Papers - Narumi, Satoshi

A study of the etiology of transient congenital hypothyroidism in Niigata Prefecture, Japan.

Nagasaki K, Narumi S, Abe K, Asami T, Sato H, Ogawa Y, Kikuchi T, Hasegawa T, Saitoh A

International journal of pediatric endocrinology  2013

Accepted

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Quantitative and Sensitive Detection of GNAS Mutations Causing McCune-Albright Syndrome with Next Generation Sequencing

Satoshi Narumi, Kumihiro Matsuo, Tomohiro Ishii, Yusuke Tanahashi, Tomonobu Hasegawa

PLOS ONE (PUBLIC LIBRARY SCIENCE)  8 ( 3 ) e60525 2013

Accepted,  ISSN  1932-6203

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Somatic activating GNAS mutations cause McCune-Albright syndrome (MAS). Owing to low mutation abundance, mutant-specific enrichment procedures, such as the peptide nucleic acid (PNA) method, are required to detect mutations in peripheral blood. Next generation sequencing (NGS) can analyze millions of PCR amplicons independently, thus it is expected to detect low-abundance GNAS mutations quantitatively. In the present study, we aimed to develop an NGS-based method to detect low-abundance somatic GNAS mutations. PCR amplicons encompassing exons 8 and 9 of GNAS, in which most activating mutations occur, were sequenced on the MiSeq instrument. As expected, our NGS-based method could sequence the GNAS locus with very high read depth (approximately 100,000) and low error rate. A serial dilution study with use of cloned mutant and wildtype DNA samples showed a linear correlation between dilution and measured mutation abundance, indicating the reliability of quantification of the mutation. Using the serially diluted samples, the detection limits of three mutation detection methods (the PNA method, NGS, and combinatory use of PNA and NGS [PNA-NGS]) were determined. The lowest detectable mutation abundance was 1% for the PNA method, 0.03% for NGS and 0.01% for PNA-NGS. Finally, we analyzed 16 MAS patient-derived leukocytic DNA samples with the three methods, and compared the mutation detection rate of them. Mutation detection rate of the PNA method, NGS and PNA-NGS in 16 patient-derived peripheral blood samples were 56%, 63% and 75%, respectively. In conclusion, NGS can detect somatic activating GNAS mutations quantitatively and sensitively from peripheral blood samples. At present, the PNA-NGS method is likely the most sensitive method to detect low-abundance GNAS mutation.

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Functional characterization of four novel PAX8 mutations causing congenital hypothyroidism: new evidence for haploinsufficiency as a disease mechanism

Satoshi Narumi, Shunsuke Araki, Naoaki Hori, Koji Muroya, Yukiyo Yamamoto, Yumi Asakura, Masanori Adachi, Tomonobu Hasegawa

EUROPEAN JOURNAL OF ENDOCRINOLOGY (BIOSCIENTIFICA LTD)  167 ( 5 ) 625 - 32 2012.11

Accepted,  ISSN  0804-4643

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Background: Individuals carrying a heterozygous inactivating PAX8 mutation are affected by congenital hypothyroidism (CH), although heterozygous Pax8 knockout mice are not. It has remained unclear whether CH in PAX8 mutation carriers is caused by haploinsufficiency or a dominant negative mechanism.
Objective: To report clinical and molecular findings of four novel PAX8 mutations, including one early-truncating frameshift mutation.
Subjects and methods: Four probands were CH patients. Two had family history of congenital or childhood hypothyroidism. Three probands were diagnosed in the frame of newborn screening for CH, while one had a negative result in screening but was diagnosed subsequently. Three had thyroid hypoplasia and one had a slightly small thyroid with low echogenicity. For these probands and their family members, we sequenced PAX8 using a standard PCR-based method. Pathogenicity of identified mutations was verified in vitro.
Results: We found four novel heterozygous PAX8 mutations in the four probands: L16P, F20S, D46SfsX24, and R133Q. Family studies showed four additional mutation carriers, who were confirmed to have high serum TSH levels. Expression experiments revealed that three mutations (L16P, F20S, and R133Q) had defects in target DNA binding, while D46fs had protein instability that was rescued by the proteasome inhibitor MG132. All four mutations had reduced transactivation on the thyroglobulin promoter, supporting that they were inactivating mutations.
Conclusion: D46fs is the first PAX8 mutation with confirmed protein instability. Our clinical and in vitro findings together suggest that pure PAX8 haploinsufficiency can cause CH in humans.

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両アレル性DUOX2変異例におけるヨード有機化能の検討

長崎 啓祐, 鳴海 覚志, 阿部 清美, 佐藤 英利, 小川 洋平, 浅見 直, 長谷川 奉延, 菊池 透

日本内分泌学会雑誌 ((一社)日本内分泌学会)  88 ( 2 ) 497 - 497 2012.09

ISSN  0029-0661

次世代シーケンシングによる先天性甲状腺機能低下症の包括的遺伝子解析

鳴海 覚志, 阿部 清美, 天野 直子, 石井 智弘, 長谷川 奉延

日本内分泌学会雑誌 ((一社)日本内分泌学会)  88 ( 2 ) 498 - 498 2012.09

ISSN  0029-0661

先天性甲状腺機能低下症発症におけるPathway Burden仮説の提唱

阿部 清美, 鳴海 覚志, 天野 直子, 石井 智弘, 室谷 浩二, 朝倉 由美, 安達 昌功, 佐々木 悟郎, 長崎 啓祐, 阿部 貴行, 長谷川 奉延

日本内分泌学会雑誌 ((一社)日本内分泌学会)  88 ( 2 ) 543 - 543 2012.09

ISSN  0029-0661

Acceleration of diabetes development in CXC chemokine receptor 3 (CXCR3)-deficient NOD mice.

Yamada Y, Okubo Y, Shimada A, Oikawa Y, Yamada S, Narumi S, Matsushima K, Itoh H

Diabetologia 55 ( 8 ) 2238 - 45 2012.08

ISSN  0012-186X

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新生児マス・スクリーニングの震災対応はどうあるべきか 東日本大震災1年を経過して 東日本大震災が新生児マス・スクリーニングシステムに及ぼした影響 内分泌疾患を中心に

南谷 幹史, 皆川 真規, 伊藤 順庸, 鬼形 和道, 鈴木 順造, 高橋 明雄, 長崎 啓祐, 鳴海 覚志, 難波 範行, 原田 正平, 藤原 幾磨, 堀川 玲子, 水野 晴夫, 横谷 進

日本マス・スクリーニング学会誌 (日本マススクリーニング学会)  22 ( 2 ) 158 - 158 2012.07

ISSN  0917-3803

東日本大震災の新生児マススクリーニングに対する影響調査

南谷 幹史, 皆川 真規, 伊藤 順庸, 鬼形 和道, 鈴木 順造, 高橋 明雄, 長崎 啓祐, 鳴海 覚志, 難波 範行, 原田 正平, 藤原 幾磨, 堀川 玲子, 水野 晴夫, 横谷 進

日本内分泌学会雑誌 ((一社)日本内分泌学会)  88 ( 1 ) 308 - 308 2012.04

ISSN  0029-0661

東日本大震災の新生児マス・スクリーニングに対する影響調査

南谷 幹史, 皆川 真規, 鈴木 順造, 高橋 明雄, 長崎 啓祐, 鳴海 覚志, 原田 正平, 藤原 幾磨, 堀川 玲子, 横谷 進, 日本小児内分泌学会震災小児甲状腺PT

日本小児科学会雑誌 ((公社)日本小児科学会)  116 ( 2 ) 278 - 278 2012.02

ISSN  0001-6543

A Family of Pseudohypoparathyroidism Type Ia With an 850-kb Submicroscopic Deletion Encompassing the Whole GNAS Locus

Toshikatsu Mitsui, Keisuke Nagasaki, Masaki Takagi, Satoshi Narumi, Tomohiro Ishii, Tomonobu Hasegawa

AMERICAN JOURNAL OF MEDICAL GENETICS PART A (WILEY-BLACKWELL)  158A ( 1 ) 261 - 4 2012.01

Accepted,  ISSN  1552-4825

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Gradual Loss of ACTH Due to a Novel Mutation in LHX4: Comprehensive Mutation Screening in Japanese Patients with Congenital Hypopituitarism

Masaki Takagi, Tomohiro Ishii, Mikako Inokuchi, Naoko Amano, Satoshi Narumi, Yumi Asakura, Koji Muroya, Yukihiro Hasegawa, Masanori Adachi, Tomonobu Hasegawa

PLOS ONE (PUBLIC LIBRARY SCIENCE)  7 ( 9 ) e46008 2012

Accepted,  ISSN  1932-6203

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Mutations in transcription factors genes, which are well regulated spatially and temporally in the pituitary gland, result in congenital hypopituitarism (CH) in humans. The prevalence of CH attributable to transcription factor mutations appears to be rare and varies among populations. This study aimed to define the prevalence of CH in terms of nine CH-associated genes among Japanese patients. We enrolled 91 Japanese CH patients for DNA sequencing of POU1F1, PROP1, HESX1, LHX3, LHX4, SOX2, SOX3, OTX2, and GLI2. Additionally, gene copy numbers for POU1F1, PROP1, HESX1, LHX3, and LHX4 were examined by multiplex ligation-dependent probe amplification. The gene regulatory properties of mutant LHX4 proteins were characterized in vitro. We identified two novel heterozygous LHX4 mutations, namely c.249-1G>A, p.V75I, and one common POU1F1 mutation, p.R271W. The patient harboring the c.249-1G>A mutation exhibited isolated growth hormone deficiency at diagnosis and a gradual loss of ACTH, whereas the patient with the p.V75I mutation exhibited multiple pituitary hormone deficiency. In vitro experiments showed that both LHX4 mutations were associated with an impairment of the transactivation capacities of POU1F1 and alpha GSU, without any dominant-negative effects. The total mutation prevalence in Japanese CH patients was 3.3%. This study is the first to describe, a gradual loss of ACTH in a patient carrying an LHX4 mutation. Careful monitoring of hypothalamic-pituitary -adrenal function is recommended for CH patients with LHX4 mutations.

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【小児内分泌学の進歩2012】骨代謝・副甲状腺 GNAS変異陰性-偽性副甲状腺機能低下症IaにおけるGNAS遺伝子領域の全てを含む欠失の同定

三井 俊賢, 長崎 啓祐, 高木 優樹, 鳴海 覚志, 石井 智弘, 長谷川 奉延

ホルモンと臨床 ((有)医学の世界社)  59 ( 12 ) 1085 - 1087 2011.12

ISSN  0045-7167

非古典型TSH不応症 奇異性123I摂取率高値を特徴とする先天性甲状腺機能低下症の新しい病型の提唱

鳴海 覚志, 長崎 啓祐, 石井 智弘, 室谷 浩二, 朝倉 由美, 安達 昌功, 長谷川 奉延

日本内分泌学会雑誌 ((一社)日本内分泌学会)  87 ( 2 ) 516 - 516 2011.09

ISSN  0029-0661

DUOX2変異による偽性優性遺伝を示した先天性甲状腺機能低下症の一家系

阿部 清美, 濱島 崇, 鳴海 覚志, 長谷川 奉延

日本内分泌学会雑誌 ((一社)日本内分泌学会)  87 ( 2 ) 509 - 509 2011.09

ISSN  0029-0661

A new rapid protocol for eyeblink conditioning to assess cerebellar motor learning.

Emi K, Kohda K, Kakegawa W, Narumi S, Yuzaki M

Neurochemical research 36 ( 7 ) 1314 - 22 2011.07

ISSN  0364-3190

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当院で経験した自律性機能性甲状腺結節(AFTN)の1男児例

神原 亜紀子, 小山 さとみ, 志村 直人, 鳴海 覚志, 長谷川 奉延, 有阪 治

日本小児科学会雑誌 ((公社)日本小児科学会)  115 ( 2 ) 434 - 434 2011.02

ISSN  0001-6543

一過性甲状腺機能低下症におけるDUOX2遺伝子解析の検討

長崎 啓祐, 鳴海 覚志, 小川 洋平, 菊池 透, 浅見 直, 長谷川 奉延, 内山 聖

日本内分泌学会雑誌 ((一社)日本内分泌学会)  86 ( 2 ) 266 - 266 2010.09

ISSN  0029-0661

甲状腺形成異常におけるCopy number variation異常解析

鳴海 覚志, 室谷 浩二, 朝倉 由美, 安達 昌功, 長崎 啓祐, 天野 直子, 石井 智弘, 石井 美穂, 江見 充, 長谷川 奉延

日本内分泌学会雑誌 ((一社)日本内分泌学会)  86 ( 2 ) 267 - 267 2010.09

ISSN  0029-0661

One Novel and Two Recurrent THRB Mutations Associated with Resistance to Thyroid Hormone: Structure-based Computational Mutation Prediction

Narumi S, Cho H, Tamada I, Kozu Y, Tsuchiya T, Nagai T, Hasegawa T.

Clinical Pediatric Endocrinology 19 ( 4 ) 91-99 - 9 2010.08

Research paper (scientific journal), Joint Work, Accepted,  ISSN  0918-5739

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Inactivating mutations of THRB, which encodes the thyroid hormone receptor β (TRβ), cause resistance to thyroid hormone (RTH
OMIM 190160). To date, more than 100 THRB mutations have been reported among RTH patients. Most mutations substitute a single amino-acid residue in the ligand-binding domain. In this report, we describe clinical and molecular findings of three families with RTH. Three families harbored one novel (p.I431M) and two recurrent (p.R320H and p.R383C) THRB mutations. To examine the pathogenicity of identified mutations, we introduced a novel computational mutation prediction method based on three-dimensional structure data of TRβ-T3 complex. First, to define the accuracy of our prediction system, we evaluated ten previously reported 'positive control' mutations, as well as 30 seemingly benign sequence variations observed among vertebral species as 'negative controls'. We found that our system had a sensitivity of 80% and a specificity of 93%. We then analyzed three mutations detected in the present study and found that all three mutations are predicted to be deleterious. Our data suggest that our structure-based prediction system would be a prompt, inexpensive and feasible method for evaluating the pathogenicity of missense THRB mutations. © 2010 by The Japanese Society for Pediatric Endocrinology.

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