研究者詳細 - 兼松　毅

Clinical course and prognosis of patients with hepatocellular carcinoma and haemophilia 査読有り国際誌

Matsuda, N; Imai, N; Yokoyama, S; Yamamoto, K; Ito, T; Ishizu, Y; Honda, T; Okamoto, S; Kanematsu, T; Suzuki, N; Matsushita, T; Ishigami, M; Kawashima, H

EUROPEAN JOURNAL OF GASTROENTEROLOGY & HEPATOLOGY 35 巻 ( 10 ) 頁： 1211 - 1215 2023年10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：European Journal of Gastroenterology and Hepatology

Introduction Although patients with haemophilia are known to develop hepatocellular carcinoma (HCC) at a lower age than patients without, there are few reports on the clinical course and prognosis of HCC. Aim We aimed to investigate the clinical course and prognosis of patients with HCC and haemophilia. Methods Twenty-two patients with haemophilia, who were initially diagnosed with HCC between 2003 and 2021, were included. Their clinical courses and prognoses were retrospectively analysed. The results were compared with those of the 24th Nationwide Follow-up Survey of Primary Liver Cancer. Results All 22 patients were male; of these, 20 patients had haemophilia A, and 2 had haemophilia B. The mean age of diagnosis was 63 years (range 45-78 years) which is lower than the mean of 72 years reported in the Nationwide Survey. The mean diameter of the largest tumour was 30 mm (range 11-70 mm), and 18 tumours (82%) were solitary at the initial diagnosis. Standard treatments for HCC were performed in all patients. Sixty-one transarterial chemoembolisation, 28 RFA, 10 hepatectomies, and 2 radiation treatments were performed, and molecular-targeted agents were administered to 5 patients during their clinical courses. No deaths were associated with complications of HCC treatments. The median survival time after initial treatment was 6.4 years (range 0.9-18.7 years) which did not differ much from the median survival time of 5.8 years in the Nationwide Survey. Conclusion Standard treatment for HCC could improve the prognosis of patients with HCC and haemophilia.

DOI： 10.1097/MEG.0000000000002628

The usefulness of tranexamic acid for bleeding symptoms of chronic consumptive coagulopathy complicated by aortic disease: a single-institute, retrospective study of 14 patients 査読有り国際誌

Suzuki, N; Suzuki, N; Kawaguchi, Y; Okamoto, S; Kanematsu, T; Katsumi, A; Suzuki, A; Tamura, S; Kojima, T; Kiyoi, H; Matsushita, T

THROMBOSIS JOURNAL 21 巻 ( 1 ) 頁： 10 - 10 2023年1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 Journal

Background: Tranexamic acid (TXA) is an antifibrinolytic drug that blocks lysine-binding sites on the profibrinolytic enzyme plasminogen. Aortic diseases with chronic consumption coagulopathy may lead to disseminated intravascular coagulation (DIC) and cause fatal bleeding. Although the use of antifibrinolytic agents in DIC is generally not recommended due to enhanced fibrin deposition risking thrombotic symptoms, the efficacy of TXA has been reported in several cases of DIC with aortic diseases. However, the efficacy and safety of TXA for bleeding symptoms of chronic consumption coagulopathy with aortic diseases have not been studied in detail. Methods: We evaluated the efficacy of TXA in 14 patients with chronic consumptive coagulopathy due to aortic disease complicated by bleeding symptoms. Changes in coagulation and fibrinolysis parameters from baseline were analyzed with Wilcoxon matched-pairs signed-rank tests, excluding missing values. Kaplan-Meier curves were used to analyze overall survival. Results: Median age was 78.5 years (range, 66–89 years) and median observation period was 448 days (range, 0–2282 days). Twelve patients had chronic renal failure and 1 patient had chronic liver failure. Before starting treatment, median Japanese Ministry of Health and Welfare DIC diagnostic criteria score was 8 (range, 4–11) and median platelet count was 64 × 109/L (range, 25–97 × 109/L). Twelve patients underwent evaluation of bleeding symptoms after introduction of TXA, and 10 of those 12 patients showed improved bleeding tendencies within 30 days (median, 5.0 days). One patient with chronic liver failure showed worsening of bleeding symptoms. Although only one patient was initiated TXA in combination with anticoagulants, no significant worsening of thrombotic events was observed within 30 days. Conclusions: TXA therapy appears effective against chronic consumptive coagulopathy with bleeding due to aortic disease, with few side effects.

DOI： 10.1186/s12959-022-00429-4

Bursitis, Bacteremia, and Disseminated Infection of Mycobacteroides (Mycobacterium) abscessus subsp. massiliense 国際誌

Oka, K; Morioka, H; Eguchi, M; Sato, Y; Tetsuka, N; Iguchi, M; Kanematsu, T; Fukano, H; Hoshino, Y; Kiyoi, H; Yagi, T

INTERNAL MEDICINE 60 巻 ( 18 ) 頁： 3041 - 3045 2021年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：一般社団法人日本内科学会

We herein report a 59-year-old woman with a 2-year history of chronic bursitis of the hand who took 50 mg/day prednisolone for several autoimmune diseases. Mycobacteroides abscessus subsp. massiliense was isolated from the abscess and blood culture. Combination therapy (imipenem/cilastatin, amikacin, and clarithromycin) was administered for a month. Two months later, M. massiliense was detected from a blood culture again, and disseminated lesions were found. Clarithromycin and sitafloxacin were administered following eight weeks of the same regimen. Six months after the diagnosis, M. massiliense was isolated from a blood culture, and she expired due to multiple organ failure.

DOI： 10.2169/internalmedicine.6189-20

[Lupus anticoagulant-hypoprothrombinemia syndrome associated with follicular lymphoma]. 査読有り国際誌

Koyama D, Hanajiri R, Kanematsu T, Ito R, Yamamoto S, Imoto N, Suzuki N, Kurahashi S, Sugiura I

[Rinsho ketsueki] The Japanese journal of clinical hematology 61 巻 ( 7 ) 頁： 745 - 749 2020年

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：一般社団法人日本血液学会

56歳女性。繰り返す四肢の腫脹，疼痛，紫斑を主訴に受診した。PTとAPTTの延長を認め，クロスミキシング試験で凝固阻害因子の存在が疑われた。ループスアンチコアグラント（LA）陽性かつプロトロンビン活性著減を認め，ループスアンチコアグラント低プロトロンビン血症症候群（LAHPS）と診断した。基礎疾患の検索にて胸骨背面の腫瘤を認め生検を予定したが，新鮮凍結血漿の投与で凝固異常を補正できず断念した。Prednisolone（PSL）1 mg/kgの投与により凝固異常は正常化し，LAと腫瘤も消失した。LAHPSの再発なくPSLを漸減したが，経過観察のCTで多発リンパ節腫大を認め，生検により濾胞性リンパ腫と診断した。LAHPSが再発する可能性を考慮しbendamustine＋rituximab療法を6サイクル行った。濾胞性リンパ腫は完全寛解となりLAHPSの再発も認めない。LAHPSはLA陽性だが出血傾向を来しうる稀な後天性凝固異常である。診断時はリンパ性腫瘍を含む基礎疾患の検索を要する。

DOI： 10.11406/rinketsu.61.745

血小板膜糖蛋白質GPIb変異体を用いたvon Willebrand因子活性測定試薬「INNOVANCE® VWF Ac」の基本性能評価国際誌

CiNii Research

J-GLOBAL

鈴木敦夫, 鈴木伸明, 兼松毅, 岡本修一, 田村彰吾, 篠原翔, 新井信夫, 菊地良介, 安藤善孝, 小嶋哲人, 松下正

日本血栓止血学会誌 31 巻 ( 4 ) 頁： 409 - 419 2020年

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：一般社団法人日本血栓止血学会

INNOVANCE® VWF Acは血小板膜糖蛋白質GPIbの変異体を用いるフォンヴィレブランド因子（von Willebrand factor: VWF）活性（VWF:GPIbM）測定試薬である．本研究では本邦で初めてその基本性能評価を行った．併行精度はCVが3％未満，室内再現性はCVが2％未満と良好な成績であった．希釈直線性も良好であり，最小検出感度は1.3 IU/dLであった．また，干渉物質や未分画ヘパリン混入による明らかな影響は見られなかった．正常検体100例を用いた相関性試験ではVWF抗原量およびVWFリストセチンコファクター活性（VWF:RCo）と良好な相関性を示した．フォンヴィレブランド病患者検体においてもVWF:RCoとの測定値に大きな差はなく，VWF:GPIbMでは特に低濃度域を評価することが可能であった．INNOVANCE® VWF AcはVWF:RCo測定試薬と比べても高い性能を有することが明らかとなった．

DOI： 10.2491/jjsth.31.409

CiNii Research

Ischemic events are rare among aging patients with hemophilia: Results of a cross-sectional study from at centers treating hemophiliacs in Japan 査読有り国際誌

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

HAEMOPHILIA 24 巻頁： 13 - 14 2018年5月

病気のはなしハイド症候群査読有り国際誌

兼松毅, 松下正

検査と技術 42 巻 ( 11 ) 頁： 1188 - 1192 2014年10月

PDPN/CLEC-2 axis modulates megakaryocyte subtypes in a hematopoietic stem cell-regulating megakaryocyte-dominant manner 査読有り国際誌

Nara, R; Notoh, H; Sasaki, T; Tsukiji, N; Shirai, T; Kamata, A; Suzuki, N; Suzuki, A; Okamoto, S; Kanematsu, T; Suzuki, N; Katsumi, A; Kojima, T; Suzuki-Inoue, K; Matsushita, T; Tamura, S

THROMBOSIS RESEARCH 245 巻頁： 109230 - 109230 2025年1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: Megakaryocytes are classified into several subtypes including LSP1-positive immune-skewed, MYLK4-positive hematopoietic stem cell (HSC)-regulating, and BMAL1-positive platelet-producing megakaryocytes. Podoplanin (PDPN)-expressing stromal cells generate a microenvironment that promotes megakaryopoiesis in the bone marrow. In this context, PDPN interacts with C-type lectin-like receptor-2 (CLEC-2) on megakaryocyte progenitors, which induces megakaryocyte proliferation. However, the megakaryocyte subtypes developed by the regulation of the PDPN/CLEC-2 axis have not yet been elucidated. Materials and methods: We established an immortalized bone marrow PDPN-expressing stromal cell line and a PDPN-knockout line (PDPN WT and KO feeder cells, respectively). Bone marrow hematopoietic progenitors were committed to megakaryocytes in co-culture with PDPN WT or KO feeder cells. The number and ploidy of megakaryocytes, resultant platelets, and the polarization of megakaryocyte subtypes were investigated. Results: The number of megakaryocytes was significantly increased in the co-culture with PDPN WT feeder cells compared to that with PDPN KO feeder cells. The megakaryocytes on the PDPN WT and KO feeders showed their main ploidy at 16 N∼32 N and 8 N∼16 N, respectively. The number of platelets was decreased in the co-culture with the PDPN WT feeder compared to that in the co-culture with the PDPN KO feeder. For each megakaryocyte subtype, the percentage of MYLK4-positive megakaryocytes significantly increased and the percentage of BMAL1-positive megakaryocytes significantly decreased when co-cultured with the PDPN WT feeder. These results were also confirmed in the co-culture of CLEC-2 conditional KO megakaryocytes with PDPN WT feeder cells. Conclusion: The PDPN/CLEC-2 axis modulates megakaryocyte subtype differentiation, with a predominance of HSC-regulating megakaryocytes.

DOI： 10.1016/j.thromres.2024.109230

Misleading antigenic von Willebrand factor levels in acquired von Willebrand syndrome secondary to monoclonal gammopathy of undetermined significance 査読有り

Okamoto, S; Suzuki, A; Tamura, S; Suzuki, N; Kanematsu, T; Suzuki, N; Kawaguchi, Y; Katsumi, A; Hayakawa, F; Kiyoi, H; Kojima, T; Matsushita, T

INTERNATIONAL JOURNAL OF HEMATOLOGY 121 巻 ( 1 ) 頁： 131 - 136 2025年1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

In the diagnosis and treatment of acquired von Willebrand syndrome (AVWS), von Willebrand factor (VWF) antigen levels (VWF:Ag) are helpful for quantifying blood VWF-protein levels. Most clinical laboratories measure VWF:Ag by latex immunoassay (LIA), but underlying diseases of AVWS may influence LIA results. A 60 year-old AVWS patient with immunoglobulin G (IgG) kappa-type monoclonal gammopathy of undetermined significance (MGUS) showed reduced VWF activity but normal levels of VWF:Ag. His VWF multimers were broadly decreased, which represented a large discrepancy with VWF:Ag. To investigate the mechanism of this discrepancy, we measured the patient’s plasma VWF:Ag by in-house enzyme-linked immunosorbent assay (ELISA) and LIA. We also purified the IgG fraction from the patient’s serum and measured VWF:Ag in VWF-deficient plasma supplemented with this fraction. VWF:Ag measured by in-house ELISA (VWF:AgELISA) was much lower than that measured by LIA (VWF:AgLIA), which indicated reduced VWF-protein volume in blood. Indeed, VWF:Ag was detected by LIA in VWF-deficient plasma spiked with a patient-derived IgG fraction. These results suggest that LIA detected a non-specific immunoreaction and overestimated the patient’s VWF:AgLIA. Clinicians should be aware that underlying diseases of AVWS could influence the LIA system, and interpret VWF:Ag cautiously.

DOI： 10.1007/s12185-024-03861-6

Prevalence Estimation of Dysfibrinogenemia Using the Clauss-CWA Approach 査読有り国際誌

Suzuki, A; Suzuki, N; Okamoto, S; Tamura, S; Kanematsu, T; Kikuchi, R; Kojima, T; Matsushita, T

INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY 2024年12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 Laboratory Hematology

Introduction: The actual prevalence of the qualitative fibrinogen abnormalities dysfibrinogenemia and hypodysfibrinogenemia is unknown. The major reasons are that patients with dysfibrinogenemia are frequently asymptomatic, and a recommended screening test, the Clauss fibrinogen assay, cannot completely distinguish qualitative from quantitative abnormalities. We previously established a high-throughput screening test (Clauss-CWA) to identify dysfibrinogenemia with high specificity and sensitivity by the Clauss fibrinogen assay alone. Aim and Methods: This was a single-center, observational study to estimate the prevalence of dysfibrinogenemia using Clauss-CWA technology. A total of 25 471 patients in Nagoya University Hospital were screened to identify patients with suspected dysfibrinogenemia. The suspected patients were investigated by further confirmatory analyses, such as antigenic fibrinogen determination, reptilase time, fibrin polymerization analysis, and scanning electron microscopy. Results and Conclusions: Of the 25 471 enrolled patients, five with suspected dysfibrinogenemia were identified. Unfortunately, one patient was not confirmed due to a lack of plasma samples. The ratio of functional to antigenic fibrinogen was decreased, and the reptilase time was prolonged in the four patients. Interestingly, two of them showed normal functional fibrinogen levels due to acute inflammatory responses. Fibrin polymerization was impaired, and structural abnormalities were found in the fibrinogen from the patients. In some cases, functional fibrinogen levels may not be effective for identifying functional fibrinogen abnormalities. Further nationwide studies are needed to more precisely understand the epidemiology of dysfibrinogenemia.

DOI： 10.1111/ijlh.14408

Newly diagnosed multiple myeloma with bleeding and coagulation abnormalities caused by a thrombin-inhibiting substance 査読有り国際誌

Ohara, F; Suzuki, A; Suzuki, N; Kanematsu, T; Okamoto, S; Tamura, S; Kiyoi, H; Matsushita, T; Katsumi, A

INTERNATIONAL JOURNAL OF HEMATOLOGY 120 巻 ( 6 ) 頁： 743 - 749 2024年12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

Approximately 7% of patients with newly diagnosed multiple myeloma (MM) experience bleeding complications with varying causes, but few reports have described these complications. Here we report the case of a patient with newly diagnosed MM who presented with a bleeding tendency and various coagulation abnormalities. Chromogenic assays, thrombin time, and reptilase time revealed the presence of a thrombin-inhibiting substance that inhibited release of fibrinopeptide A from fibrinogen. The coagulation abnormalities improved after treatment with daratumumab, lenalidomide, and dexamethasone. As the thrombin inhibition mechanism remains unclear, no previous studies have reported recent treatment outcomes in older patients producing thrombin-inhibiting substances, which can hinder clinical treatment. Therefore, we believe that the diagnosis and the treatment course of this case provide valuable information. Moreover, such case reports provide significant insights into the pathophysiology of bleeding complications associated with MM.

DOI： 10.1007/s12185-024-03849-2

その他リンク： https://link.springer.com/article/10.1007/s12185-024-03849-2/fulltext.html

Variability in combinations of APTT reagent and substrate plasma for a one-stage clotting assay to measure factor VIII products 査読有り国際誌

Suzuki, A; Suzuki, N; Kanematsu, T; Okamoto, S; Suzuki, N; Tamura, S; Kikuchi, R; Katsumi, A; Kojima, T; Matsushita, T

INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY 46 巻 ( 4 ) 頁： 705 - 713 2024年8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 Laboratory Hematology

Introduction: An investigation of the suitability of reagents for measuring FVIII products in a one-stage clotting assay (OSA) showed variations in their FVIII activity (FVIII:C). Most studies have focused on the activated partial thromboplastin time (APTT) reagent rather than FVIII-deficient plasma (F8DP), even though the APTT-based OSA is comprised of APTT reagents and factor-deficient plasma. Aim: A single-centre study was conducted to clarify variations in measurements of FVIII products in an OSA using a total of 12 reagent combinations, including four APTT reagents and three types of F8DP. Methods: FVIII:C in nine types of FVIII product-spiked plasma was measured using an OSA with four different APTT reagents and three types of F8DP. Results: F8DP-dependent variations were found in addition to differences derived from APTT reagents. Variations in target recovery (TR) were observed for NovoEight®, Eloctate®, and Jivi®. Reduced TR for Jivi was found only for Pathromtin SL in combination with congenital F8DP (F8DP-3). This lower TR was not observed with alternative manufacturing lots of F8DP-3. The reduced TR for Jivi might be related to impaired contact activation due to lower factor XI activity in F8DP-3. Conclusion: In addition to APTT reagents, variations in F8DPs used for OSAs can also affect FVIII:C results. F8DPs as well as the APTT reagent used for OSA should be chosen with caution, and laboratories should evaluate reagents for F8DPs as they currently do for APTT reagents, especially when lot changes occur.

DOI： 10.1111/ijlh.14258

盲腸がん発症を契機に偶然診断された高齢者先天性第V因子欠乏症の1例査読有り国際誌

横田裕史, 兼松毅, 鈴木奈瑠子, 岡本修一, 鈴木敦夫, 田村彰吾, 鈴木伸明, 早川文彦, 清井仁, 松下正

臨床血液 65 巻 ( 8 ) 頁： 842 - 842 2024年8月

Clauss-CWA法による鑑別を逃避する異常フィブリノゲン血症 Fibrinogen Milano VII 査読有り国際誌

鈴木敦夫, 鈴木伸明, 田村彰吾, 岡本修一, 兼松毅, 小嶋哲人, 松下正

日本検査血液学会雑誌 25 巻 ( 学術集会 ) 頁： S152 - S152 2024年6月

異常フィブリノゲン血症の有病率推定を目的とした単施設pilot study 査読有り国際誌

鈴木敦夫, 鈴木伸明, 兼松毅, 鈴木奈瑠子, 岡本修一, 田村彰吾, 小嶋哲人, 松下正

日本血栓止血学会誌 35 巻 ( 2 ) 頁： 328 - 328 2024年5月

血友病Aにおける半減期延長型製剤による持続輸注の有効性と安全性の検討査読有り国際誌

鈴木奈瑠子, 鈴木伸明, 岡本修一, 兼松毅, 鈴木敦夫, 田村彰吾, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 35 巻 ( 2 ) 頁： 268 - 268 2024年5月

血中VWF抗原量が偽高値を呈したMGUSを背景とするAVWSの1例査読有り国際誌

岡本修一, 鈴木敦夫, 田村彰吾, 鈴木伸明, 兼松毅, 鈴木奈瑠子, 早川文彦, 清井仁, 小嶋哲人, 松下正

日本血栓止血学会誌 35 巻 ( 2 ) 頁： 354 - 354 2024年5月

Basement membrane extract potentiates the endochondral ossification phenotype of bone marrow-derived mesenchymal stem cell-based cartilage organoids 査読有り国際誌

Notoh, H; Yamasaki, S; Suzuki, N; Suzuki, A; Okamoto, S; Kanematsu, T; Suzuki, N; Katsumi, A; Kojima, T; Matsushita, T; Tamura, S

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 701 巻頁： 149583 - 149583 2024年3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：Biochemical and Biophysical Research Communications

Endochondral ossification is a developmental process in the skeletal system and bone marrow of vertebrates. During endochondral ossification, primitive cartilaginous anlages derived from mesenchymal stem cells (MSCs) undergo vascular invasion and ossification. In vitro regeneration of endochondral ossification is beneficial for research on the skeletal system and bone marrow development as well as their clinical aspects. However, to achieve the regeneration of endochondral ossification, a stem cell-based artificial cartilage (cartilage organoid, Cart-Org) that possesses an endochondral ossification phenotype is required. Here, we modified a conventional 3D culture method to create stem cell-based Cart-Org by mixing it with a basement membrane extract (BME) and further characterized its chondrogenic and ossification properties. BME enlarged and matured the bone marrow MSC-based Cart-Orgs without any shape abnormalities. Histological analysis using Alcian blue staining showed that the production of cartilaginous extracellular matrices was enhanced in Cart-Org treated with BME. Transcriptome analysis using RNA sequencing revealed that BME altered the gene expression pattern of Cart-Org to a dominant chondrogenic state. BME triggered the activation of the SMAD pathway and inhibition of the NK-κB pathway, which resulted in the upregulation of SOX9, COL2A1, and ACAN in Cart-Org. BME also facilitated the upregulation of genes associated with hypertrophic chondrocytes (IHH, PTH1R, and COL10A1) and ossification (SP7, ALPL, and MMP13). Our findings indicate that BME promotes cartilaginous maturation and further ossification of bone marrow MSC-based Cart-Org, suggesting that Cart-Org treated with BME possesses the phenotype of endochondral ossification.

DOI： 10.1016/j.bbrc.2024.149583

Dアイマーモニタリングによる血栓ハイリスク妊婦に対する抗血栓療法査読有り国際誌

川口裕佳, 鈴木伸明, 鈴木奈瑠子, 岡本修一, 兼松毅, 鈴木敦夫, 田村彰吾, 小嶋哲人, 牛田貴文, 今井健史, 小谷友美, 清井仁, 松下正

日本血液学会学術集会 85回巻頁： 1370 - 1370 2023年10月

VWF欠損患者における血管新生能の異常査読有り国際誌

岡本修一, 鈴木伸明, 田村彰吾, 鈴木敦夫, 兼松毅, 鈴木奈瑠子, 川口裕佳, 早川文彦, 清井仁, 小嶋哲人, 松下正

日本血液学会学術集会 85回巻頁： 57 - 57 2023年10月

周術期の止血に難渋した先天性第V因子欠乏症の1例査読有り国際誌

兼松毅, 鈴木伸明, 鈴木奈瑠子, 岡本修一, 鈴木敦夫, 田村彰吾, 早川文彦, 小嶋哲人, 清井仁, 松下正

日本血液学会学術集会 85回巻頁： 1548 - 1548 2023年10月

Clauss-CWA法による異常フィブリノゲン血症スクリーニング 1年間の単施設前向き観察研究査読有り国際誌

鈴木敦夫, 鈴木伸明, 兼松毅, 鈴木奈瑠子, 岡本修一, 田村彰吾, 小嶋哲人, 松下正

日本検査血液学会雑誌 24 巻 ( 学術集会 ) 頁： S157 - S157 2023年7月

Dダイマーモニタリングによる血栓ハイリスク妊婦に対する抗血栓療法当院における35症例査読有り国際誌

川口裕佳, 鈴木伸明, 鈴木奈瑠子, 岡本修一, 兼松毅, 鈴木敦夫, 田村彰吾, 小嶋哲人, 牛田貴文, 今井健史, 小谷友美, 清井仁, 松下正

日本血栓止血学会誌 34 巻 ( 2 ) 頁： 196 - 196 2023年5月

フィブリノゲン製剤の定期補充療法が有効であった先天性無フィブリノゲン血症の1例査読有り国際誌

鈴木奈瑠子, 鈴木伸明, 鈴木敦夫, 田村彰吾, 川口裕佳, 岡本修一, 兼松毅, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 34 巻 ( 2 ) 頁： 251 - 251 2023年5月

マウス下大静脈結紮モデルを用いたアンチトロンビンレジスタンスに対する抗血栓療法の検討査読有り国際誌

徳丸星奈, 鈴木伸明, 田村彰吾, 鈴木敦夫, 兼松毅, 岡本修一, 鈴木奈瑠子, 川口裕佳, 小嶋哲人, 早川文彦, 松下正

日本血栓止血学会誌 34 巻 ( 2 ) 頁： 258 - 258 2023年5月

患者由来endothelial colony forming cell(ECFC)を用いたVWF欠損状態における血管新生能の評価査読有り国際誌

岡本修一, 鈴木伸明, 田村彰吾, 兼松毅, 鈴木敦夫, 鈴木奈瑠子, 川口裕佳, 清井仁, 小嶋哲人, 松下正, 早川文彦

日本血栓止血学会誌 34 巻 ( 2 ) 頁： 275 - 275 2023年5月

血友病患者における抗血小板療法の現状当施設での経験査読有り国際誌

兼松毅, 鈴木伸明, 川口裕佳, 鈴木奈瑠子, 岡本修一, 鈴木敦夫, 田村彰吾, 早川文彦, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 34 巻 ( 2 ) 頁： 266 - 266 2023年5月

VWF-Gly2752Ser, a novel non-cysteine substitution variant in the CK domain, exhibits severe secretory impairment by hampering C-terminal dimer formation 査読有り国際誌

Okamoto, S; Tamura, S; Sanda, N; Odaira, K; Hayakawa, Y; Mukaide, M; Suzuki, A; Kanematsu, T; Hayakawa, F; Katsumi, A; Kiyoi, H; Kojima, T; Matsushita, T; Suzuki, N

JOURNAL OF THROMBOSIS AND HAEMOSTASIS 20 巻 ( 8 ) 頁： 1784 - 1796 2022年8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：Journal of Thrombosis and Haemostasis

Background: Von Willebrand factor (VWF) is a multimeric glycoprotein that plays important roles in hemostasis and thrombosis. C-terminal interchain-disulfide bonds in the cystine knot (CK) domain are essential for VWF dimerization. Previous studies have reported that missense variants of cysteine in the CK domain disrupt the intrachain-disulfide bond and cause type 3 von Willebrand disease (VWD). However, type 3 VWD-associated noncysteine substitution variants in the CK domain have not been reported. Objective: To investigate the molecular mechanism of a novel non-cysteine variant in the CK domain, VWF c.8254 G>A (p.Gly2752Ser), which was identified in a patient with type 3 VWD as homozygous. Methods: Genetic analysis was performed by whole exome sequencing and Sanger sequencing. VWF multimer analysis was performed using SDS-agarose electrophoresis. VWF production and subcellular localization were analyzed using ex vivo endothelial colony forming cells (ECFCs) and an in vitro recombinant VWF (rVWF) expression system. Results: The patient was homozygous for VWF-Gly2752Ser. Plasma VWF enzyme-linked immunosorbent assay showed that the VWF antigen level of the patient was 1.2% compared with healthy subjects. A tiny amount of VWF was identified in the patient's ECFC. Multimer analysis revealed that the circulating VWF-Gly2752Ser presented only low molecular weight multimers. Subcellular localization analysis of VWF-Gly2752Ser-transfected cell lines showed that rVWF-Gly2752Ser was severely impaired in its ER-to-Golgi trafficking. Conclusion: VWF-Gly2752Ser causes severe secretory impairment because of its dimerization failure. This is the first report of a VWF variant with a noncysteine substitution in the CK domain that causes type 3 VWD.

DOI： 10.1111/jth.15746

CN-6000に搭載したフィブリノゲン異常症検出ソフトウェアのバリデーションと日常検査における活用査読有り国際誌

鈴木敦夫, 鈴木伸明, 兼松毅, 岡本修一, 鈴木奈瑠子, 田村彰吾, 松下正

日本検査血液学会雑誌 23 巻 ( 学術集会 ) 頁： S145 - S145 2022年7月

血液凝固第IX因子濃縮製剤へのアレルギーに対する減感作療法の有効性査読有り国際誌

鈴木伸明, 兼松毅, 岸本磨由子, 鈴木奈瑠子, 岡本修一, 田村彰吾, 清井仁, 松下正

日本輸血細胞治療学会誌 68 巻 ( 3 ) 頁： 422 - 427 2022年6月

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：一般社団法人日本輸血・細胞治療学会

血液凝固第IX因子（FIX）濃縮製剤に対する重度なアレルギーに対し，減感作療法が有効であった血友病B 2例を報告する．【症例1】9カ月男児．重症．遺伝子組換え血液凝固第IX因子（rFIX）製剤の定期補充開始後，10回投与目にアナフィラキシーを発症．【症例2】55歳男性．中等症．出血症状に対して，通算4回目の血漿由来血液凝固第IX因子濃縮製剤を投与したところ，アナフィラキシーを発症．以上の2症例に対し，rFIXによる減感作療法を実施した．【結果】2症例とも減感作療法により，FIXに対するアレルギーが消失した．減感作療法中はアレルギー反応を始めとする有害事象を認めなかった．【考察】症例2は中等症であり，アレルギーの発症リスクが低い症例であったが，アナフィラキシーを発症し，中等症でも注意が必要であると考えられた．2症例共にアナフィラキシー発症前後に感度以下の弱力価インヒビターの存在が疑われたことから，インヒビターの発症に注意することはアナフィラキシーの発症予測につながる可能性が示唆された．減感作療法の治療プロトコールについては最適化されておらず，更なる検討が必要であると考えられた．

DOI： 10.3925/jjtc.68.422

CiNii Research

F9mRNA splicing aberration due to a deep Intronic structural variation in a patient with moderate hemophilia B 査読有り国際誌

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

THROMBOSIS RESEARCH 213 巻頁： 91 - 96 2022年5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 (HB) is a hereditary bleeding disorder caused by the genetic variation of the coagulation factor IX (FIX) gene (F9). Several F9 structural abnormalities, including large deletion and/or insertion, have been observed to cause HB development. However, there is limited information available on F9 deep intronic variations. In this study, we report about a novel large deletion/insertion observed in a deep region of F9 intron 1 that causes mRNA splicing abnormalities. Patient and methods: The patient was a Japanese male diagnosed with moderate HB (FIX:C = 3.0 IU/dL). The genomic DNA of the patient was isolated from peripheral blood leukocytes. DNA sequences of F9 exons and splice donor/acceptor sites were analyzed via polymerase chain reaction and Sanger sequencing. Variant-affected F9 mRNA aberration and FIX protein production, secretion, and coagulant activity were analyzed by cell-based exon trap and splicing-competent FIX expression vector systems. Results: A 28-bp deletion/476-bp insertion was identified in the F9 intron 1 of a patient with moderate HB. A DNA sequence identical to a part of the inverted HNRNPA1 exon 12 was inserted. Cell-based transcript analysis revealed that this large intronic deletion/insertion disrupted F9 mRNA splicing pattern, resulting in reduction of protein-coding F9 mRNA. Conclusion: A novel deep intronic F9 rearrangement was identified in a Japanese patient with moderate HB. Abnormal F9 mRNA splicing pattern due to this deep intronic structural variation resulted in a reduction of protein-coding F9 mRNA, which probably caused the moderate HB phenotype.

DOI： 10.1016/j.thromres.2022.03.010

Periosteum-derived podoplanin-expressing stromal cells regulate nascent vascularization during epiphyseal marrow development 査読有り国際誌

Tamura, S; Mukaide, M; Katsuragi, Y; Fujii, W; Odaira, K; Suzuki, N; Tsukiji, N; Okamoto, S; Suzuki, A; Kanematsu, T; Katsumi, A; Takagi, A; Ikeda, K; Ueyama, J; Hirayama, M; Suzuki-Inoue, K; Matsushita, T; Kojima, T; Hayakawa, F

JOURNAL OF BIOLOGICAL CHEMISTRY 298 巻 ( 5 ) 頁： 101833 - 101833 2022年5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：Journal of Biological Chemistry

Bone marrow development and endochondral bone formation occur simultaneously. During endochondral ossification, periosteal vasculatures and stromal progenitors invade the primary avascular cartilaginous anlage, which induces primitive marrow development. We previously determined that bone marrow podoplanin (PDPN)-expressing stromal cells exist in the perivascular microenvironment and promote megakaryopoiesis and erythropoiesis. In this study, we aimed to examine the involvement of PDPN-expressing stromal cells in postnatal bone marrow generation. Using histological analysis, we observed that periosteum-derived PDPN-expressing stromal cells infiltrated the cartilaginous anlage of the postnatal epiphysis and populated on the primitive vasculature of secondary ossification center. Furthermore, immunophenotyping and cellular characteristic analyses indicated that the PDPN-expressing stromal cells constituted a subpopulation of the skeletal stem cell lineage. In vitro xenovascular model cocultured with human umbilical vein endothelial cells and PDPN-expressing skeletal stem cell progenies showed that PDPN-expressing stromal cells maintained vascular integrity via the release of angiogenic factors and vascular basement membrane-related extracellular matrices. We show that in this process, Notch signal activation committed the PDPN-expressing stromal cells into a dominant state with basement membrane-related extracellular matrices, especially type IV collagens. Our findings suggest that the PDPN-expressing stromal cells regulate the integrity of the primitive vasculatures in the epiphyseal nascent marrow. To the best of our knowledge, this is the first study to comprehensively examine how PDPN-expressing stromal cells contribute to marrow development and homeostasis.

DOI： 10.1016/j.jbc.2022.101833

Multimerの経時的変化が止血効果の指標となり得た後天性von Willebrand症候群の1例査読有り国際誌

岡本修一, 鈴木伸明, 兼松毅, 鈴木奈瑠子, 鈴木敦夫, 清井仁, 松下正

日本血栓止血学会誌 33 巻 ( 2 ) 頁： 264 - 264 2022年5月

凝固波形解析を用いたフィブリノゲン質的分析法の自動解析ソフトウェア構築とバリデーション査読有り国際誌

鈴木敦夫, 鈴木伸明, 篠原翔, 黒野浩司, 新井信夫, 兼松毅, 岡本修一, 鈴木奈瑠子, 田村彰吾, 小嶋哲人, 松下正

日本血栓止血学会誌 33 巻 ( 2 ) 頁： 273 - 273 2022年5月

先天性アンチトロンビン欠乏症に同定した新規SERPINC1変異(AT-I218F)の分子病態解析査読有り国際誌

藤井渉, 田村彰吾, 鈴木伸明, 向出将人, 桂木裕実, 鈴木敦夫, 兼松毅, 岡本修一, 鈴木奈瑠子, 勝見章, 小嶋哲人, 松下正, 早川文彦

日本血栓止血学会誌 33 巻 ( 2 ) 頁： 240 - 240 2022年5月

Endothelial colony forming cells(ECFCs)の安定した樹立を目指して査読有り国際誌

岡本修一, 鈴木伸明, 田村彰吾, 鈴木敦夫, 大平晃也, 兼松毅, 鈴木奈瑠子, 清井仁, 松下正

日本輸血細胞治療学会誌 68 巻 ( 2 ) 頁： 376 - 376 2022年4月

フィブリノゲン製剤の定期補充療法が有効であった先天性無フィブリノゲン血症の1例査読有り国際誌

鈴木奈瑠子, 鈴木伸明, 鈴木敦夫, 田村彰吾, 荒木芳生, 泉孝嗣, 岡本修一, 兼松毅, 松下正

日本輸血細胞治療学会誌 68 巻 ( 2 ) 頁： 307 - 307 2022年4月

フィブリノゲンの新しい質的分析法の構築とバリデーション査読有り国際誌

鈴木敦夫, 鈴木伸明, 篠原翔, 黒野浩司, 新井信夫, 兼松毅, 岡本修一, 鈴木奈瑠子, 田村彰吾, 加藤千秋, 小嶋哲人, 松下正

日本輸血細胞治療学会誌 68 巻 ( 2 ) 頁： 308 - 308 2022年4月

異常フィブリノゲン血症の臨床症状と分子遺伝学的特徴の関連査読有り国際誌

田村彰吾, 向出将人, 鈴木伸明, 藤井渉, 桂木裕実, 鈴木敦夫, 兼松毅, 岡本修一, 鈴木奈瑠子, 松下正, 小嶋哲人, 早川文彦

日本輸血細胞治療学会誌 68 巻 ( 2 ) 頁： 333 - 333 2022年4月

Protein S-Leu17Pro disrupts the hydrophobicity of its signal peptide causing a proteasome-dependent degradation 査読有り国際誌

Okada, K; Tamura, S; Suzuki, N; Odaira, K; Mukaide, M; Fujii, W; Katsuragi, Y; Suzuki, A; Kanematsu, T; Okamoto, S; Suzuki, N; Katsumi, A; Matsushita, T; Kojima, T; Hayakawa, F

THROMBOSIS RESEARCH 210 巻頁： 26 - 32 2022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

Introduction: Protein S is a vitamin K-dependent glycoprotein with important anticoagulant, fibrinolytic, anti-inflammatory, anti-apoptotic, and cytoprotective functions. Congenital protein S deficiency is an autosomal dominant thrombophilia due to protein S gene (PROS1) variations. Our group identified a variation in PROS1 that translates into protein S deficiency: c.50 T > C (p.Leu17Pro). Here, we investigated the mechanisms by which this variation results in protein S deficiency. Materials and methods: The effect of L17P substitution on protein S signal peptide was predicted by in silico (a computational prediction technique) analysis of hydrophobicity and signal peptide cleavage. Recombinant protein S was overexpressed in HEK293 and COS-7 cells. Intracellular kinetics and extracellular secretion of recombinant protein S-L17P were analyzed by western blotting and immunocytochemistry. Results: In silico hydrophobicity analysis showed that protein S-L17P had disrupted hydrophobic status in the h-region of its signal peptide. Under normal culture conditions, recombinant protein S -L17P was not detected in either transfectant cell lysates or medium. Upon treatment with a proteasome inhibitor, recombinant protein S-L17P was clearly detected in the cell lysate, but not in the culture medium. Recombinant protein S-L17P did not undergo post-translational modification with N-glycosylation, suggesting that the nascent polypeptide of recombinant protein S-L17P is not transported to the endoplasmic reticulum lumen, but is mislocalized to the cytosol. Conclusion: PROS1-L17P variation translates into protein S deficiency. Protein S-L17P causes its cytosolic mislocalization resulting in its proteasome-dependent degradation.

DOI： 10.1016/j.thromres.2021.12.014

Development and validation of a novel qualitative test for plasma fibrinogen utilizing clot waveform analysis 査読有り国際誌

Suzuki, A; Suzuki, N; Kanematsu, T; Shinohara, S; Kurono, H; Arai, N; Okamoto, S; Suzuki, N; Tamura, S; Kikuchi, R; Katsumi, A; Kojima, T; Matsushita, T

SCIENTIFIC REPORTS 12 巻 ( 1 ) 頁： 434 2022年1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：Scientific Reports

Plasma fibrinogen is commonly examined by Clauss fibrinogen assay, which cannot distinguish between quantitative and qualitative fibrinogen anomalies. However, our previously reported Clauss fibrinogen assay utilizing clot waveform analysis (Clauss-CWA) provides additional information that contributes to the classification of fibrinogen anomalies. In this study, we adopted the Clauss-CWA method for an autoanalyzer to automatically measure the antigenic estimate (eAg) of fibrinogen in addition to the functional amount (Ac), and to thus provide the Ac/eAg ratio as a qualitative indicator. Performance was validated by receiver operating characteristics (ROC) and precision recall (PR) curve analyses using a patient cohort, consisting of a training cohort (n = 519) and a validation cohort (n = 523), both of which contained cases of congenital (hypo)dysfibrinogenemia as qualitative defects. We obtained an optimal cutoff of 0.65 for Ac/eAg by ROC curve analysis of the training cohort, offering superior sensitivity (> 0.9661) and specificity (1.000). This cutoff was validated in the validation cohort, providing positive predictive value > 0.933 and negative predictive value > 0.998. PR curve analysis also showed that Clauss-CWA provided excellent performance for detecting qualitative fibrinogen anomalies. The Clauss-CWA method may represent a useful approach for detecting qualitative fibrinogen abnormalities in routine laboratory testing.

DOI： 10.1038/s41598-021-04464-5

その他リンク： https://www.nature.com/articles/s41598-021-04464-5

The influence of hepatitis C virus eradication on hepatocarcinogenesis in patients with hemophilia HCC after HCV eradication in hemophilia 査読有り国際誌

Inukai, Y; Imai, N; Yamamoto, K; Ito, T; Ishizu, Y; Honda, T; Okamoto, S; Kanematsu, T; Suzuki, N; Matsushita, T; Ishigami, M; Fujishiro, M

ANNALS OF HEPATOLOGY 27 巻 ( 1 ) 頁： 100545 - 100545 2022年1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：Annals of Hepatology

Introduction and objectives: Hepatitis C virus (HCV) infections in patients with hemophilia lead to the development of hepatocellular carcinoma (HCC) at a relatively younger age than that in patients without hemophilia. Although recent progress in direct-acting-antivirals has facilitated a high rate of sustained virological response (SVR), the clinical influence of HCV eradication in hemophilia patients remains unclear. This study aimed to compare the clinical outcomes of SVR against HCV in patients with and without hemophilia. Patients and methods: The study enrolled 699 patients who achieved SVR after HCV antiviral treatment. Patients were divided into two groups: 78 patients with hemophilia (H group) and 621 patients without hemophilia (NH group). We evaluated patient characteristics, clinical outcomes, and the cumulative incidence of HCC after SVR. Results: Compared with the NH group, patients in the H-group were significantly younger and had a lower hepatic fibrosis score. No difference was found in the incidence of liver-related disease or overall death between the two groups over a mean follow-up period of 7 years. Four patients in the H group and 36 patients in the NH group were diagnosed with HCC after SVR. Multivariate analysis showed that male sex, age, and cirrhosis were significant risk factors for HCC incidence. There was no significant difference in the cumulative incidence of HCC after propensity-score matching adjusting for the risk factors of HCC between the two groups. Conclusion: Hemophilia is not a significant risk factor for hepatocarcinogenesis after SVR against HCV.

DOI： 10.1016/j.aohep.2021.100545

21ヵ月の経過で自然寛解した高力価インヒビター保有自己免疫性第V因子欠乏症(AiF5D)の一例(Spontaneous remission of autoimmune factor V deficiency(AiF5D) with high titer inhibitor) 査読有り国際誌

兼松毅, 鈴木伸明, 岡本修一, 尾崎司, 惣宇利正善, 鈴木敦夫, 田村彰吾, 早川文彦, 小嶋哲人, 清井仁, 一瀬白帝, 松下正

日本血液学会学術集会 83回巻頁： PS - 6 2021年9月

Tolerability of Molecular-targeted Agents for Hepatocellular Carcinoma Treatment in Haemophiliacs 査読有り国際誌

Yamamoto, T; Imai, N; Yamamoto, K; Ito, T; Ishizu, Y; Honda, T; Okamoto, S; Kanematsu, T; Suzuki, N; Matsushita, T; Ishigami, M; Fujishiro, M

ANTICANCER RESEARCH 41 巻 ( 5 ) 頁： 2569 - 2573 2021年5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：Anticancer Research

Background: Hepatocellular carcinoma (HCC) is considered a leading cause of death in patients with haemophilia. Recent advances in the treatment of unresectable HCC with molecular-Targeted agents (MTAs) have led to better clinical outcomes. However, the tolerability of MTAs by haemophilic patients with HCC remains unclear. Aim: This study aimed to compare the tolerability of MTAs in such patients. Patients and Methods: From January 2011 to October 2020, five haemophilic patients with HCC were treated with MTAs. Adverse events were assessed in comparison with 265 non-haemophilic patients with HCC. Results: The prevalence of hand foot skin reaction was not higher in the haemophiliacs than in the non-haemophiliacs, whereas the rate of haemorrhagic events was higher in the haemophiliacs (6.0% versus 40.0%, p=0.037). Conclusion: Haemophiliacs tolerate long-Term MTA use, without the occurrence of life-Threatening complications. However, careful observation and prevention are needed for MTA-related gastrointestinal bleeding in haemophiliacs.

DOI： 10.21873/anticanres.15035

第VIII因子製剤の活性測定における試薬バリエーションを把握するための単施設研究査読有り国際誌

鈴木敦夫, 鈴木伸明, 兼松毅, 岡本修一, 田村彰吾, 小嶋哲人, 松下正

日本血栓止血学会誌 32 巻 ( 2 ) 頁： 198 - 198 2021年5月

F8 Int22h-1が関与するF8大規模欠失挿入変異による重症血友病A症例査読有り国際誌

大平晃也, 早川友梨, 田村彰吾, 鈴木伸明, 所真昼, 河島史華, 向出将人, 岡田健太郎, 桂木裕実, 藤井渉, 岡本修一, 兼松毅, 鈴木敦夫, 松下正, 小嶋哲人, 早川文彦

日本血栓止血学会誌 32 巻 ( 2 ) 頁： 228 - 228 2021年5月

周術期の止血に難渋した先天性第V因子欠乏症の1例査読有り国際誌

兼松毅, 鈴木伸明, 岡本修一, 鈴木敦夫, 田村彰吾, 早川文彦, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 32 巻 ( 2 ) 頁： 232 - 232 2021年5月

マウス血管結紮モデルによるアンチトロンビンレジスタンスの血栓形成解析査読有り国際誌

鈴木伸明, 鈴木敦夫, 田村彰吾, 向出将人, 岡本修一, 兼松毅, 小嶋哲人, 松下正

日本血栓止血学会誌 32 巻 ( 2 ) 頁： 202 - 202 2021年5月

プロテインS欠乏症に同定したシグナルペプチド内変異プロテインSの分子病態解析査読有り国際誌

岡田健太郎, 田村彰吾, 鈴木伸明, 大平晃也, 早川友梨, 向出将人, 藤井渉, 桂木裕実, 兼松毅, 岡本修一, 鈴木敦夫, 松下正, 小嶋哲人, 早川文彦

日本血栓止血学会誌 32 巻 ( 2 ) 頁： 206 - 206 2021年5月

フィブリノゲン強制発現系の最適化に向けた基礎検討査読有り国際誌

向出将人, 田村彰吾, 鈴木伸明, 大平晃也, 早川友梨, 岡田健太郎, 藤井渉, 桂木裕実, 鈴木敦夫, 兼松毅, 岡本修一, 松下正, 小嶋哲人, 早川文彦

日本血栓止血学会誌 32 巻 ( 2 ) 頁： 228 - 228 2021年5月

VWF G2752S変異体の分子病態解析査読有り国際誌

岡本修一, 大平晃也, 早川友梨, 河島史華, 田村彰吾, 鈴木伸明, 兼松毅, 鈴木敦夫, 早川文彦, 清井仁, 小嶋哲人, 松下正

日本血栓止血学会誌 32 巻 ( 2 ) 頁： 248 - 248 2021年5月

Essential role of a carboxyl-terminal α-helix motif in the secretion of coagulation factor XI 査読有り国際誌

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

JOURNAL OF THROMBOSIS AND HAEMOSTASIS 19 巻 ( 4 ) 頁： 920 - 930 2021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：Journal of Thrombosis and Haemostasis

Background: Coagulation factor XI (FXI) is a plasma serine protease zymogen that contributes to hemostasis. However, the mechanism of its secretion remains unclear. Objective: To determine the molecular mechanism of FXI secretion by characterizing a novel FXI mutant identified in a FXI-deficient Japanese patient. Patient/Methods: The FXI gene (F11) was analyzed by direct sequencing. Mutant recombinant FXI (rFXI) was overexpressed in HEK293 or COS-7 cells. Western blotting and enzyme-linked immunosorbent assay were performed to examine the FXI extracellular secretion profile. Immunofluorescence microscopy was used to investigate the subcellular localization of the rFXI mutant. Results: We identified a novel homozygous frameshift mutation in F11 [c.1788dupC (p.E597Rfs*65)], resulting in a unique and extended carboxyl-terminal (C-terminal) structure in FXI. Although rFXI-E597Rfs*65 was intracellularly synthesized, its extracellular secretion was markedly reduced. Subcellular localization analysis revealed that rFXI-E597Rfs*65 was abnormally retained in the endoplasmic reticulum (ER). We generated a series of C-terminal–truncated rFXI mutants to further investigate the role of the C-terminal region in FXI secretion. Serial rFXI experiments revealed that a threonine at position 622, the fourth residue from the C-terminus, was essential for secretion. Notably, Thr622 engages in the formation of an α-helix motif, indicating the importance of the C-terminal α-helix in FXI intracellular behavior and secretion. Conclusion: FXI E597Rfs*65 results in the pathogenesis of a severe secretory defect resulting from aberrant ER-to-Golgi trafficking caused by the lack of a C-terminal α-helix motif. This study demonstrates the impact of the C-terminal structure, especially the α-helix motif, on FXI secretion.

DOI： 10.1111/jth.15242

Impact of variation in reagent combinations for one-stage clotting assay on assay discrepancy in nonsevere haemophilia A 査読有り国際誌

Suzuki, A; Suzuki, N; Kanematsu, T; Okamoto, S; Tamura, S; Kikuchi, R; Katsumi, A; Kiyoi, H; Kojima, T; Matsushita, T

INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY 43 巻 ( 1 ) 頁： 131 - 138 2021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：International Journal of Laboratory Hematology

Introduction: Factor VIII activity (FVIII:C) is measured by one-stage clotting assay (OSA) or chromogenic substrate assay (CSA). Significant differences in FVIII:C between OSA (FVIII:C1st) and CSA (FVIII:CChr) are described as assay discrepancy in nonsevere haemophilia A (HA). A large number of reagent combinations (APTT reagent and FVIII-deficient plasma) are used for OSA, but the impact of variations in reagent combinations on assay discrepancy has not been fully characterized. Aim: To clarify the variations in FVIII:C1st/FVIII:CChr ratios according to OSA reagent combination in HA subjects with/without assay discrepancy. Methods: Thirty-nine patients previously diagnosed with nonsevere HA were enrolled, and their FVIII genes were investigated and FVIII:C levels were assessed by a single CSA reagent and 11 OSA reagent combinations. Receiver operating characteristic (ROC) curve analysis was used to predict possible cut-off values of the FVIII:C1st/FVIII:CChr ratio to define FVIII assay discrepancy for each reagent combination. Results: Patients were categorized into nondiscrepant (n = 25), discrepant (n = 5) and unclassified (n = 9) groups according to their genotypes and information in the database. The FVIII:C1st/FVIII:CChr ratio in nondiscrepant HA varied widely, depending on the APTT reagents and FVIII-deficient plasma used. The ratio in discrepant HA patients differed with respect to their genotype and the reagent combination used. ROC curve analyses revealed that cut-off values to distinguish the assay discrepancy differed depending on the reagents used, but revealed two novel genotype variants, p.Cys573Gly and p.Gly582Arg, associated with FVIII assay discrepancy. Conclusion: Our findings showed that the FVIII:C1st/FVIII:CChr ratio is dependent on the reagent combination used for OSA.

DOI： 10.1111/ijlh.13335

Myh9 R702C is associated with erythroid abnormality with splenomegaly in mice 査読有り

Kanematsu, T; Suzuki, N; Tamura, S; Suzuki, A; Ishikawa, Y; Katsumi, A; Kiyoi, H; Saito, H; Kunishima, S; Kojima, T; Matsushita, T

NAGOYA JOURNAL OF MEDICAL SCIENCE 83 巻 ( 1 ) 頁： 75 - 86 2021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：Nagoya Journal of Medical Science

MYH9 disorders are characterized by giant platelets, thrombocytopenia, and Döhle body-like cytoplasmic inclusion bodies in granulocytes. However, whether these disorders cause any changes in erythroid cells has yet to be determined. This study analyzed the influence of Myh9 R702C, as one of the most commonly detected MYH9 disorders, on erythroid cells in a mouse model. Knock-in mice expressing Myh9 R702C mutation either systemically or specific to hematological cells (R702C and R702C vav1 mice, respectively) were used in this study. Both displayed lower hemoglobin and higher erythropoietin levels than wild-type (WT) mice, along with significant splenomegaly. Flow cytometric analysis revealed erythroblasts present at a higher rate than WT mice in the spleen. However, no obvious abnormalities were seen in erythroid differentiation from megakaryocyte/erythroid progenitor to erythrocyte. Cell culture assay by fetal liver and colony assay also showed normal progression of erythroid differentiation from erythroid burst-forming unit to red blood cell. In conclusion, R702C and R702C vav1 mice displayed erythroid abnormality with splenomegaly. However, erythroid differentiation showed no obvious abnormality. Further research is required to elucidate the underlying mechanisms.

DOI： 10.18999/nagjms.83.1.75

Safety and efficacy of percutaneous radiofrequency ablation for hepatocellular carcinoma patients with haemophilia 査読有り国際誌

Yamamoto, T; Imai, N; Yamamoto, K; Ito, T; Ishizu, Y; Honda, T; Okamoto, S; Kanematsu, T; Suzuki, N; Matsushita, T; Ishigami, M; Fujishiro, M

HAEMOPHILIA 27 巻 ( 1 ) 頁： 100 - 107 2021年1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

Haemophilia is an X-linked inherited bleeding disorder caused by coagulation factor deficiency. Hepatocellular carcinoma (HCC) is a major complication associated with the disease. No study thus far has investigated the safety and efficacy of percutaneous radiofrequency ablation (RFA) for HCC in patients with haemophilia. Aim: This study aimed to evaluate the safety and efficacy of RFA for HCC in haemophilia patients. Methods: From July 2008 to June 2019, 217 patients with HCC underwent 300 RFA sessions. Of these, 18 sessions were performed in ten haemophilia patients (H group) and 282 in 207 non-haemophilia patients (NH group). The patients' characteristics, incidence of haemorrhagic complications and rates of local tumour recurrence were compared between the groups. Results: A majority of the haemophilia patients received clotting factor concentrate replacement therapy before and after RFA treatment, with the aim of reaching a plasma clotting factor level of higher than 60%–80%. Twelve haemorrhagic complications were observed in the NH group (4.2%; 12/282). Major bleeding requiring control procedures was observed in two patients and minor bleeding with careful observation was noted in ten patients. No bleeding complications were observed in the H group (0/18). There were no significant differences in the 5-year local tumour recurrence rates after RFA treatment between the groups (35.0% in the H group and 32.1% in the NH group). Conclusion: RFA could be an effective and a safe method for HCC treatment in patients with haemophilia.

DOI： 10.1111/hae.14220

Aberrant X chromosomal rearrangement through multi-step template switching during sister chromatid formation in a patient with severe hemophilia A 査読有り国際誌

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

MOLECULAR GENETICS & GENOMIC MEDICINE 8 巻 ( 9 ) 頁： e1390 2020年9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：Molecular Genetics and Genomic Medicine

Background: Hemophilia A (HA) is an X-linked recessive bleeding disorder caused by pathogenic variants of the coagulation factor VIII gene (F8). Half of the patients with severe HA have a recurrent inversion in the X chromosome, that is, F8 intron 22 or intron 1 inversion. Here, we characterized an abnormal F8 due to atypical complex X chromosome rearrangements in a Japanese patient with severe HA. Methods: Recurrent F8 inversions were tested with inverse shifting-PCR. The genomic structure was investigated using PCR-based direct sequencing or quantitative PCR. Results: The proband's X chromosome had a 119.5 kb insertion, a reverse duplex of an extragenic sequence on the F8 telomere region into the F8 intron 1 with two breakpoints. The telomeric breakpoint was a joining from the F8 intron 1 to the inverted FUNDC2 via a two-base microhomology, and the centromeric breakpoint was a recombination between F8 intron 1 homologous sequences. The rearrangement mechanism was suggested as a multi-step rearrangement with template switching such as fork stalling and template switching (FoSTeS)/microhomology-mediated break-induced replication (MMBIR) and/or homologous sequence-associated recombination during a sister chromatid formation. Conclusion: We identified the aberrant X chromosome with a split F8 due to a multi-step rearrangement in a patient with severe HA.

DOI： 10.1002/mgg3.1390

その他リンク： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2020&ichushi_jid=J02561&link_issn=&doc_id=20200812220008&doc_link_id=10.2491%2Fjjsth.31.409&url=https%3A%2F%2Fdoi.org%2F10.2491%2Fjjsth.31.409&type=J-STAGE&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00007_3.gif

血小板膜糖蛋白質GPIb変異体を用いたvon Willebrand因子活性測定試薬「INNOVANCE VWF Ac」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, 田村彰吾, 篠原翔, 新井信夫, 菊地良介, 安藤善孝, 小嶋哲人, 松下正

日本血栓止血学会誌 31 巻 ( 4 ) 頁： 409 - 419 2020年8月

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：(一社)日本血栓止血学会

INNOVANCE VWF Acは血小板膜糖蛋白質GPIbの変異体を用いるフォンヴィレブランド因子(von Willebrand factor:VWF)活性(VWF:GPIbM)測定試薬である。本研究では本邦で初めてその基本性能評価を行った。併行精度はCVが3%未満、室内再現性はCVが2%未満と良好な成績であった。希釈直線性も良好であり、最小検出感度は1.3IU/dLであった。また、干渉物質や未分画ヘパリン混入による明らかな影響は見られなかった。正常検体100例を用いた相関性試験ではVWF抗原量およびVWFリストセチンコファクター活性(VWF:RCo)と良好な相関性を示した。フォンヴィレブランド病患者検体においてもVWF:RCoとの測定値に大きな差はなく、VWF:GPIbMでは特に低濃度域を評価することが可能であった。INNOVANCE VWF AcはVWF:RCo測定試薬と比べても高い性能を有することが明らかとなった。(著者抄録)

凝固一段法による第VIII因子活性測定における試薬の組み合わせに関する検討査読有り国際誌

鈴木敦夫, 鈴木伸明, 兼松毅, 岡本修一, 田村彰吾, 安藤善孝, 清井仁, 松下正

日本血栓止血学会誌 31 巻 ( 2 ) 頁： 231 - 231 2020年5月

関節内出血を未発症であるにもかかわらず血友病性関節症を発症した症例の検討査読有り国際誌

鈴木伸明, 高橋伸典, 鈴木敦夫, 田村彰吾, 鈴木幸子, 服部有那, 垣原美沙樹, 小嶋俊久, 小川実加, 兼松毅, 清井仁, 小嶋哲人, 石黒直樹, 松下正

日本血栓止血学会誌 31 巻 ( 2 ) 頁： 233 - 233 2020年5月

C末端伸長変異型Factor XIにおける分泌異常の解析査読有り国際誌

早川友梨, 田村彰吾, 鈴木伸明, 大平晃也, 所真昼, 河島史華, 早川文彦, 高木明, 鈴木敦夫, 岡本修一, 兼松毅, 松下正, 小嶋哲人

日本血栓止血学会誌 31 巻 ( 2 ) 頁： 220 - 220 2020年5月

Endothelial colony forming cells(ECFCs)樹立法の標準化への試み査読有り国際誌

岡本修一, 鈴木伸明, 兼松毅, 鈴木敦夫, 大平晃也, 所真昼, 早川友梨, 田村彰吾, 早川文彦, 清井仁, 小嶋哲人, 松下正

日本血栓止血学会誌 31 巻 ( 2 ) 頁： 284 - 284 2020年5月

当施設における新生児・乳幼児に対するエミシズマブの使用経験査読有り国際誌

兼松毅, 鈴木伸明, 岡本修一, 鈴木敦夫, 田村彰吾, 早川文彦, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 31 巻 ( 2 ) 頁： 248 - 248 2020年5月

Higher FVIII:C measured by chromogenic substrate assay than by one-stage assay is associated with silent hemophilic arthropathy 査読有り国際誌

Ogawa, M; Suzuki, N; Takahashi, N; Tamura, S; Suzuki, A; Suzuki, S; Hattori, Y; Kakihara, M; Kanematsu, T; Kojima, T; Katsumi, A; Hayakawa, F; Kojima, T; Ishiguro, N; Kiyoi, H; Matsushita, T

THROMBOSIS RESEARCH 188 巻頁： 103 - 105 2020年4月

Successful Perioperative Combination of High-Dose FVIII Therapy Followed by Emicizumab in a Patient with Hemophilia A with Inhibitors. 査読有り国際誌

Okamoto S, Suzuki N, Suzuki A, Suzuki S, Tamura S, Suzuki M, Takahashi N, Kojima T, Kanematsu T, Kojima T, Kiyoi H, Ishiguro N, Matsushita T

TH open : companion journal to thrombosis and haemostasis 3 巻 ( 4 ) 頁： e364 - e366 2019年10月

Performance evaluation of Revohem™ FVIII chromogenic and Revohem™ FIX chromogenic in the CS-5100 autoanalyser 査読有り国際誌

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

INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY 41 巻 ( 5 ) 頁： 664 - 670 2019年10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 Laboratory Hematology

Introduction: Chromogenic substrate assay (CSA) reagents Revohem™ FVIII and Revohem™ FIX are now available as in vitro diagnostic reagents for autoanalysers in Japan. In this study, we evaluated the performance of these reagents in the CS-5100 automated coagulation analyser. Methods: We assessed within-run and between-day imprecision, on-board stability and frozen-storage stability of Revohem FVIII and FIX. Sensitivity to lupus anticoagulant (LA) was examined using LA-positive patient plasma. Correlations were analysed using plasma samples from normal individuals and patients with haemophilia A (HA) or B (HB) or von Willebrand disease (VWD). Results: Imprecision was <2% for Revohem FVIII and <6.5% for Revohem FIX. On-board storage of Revohem FVIII resulted in a <10% decrease in FVIII levels from baseline at 24 hours, whereas Revohem FIX showed a >10% decrease at 8 hours. Revohem FVIII showed good stability while frozen for 22 days. Although Revohem FIX showed degradation due to freeze-thawing, a new calibration improved stability up to 22 days. Interference from LA was not observed with Revohem FVIII or FIX. The FVIII CSA-CSA correlation was excellent in normal (r = 0.9924), HA (r = 0.9945) and VWD (r = 0.9914). The FVIII CSA-OSA correlation was good in normal (r = 0.8468) and excellent in HA (r = 0.975) and VWD (r = 0.9936). The FIX CSA-OSA correlation was fair in normal (r = 0.4791) and excellent in HB (r = 0.9501). Conclusion: Revohem FVIII and FIX both showed excellent performance in the CS-5100 analyser. These reagents could be useful in routine laboratory testing for diagnosing and treating haemophilia.

DOI： 10.1111/ijlh.13083

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

Vwf K1362A resulted in failure of protein synthesis in mice

凝固波形解析を利用した新しい異常フィブリノゲン血症検出法査読有り国際誌

鈴木敦夫, 篠原翔, 新井信夫, 兼松毅, 鈴木伸明, 安藤善孝, 松下正

日本検査血液学会雑誌 19 巻 ( 学術集会 ) 頁： S166 - S166 2018年6月

Ischemic events are rare among aging patients with hemophilia: Results of a cross-sectional study from at centers treating hemophiliacs in Japan 査読有り国際誌

Suzuki Nobuaki, Takedani Hideyuki, Yamasaki Naoya, Chikasawa Yushi, Sawada Akihiro, Kanematsu Takeshi, Higasa Satoshi, Amano Kagehiro, Fukutake Katsuyuki, Nagao Azusa

HAEMOPHILIA 24 巻頁： 13-14 2018年5月

凝固一段法と合成基質法による第VIII因子活性測定において乖離を示した血友病A症例査読有り国際誌

鈴木敦夫, 鈴木伸明, 兼松毅, 岸本磨由子, 田村彰吾, 高木明, 川上萌, 梶浦容子, 小嶋哲人, 松下正

日本血栓止血学会誌 29 巻 ( 2 ) 頁： 210 - 210 2018年5月

GPIb変異体を用いたVWF活性とリストセチンコファクター活性の比較検討査読有り国際誌

川上萌, 鈴木敦夫, 鈴木伸明, 兼松毅, 岸本磨由子, 篠原翔, 新井信夫, 梶浦容子, 松下正

日本血栓止血学会誌 29 巻 ( 2 ) 頁： 225 - 225 2018年5月

ループスアンチコアグラント低プロトロンビン血症症候群(LAHPS)の2例査読有り国際誌

兼松毅, 鈴木伸明, 鈴木敦夫, 高木明, 岸本磨由子, 川上萌[村田], 三田直美, 小山大輔, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 29 巻 ( 2 ) 頁： 220 - 220 2018年5月

VWF K1362Aノックインマウスの解析査読有り国際誌

三田直美, 鈴木伸明, 鈴木敦夫, 兼松毅, 岸本磨由子, 高木明, 小嶋哲人, 松下正, 中村栄男

日本血栓止血学会誌 29 巻 ( 2 ) 頁： 224 - 224 2018年5月

血友病Aにおいて関節症の進行抑制には出血抑制よりも高いFVIII活性が必要である査読有り国際誌

鈴木伸明, 鈴木敦夫, 高橋伸典, 田村彰吾, 兼松毅, 高木明, 小嶋俊久, 清井仁, 小嶋哲人, 石黒直樹, 松下正

日本血栓止血学会誌 29 巻 ( 2 ) 頁： 191 - 191 2018年5月

Sanda, N; Suzuki, N; Suzuki, A; Kanematsu, T; Kishimoto, M; Hasuwa, H; Takagi, A; Kojima, T; Matsushita, T; Nakamura, S

INTERNATIONAL JOURNAL OF HEMATOLOGY 107 巻 ( 4 ) 頁： 428 - 435 2018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

Von Willebrand factor (VWF) is synthesized in megakaryocytes and endothelial cells (ECs) and has two main roles: to carry and protect coagulation factor VIII (FVIII) from degradation by forming VWF–FVIII complex; and to mediate platelet adhesion and aggregation at sites of vascular injury. Previous research using the HEK293 cell line revealed that the VWF K1362 mutation interacted directly with platelet glycoprotein Ib (GPIb). Vwf K1362A knock-in (KI) mice were therefore generated to verify the in vivo function of residue 1362 in binding to platelet GPIb. The Cre-loxP system was employed to introduce the Vwf K1362A mutation systemically in mice. In blood coagulation analysis, the VWF antigen (VWF:Ag) of Lys1362Ala KI homozygous (homo) mice was below the sensitivity of detection by enzyme-linked immunosorbent assay. FVIII activities (FVIII:C) were 47.9 ± 0.3 and 3.3 ± 0.3% (K1362A heterozygous (hetero) and K1362A KI homo mice, respectively) compared to wild-type mice. Immunohistochemical staining analysis revealed that VWF protein did not exist in ECs of K1362A KI homo mice. These results indicated that VWF protein synthesis of K1362A was impaired after transcription in mice. K1362 seems to represent a very important position not only for VWF function, but also for VWF synthesis in mice.

DOI： 10.1007/s12185-017-2394-y

Combined deficiency of factors V and VIII by chance coinheritance of parahaemophilia and haemophilia A, but not by mutations of either LMAN1 or MCFD2, in a Japanese family 国際誌

Suzuki, S; Nakamura, Y; Suzuki, N; Yamazaki, T; Takagi, Y; Tamura, S; Takagi, A; Kanematsu, T; Matsushita, T; Kojima, T

HAEMOPHILIA 24 巻 ( 1 ) 頁： e13 - e16 2018年1月

その他の凝固因子濃縮製剤の使い方（フィブリノゲン製剤など）国際誌

兼松毅

日本血栓止血学会誌 29 巻 ( 6 ) 頁： 744-747 - 747 2018年

凝固波形解析による異常フィブリノゲン血症の新規検出法査読有り国際誌

鈴木敦夫, 岸本磨由子, 兼松毅, 小川実加, 鈴木伸明, 松下正

日本血栓止血学会誌 28 巻 ( 2 ) 頁： 212 - 212 2017年4月

合成基質法と凝固一段法による凝固第VIII因子製剤測定値の比較検討査読有り国際誌

鈴木敦夫, 鈴木伸明, 岸本磨由子, 兼松毅, 小川実加, 松下正

日本血栓止血学会誌 28 巻 ( 2 ) 頁： 185 - 185 2017年4月

インヒビター保有von Willebrand病type3の止血治療の検討査読有り国際誌

兼松毅, 鈴木伸明, 小川実加, 岸本磨由子, 村田萌, 三田直美, 鈴木敦夫, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 28 巻 ( 2 ) 頁： 202 - 202 2017年4月

原発性マクログロブリン血症に合併した後天性von Willebrand症候群の1例査読有り国際誌

兼松毅, 鈴木伸明, 小川実加, 岸本磨由子, 村田萌, 三田直美, 鈴木敦夫, 後藤辰徳, 入山智沙子, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 28 巻 ( 2 ) 頁： 201 - 201 2017年4月

第一世代ピルが止血管理に有効だったAcquired von Willebrand syndrome(AvWS)の一例査読有り国際誌

村田萌, 鈴木伸明, 鈴木敦夫, 兼松毅, 岸本磨由子, 小川実加, 清井仁, 松下正

日本血栓止血学会誌 28 巻 ( 2 ) 頁： 258 - 258 2017年4月

血友病患者の出血頻度と相関する因子の検討査読有り国際誌

小川実加, 鈴木伸明, 鈴木敦夫, 垣原美紗樹, 村田萌, 田村彰吾, 兼松毅, 岸本磨由子, 小嶋哲人, 清井仁, 松下正

日本血栓止血学会誌 28 巻 ( 2 ) 頁： 223 - 223 2017年4月

Retrospective analysis of in vivo recovery and clearance during continuous infusion of recombinant factor VIII products: a single-institution study 国際誌

Suzuki, N; Hirakawa, A; Kishimoto, M; Kanematsu, T; Ogawa, M; Kiyoi, H; Matsushita, T

HAEMOPHILIA 23 巻 ( 2 ) 頁： 215 - 221 2017年3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

Background: Continuous infusion (CI) of recombinant FVIII (rFVIII) concentrates has been reported as an effective and safe method to achieve haemostasis during major surgeries or severe bleeding events. For more effective and safer CI, better understanding of in vivo recovery (IVR) and clearance (CL) issues is imperative. Objective: We investigated the following factors affecting IVR and CL using univariate and multivariate regression analyses during 47 CIs in 34 patients: rFVIII concentrate type, haemophilia severity, blood type, the presence of hepatitis C virus (HCV) or human immunodeficiency virus (HIV), age and body mass index (BMI). Results: The mean IVR was 1.64 ± 0.49 IU dL−1 per IU kg−1, and the mean CL during CI was 3.56 ± 1.57 mL h−1 kg−1. The univariate and multivariate regression analyses showed that the CL of octocog alfa was significantly lower than that of rurioctocog alfa (P = 0.043 and 0.0034, respectively). There was a significant difference in BMI in the univariate and multivariate regression analyses (P = 0.0403 and 0.0376, respectively). Conclusions: This study indicated that CL during CI was potentially affected by the type of rFVIII concentrate used and BMI.

DOI： 10.1111/hae.13082