Denise Lasigliè1,2, Anna Mensa-Vilaro1,2, Denise Ferrera1,2, Roberta Caorsi1,2, Federica Penco1,2, Giuseppe Santamaria1,2, Marco Di Duca1,2, Giulia Amico1,2, Kenji Nakagawa1,2, Francesca Antonini1,2, Alberto Tommasini1,2, Rita Consolini1,2, Antonella Insalaco1,2, Marco Cattalini1,2, Laura Obici1,2, Romina Gallizzi1,2, Francesca Santarelli1,2, Genny Del Zotto1,2, Mariasavina Severino1,2, Anna Rubartelli1,2, Roberto Ravazzolo1,2, Alberto Martini1,2, Isabella Ceccherini1,2, Ryuta Nishikomori1,2, Marco Gattorno1,2, Juan I Arostegui1,2, Silvia Borghini3,4. 1. From these departments of the Istituto Giannina Gaslini, Genoa: Unità Operativa Complessa (UOC) Genetica Medica, UO Pediatria II, Laboratorio Fisiopatologia dell'Uremia, Core facilities, and Neuroradiology Unit; Institute for Maternal and Child Health, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Burlo Garofolo, Trieste; Department of Pediatrics, Immunology and Rheumatology Section, University of Pisa, Pisa; Division of Rheumatology, Department of Pediatric Medicine, Bambino Gesù Children's Hospital, IRCCS, Rome; Pediatric Clinic, Azienda Socio Sanitaria Territoriale (ASST) Spedali Civili and University of Brescia, Brescia; Amyloidosis Research and Treatment Centre, Biotechnology Research Laboratories, Fondazione IRCCS Policlinico San Matteo, Pavia; Department of Pediatrics, Azienda G. Martino, University of Messina, Messina; Department of Pediatrics, Immunology and Rheumatology, University of Torino, Torino; Cell Biology Unit, IRCCS L'Azienda Ospedaliera Universitaria (AOU) San Martino, San Martino, Italy; Department of Immunology, Hospital Clínic-Institut d'Investigacions Biomdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan. 2. D. Lasigliè, PhD, UOC Genetica Medica, Istituto Giannina Gaslini; D. Ferrera, PhD, UOC Genetica Medica, Istituto Giannina Gaslini; G. Santamaria, BSc, UOC Genetica Medica, Istituto Giannina Gaslini; G. Amico, MSc, UOC Genetica Medica, Istituto Giannina Gaslini; R. Ravazzolo, MD, UOC Genetica Medica, Istituto Giannina Gaslini; I. Ceccherini, PhD, UOC Genetica Medica, Istituto Giannina Gaslini; S. Borghini, PhD, UOC Genetica Medica, Istituto Giannina Gaslini; M. Di Duca, BSc, Laboratorio Fisiopatologia dell'Uremia, Istituto Giannina Gaslini; F. Penco, PhD, UO Pediatria II Istituto Giannina Gaslini; R. Caorsi, MD, UO Pediatria II Istituto Giannina Gaslini; A. Martini, MD, UO Pediatria II Istituto Giannina Gaslini; M. Gattorno, MD, UO Pediatria II Istituto Giannina Gaslini; A. Mensa-Vilaro, MD, Hospital Clínic-IDIBAPS; J.I. Arostegui, MD, Department of Immunology, Hospital Clínic-IDIBAPS; A. Tommasini, MD, PhD, Institute for Maternal and Child Health, IRCCS Burlo Garofolo; F. Antonini, MSc, Core facilities, Istituto Giannina Gaslini; G. del Zotto, PhD, Core facilities, Istituto Giannina Gaslini; R. Consolini, MD, Department of Pediatrics, Immunology and Rheumatology Section, University of Pisa; A. Insalaco, MD, Division of Rheumatology, Department of Pediatric Medicine, Bambino Gesù Children's Hospital, IRCCS; M. Cattalini, MD, Pediatric Clinic, ASST Spedali Civili and University of Brescia; L. Obici, MD, Amyloidosis Research and Treatment Centre, Biotechnology Research Laboratories, Fondazione IRCCS Policlinico San Matteo; R. Gallizzi, MD, Department of Pediatrics, Azienda G. Martino, University of Messina; F. Santarelli, MD, Department of Pediatrics, Immunology and Rheumatology, University of Torino; A. Rubartelli, MD, Cell Biology Unit, IRCSS AOU San Martino; M. Severino, MD, Neuroradiology Unit, Istituto Giannina Gaslini; R. Nishikomori, MD, PhD, Department of Pediatrics, Graduate School of Medicine, Kyoto University; K. Nakagawa, MD, PhD, Department of Pediatrics, Graduate School of Medicine, Kyoto University. 3. From these departments of the Istituto Giannina Gaslini, Genoa: Unità Operativa Complessa (UOC) Genetica Medica, UO Pediatria II, Laboratorio Fisiopatologia dell'Uremia, Core facilities, and Neuroradiology Unit; Institute for Maternal and Child Health, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Burlo Garofolo, Trieste; Department of Pediatrics, Immunology and Rheumatology Section, University of Pisa, Pisa; Division of Rheumatology, Department of Pediatric Medicine, Bambino Gesù Children's Hospital, IRCCS, Rome; Pediatric Clinic, Azienda Socio Sanitaria Territoriale (ASST) Spedali Civili and University of Brescia, Brescia; Amyloidosis Research and Treatment Centre, Biotechnology Research Laboratories, Fondazione IRCCS Policlinico San Matteo, Pavia; Department of Pediatrics, Azienda G. Martino, University of Messina, Messina; Department of Pediatrics, Immunology and Rheumatology, University of Torino, Torino; Cell Biology Unit, IRCCS L'Azienda Ospedaliera Universitaria (AOU) San Martino, San Martino, Italy; Department of Immunology, Hospital Clínic-Institut d'Investigacions Biomdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan. s.borghini@unige.it. 4. D. Lasigliè, PhD, UOC Genetica Medica, Istituto Giannina Gaslini; D. Ferrera, PhD, UOC Genetica Medica, Istituto Giannina Gaslini; G. Santamaria, BSc, UOC Genetica Medica, Istituto Giannina Gaslini; G. Amico, MSc, UOC Genetica Medica, Istituto Giannina Gaslini; R. Ravazzolo, MD, UOC Genetica Medica, Istituto Giannina Gaslini; I. Ceccherini, PhD, UOC Genetica Medica, Istituto Giannina Gaslini; S. Borghini, PhD, UOC Genetica Medica, Istituto Giannina Gaslini; M. Di Duca, BSc, Laboratorio Fisiopatologia dell'Uremia, Istituto Giannina Gaslini; F. Penco, PhD, UO Pediatria II Istituto Giannina Gaslini; R. Caorsi, MD, UO Pediatria II Istituto Giannina Gaslini; A. Martini, MD, UO Pediatria II Istituto Giannina Gaslini; M. Gattorno, MD, UO Pediatria II Istituto Giannina Gaslini; A. Mensa-Vilaro, MD, Hospital Clínic-IDIBAPS; J.I. Arostegui, MD, Department of Immunology, Hospital Clínic-IDIBAPS; A. Tommasini, MD, PhD, Institute for Maternal and Child Health, IRCCS Burlo Garofolo; F. Antonini, MSc, Core facilities, Istituto Giannina Gaslini; G. del Zotto, PhD, Core facilities, Istituto Giannina Gaslini; R. Consolini, MD, Department of Pediatrics, Immunology and Rheumatology Section, University of Pisa; A. Insalaco, MD, Division of Rheumatology, Department of Pediatric Medicine, Bambino Gesù Children's Hospital, IRCCS; M. Cattalini, MD, Pediatric Clinic, ASST Spedali Civili and University of Brescia; L. Obici, MD, Amyloidosis Research and Treatment Centre, Biotechnology Research Laboratories, Fondazione IRCCS Policlinico San Matteo; R. Gallizzi, MD, Department of Pediatrics, Azienda G. Martino, University of Messina; F. Santarelli, MD, Department of Pediatrics, Immunology and Rheumatology, University of Torino; A. Rubartelli, MD, Cell Biology Unit, IRCSS AOU San Martino; M. Severino, MD, Neuroradiology Unit, Istituto Giannina Gaslini; R. Nishikomori, MD, PhD, Department of Pediatrics, Graduate School of Medicine, Kyoto University; K. Nakagawa, MD, PhD, Department of Pediatrics, Graduate School of Medicine, Kyoto University. s.borghini@unige.it.
Abstract
OBJECTIVE: To evaluate the rate of somatic NLRP3 mosaicism in an Italian cohort of mutation-negative patients with cryopyrin-associated periodic syndrome (CAPS). METHODS: The study enrolled 14 patients with a clinical phenotype consistent with CAPS in whom Sanger sequencing of the NLRP3 gene yielded negative results. Patients' DNA were subjected to amplicon-based NLRP3 deep sequencing. RESULTS: Low-level somatic NLRP3 mosaicism has been detected in 4 patients, 3 affected with chronic infantile neurological cutaneous and articular syndrome and 1 with Muckle-Wells syndrome. Identified nucleotide substitutions encode for 4 different amino acid exchanges, with 2 of them being novel (p.Y563C and p.G564S). In vitro functional studies confirmed the deleterious behavior of the 4 somatic NLRP3 mutations. Among the different neurological manifestations detected, 1 patient displayed mild loss of white matter volume on brain magnetic resonance imaging. CONCLUSION: The allele frequency of somatic NLRP3 mutations occurs generally under 15%, considered the threshold of detectability using the Sanger method of DNA sequencing. Consequently, routine genetic diagnostic of CAPS should be currently performed by next-generation techniques ensuring high coverage to identify also low-level mosaicism, whose actual frequency is yet unknown and probably underestimated.
OBJECTIVE: To evaluate the rate of somatic NLRP3 mosaicism in an Italian cohort of mutation-negative patients with cryopyrin-associated periodic syndrome (CAPS). METHODS: The study enrolled 14 patients with a clinical phenotype consistent with CAPS in whom Sanger sequencing of the NLRP3 gene yielded negative results. Patients' DNA were subjected to amplicon-based NLRP3 deep sequencing. RESULTS: Low-level somatic NLRP3 mosaicism has been detected in 4 patients, 3 affected with chronic infantile neurological cutaneous and articular syndrome and 1 with Muckle-Wells syndrome. Identified nucleotide substitutions encode for 4 different amino acid exchanges, with 2 of them being novel (p.Y563C and p.G564S). In vitro functional studies confirmed the deleterious behavior of the 4 somatic NLRP3 mutations. Among the different neurological manifestations detected, 1 patient displayed mild loss of white matter volume on brain magnetic resonance imaging. CONCLUSION: The allele frequency of somatic NLRP3 mutations occurs generally under 15%, considered the threshold of detectability using the Sanger method of DNA sequencing. Consequently, routine genetic diagnostic of CAPS should be currently performed by next-generation techniques ensuring high coverage to identify also low-level mosaicism, whose actual frequency is yet unknown and probably underestimated.
Authors: Micol Romano; Z Serap Arici; David Piskin; Sara Alehashemi; Daniel Aletaha; Karyl Barron; Susanne Benseler; Roberta A Berard; Lori Broderick; Fatma Dedeoglu; Michelle Diebold; Karen Durrant; Polly Ferguson; Dirk Foell; Jonathan S Hausmann; Olcay Y Jones; Daniel Kastner; Helen J Lachmann; Ronald M Laxer; Dorelia Rivera; Nicola Ruperto; Anna Simon; Marinka Twilt; Joost Frenkel; Hal M Hoffman; Adriana A de Jesus; Jasmin B Kuemmerle-Deschner; Seza Ozen; Marco Gattorno; Raphaela Goldbach-Mansky; Erkan Demirkaya Journal: Arthritis Rheumatol Date: 2022-05-27 Impact factor: 15.483
Authors: Keith A Sikora; Kristina V Wells; Ertugrul Cagri Bolek; Adrianna I Jones; Peter C Grayson Journal: Rheumatology (Oxford) Date: 2022-08-03 Impact factor: 7.046