Study of reproductive biological functions of bisecting GlcNAc using glycotechnology and immunological methods

• Project/Area Number: 16K11111
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Obstetrics and gynecology
• Research Institution: Juntendo University
• Principal Investigator: Yoshitake Hiroshi 順天堂大学, 医学(系)研究科(研究院), 非常勤講師 (00396574)
• Co-Investigator(Kenkyū-buntansha): 荒木 慶彦 順天堂大学, 医学(系)研究科(研究院), 先任准教授 (70250933)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 抗精子抗体 / バイセクト型糖鎖 / NUP62 / 精巣 / CD73 / 精細胞 / α-N-アセチルグルコサミニダーゼ / 糖鎖 / 糖タンパク質

Outline of Final Research Achievements

We previously produced an anti-mouse sperm auto-monoclonal antibody, Ts4, which immunoreacts against some kinds of glycoproteins via binding to an epitope containg a fucosylated agalacto-complex-type biantennary glycan carrying bisecting N-acetylglucosamine. We have already identified α-N-acetylglucosaminidase (a degradation enzyme of heparan sulfate) and TEX101 (a germ cell-specific glycosylphosphatidyl inositol-anchored glycoprotein) as the Ts4-recognized molecules in the mature spermatozoa and the mature testis, respectively. Furthermore, in this study, we demonstrated that Ts4 exhibits immunoreactivity against nuclear pore glycoprotein p62 (NUP62) in the immature testis. Our previous and present results show that the glycoproteins modified with the Ts4-reactive oligosaccharide chain vary across different stages of male germ cell differentiation and proliferation and may play roles in fertilization and development of male gonadal organs.

Academic Significance and Societal Importance of the Research Achievements

本邦で深刻化している少子化問題は、社会科学的に議論されることが多いが、不妊症の増加など医学的な原因も無視できない。不妊症患者血清中に抗精子抗体が検出されることがあるが、実際に受精を阻害する免疫学的作用を示すことがあり、不妊の原因になる可能性は以前より指摘されていた。しかしそのほとんどが不妊症患者血清を用いた実験で判明したものであり、その対応抗原や生物学的機能について検討することは困難であった。Ts4が認識する糖鎖とその修飾糖タンパク質の実態が明らかになれば、本分子を介した全く新しい受精の分子メカニズムの解明が期待でき、これまで原因不明とされてきた不妊症の一部の分子基盤を構築できる可能性がある。

Research Products

• [Journal Article] Promoting Roles of Embryonic Signals in Embryo Implantation and Placentation in Cooperation with Endocrine and Immune Systems (2020)
  • Author(s): Fujiwara Hiroshi、Ono Masanori、Sato Yukiyasu、Imakawa Kazuhiko、Iizuka Takashi、Kagami Kyosuke、Fujiwara Tomoko、Horie Akihito、Tani Hirohiko、Hattori Akira、Daikoku Takiko、Araki Yoshihiko
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 5 Pages: 1885-1885
  • DOI: 10.3390/ijms21051885
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hypertensive disorders of pregnancy: Strategy to develop clinical peptide biomarkers for more accurate evaluation of the pathophysiological status of this syndrome (2020)
  • Author(s): Araki Yoshihiko、Yanagida Mitsuaki
  • Journal Title: Advances in Clinical Chemistry Volume: 94 Pages: 1-30
  • DOI: 10.1016/bs.acc.2019.07.007
  • ISBN: 9780128208014
  • Peer Reviewed
• [Journal Article] Knockdown of DEAD-box RNA helicase DDX5 selectively attenuates serine 311 phosphorylation of NF-κB p65 subunit and expression level of anti-apoptotic factor Bcl-2. (2020)
  • Author(s): Tanaka K, Tanaka T*, Nakano T, Hozumi Y, Yanagida M, Araki Y, Iwazaki K, Takagi M, Goto K
  • Journal Title: Cellular Signalling Volume: 65 Pages: 1-8
  • DOI: 10.1016/j.cellsig.2019.109428
  • Peer Reviewed
• [Journal Article] NUP62: the target of an anti-sperm auto-monoclonal antibody during testicular development (2019)
  • Author(s): Oda-Sakurai Risako、Yoshitake Hiroshi、Miura Yoshiki、Kazuno Saiko、Ueno Takashi、Hasegawa Akiko、Yamatoya Kenji、Takamori Kenji、Itakura Atsuo、Fujiwara Hiroshi、Takeda Satoru、Araki Yoshihiko
  • Journal Title: Reproduction Volume: 158 Issue: 6 Pages: 503-516
  • DOI: 10.1530/rep-19-0333
  • Peer Reviewed
• [Journal Article] Cleavage of SPACA1 regulates assembly of sperm?egg membrane fusion machinery in mature spermatozoa† (2019)
  • Author(s): Yamatoya Kenji、Kousaka Marika、Ito Chizuru、Nakata Kazuya、Hatano Masahiko、Araki Yoshihiko、Toshimori Kiyotaka
  • Journal Title: Biology of Reproduction Volume: 102 Issue: 3 Pages: 750-757
  • DOI: 10.1093/biolre/ioz223
  • Peer Reviewed / Open Access
• [Journal Article] The simultaneous quantification of candidate serum biomarker peptides for hypertensive disorders of pregnancy (2019)
  • Author(s): Yanagida Mitsuaki、Hamamura Kensuke、Takamori Kenji、Araki Yoshihiko
  • Journal Title: Annals of Clinical Biochemistry: International Journal of Laboratory Medicine Volume: 56 Issue: 4 Pages: 457-465
  • DOI: 10.1177/0004563219839084
  • Peer Reviewed
• [Journal Article] Quantitative measurement of a candidate serum biomarker peptide derived from α2-HS-glycoprotein, and a preliminary trial of multidimensional peptide analysis in females with pregnancy-induced hypertension. (2018)
  • Author(s): Hamamura K, Yanagida M, Ishikawa H, Banzai M, Yoshitake H, Nonaka D, Tanaka K, Sakuraba M, Miyakuni Y, Takamori K, Nojima M, Yoshida K, Fujiwara H, Takeda S, Araki Y.
  • Journal Title: Ann Clin Biochem Volume: 55 Issue: 2 Pages: 287-295
  • DOI: 10.1177/0004563217717748
  • Peer Reviewed
• [Journal Article] Factors Regulating Human Extravillous Trophoblast Invasion: Chemokine-peptidase and CD9-integrin Systems (2018)
  • Author(s): Fujiwara Hiroshi、Matsumoto Hisanori、Sato Yukiyasu、Horie Akihito、Ono Masanori、Nakamura Mitsuhiro、Mizumoto Yasunari、Kagami Kyosuke、Fujiwara Tomoko、Hattori Akira、Maida Yoshiko、Daikoku Takiko、Imakawa Kazuhiko、Araki Yoshihiko
  • Journal Title: Current Pharmaceutical Biotechnology Volume: 19 Issue: 10 Pages: 764-770
  • DOI: 10.2174/1389201019666181029164906
  • Peer Reviewed
• [Journal Article] Involvement of cross-reactive carbohydrate determinants-specific IgE in pollen allergy testing. (2017)
  • Author(s): Yokoi H, Yoshitake H, Matsumoto Y, Kawada M, Takato Y, Shinagawa K, Sakurai H, Saito K.
  • Journal Title: Asia Pac Allergy Volume: 7 Issue: 1 Pages: 29-36
  • DOI: 10.5415/apallergy.2017.7.1.29
  • Peer Reviewed / Open Access
• [Journal Article] Platelet-derived microparticles and soluble factors differentially regulate human endometrial epithelial cell movement (2017)
  • Author(s): Suginami K, Sato Y, Horie A, Matsumoto H, Tani H, Mizumoto Y, Ono M, Matsuoka A, Kyo S, Araki Y, Konishi I, Fujiwara H
  • Journal Title: Am J Reprod Immunol Volume: Feb 7 Issue: 4 Pages: 12641-12641
  • DOI: 10.1111/aji.12641
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Platelets are a possible regulator of human endometrial re-epithelialization during menstruation (2016)
  • Author(s): Suginami K, Sato Y, Horie A, Matsumoto H, Kyo S, Araki Y, Konishi I, Fujiwara H
  • Journal Title: Am J Reprod Immunol Volume: 77 Issue: 1 Pages: 12609-12609
  • DOI: 10.1111/aji.12609
  • Peer Reviewed
• [Journal Article] Identification of an anti-sperm auto-monoclonal antibody (Ts4)-recognized molecule in the mouse sperm acrosomal region and its inhibitory effects on fertilization in vitro. (2016)
  • Author(s): Yoshitake H., Oda R., Yanagida M., Kawasaki Y., Sakuraba M., Takamori K., Hasegawa A., Fujiwara H., Araki Y.
  • Journal Title: J.Reprod.Immunol. Volume: 115 Pages: 6-13
  • DOI: 10.1016/j.jri.2016.04.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] TEX101, a glycoprotein essential for sperm fertility, is required for stable expression of Ly6k on testicular germ cells (2016)
  • Author(s): Endo S, Yoshitake H, Tsukamoto H, Matsuura H, Kato K, Sakuraba M, Takamori K, Fujiwara H, Takeda S, Araki Y
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 23616-23616
  • DOI: 10.1038/srep23616
  • Peer Reviewed / Open Access
• [Presentation] CD73: バイセクト型N-アセチルグルコサミン糖鎖構造を認識する抗精子自己抗体Ts4の精巣内生殖細胞における標的候補分子 (2018)
  • Author(s): 小田理沙子、吉武洋、數野彩子、三浦芳樹、上野隆、櫻庭真弓、髙森建二、長谷川昭子、藤原浩、竹田省、荒木慶彦
  • Organizer: 第33回日本生殖免疫学会総会・学術集会
• [Presentation] Bisecting GlcNAc, the molecular epitope of an anti-sperm auto-antibody, Ts4: Its morphological characteristics and identification of the candidate glycoprotein(s) possessing the sugar moiety in mice (2018)
  • Author(s): Risako Oda, Hiroshi Yoshitake, Saiko Kazuno, Yoshiki Miura, Takashi Ueno, Mayumi Sakuraba, Kenji Takamori, Akiko Hasegawa, Hiroshi Fujiwara, Satoru Takeda, Yoshihiko Araki
  • Organizer: Society for the Study of Reproduction, 51th Annual Meeting
  • Int'l Joint Research
• [Presentation] 単クローン抗体RP-3の反応性を指標としたラット末梢血多形核白血球の性差と多様性 (2016)
  • Author(s): 吉武洋、有井大介、濱村憲佑、松田浩則、櫻庭真弓、川崎優、髙森建二、武田裕司、仙道富士郎、藤原浩、野島美知夫、荒木慶彦
  • Organizer: 日本生殖免疫学会
  • Place of Presentation: 兵庫県神戸市
  • Year and Date: 2016-12-02
• [Patent(Industrial Property Rights)] Laeverinを用いたPSTT（胎盤部トロホブラスト腫瘍）の検出 (2018)
  • Inventor(s): 藤原浩、荒木慶彦
  • Industrial Property Rights Holder: 藤原浩、荒木慶彦
  • Industrial Property Rights Type: 特許
  • Filing Date: 2018