意大利患者的 Cryopyrin 相关周期性综合征：体细胞 NLRP3 镶嵌率的评估和表型特征分析。
Cryopyrin-associated Periodic Syndromes in Italian Patients: Evaluation of the Rate of Somatic NLRP3 Mosaicism and Phenotypic Characterization.
机构信息
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.
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.
出版信息
J Rheumatol. 2017 Nov;44(11):1667-1673. doi: 10.3899/jrheum.170041. Epub 2017 Sep 15.
OBJECTIVE
To evaluate the rate of somatic 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 gene yielded negative results. Patients' DNA were subjected to amplicon-based deep sequencing.
RESULTS
Low-level somatic 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). 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 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.
目的
评估阴性突变的 Cryopyrin 相关周期性综合征（CAPS）意大利患者队列中的体细胞镶嵌率。
方法
本研究纳入了 14 名临床表型符合 CAPS 的患者，这些患者的基因测序结果为阴性。对患者的 DNA 进行基于扩增子的深度测序。
结果
在 4 名患者中检测到低水平的体细胞镶嵌，其中 3 名患有慢性婴儿神经皮肤关节综合征，1 名患有 Muckle-Wells 综合征。鉴定出的核苷酸取代编码了 4 种不同的氨基酸交换，其中 2 种是新的（p.Y563C 和 p.G564S）。功能研究证实了这 4 种体细胞 NLRP3 突变的有害行为。在检测到的不同神经表现中，1 名患者的大脑磁共振成像显示出轻度的白质体积丢失。
结论
体细胞突变的等位基因频率通常低于 15%，这是使用 DNA 测序的 Sanger 方法检测的阈值。因此，目前 CAPS 的常规基因诊断应通过确保高覆盖率的下一代技术来进行，以识别低水平的镶嵌体，其实际频率尚不清楚，可能被低估了。
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