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Clinical, Pathological and Genetic Characteristics of Autosomal Dominant Inherited Dynamin 2 Centronuclear Myopathy

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Clinical, Pathological and Genetic Characteristics of Autosomal Dominant Inherited Dynamin 2 Centronuclear Myopathy MOLECULAR MEDICINE REPORTS 13: 4273-4278, 2016 Clinical, pathological and genetic characteristics of autosomal dominant inherited dynamin 2 centronuclear myopathy XINHONG LIU1,2, HUAMIN WU3, JIAN GONG4, TAO WANG2 and CHUANZHU YAN1 1Department of Neurology, Qilu Hospital of Shandong University, Jinan, Shandong 250012; Departments of 2Neurology, 3Radiology and 4Digestive System, Tai'an Central Hospital, Tai'an, Shandong 271000, P.R. China Received February 21, 2015; Accepted December 18, 2015 DOI: 10.3892/mmr.2016.5047 Abstract. The aim of the present study was to report on a the myotubularin 1 gene (MTM1) (2). A third gene, associ- family with pathologically and genetically diagnosed auto- ated with autosomal recessive inherited CNM, is bridging somal dominant inherited centronuclear myopathy (CNM). In integrator 1 (3). addition, this study aimed to investigate the clinical, patho- Previous studies on DNM2-associated CNM logical and molecular genetic characteristics of the disease. (DNM2 -CNM) cases have demonstrated that it can range from This pedigree was traced back three generations, four patients severe neonatal onset to mild adult onset, and the severity of underwent neurological examination, two patients underwent the clinical manifestations vary (4-7). The majority of patients muscle biopsy, and eight family members were subjected to exhibit distal or proximal muscle weakness as a first symptom, dynamin 2 (DNM2) gene mutation analysis. DNM2 mutations presenting as general muscle weakness, eyelid drooping, were detected in seven family members, of which four patients extraocular muscle paralysis, high arch palate, contracture of exhibited DNM2 mutation‑specific clinical and pathological the Achilles tendon, trismus, and reduction or lack of tendon features. Lower extremity weakness was the predominant reflexes, among which the weakness of the distal or lower limbs symptom of these patients, however, proximal and distal lower is most notable (4,8). Disease progression is predominantly extremity involvement was inconsistent. All patients exhibited slow and few patients develop independent‑walking difficulties marked systematic muscle atrophy and various degrees of in middle age or restrictive ventilatory disorders (9). Typical facial muscle involvement. The patients presented the typical pathological features of DNM2‑CNM are type I fiber predom- pathological changes of CNM, and their muscle tissues were inance and type I atrophy, increased proportions of radial heavily replaced by adipose tissue, with clustered distribution fibers and central nuclei in the sarcoplasmic reticulum (1). of muscle fibers as another notable feature. DNM2-CNM Approximately 100 DNM2-CNM families have exhibited patients of this pedigree exhibited heterogeneous clinical and 18 different DNM2 mutations (10). Patients with mutations pathological features, providing a basis for further molecular affecting the intermediate domains of DNM2 usually present genetic analysis. with relatively mild clinical features (5,11), while mutations impacting the pleckstrin protein family homology (PH) Introduction domain and GED structures result in more serious clinical manifestations (4). Centronuclear myopathies (CNMs) are a group of rare slowly To the best of our knowledge, this study is the first to progressive congenital myopathies, which are character- report on a Chinese family with pathologically and genetically ized by centronuclear dysplasia in muscle fibers. Autosomal confirmed DNM2‑associated autosomal dominant inherited dominant inherited mutations of the dynamin 2 (DNM2) CNM. This study aimed to investigate the clinical, patholog- gene are the cause of 50% of CNM cases (1). Clinical symp- ical and genetic characteristics of this disease, and contribute toms of DNM2-associated CNM are milder than those of to further molecular genetic studies of autosomal dominant X -linked recessive inheritedRETRACTED CNM caused by mutations in inherited CNM. Materials and methods Subjects. Four patients, two males and two females, belonging Correspondence to: Professor Chuanzhu Yan, Department of Neurology, Qilu Hospital of Shandong University, 107 Culture West to a family with three generations of autosomal dominant Road, Jinan, Shandong 250012, P.R. China inherited CNM (pedigree presented in Fig. 1), were investi- E -mail: [email protected] gated. The patients were recruited from the Qilu Hospital of Shandong University (Jinan, China) and presented muscle Key words: autosomal dominant inheritance, dynamin 2, centronuclear weakness. All patients underwent physical examination, myopathy genetic testing, and two of the patients further provided muscle biopsies. Detailed case history enquiries and clinical examina- tion were conducted by two neurologists. The present study 4274 LIU et al: CHARACTERISTICS OF DYNAMIN 2-CENTRONUCLEAR MYOPATHY Table I. Clinical patient data from d=four cases of dynamin 2-centronuclear myopathy. Variable I1 II2 II4 II5 Gender/age Female Male Female Male Age (years) 59 38 33 30 Onset Adolescence Adolescence Adolescence Adolescence Developmental delay Normal Normal Could walk at 2-years-old Unknown First symptom Lower extremity Lower extremity Lower extremity Lower extremity weakness weakness weakness weakness Muscle involvement Neck + - - - Upper limb Proximal 3 5 3-4 5 Distal 4 5 5 5 Lower limb Proximal 3 4-5 4 5 Distal 4 3 3 5 Limb-girdle muscular Upper limb 3-4 4 4 5 Lower limb 3-4 4 4 4 Facial muscle involvement + + + + Eyelid drooping + - - - Ophthalmoplegia ‑ ‑ ‑ ‑ High-vaulted arch - + - + Joint contractures - - - + Arched feet - - - - Curvature of spine - - - - Muscle atrophy + + + + Tendon reflexes + ‑ + ‑ + ‑ + ‑ Serum CK 126 Not tested Not tested Not tested EMG Myogenic damage Myogenic damage Not tested Not tested Dysphagia ‑ ‑ ‑ ‑ Breathing difficulty ‑ ‑ ‑ ‑ CK, creatine kinase; EMG, electromyography. -, negative; +, positive; + -, likely positive. Muscle strength grade: level 0, muscle paralysis, no contraction; level 1, slight contraction of the muscles, but no action; level 2, muscles only function when not subjected to excessive weight; level 3, muscles can carry out actions against gravity but not with added resistance; level 4, able to counteract some resistance against the muscle; level 5, normal muscle strength. was conducted in accordance with the declaration of Helsinki and with approval from the Ethics Committee of Shandong University (Jinan, China). Written informed consent was obtained from all participants.RETRACTED Pathological specimens. Muscle specimens of two patients were obtained from the left biceps by open biopsy. Fresh muscle samples were snap-frozen in liquid nitrogen-cooled isopentane (Sinopharm Chemical Reagent Co., Ltd., Beijing, China), and 8-µm tissue sections were cut using a Figure 1. Dynamin 2-associated autosomal dominant central nucleus myopathy cryo -ultramicrotome (Leica 1900; Leica Microsystems GmbH, family pedigree. Circle, female; square, male; black, affected; white, unaffected. Wetzlar, Germany) at ‑20 to ‑25˚C. Tissues were then stained with hematoxylin-eosin (H&E), modified Gomori's trichrome, NADH -tetrazolium reductase, cytochrome c oxidase, succinate routine use of dynamin-N, R, and C monoclonal antibody dehydrogenase, lactate dehydrogenase, periodic acid-Schiff, immunohistochemical staining in order to analyze dynamin and oil red O. Dystrophin staining was carried out with the protein expression levels in the sarcolemma using an optical MOLECULAR MEDICINE REPORTS 13: 4273-4278, 2016 4275 A B C D E F G H Figure 2. (A and B) I1, H&E staining, visible adipose tissue, muscle fibers distributed in clusters, fiber size is different in muscle tissue, the fibers are homo- geneously small, round and polygonal, with no obvious hyperplasia of the endomysium, >90% of the muscle fibers contain a central nucleus. (C) I1, NADH was used to distinguish between two types of fiber. (D‑I) II2, eldest son of I1 muscle pathology, (D and E) H&E staining, >90% of muscle fibers contain a central nucleus and exhibit visible adipose infiltration between the fibers. (F) NADH staining, typical appearance of fibrin observed. (G) Trophin staining demonstrates the membrane integrity with fat infiltration between the muscle fibers. (H) ATP staining demonstrated type I fiber predominated, however, a number of type II fibers were observed with normal ATP staining. microscope (Olympus BH-2; Olympus Corporation, Tokyo, involvement and various degrees of systematic muscle Japan). Remaining samples were stored at ‑80˚C until further atrophy. Symptoms are first identified at ~20 years of age, use. All specimens were reviewed and confirmed by two experi- while they were capable of taking care of themselves and enced clinical neurologists. living normal lives. The grandchildren of proband I1, III1 (13 years old), III2 (4 years old), III3 (4 years old) and III4 Gene sequencing analysis. Based on the DNM2 genome (5 years old), did not exhibit symptoms of muscle weakness, sequence, primers were designed to cover 22 exons, including and their sports activities were the same as healthy children alternate exons, exon 10b and exon 13b. DNM2 sequences of of the same age. However, III3 exhibited general features of eight family members were determined and compared with congenital muscular disease, including emaciation, narrow the human gene mutation database (http://www.hgmd.cf.ac. face and high-vaulted arch. Judging from the combined uk/ac/index.php) by Beijing ACCB Biotech, Ltd. (Beijing, inheritance mode and clinical
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