HIV-1 RNA AND DNA MUTATIONAL LOAD IN HTE PATIENTS WITH MDR VIRUS: A TOOL FOR DECIDING OBR? Maria Mercedes Santoro 1, Daniele Armenia 1,2, Maria Concetta Bellocchi 1, Luca Carioti 1, Laura Galli 3, Andrea Galli 3, Rossana Scutari 1, Eleonora Salsi 4, Cristina Mussini 5, Gaetana Sterrantino 6, Leonardo Calza 7, Barbara Rossetti 8, Maurizio Zazzi 9, Antonella Castagna 3,10, on behalf of the PRESTIGIO Registry Study Group. 1 Department of Experimental Medicine, University of Rome "Tor Vergata", Rome; 2 Saint Camillus International University of Health Sciences, Rome; 3 Clinic of Infectious Diseases, Istituto Scientifico San Raffaele, Milano; 4 Azienda Ospedaliero-Universitaria Parma; 5 Malattie Infettive, Policlinico di Modena, Modena; 6 Department of Experimental and Clinical Medicine, University of Florence; 7 Policlinico Sant`Orsola-Malpighi, Bologna, Italy; 8 Azienda Ospedaliera-Universitaria Senese, Siena, Italy; 9 University of Siena, Siena, Italy; 10 Clinic of Infectious Diseases, Vita-Salute San Raffaele University, Milano.
Disclosure No conflict of interest to declare
Introduction/Summary Mutational load in isolates from plasma and peripheral blood mononuclear cells (PBMCs) could impact on virological outcome and might be a useful tool to identify heavily treatment-experienced patients at higher risk of virological failure. Study Design and methods Patients information was retrieved from the database of the PRESTIGIO Registry. Inclusion criteria: age 18 years; Previous evidence of 4-class drug resistance; availability of paired plasma and PBMC samples; HIV plasma viral load >50 copies/ml at sample collection (baseline = BL). Analyses: Resistance evaluation as intra-patients frequency and mutational load, according to through deepsequencing (NGS) on MiSeq platform (Illumina Inc) according to Stanford drug-resistance list 2019. For the resistant variants detected by NGS, mutational load was calculated by multiplying the mutant frequency in plasma and proviral DNA for the HIV-1 DNA and RNA levels detected in the same sample, respectively. Evaluation of APOBEC activity according to HIVdb 8.9-1 algorithm as the presence of signature APOBEC mutations, defined as amino acid mutations at highly conserved positions that result from a GG=>AG (APOBEC3G context) and GA=>AA (APOBEC3F context). Comparison between resistance detected through Illumina-NGS at BL and through Sanger techniques in historical GRT collected in PRESTIGIO database. Evaluation of virological outcome according to Mutation load levels and APOBEC activity.
Results P<0.001 P<0.001
By NGS, 4-class resistance was detected in 26% and 68% of patients in plasma and PBMC, respectively. Complex resistance patterns were detected in all individuals; 12 (60%) persons harboured an X4-tropic virus in at least one compartment.
Conclusions APOBEC activity can be detected in HIV-1 proviral DNA in most HTE individuals with 4-drug class resistance, with a low viral burden, and seems to have a lower variant frequency in failing patients compared to responders. HIV-1 RNA/DNA ML might help to identify individuals more prone to experience virological failure and guide salvage therapy optimization.
Registry STEERING COMMITTEE: Antonella Castagna (Coordinator), Nicola Gianotti, Laura Galli, Franco Maggiolo, Leonardo Calza, Emanuele Focà, Gaetana Sterrantino, Giovanni Cenderello, Antonio Di Biagio, Stefano Rusconi, Cristina Mussini, Marianna Menozzi, Andrea Antinori, Roberta Gagliardini, Stefano Bonora, Micol Ferrara, Maurizio Zazzi, Maria Santoro, GiulioMaria Corbelli. VIROLOGY TEAM AND BIOLOGICAL BANK: Maurizio Zazzi, Maria Santoro, Andrea Galli. STUDY COORDINATORS: ElisabettaCarini,Maria RitaParisi. STATISTICAL AND MONITORING TEAM: Laura Galli, Andrea Poli, Alba Bigoloni. PARTICIPATING CENTERS: Ancona: Marcello Tavio, Luca Butini, Andrea Giacometti; Aviano: Emanuela Vaccher, Ferdinando Martellotta, Valentina Da Ros; Bari: Gioacchino Angarano, Annalisa Saracino, Flavia Balena; Bergamo: Franco Maggiolo, Laura Comi, Elisa Di Filippo, Daniela Valenti, Claudia Suardi, Barbara Mazzola; Bologna: Pierluigi Viale, Leonardo Calza, Elena Rosselli del Turco, Marta Vacas Ramirez; Brescia: Francesco Castelli, Emanuele Focà, Anna Celotti, Francesca Brognoli; Busto Arsizio: Guido Bonoldi, Barbara Menzaghi, Clara Abeli, Maddalena Farinazzo; Cagliari: Francesco Ortu, Marco Campus; Catania: Bruno Cacopardo, Maurizio Celesia; Cremona: Angelo Pan, Chiara Fornabaio; Firenze: Alessandro Bartoloni, Gaetana Sterrantino, Francesca Rinaldi, Susanna Giachè, Blanc Pierluigi, Francesca Vichi, Francesco Maria Fusco; Foggia: Teresa Santantonio, Sergio Ferrara, Serena Rita Bruno; Genova: Giovanni Cassola, Giovanni Cenderello, Feasi Marcello, Francesca Calautti, Claudio Viscoli, Antonio Di Biagio, Bianca Bruzzone; La Spezia: Stefania Artioli; Milano: Adriano Lazzarin, Antonella Castagna, Nicola Gianotti, Elisabetta Carini, Maria Rita Parisi, Laura Galli, Andrea Poli, Andrea Galli, Diana Canetti, Massimo Galli, Stefano Rusconi, Tiziana Formenti, Valentina Morena, Arianna Gabrieli, Antonella d'arminio Monforte, Lidia Gazzola, Esther Merlini, Valentina Minieri, Andrea Gori, Alessandra Bandera, Valeria Pastore, Valentina Ferroni, Massimo Puoti, Cristina Moioli; Sara Vassalli; Modena: Cristina Mussini, Marianna Menozzi, Roncaglia Enrica, Nardini Giulia, Barbara Beghetto; Napoli: Elio Manzillo, Alfredo Franco; Padova: Anna Maria Cattelan, Serena Marinello, Silvia Cavinato, Annamaria Macario; Palermo: AntonioCascio, Giovanni Mazzola; Parma: Anna Maria degli Antoni, CarloFerrari, Diletta Laccabue; Pavia: GaetanoFilice, RobertoGulminetti, LaylaPagnucco, Annalia Asti; Perugia/Terni: Daniela Francisci, Pasticci, Elisabetta Schiaroli, Chiara Papalini, Francesca Italiani; Pistoia: Massimo Di Pietro; Reggio Emilia: Giacomo Magnani, Garlassi Elisa, Enrico Barchi, Romina Corsini; Roma: Andrea Antinori, Roberta Gagliardini, Alessandra Vergori, Stefania Cicalini, Giovanna Onnelli, Alberto Giannetti, Roberto Cauda, Arturo Ciccullo, Silvia La Monica, Vincenzo Vullo, Gabriella Dettorre, Eugenio Nelson Cavallari, Massimo Andreoni, Vincenzo Malagnino, Laura Ceccarelli; Rovigo: Filippo Viviani, Lolita Sasset; Sanremo: Chiara Dentone; Siena: Barbara Rossetti, Sara Modica, Valentina Borgo; Torino: Giovanni Di Perri, Stefano Bonora, Micol Ferrara, Chiara Carcieri; Verona: Marina Malena, Marta Fiscon, Barbara Padovani; Trieste: Roberto Luzzati, Sandro Centonze,Romina Valentinotti. Acknowledgements The PRESTIGIO Registry is supported by ViiV Healthcare.