ミヤタ　フミ   Miyata Fumi   宮田　文    所属   医学部 医学科（東京女子医科大学病院）    職種   助教
論文種別	原著
言語種別	英語
査読の有無	査読あり
表題	Analysis of the diagnosis of Japanese patients with primary ciliary dyskinesia using a conditional reprogramming culture.
掲載誌名	正式名：Respiratory investigation 略　 称：Respir Investig ISSNコード：22125353/22125345
掲載区分	国外
巻・号・頁	60(3),pp.407-417
著者・共著者	KUROKAWA Atsushi†, KONDO Mitsuko, HONDA Nahoko, ORIMO Mami, MIYOSHI Azusa, KOBAYASHI Fumi, ABE Kazuhiro, AKABA Tomohiro, TSUJI Mayoko, ARIMURA Ken, NAKATANI Kaname, IKEJIRI Makoto, YAGI Osamitsu, TAKEYAMA Kiyoshi, KATSURA Hideki, TAKEUCHI Kazuhiko, TAGAYA Etsuko
発行年月	2022/05
概要	BACKGROUND:Primary ciliary dyskinesia (PCD) is diagnosed through multiple methods, including transmission electron microscopy (TEM), a high-speed video microscopy analysis (HSVA), immunofluorescence (IF), and genetic testing. A primary cell culture has been recommended to avoid the misdiagnosis of secondary ciliary dyskinesia derived from infection or inflammation and improve diagnostic accuracy. However, primary cells fail to differentiate into ciliated cells through repeated passages. The conditional reprogramming culture (CRC) method, a combination of a Rho-kinase inhibitor and fibroblast feeder cells, has been applied to cystic fibrosis. The goal of this study was to evaluate the value of CRC in diagnosing PCD in Japanese patients.METHODS:Eleven patients clinically suspected of having PCD were included. Airway epithelial cells were obtained from an endobronchial forceps biopsy and cultured at the air-liquid interface (ALI) combined with CRC. Ciliary movement, ultrastructure, and mutated ciliary protein evaluation were performed using HSVA, TEM, and IF, respectively. Genetic testing was performed on some patients.RESULTS:CRC yielded dense and well-differentiated ciliated cells with a high success rate (∼90%). In patients with PCD, the ciliary ultrastructure phenotype (outer dynein arm defects or normal ultrastructure) and IF findings (absence of the mutated ciliary protein) were confirmed after CRC. In DNAH11-mutant cases with normal ultrastructure by TEM, the HSVA revealed stiff and hyperfrequent ciliary beating with low bending capacity in CRC-expanded cells, thereby supporting the diagnosis.CONCLUSIONS:CRC could be a potential tool for improving diagnostic accuracy and contributing to future clinical and basic research in PCD.
DOI	10.1016/j.resinv.2022.02.003
PMID	35305968