研究者詳細 - 兼松　毅

血友病Aにおけるassay discrepancy評価のための凝固一段法試薬のバリデーション査読有り国際誌

鈴木敦夫, 鈴木伸明, 兼松毅, 安藤善孝, 松下正

臨床病理 67 巻 ( 補冊 ) 頁： 251 - 251 2019年10月

Apparent synonymous mutation F9 c.87A > G causes secretion failure by in-frame mutation with aberrant splicing 査読有り国際誌

Odaira, K; Tamura, S; Suzuki, N; Kakihara, M; Hattori, Y; Tokoro, M; Suzuki, S; Takagi, A; Katsumi, A; Hayakawa, F; Okamoto, S; Suzuki, A; Kanematsu, T; Matsushita, T; Kojima, T

THROMBOSIS RESEARCH 179 巻頁： 95 - 103 2019年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Thrombosis Research

Introduction: Hemophilia B is an X-linked recessive bleeding disorder caused by coagulation factor IX (FIX) gene (F9) mutations. Several F9 synonymous mutations have been known to cause hemophilia B; however, the deleterious mechanisms underlying the development of hemophilia B have not been completely understood. To elucidate the molecular pathogenesis causing hemophilia B, we investigated the synonymous F9 mutation: c.87A>G, p.(Thr29=). Materials and methods: The influence of F9 c.87A>G on mRNA splicing was analyzed by exon-trap assay and in silico prediction. We prepared FIX expression vectors using mutant F9 cDNA and transfected HepG2 cells to investigate intracellular transport and extracellular secretion of FIX. Intracellular kinetics of the expressed FIX was examined by treatment with the proteasome inhibitor MG132. Results: Exon-trap analysis revealed that F9 c.87A>G resulted in almost (99.1%) aberrant splicing (r.83_88del). In silico analysis predicted that F9 c.87A>G influenced the splicing pattern by generating an available aberrant 5′ splice site. The aberrant F9 mRNA (r.83_88del) was translated to a mutant FIX p.Cys28_Val30delinsPhe with an in-frame mutation at the signal peptide cleavage site. Simultaneously, a small amount (0.9%) of mutant F9 r.87A>G translating into WT FIX p.Thr29 = was also observed. The mutant FIX was abnormally retained in the endoplasmic reticulum (ER) and was not extracellularly secreted. It appeared to be intracellularly degraded via proteasome-dependent degradation machinery. Conclusion: F9 c.87A>G was found to cause abnormal mRNA splicing, r.83_88del, and produce the mutant FIX p.Cys28_Val30delinsPhe. The mutant FIX is an abnormal protein with extracellular secretory defects and is intracellularly eliminated via proteasome-dependent ER-associated degradation.

DOI： 10.1016/j.thromres.2019.04.022

Thalidomide maintenance therapy in Japanese myeloma patients: a multicenter, phase II clinical trial (COMET study) (vol 109, pg 409, 2019) 査読有り

Murakami, H; Kasamatsu, T; Murakami, J; Kiguchi, T; Kanematsu, T; Ogawa, D; Takamatsu, H; Handa, H; Ozaki, S; Miki, H; Takahashi, T; Takeo, T; Yamauchi, T; Morishita, T; Kosugi, H; Shimizu, K

INTERNATIONAL JOURNAL OF HEMATOLOGY 110 巻 ( 1 ) 頁： 125 - 126 2019年7月

Ischaemic events are rare, and the prevalence of hypertension is not high in Japanese adults with haemophilia: First multicentre study in Asia 査読有り国際誌

Nagao, A; Suzuki, N; Takedani, H; Yamasaki, N; Chikasawa, Y; Sawada, A; Kanematsu, T; Nojima, M; Higasa, S; Amano, K; Fukutake, K; Fujii, T; Matsushita, T; Suzuki, T

HAEMOPHILIA 25 巻 ( 4 ) 頁： E223 - E230 2019年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Haemophilia

Introduction: With the increasing life expectancy of patients with haemophilia (PWH), the number of PWH with age-related comorbidities, such as ischaemic events, is increasing. Aim: We conducted this multicentre observational study to identify the risk factors for major ischaemic events in PWH. Methods: This study was the first multicentre observational study, conducted with the participation of five haemophilia treatment centres in Japan, conducted in ≥30-year-old adult PWH. The latest data recorded in the medical charts between 1 January and 31 December 2016 were reviewed. Healthcare data collected from the National Health and Nutrition Survey were used as the control data. Results: Data of a total of 711 patients were collected. Only two PWH (0.3%) had a history of ischaemic events. Age-adjusted analysis indicated that the prevalence of hypertension defined as a blood pressure of 140/90 mm Hg or over was similar in the PWH to that in the males of the general population. However, when hypertension was defined more strictly (≥130/85 mm Hg), the prevalence was significantly lower in PWH than in the general male population. The hypertension in PWH was associated with the age, BMI, CKD, HIV infection and inhibitors. In particular, the odds ratio for the presence of inhibitors was high (odds ratio = 7.529). Conclusion: Whether the present results can be attributed to Japanese ethnicity or to the presence of haemophilia per se remains uncertain. We propose to initiate a prospective study for further investigation.

DOI： 10.1111/hae.13749

Molecular basis of SERPINC1 mutations in Japanese patients with antithrombin deficiency 査読有り国際誌

Tamura, S; Hashimoto, E; Suzuki, N; Kakihara, M; Odaira, K; Hattori, Y; Tokoro, M; Suzuki, S; Takagi, A; Katsumi, A; Hayakawa, F; Suzuki, A; Okamoto, S; Kanematsu, T; Matsushita, T; Kojima, T

THROMBOSIS RESEARCH 178 巻頁： 159 - 170 2019年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Thrombosis Research

Background: Congenital antithrombin (AT) deficiency, which arises from various SERPINC1 defects, is an autosomal-dominant thrombophilic disorder associated with a high risk of recurrent venous thromboembolism. Patients/methods: We investigated SERPINC1 defects in Japanese patients with congenital AT deficiency who developed venous thromboembolism or had a family history of deep vein thrombosis. We analyzed the full DNA sequences of SERPINC1 exons and exon-intron junctions by PCR-mediated direct sequencing. If no mutation was found, multiplex ligation-dependent probe amplification (MLPA) was conducted for the relative quantification of the copy number of all exons in SERPINC1. If splice-site mutations were detected, mRNA splicing abnormalities were further investigated using an in vitro cell-based exontrap assay. Results: We identified 19 different SERPINC1 abnormalities, including 8 novel mutations, in 21 Japanese patients with AT deficiency. These abnormalities were distributed as follows: 9 missense mutations (42.9%), 3 nonsense mutations (14.3%), 1 splice-site mutation (4.8%), 2 small insertions (9.5%), 2 deletion mutations (9.5%) and 4 large deletions (19.0%). Cases with large deletions of SERPINC1 included Alu-mediated gene rearrangements and non-Alu-mediated complex gene rearrangements; the latter could conceivably be explained using the fork stalling and template switching (FoSTeS) model. Conclusions: We identified a variety of SERPINC1 defects in Japanese patients with AT deficiency. The SERPINC1 mutations detected in patients with type I AT deficiency included single nucleotide missense or nonsense mutations, small intragenic insertions or deletions, and large genomic structural deletions. Large deletions of SERPINC1 were caused by various recurrent or non-recurrent complex genomic rearrangement mutations.

DOI： 10.1016/j.thromres.2019.04.004

Thalidomide maintenance therapy in Japanese myeloma patients: a multicenter, phase II clinical trial (COMET study)

GPIb変異体を用いたvon Willebrand因子活性測定試薬の基本性能評価査読有り国際誌

鈴木敦夫, 鈴木伸明, 兼松毅, 岡本修一, 篠原翔, 新井信夫, 安藤善孝, 松下正

日本血栓止血学会誌 30 巻 ( 2 ) 頁： 470 - 470 2019年5月

特発性血小板減少性紫斑病の病勢に対し、サイトメガロウイルス再活性化の関与が疑われた2例査読有り国際誌

兼松毅, 鈴木伸明, 岡本修一, 鈴木敦夫, 川上萌, 三田直美, 田村彰吾, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 30 巻 ( 2 ) 頁： 471 - 471 2019年5月

Murakami, H; Kasamatsu, T; Murakami, J; Kiguchi, T; Kanematsu, T; Ogawa, D; Takamatsu, H; Handa, H; Ozaki, S; Miki, H; Takahashi, T; Takeo, T; Yamauchi, T; Morishita, T; Kosugi, H; Shimizu, K

INTERNATIONAL JOURNAL OF HEMATOLOGY 109 巻 ( 4 ) 頁： 409 - 417 2019年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：International Journal of Hematology

A prospective, multicenter, phase II study was performed to assess the efficacy and safety of thalidomide maintenance therapy at different doses in Japanese multiple myeloma (MM) patients. This study included 34 patients (median age, 74 years) who were previously treated with not more than three prior therapies and whose response status was evaluated as at least stable disease. They were randomized into Group A (no maintenance; 12 patients), Group B (50 mg thalidomide maintenance; 12 patients), and Group C (100 mg thalidomide maintenance; 10 patients), respectively. Thalidomide maintenance therapy resulted in improved depth of response in three cases (13.6%) and sustained response after induction therapy in eight cases (36.4%). Two-year progression-free survival (PFS) was 25.0%, 33.3%, and 77.8% in Groups A, B, and C, respectively, and was significantly higher in Group C than in Group A (p = 0.005). There was no difference in the incidence of hematological or non-hematological adverse events between Groups B and C. The current study demonstrates that maintenance with daily thalidomide at 100 mg, but not 50 mg, improved depth of response and prolonged PFS, and that this treatment was feasible for use in Japanese MM patients.

DOI： 10.1007/s12185-019-02607-z

Clot waveform analysis in Clauss fibrinogen assay contributes to classification of fibrinogen disorders 国際誌

Fc融合遺伝子組換え第VIII因子製剤(rFVIIIFc)の持続輸注にて周術期止血管理をおこなった血友病Aの2症例査読有り国際誌

鈴木伸明, 兼松毅, 鈴木敦夫, 岡本修一, 松下正

日本輸血細胞治療学会誌 65 巻 ( 2 ) 頁： 477 - 477 2019年4月

Suzuki, A; Suzuki, N; Kanematsu, T; Shinohara, S; Arai, N; Kikuchi, R; Matsushita, T

THROMBOSIS RESEARCH 174 巻頁： 98 - 103 2019年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Thrombosis Research

Background: Clauss fibrinogen assay (CFA) is widely used as a screening test to detect fibrinogen disorders. However, CFA alone cannot distinguish quantitative and qualitative defects because it depends on functional fibrinogen activity (Ac), and fibrinogen antigen (Ag) determination is required to classify fibrinogen disorders. Objectives: To establish a novel approach to classify fibrinogen disorders, we investigated the potential of clot waveform analysis (CWA) of CFA and searched for a surrogate marker for fibrinogen Ag. Materials and methods: We analyzed CWA parameters obtained from CFA using plasma from normal patients (n = 91) and those with fibrinogen disorders (n = 27, including 15 hypofibrinogenemia, 6 dysfibrinogenemia and 6 hypodysfibrinogenemia) with a CS-5100 autoanalyzer. Results: We found that maximum coagulation velocity (Min1) levels were most strongly correlated with fibrinogen Ag in both normal and fibrinogen disorders. Hence, Min1 appeared to function as a surrogate for fibrinogen Ag. Although the Ac/Min1 ratio did not simply reflect the measured Ac/Ag ratio, we found that the Ac/Min1 ratio was significantly higher than normal in hypofibrinogenemia and hypodysfibrinogenemia, but not in dysfibrinogenemia. On the other hand, we could distinguish type II deficiency from type I using estimated fibrinogen Ag (eAg) predicted from Min1. The Ac/eAg ratios of dysfibrinogenemia and hypodysfibrinogenemia were significantly lower than those of normal and hypofibrinogenemia. Conclusion: The CWA of CFA could distinguish fibrinogen disorders using a combination of Ac/Min1 and Ac/eAg values. This analysis allows the qualitative detection of fibrinogen disorder easily and represents a novel screening test for fibrinogen disorders.

DOI： 10.1016/j.thromres.2018.12.018