Identification of cytotoxic factors for eggs and development of neutralizing drugs

• Project/Area Number: 26293363
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Obstetrics and gynecology
• Research Institution: National Center for Child Health and Development
• Principal Investigator: MIYADO Kenji 国立研究開発法人国立成育医療研究センター, 細胞医療研究部, 室長 (60324844)
• Co-Investigator(Kenkyū-buntansha): 河野 菜摘子 明治大学, 農学部, 講師 (00451691)
• Co-Investigator(Renkei-kenkyūsha): INOUE Naokazu (SAITO Hidekazu) 福島県立医科大学, 医学部, 准教授 (90125766) ; HAMATANI Toshio 慶應義塾大学, 医学部, 講師 (60265882) ; MIYAMOTO YOshitaka 国立成育医療研究センター, 細胞医療研究部, 研究員 (20425705)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥15,860,000 (Direct Cost: ¥12,200,000、Indirect Cost: ¥3,660,000); Fiscal Year 2016: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2015: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2014: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: 卵 / 受精 / 膜融合 / リン脂質 / CD9 / マイクロエクソソーム / エクソソーム / 細胞毒性 / テトラスパニン / ストレス応答 / 再生医学 / 発生 / 細胞 / 脂質 / ストレス / エキソソーム / 顆粒膜細胞 / 老化

Outline of Final Research Achievements

Since CD9 is involved in sperm-egg fusion, this protein may organize membrane architectures. Immunoelectron microscopic analysis revealed that CD9 is incorporated into microexosomes. Since microexosomes have no overt lipid bilayers and seem to be small units, less than 5 nm in diameter, they are structurally different from exosomes. Furthermore, Cd9-deficient eggs could not fuse with sperm, but in the presence of wild-type eggs, 70% of the Cd9-deficient eggs fused with the sperm. Notably, the microexosomes rendered sperm fusion competent with Cd9-deficient eggs, which means that microexosomes, but not the egg microvilli, are essential for sperm-egg fusion. In addition, microexosomes are observed inside the uterus. Taken together, these results indicate that there are two forms of tetraspanin, namely the intercellular transportation-related forms and membrane fusion-related forms.

Research Products

• [Journal Article] Birth weights and Down syndrome in neonates delivered after frozen-thawed embryo transfer: Six-year study (2007-2012) on the national assisted reproduction registry data in Japan (2017)
  • Author(s): Yamatoya K, Saito K, Saito T, Kang W, Nakamura A, Miyado M, Kawano N, Miyamoto Y, Umezawa A, Miyado K, Saito H
  • Journal Title: Reproductive Medicine and Biology Volume: 16 Issue: 2 Pages: 228-234
  • DOI: 10.1002/rmb2.12033
  • Peer Reviewed / Open Access
• [Journal Article] Exosomes versus microexosomes: Shared components but distinct functions (2017)
  • Author(s): Miyado K, Kang W, Yamatoya K, Hanai M, Nakamura A, Mori T, Miyado M, Kawano N
  • Journal Title: J Plant Res Volume: 130 Issue: 3 Pages: 479-483
  • DOI: 10.1007/s10265-017-0907-7
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Complementary critical functions of Zfy1 and Zfy2 in mouse spermatogenesis and reproductiontran . (2017)
  • Author(s): Nakasuji T, Ogonuki N, Chiba T, Kato T, Shiozawa K, Yamatoya K, Tanaka H, Kondo T, Miyado K, Miyasaka N, Kubota T, Ogura A, Asahara H*.
  • Journal Title: PLoS Genetics. Volume: 13 Issue: 1 Pages: e1006578-e1006578
  • DOI: 10.1371/journal.pgen.1006578
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Expression patterns of Fgf8 and Shh in the developing external genitalia of Suncus murinus (2017)
  • Author(s): Miyado M, Miyado K, Nakamura A, Fukami M, Yamada G, Oda SI
  • Journal Title: Reproduction Volume: 153 Issue: 2 Pages: 187-195
  • DOI: 10.1530/rep-16-0231
  • Peer Reviewed / Open Access
• [Journal Article] Increased incidence of post-term delivery and Cesarean section after frozen-thawed embryo transfer during a hormone replacement cycle (2016)
  • Author(s): Saito K, Miyado K, Yamatoya K, Kuwahara A, Inoue E, Miyado M, Fukami M, Ishikawa T, Saito T, Kubota T, Saito H
  • Journal Title: J Assist Reprod Genet Volume: 34 Issue: 4 Pages: 465-470
  • DOI: 10.1007/s10815-017-0869-7
  • Peer Reviewed / Open Access
• [Journal Article] The p.R92W variant of NR5A1/Nr5a1 induces testicular development of 46,XX gonads in humans, but not in mice: phenotypic comparison of human patients and mutation-induced mice (2016)
  • Author(s): Miyado M, Inui M, Igarashi M, Katoh-Fukui Y, Takasawa K, Hakoda A, Kanno J, Kashimada K, Miyado K, Tamano M, Ogata T, Takada S, Fukami M
  • Journal Title: Biol Sex Differ Volume: 7 Issue: 1 Pages: 56-56
  • DOI: 10.1186/s13293-016-0114-6
  • Peer Reviewed / Open Access
• [Journal Article] <b>The role of tetraspanin CD9 in osteoarthritis using three different mouse </b><b>models </b> (2016)
  • Author(s): Sumiyoshi N, Ishitobi H, Miyaki S, Miyado K, Adachi N, Ochi M.
  • Journal Title: Biomedical Research Volume: 37 Issue: 5 Pages: 283-291
  • DOI: 10.2220/biomedres.37.283
  • NAID: 130005432956
  • ISSN: 0388-6107, 1880-313X
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Seminal vesicle proteins SVS3 and SVS4 facilitate SVS2 effect on sperm capacitation (2016)
  • Author(s): Araki N, Yoshida K, Kawano N, Woodin K, Miyado K, Yoshida M
  • Journal Title: Reproduction Volume: 152 Issue: 4 Pages: 313-321
  • DOI: 10.1530/rep-15-0551
  • Peer Reviewed / Open Access
• [Journal Article] Mesenchymal Stem Cell-Derived Exosomes Promote Fracture Healing in a Mouse Model (2016)
  • Author(s): TAISUKE FURUTA, SHIGERU MIYAKI, HIROYUKI ISHITOBI, TOSHIHIKO OGURA, YOSHIO KATO, NAOSUKE KAMEI, KENJI MIYADO, YUKIHITO HIGASHI, MITSUO OCHI
  • Journal Title: STEM CELLS TRANSLATIONALMEDICINE Volume: 5 Issue: 12 Pages: 1620-1630
  • DOI: 10.5966/sctm.2015-0285
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Critical roles of seminal plasma on sperm migration in the female reproductive tract (2016)
  • Author(s): Kang W, Kawano N, Yamatoya K, Yoshida M, Yoshida K, Miyado K
  • Journal Title: J Reprod Eng Volume: 18 Pages: 1-19
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Conditional deletion of CD98hc inhibits osteoclast development. (2016)
  • Author(s): Tsumura H, Ito M, Takami M, Arai M, Li XK, Hamatani T, Igarashi A, Takada S, Miyado K, Umezawa A, Ito Y.
  • Journal Title: Biochemistry and Biophysics Reports Volume: 5 Pages: 203-210
  • DOI: 10.1016/j.bbrep.2015.11.023
  • Peer Reviewed
• [Journal Article] Epididymal C4b-binding protein is processed and degraded during transit through the duct and is not essential for fertility (2015)
  • Author(s): Mayumi I Nonaka, Eva Zsigmond, Akihiko Kudo, Hayato Kawakami, Kaoru Yoshida, Manabu Yoshida, Natsuko Kawano, Kenji Miyado, Masaru Nonaka, Rick A Wetsel
  • Journal Title: Immunobiology Volume: 220(4) Pages: 467-475
  • DOI: 10.1016/j.reth.2015.11.001
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Parturition failure in mice lacking Mamld1. (2015)
  • Author(s): Miyado M, Miyado K, Katsumi M, Saito K, Nakamura A, Shihara D, Ogata T, Fukami M.
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 14705-14705
  • DOI: 10.1038/srep14705
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Xenogeneic-free defined conditions for derivation and expansion of human embryonic stem cells with mesenchymal stem cells. (2015)
  • Author(s): Akutsu H, Machida M, Kanzaki S, Sugawara T, Ohkura T, Nakamura N, Yamazaki-Inoue M, Miura T, Vemuri MC, Rao MS, Miyado K, Umezawa A.
  • Journal Title: Regenerative Therapy Volume: 1 Pages: 18-29
  • Peer Reviewed / Open Access
• [Journal Article] Staphylococcus epidermidis is involved in a mechanism for female reproduction in mice. (2015)
  • Author(s): Ono C, Yoshida M, Kawano N, Miyado K, Umezawa A.
  • Journal Title: Regenerative Therapy Volume: 1 Pages: 11-17
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Phosphorus-insufficient maternal milk is associated with ectopic expression of collagen I and female-specific bony changes in infant mouse cartilages. (2015)
  • Author(s): Nakamura A, Miyado K, Nasu M, Kono T, Umezawa A.
  • Journal Title: Regenerative Therapy Volume: 1 Pages: 5-10
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Seminal Vesicle Secretion 2 Acts as a Protectant of Sperm Sterols and Prevents Ectopic Sperm Capacitation in Mice (2014)
  • Author(s): N Araki, G Trencsenyi, ZT Krasznai, E Nizsaloczki, A Sakamoto, N Kawano, Kenji Miyado, Kaoru Yoshida, Manabu Yoshida
  • Journal Title: Biology of Reproduction Volume: 114 Issue: 1 Pages: 120642-120642
  • DOI: 10.1095/biolreprod.114.120642
  • Peer Reviewed / Open Access
• [Journal Article] Epididymal C4b-binding protein is processed and degraded during transit through the duct and is not essential for fertility (2014)
  • Author(s): MI Nonaka, E Zsigmond, A Kudo, H Kawakami, K Yoshida, N Kawano, K Miyado, M Nonaka, RA Wetsel
  • Journal Title: Immunobiology Volume: 220 Issue: 4 Pages: 467-475
  • DOI: 10.1016/j.imbio.2014.11.001
  • Peer Reviewed / Open Access
• [Journal Article] Mitochondrial Fission Factor Drp1 Maintains Oocyte Quality via Dynamic Rearrangement of Multiple Organelles (2014)
  • Author(s): O Udagawa, T Ishihara, M Maeda, Y Matsunaga, S Tsukamoto, N Kawano, K Miyado, H Shitara, S Yokota, M Nomura, K Mihara, N Mizushima, N Ishihara
  • Journal Title: Current Biology Volume: 24 Issue: 20 Pages: 2451-2458
  • DOI: 10.1016/j.cub.2014.08.060
  • Peer Reviewed / Open Access
• [Journal Article] Mitochondrial dynamics controlled by mitofusins define organelle positioning and movement during mouse oocyte maturation (2014)
  • Author(s): Takuya Wakai, Yuichirou Harada, Kenji Miyado, Tomohiro Kono
  • Journal Title: Molecular Human Reproduction Volume: 20 Issue: 11 Pages: 1090-1100
  • DOI: 10.1093/molehr/gau064
  • Peer Reviewed / Open Access
• [Journal Article] Absence of CD9 reduces endometrial VEGF secretion and impairs uterine repair after parturition (2014)
  • Author(s): N Kawano, K Miyado, N Yoshii, S Kanai, H Saito, M Miyado, N Inagaki, Y Odawara, T Hamatani, A Umezawa
  • Journal Title: Scientific reports Volume: 4 Issue: 1 Pages: 4701-4701
  • DOI: 10.1038/srep04701
  • Peer Reviewed / Open Access
• [Journal Article] Derivation of human decidua-like cells from amnion and menstrual blood (2014)
  • Author(s): Sugawara K, Hamatani T, Yamada M, Ogawa S, Kamijo S, Kuji N, Akutsu H, Miyado K, Yoshimura Y, Umezawa A
  • Journal Title: Sci Rep Volume: 4 Issue: 1 Pages: 4599-4599
  • DOI: 10.1038/srep04599
  • Peer Reviewed / Open Access
• [Presentation] 卵に対する細胞障害性因子の同定と対処法の開発 (2016)
  • Author(s): 宮戸健二
  • Organizer: 第34回日本受精着床学会総会・学術講演会
  • Place of Presentation: 軽井沢
  • Year and Date: 2016-09-15
  • Invited
• [Presentation] 精漿タンパク質SVS2欠損マウスから見えて来た、精子を殺すメスの免疫機構 (2015)
  • Author(s): 河野菜摘子、康宇鎭、吉田薫、吉田学、宮戸健二
  • 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、ワークショップ：「生殖」から読み解く哺乳類の生命現象（オーガナイザー：深見真紀、宮戸健二）
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] テトラスパニン／エキソソームによる膜融合の制御機構 (2015)
  • Author(s): 宮戸健二、大和屋健二、河野菜摘子
  • Organizer: 日本動物学会第86回新潟大会
  • Place of Presentation: 朱鷺メッセ：新潟コンベンションセンター
  • Year and Date: 2015-09-17
• [Presentation] 哺乳類における配偶子融合の分子機構 (2015)
  • Author(s): 大和屋健二、宮戸健二、河野菜摘子
  • Organizer: 日本動物学会第86回新潟大会
  • Place of Presentation: 朱鷺メッセ：新潟コンベンション
  • Year and Date: 2015-09-17
• [Presentation] 卵由来エキソソームによる配偶子融合の制御機構 (2015)
  • Author(s): 宮戸健二
  • Organizer: 日本獣医解剖学会シンポジウム
  • Place of Presentation: 北里大学獣医学部
  • Year and Date: 2015-09-07
  • Invited
• [Presentation] Genesis of life: A mechanism of sperm-egg fusion in mammals. (2015)
  • Author(s): Kenji Miyado, Kenji Yamatoya, Natsuko Kawano.
  • Organizer: 日本植物学会第79回大会 シンポジウム: “Fusion” in Fertilization: Interdisciplinary Collaboration among Plant and Animal Scientists
  • Place of Presentation: 朱鷺メッセ：新潟コンベンションセンター
  • Year and Date: 2015-09-06
  • Invited
• [Presentation] 受精における副生殖腺の役割 (2015)
  • Author(s): 宮戸健二
  • Organizer: 日本生殖医学会学術講演会 シンポジウム3受精に関する最近の話題
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-04-27
  • Invited
• [Presentation] メス生殖器における精子サバイバル術 (2014)
  • Author(s): 河野菜摘子、荒木直也、吉田薫、吉田学、宮戸健二
  • Organizer: 日本動物学会
  • Place of Presentation: 東北大学川内北キャンパス
  • Year and Date: 2014-09-11 – 2014-09-13
  • Invited
• [Presentation] 男と女の生存競争-子宮内での精子の細胞死と防御システム (2014)
  • Author(s): 宮戸健二
  • Organizer: 日本アンドロロジー学会
  • Place of Presentation: 軽井沢プリンスホテルウエスト
  • Year and Date: 2014-06-12 – 2014-06-13
  • Invited
• [Presentation] 河野菜摘子、宮戸健二、吉田薫、吉田学 (2014)
  • Author(s): 新たに見つかった子宮内での精子保護機構
  • Organizer: 日本アンドロロジー学会
  • Place of Presentation: 軽井沢プリンスホテルウエスト
  • Year and Date: 2014-06-12 – 2014-06-13
  • Invited