Function as well as regulation on expression of a novel hyaluronidase in cancer and inflammation

• Project/Area Number: 15K07951
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Meijo University
• Principal Investigator: Yamada Shuhei 名城大学, 薬学部, 教授 (70240017)
• Co-Investigator(Renkei-kenkyūsha): MIZUMOTO Shuji 名城大学, 薬学部, 助教 (40443973)
• Research Collaborator: SUGAHARA Kazuyuki 北海道大学院, 生命科学院, 名誉教授 ; BASAPPA バンガロール大学, 助教 ; YIP George W.C. シンガポール国立大学, 准教授
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ヒアルロニダーゼ / コンドロイチン硫酸 / ヒアルロン酸 / 癌

Outline of Final Research Achievements

A novel enzyme hyaluronidase-4 (HYAL4) specifically acts on chondroitin sulfate (CS), and may be involved in cancer metastasis. To investigate the role of HYAL4 in metastasis as well as inflammation, several experiments have been performed. HYAL4 gene was stably transfected into a cancer cell line and HYAL4-overexpressing cell lines have been established. HYAL4 shRNA was also stably transfected into a cell line which highly expresses HYAL4, and the knockdown cells have been prepared. The amount of CS in these cell lines has been determined. Influences of the addition of CS oligosaccharides on cell proliferation as well as cell migration of Lewis lung carcinoma cells, have been evaluated. HYAL4-deficient mice have been generated using a CRISPR/CAS9 system. High level expression of HYAL4 in various cancer tissues has been investigated by real-time PCR.

Research Products

• [Int'l Joint Research] バンガロール大学(インド)
• [Journal Article] Defect in dermatan sulfate in urine of patients with Ehlers-Danlos syndrome caused by a CHST14/D4ST1 deficiency (2018)
  • Author(s): Shuji Mizumoto, Tomoki Kosho, Atsushi Hatamochi, Tomoko Honda, Tomomi Yamaguchi, Nobuhiko Okamoto, Noriko Miyake, Shuhei Yamada, and Kazuyuki Sugahara
  • Journal Title: Clinical Biochemistry Volume: 50 Issue: 12 Pages: 670-677
  • DOI: 10.1016/j.clinbiochem.2017.02.018
  • NAID: 120006398760
  • Peer Reviewed
• [Journal Article] Roles of two types of heparan sulfate clusters in Wnt distribution and signaling in Xenopus (2018)
  • Author(s): Yusuke Mii, Takayoshi Yamamoto, Ritsuko Takada, Shuji Mizumoto, Makoto Matsuyama, Shuhei Yamada, Shinji Takada, and Masanori Taira
  • Journal Title: Nature Communications Volume: 8 Issue: 1 Pages: 1973-1973
  • DOI: 10.1038/s41467-017-02076-0
  • Peer Reviewed / Open Access
• [Journal Article] The reduction of heparan sulphate in the glomerular basement membrane does not augment urinary albumin excretion (2018)
  • Author(s): Satoshi Aoki, Akiko Saito-Hakoda, Takeo Yoshikawa, Kyoko Shimizu, Kiyomi Kisu, Susumu Suzuki, Kiyoshi Takagi, Shuji Mizumoto, Shuhei Yamada, Toin H. van Kuppevelt, Atsushi Yokoyama, Taiji Matsusaka, Hiroshi Sato, Sadayoshi Ito, and Akira Sugawara
  • Journal Title: Nephrology Dialysis Transplantation Volume: 33 Issue: 1 Pages: 26-33
  • DOI: 10.1093/ndt/gfx218
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Vascular abnormalities in the placenta of Chst14-/- fetuses: implications in the pathophysiology of perinatal lethality of the murine model and vascular lesions in human CHST14/D4ST1 deficiency (2018)
  • Author(s): Takahiro Yoshizawa, Shuji Mizumoto, Yuki Takahashi, Shin Shimada, Kazuyuki Sugahara, Jun Nakayama, Shin’ichi Takeda, Yoshihiro Nomura, Yuko Nitahara-Kasahara, Takashi Okada, Kiyoshi Matsumoto, Shuhei Yamada, and Tomoki Kosho
  • Journal Title: Glycobiology Volume: 28 Issue: 2 Pages: 80-89
  • DOI: 10.1093/glycob/cwx099
  • Peer Reviewed
• [Journal Article] Identification of biallelic EXTL3 mutations in a novel type of spondylo-epi-metaphyseal dysplasia (2017)
  • Author(s): *Long Guo, *Nursel H Elcioglu, *Shuji Mizumoto, Zheng Wang, Bilge Noyan, Hatice M Albayrak, Shuhei Yamada, Naomichi Matsumoto, Noriko Miyake, Gen Nishimura, Shiro Ikegawa (equal contribution)
  • Journal Title: Journal of Human Genetics Volume: － Issue: 8 Pages: 797-801
  • DOI: 10.1038/jhg.2017.38
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Chondroitin sulfate N-acetylgalactosaminyltransferase-1 (CSGalNAcT-1) deficiency results in a mild skeletal dysplasia and joint laxity. (2017)
  • Author(s): Julia Vodopiutz, Shuji Mizumoto, Ekkehard Lausch, Antonio Rossi, Sheila Unger, Nikolaus Janocha, Rainer Seidl, Susanne Greber-Platzer, Shuhei Yamada, Thomas Muller, Bernd Jilma, Rudolf Ganger, Andrea Superti-Furga, Shiro Ikegawa, Kazuyuki Sugahara, Andreas R. Janecke.
  • Journal Title: Human Mutation Volume: 38 Issue: 1 Pages: 34-38
  • DOI: 10.1002/humu.23070
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Functional validation of novel compound heterozygous variants in B3GAT3 resulting in severe osteopenia and fractures: expanding the disease phenotype. (2016)
  • Author(s): *Florian Job, *Shuji Mizumoto, Laurie Smith, Natario Couser, Ashley Brazil, Howard Saal, Melanie Patterson, Margaret I. Gibson, Sarah Soden, Neil Miller, Isabelle Thiffault, Carol Saunders, Shuhei Yamada, Katrin Hoffmann, Kazuyuki Sugahara, Emily Farrow. (*equal contribution)
  • Journal Title: BMC Medical Genetics Volume: 17 Issue: 1 Pages: 86-86
  • DOI: 10.1186/s12881-016-0344-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Molecular interactions between chondroitin-dermatan sulfate and growth factors/receptors/matrix proteins (2016)
  • Author(s): Shuji Mizumoto, Shuhei Yamada, Kazuyuki Sugahara
  • Journal Title: Curr. Opin. Struct. Biol. Volume: 34 Pages: 35-42
  • DOI: 10.1016/j.sbi.2015.06.004
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Mutations in biosynthetic enzymes for the protein linker region of chondroitin/dermatan/heparan sulfate cause skeletal and skin dysplasias (2015)
  • Author(s): Shuji Mizumoto, Shuhei Yamada, Kazuyuki Sugahara
  • Journal Title: BioMed Res. Int. Volume: 2015 Pages: 861752-861752
  • DOI: 10.1155/2015/861752
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] コンドロイチン硫酸／デルマタン硫酸生合成酵素の先天的欠損による遺伝性疾患 (2018)
  • Author(s): 山田修平
  • Organizer: 第31回日本軟骨代謝学会
  • Invited
• [Presentation] 発達段階の脳におけるコンドロイチン硫酸/ヒアルロン酸分解酵素の発現解析 (2017)
  • Author(s): 山田修平、三輪一貴、水本秀二
  • Organizer: 第49回日本結合組織学会学術大会
• [Presentation] 脊髄損傷モデルマウスへのヒアルロニダーゼの投与 (2017)
  • Author(s): 直井友弥、柴本秀太、水本秀二、勝又優子、森岡 幸、武内恒成、山田修平
  • Organizer: 第63回日本薬学会東海支部総会・大会
• [Presentation] ヒアルロニダーゼ1および４の基質認識に関与するアミノ酸残基の解析 (2017)
  • Author(s): 小林智恵里、白水仁子、稲葉史恵、安部優貴子、水本秀二、山田修平
  • Organizer: 第63回日本薬学会東海支部総会・大会
• [Presentation] コンドロイチン硫酸を特異的に加水分解するヒアルロニダーゼ４の細胞内局在の解析 (2017)
  • Author(s): 牧野冴紀、水本秀二、山田修平
  • Organizer: 第36回日本糖質学会年会
• [Presentation] 発生段階の異なるマウス脳におけるグリコサミノグリカン分解酵素の発現解析 (2017)
  • Author(s): 禰津瑠里子、三輪一貴、水本秀二、山田修平
  • Organizer: 第36回日本糖質学会年会
• [Presentation] ヒアルロニダーゼ1および４の基質認識に関与するアミノ酸残基の解析 (2017)
  • Author(s): 白水仁子、小林智恵里、稲葉史恵、安部優貴子、水本秀二、山田修平
  • Organizer: 第36回日本糖質学会年会
• [Presentation] The mammalian chondroitinase, hyaluronidase 4, is produced by human mast cells and generates CS cleavage neo-epitopes (2017)
  • Author(s): Brooke L. Farrugia, Shuji Mizumoto, Megan S. Lord, Shuhei Yamada, Bruce Caterson, John M. Whitelock
  • Organizer: 7 Lakes Proteoglycan Conference
  • Int'l Joint Research
• [Presentation] マウス脳の各発達過程におけるヒアルロン酸代謝関連酵素の発現の定量的解析 (2017)
  • Author(s): 山田修平、三輪一貴、水本秀二
  • Organizer: 平成28年度 新学術領域「神経糖鎖生物学」最終シンポジウム
  • Place of Presentation: 名古屋
• [Presentation] 中枢神経系におけるコンドロイチン硫酸/ヒアルロン酸の分解機構に関する研究 (2017)
  • Author(s): 永井冴、藤井貴夢、水本秀二、山田修平
  • Organizer: 平成28年度 新学術領域「神経糖鎖生物学」最終シンポジウム
  • Place of Presentation: 名古屋
• [Presentation] コンドロイチン硫酸に特異的な加水分解酵素であるHYAL4の酵母（Pichia pastoris）での発現および精製 (2016)
  • Author(s): 柴本秀太、杉田真千子、水本秀二、菅原一幸、山田修平
  • Organizer: 日本薬学会第136年会
  • Place of Presentation: 横浜
  • Year and Date: 2016-03-29
• [Presentation] 脳・神経系におけるコンドロイチン硫酸/ヒアルロン酸分解機構に関する研究 (2016)
  • Author(s): 山田修平、永井冴、三輪一貴、水本秀二
  • Organizer: 第48回日本結合組織学会学術大会
  • Place of Presentation: 長崎
• [Presentation] ヒアルロニダーゼ1および4におけるコンドロイチン硫酸およびヒアルロン酸の認識に関わるアミノ酸残基の解析 (2016)
  • Author(s): 山田修平、安倍優貴子、山極健太郎、水本秀二
  • Organizer: 第48回日本結合組織学会学術大会
  • Place of Presentation: 長崎
• [Presentation] コンドロイチン硫酸を特異的に加水分解するヒアルロニダーゼ４の細胞内局在の解析 (2016)
  • Author(s): 三輪茜音、水本秀二、山田修平
  • Organizer: 第62回日本薬学会東海支部総会 大会
  • Place of Presentation: 名古屋
• [Presentation] マウス脳におけるコンドロイチン硫酸とヒアルロン酸の分解活性の検出 (2016)
  • Author(s): 石川輝、水本秀二、山田修平
  • Organizer: 第62回日本薬学会東海支部総会 大会
  • Place of Presentation: 名古屋
• [Presentation] Quantitative analysis of the expression of hyaluronan-chondroitin sulfate-degrading enzymes in the mouse brain during development. (2016)
  • Author(s): Kazutaka Miwa, Shuji Mizumoto, Shuhei Yamada
  • Organizer: XXVIII International Carbohydrate Symposium
  • Place of Presentation: New Orleans, USA
• [Presentation] Investigation of the amino acid residues in hyaluronidases-1 and -4 responsible for their substrate recognition (2016)
  • Author(s): Yukiko Abe, Kentaro Yamagiwa, Shuji Mizumoto, Shuhei Yamada
  • Organizer: XXVIII International Carbohydrate Symposium
  • Place of Presentation: New Orleans, USA
• [Presentation] Studies on the mechanism of degradation of chondroitin sulfate/hyaluronan in central nervous system (2016)
  • Author(s): Shuhei Yamada, Sae Nagai, Shuji Mizumoto
  • Organizer: XXVIII International Carbohydrate Symposium
  • Place of Presentation: New Orleans, USA
• [Presentation] ヒアルロニダーゼ1および4におけるコンドロイチン硫酸およびヒアルロン酸の認識に関わるアミノ酸残基の解析 (2015)
  • Author(s): 安倍優貴子、山極健太郎、岩月遥奈、水本秀二、山田修平
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-02
• [Presentation] Investigation of the Amino Acid Residue Responsible for Diferrent Substrate Specificity of Hyaluronidases-1 and -4 (2015)
  • Author(s): Kentaro Yamagiwa, Haruna Iwatsuki, Yukiko Abe, Shuji Mizumoto, and Shuhei Yamada
  • Organizer: 9th International Conference on Proteoglycans and 10th Pan-Pacific Connective Tissue Societies Symposium
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2015-08-25
  • Int'l Joint Research
• [Presentation] Quantitative Analysis of the Expression of Hyaluronidases in the Mouse Brain during Development (2015)
  • Author(s): Mayu Mitani, Shuji Mizumoto, Shuhei Yamada
  • Organizer: 9th International Conference on Proteoglycans and 10th Pan-Pacific Connective Tissue Societies Symposium
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2015-08-25
  • Int'l Joint Research
• [Presentation] Cellular localization of chondroitin sulfate hydrolase, hyaluronidase－4 (2015)
  • Author(s): Yumiko Nomura, Shuji Mizumoto, and Shuhei Yamada
  • Organizer: 9th International Conference on Proteoglycans and 10th Pan-Pacific Connective Tissue Societies Symposium
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2015-08-25
  • Int'l Joint Research
• [Presentation] 発生段階の異なるマウス脳におけるヒアルロニダーゼファミリーメンバーの発現解析 (2015)
  • Author(s): 三谷真夕、水本秀二、山田修平
  • Organizer: 第34回日本糖質学会年会
  • Place of Presentation: 東京
  • Year and Date: 2015-08-02
• [Presentation] 酵母（Pichia pastoris）を用いたヒアルロニダーゼ４の発現及び精製 (2015)
  • Author(s): 杉田真千子、柴本秀太、水本秀二、菅原一幸、山田修平
  • Organizer: 第34回日本糖質学会年会
  • Place of Presentation: 東京
  • Year and Date: 2015-08-02
• [Presentation] ヒアルロニダーゼ1と4の基質認識の違いに関与するアミノ酸残基の解析 (2015)
  • Author(s): 山極健太郎、岩月遥奈、安倍優貴子、水本秀二、山田修平
  • Organizer: 第34回日本糖質学会年会
  • Place of Presentation: 東京
  • Year and Date: 2015-08-02
• [Presentation] ヒトの癌細胞株や癌組織におけるヒアルロニダーゼ4の発現解析 (2015)
  • Author(s): 西島大樹、水本秀二、山田修平
  • Organizer: 第61回日本薬学会東海支部大会
  • Place of Presentation: 名古屋
  • Year and Date: 2015-07-04
• [Book] Encyclopedia of Polymeric Nanomaterials (2015)
  • Author(s): Kazuyuki Sugahara, Shuji Mizumoto, and Shuhei Yamada
  • Total Pages: 2672
  • Publisher: Springer
• [Remarks] 名城大学薬学部病態生化学薬学部病態生化学
  • URL: http://www-yaku.meijo-u.ac.jp/Research/Laboratory/pathobio/index.html
• [Remarks] 名城大学薬学部病態生化学研究室ホームページ
  • URL: http://www-yaku.meijo-u.ac.jp/Research/Laboratory/pathobio/index.html
• [Remarks] 名城大学薬学部病態生化学
  • URL: http://www-yaku.meijo-u.ac.jp/Research/Laboratory/pathobio/index.html