, Palermo, Italy, (18)Department of Clinical Medicine, Clinic of Tropical Medicine, Sapienza University of Rome, Rome, Italy, (19)Division of Infectious Diseases, Asst Grande Ospedale Metropolitano Niguarda, Milan, Italy, (20) Infectious Diseases Unit, Ente Ospedaliero Galliera Hospital,

HEPATOLOGY, VOLUME 70, NUMBER 1 (SUPPL) AASLD ABSTRACTS (Poster) 997A

 Denotes AASLD Presidential Poster of Distinction  Denotes AASLD Foundation Abstract Award Recipient Genoa, Italy, (21)Gastroenterology Unit, “La Sapienza”

University of Rome, Rome, Italy, (22)Infectious Diseases Unit, Sapienza University, Polo Pontino, Latina, Italy, (23)Residency Program in Microbiology and Virology, Università Degli Studi Di Milano, Milan, Italy, (24)Department of Emergency and Organ Transplantation, Section of Gastroenterology, University Hospital, Bari, Italy, (25)Infectious Diseases Unit, University Hospital of Rome Tor Vergata, Rome, Italy, (26)Infectious Disease Unit, Pescara General Hospital, Pescara, Italy, (27)Laboratory of Microbiology and Virology, Amedeo Di Savoia Hospital, ASL Città Di Torino, Turin, Italy, (28)Infectious Diseases Unit, Sant’Andrea Hospital – Sapienza University, Rome, Italy, (29)Department of Health Sciences (DISSAL), University of Genoa, Genoa, Italy, (30)Department of Public Health and Infectious Diseases, Sapienza University, Rome, Italy, (31)Hepatology Unit, San Camillo Forlanini Hospital, Rome, Italy, (32)Infectious Diseases Unit, Arnas Civico-Dicristina-Benefratelli, Palermo, Italy, (33)Division of Gastroenterology, National Institute of Gastroenterology S De Bellis, Castellana Grotte, Bari, Italy, (34)Hospital Cotugno, Naples, Italy, (35)Liver Transplant Medicine, Fondazione Policlinico Universitario a. Gemelli Irccs, Università Cattolica Del Sacro Cuore, Rome, Italy, (36) Internal Medicine and Hepatology, Di.Bi.M.I.S, University of Palermo, Palermo, Italy, (37)Unit of Gastroenterology, Department of Medicine, Hospital San Gerardo, Monza, Italy, (38)Hepatology Unit, Pescara General Hospital, Pescara, Italy, (39)Gastroenterology and Hepatology, Asst Santi Paolo e Carlo, Milan, Italy, (40)Department of Medical Sciences, University of Turin, Amedeo Di Savoia Hospital, Turin, Italy, (41)Unit of Gastroenterology, University of Turin, Department of Medical Sciences, City of Health and Science of Molinette Turin Hospital, Turin, Italy, (42)Department of Biomedical Sciences, Humanitas University, Pieve Emanuele (MI), Italy, (43)Department of Biomedical Sciences, University of Modena School of Medicine, Modena, Italy, (44)Department of Medical and Surgical Sciences, Maternal-Infantile and Adult Sciences, University of Modena and Reggio Emilia, Modena, Italy, (45)Hepatology Unit, University Hospital of Pisa, Pisa, Italy, (46)Department of Internal Medicine, University Hospital of Messina, Messina, Italy, (47)Infectious Diseases Unit, University of Foggia, Foggia, Italy, (48)Microbiology and Virology, Azienda Ospedaliera Specialistica Dei Colli Monaldi - Cotugno - C.T.O., Naples, Italy, (49)Microbiology and Virology, Siena University Hospital, Siena, Italy, (50) Department of Oncology and Oncohematology, University of Milan, Milan, Italy.

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resistance-associated substitutions (RASs) are commonly detected at failure of interferon-free DAA regimens. 0HWKRGV :LWKLQ WKH ,WDOLDQ QHWZRUN 9,521(7& WKH SUHYDOHQFH RI NS3/NS5A/NS5B RASs was retrospectively evaluated in patients who failed an EASL recommended DAA-regimen in 2015-2018. The geno2pheno system and Sorbo MC et DO'UXJ5HVLVWDQFH8SGDWHVZHUHXVHGWRLQIHU+&9 genotype/subtype and predict drug resistance. The changes in prevalence of RASs over time were evaluated by chi-square test for trend, predictors of RASs at failure were analysed by logistic regression. 5HVXOWV:HLQFOXGHG+&9LQIHFWHG SDWLHQWVPDOHVPHGLDQDJHZDV\HDUV,45 PHWDYLU¿EURVLVVWDJH)LQZHUHWUHDWPHQW H[SHULHQFHGZLWK,)1EDVHGWUHDWPHQWV ZLWK'$$V3DWLHQWVZLWK+,9DQG+%9FRLQIHFWLRQZHUH DQGUHVSHFWLYHO\+&9JHQRW\SHZDVE in 122 pts (32%), 3 in 109 (28%), 1a in 97 (25%), 4 in 37 LQ'$$UHJLPHQVZHUH/'962)LQ '&962)LQ'LQ(%5*5= LQ   9(/62) LQ   */(3,% LQ   DQG 2D in 6 (2%); ribavirin was administered in 123 (32%). The NS5A fasta-sequence was available for all patients, NS5B for 361 (94%), NS3 for 365 (95%). According to the DAA failed the prevalence of any RASs was 90%, namely 80/135 (59%) in NS3, 313/359 (87%) in NS5A, 114/286 (40%) in NS5B. 7KHSUHYDOHQFHRIDQ\5$6VVLJQL¿FDQWO\GHFOLQHGIURP to 2018 (93% vs  S  16$ 5$6V IURP  WR 72% (p=0.29), NS3 RASs from 74% to 18% (p<0.001), while NS5B RASs remained stable. Independent predictors of any 5$6VLQFOXGHGDGYDQFHG¿EURVLV$25&, p=0.004) and genotype (G2 vs G1a AOR 0.03, CI 95% 0.002-0.31, p=0.004; G3 vs G1a AOR 0.08, CI 95% 0.01-0.62, p=0.02; G4 vs G1a AOR 0.05, CI 95% 0.006-0.46, p=0.008), DIWHUDGMXVWLQJIRUDJHSUHYLRXV+&9WUHDWPHQWDQG\HDURI genotype. Notably, full activity was predicted for GLE/PIB in RIFDVHVDQGIRUDWOHDVWWZRFRPSRQHQWVRI9(/62)92; in 53% of cases, no case with full-resistance to either regimen was found. &RQFOXVLRQDespite decreasing prevalence over the years, RASs remain common at virological failure of DAA treatment, particularly in patients with the highest grade of OLYHU¿EURVLV7KHLGHQWL¿FDWLRQRI5$6VDIWHUIDLOXUHFRXOGSOD\ a crucial role in optimizing retreatment strategies.

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