Roles of exosomes in reproductive system

• Project/Area Number: 16H05475
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Obstetrics and gynecology
• Research Institution: Keio University
• Principal Investigator: HAMATANI TOSHIO 慶應義塾大学, 医学部(信濃町), 講師 (60265882)
• Co-Investigator(Kenkyū-buntansha): 宮戸 健二 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) ; 阿久津 英憲 国立研究開発法人国立成育医療研究センター, 生殖医療研究部, 部長 (50347225) ; 山田 満稔 慶應義塾大学, 医学部(信濃町), 助教 (40383864)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2019: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2018: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2017: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2016: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
• Keywords: CD9 / 子宮内膜 / 胚 / 細胞外分泌顆粒 / エクソソーム / miRNA / 卵胞 / 胚盤胞 / 培養液 / 生殖医療 / 生殖医学

Outline of Final Research Achievements

Tetraspanin membrane protein CD9 was localized predominantly on the basal region of uterine epithelial cells at estrus-stage mice, but relocated to the apical region upon embryo implantation. Extracellular CD9 was most detected in uterine cavity at the estrus stage. In uterine epithelial cells lacking Cd9 genes, membrane projections were shortened and the number of mitochondria was reduced. CD9 repositioning and release may affect both membrane structures and mitochondrial state in the uterus, and contribute to female fertility.
miRNAs secreted via exosome from human blastocysts were profiled using RNA-seq and qRT-PCR. The spent medium in which a good-qualty blastocyst was harvested was analyzed. Some of miRNAs were significantly highly expressed in the spent media derived from blastocysts which resulted in implantation failure, compared to those in pregnancy. The embryo selection based on the detection of the specific miRNAs in spent medium may improve the pregnancy rate.

Academic Significance and Societal Importance of the Research Achievements

生殖医療における細胞外分泌顆粒（EMV）に関する報告は、我々の論文も含め未だ数報しかない。テトラスパニンCD9はEMVの表面に存在しEMVマーカーとなるため、本研究ではCD9に注目して子宮内膜上皮細胞からのEMV分泌機構を研究する一方、EMVを介して胚から分泌されるmiRNAにも網羅的検討を加えた。EMVの観点から内膜側と胚側の両側の着床因子を検討し、着床の分子機構におけるEMVの寄与が示唆された。これらの結果は、反復着床不全を克服するための胚の質的評価（移植胚の選択）法やEMV（e.g. 自己多血小板血漿）の子宮内投与による着床促進療法などへの臨床応用に繋がることが期待される。

Research Products

• [Journal Article] Impact of Oxidative Stress on Age-Associated Decline in Oocyte Developmental Competence (2019)
  • Author(s): Sasaki Hiroyuki、Hamatani Toshio、Kamijo Shintaro、Iwai Maki、Kobanawa Masato、Ogawa Seiji、Miyado Kenji、Tanaka Mamoru
  • Journal Title: Frontiers in Endocrinology Volume: 10 Pages: 811-811
  • DOI: 10.3389/fendo.2019.00811
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Assisted reproductive technology in Japan: A summary report for 2017 by the Ethics Committee of the Japan Society of Obstetrics and Gynecology. (2019)
  • Author(s): Ishihara O, Jwa SC, Kuwahara A, Katagiri Y, Kuwabara Y, Hamatani T, Harada M, Ichikawa T.
  • Journal Title: Reprod Med Biol Volume: 19(1) Issue: 1 Pages: 3-12
  • DOI: 10.1002/rmb2.12307
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Zscan5b Deficiency Impairs DNA Damage Response and Causes Chromosomal Aberrations during Mitosis. (2019)
  • Author(s): Ogawa S, Yamada M, Nakamura A, Sugawara T, Nakamura A, Miyajima S, Harada Y, Ooka R, Okawa R, Miyauchi J, Tsumura H, Yoshimura Y, Miyado K, Akutsu H, Tanaka M, Umezawa A, Hamatani T.
  • Journal Title: Stem Cell Reports Volume: 12 Issue: 6 Pages: 1366-1379
  • DOI: 10.1016/j.stemcr.2019.05.002
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Membrane protein CD9 is repositioned and released to enhance uterine function. (2019)
  • Author(s): Iwai M, Hamatani T, Nakamura A, Kawano N, Kanai S, Kang W, Yoshii N, Odawara Y, Yamada M, Miyamoto Y, Saito T, Saito H, Miyado M, Umezawa A, Miyado K, Tanaka M.
  • Journal Title: Laboratory Investigation Volume: 99 Issue: 2 Pages: 200-209
  • DOI: 10.1038/s41374-018-0145-1
  • Peer Reviewed / Open Access
• [Journal Article] Autophagy-disrupted LC3 abundance leads to death of supporting cells of human oocytes. (2018)
  • Author(s): Kang W, Ishida E, Yamatoya K, Nakamura A, Miyado M, Miyamoto Y, Iwai M, Tatsumi K, Saito T, Saito K, Kawano N, Hamatani T, Umezawa A, Miyado K, Saito H.
  • Journal Title: Biochem Biophys Rep. Volume: 21(15) Pages: 107-114
  • DOI: 10.1016/j.bbrep.2018.08.002
  • Peer Reviewed / Open Access
• [Journal Article] Degradation of phosphate polymer polyP enhances lactic fermentation in mice. (2018)
  • Author(s): Nakamura A, Kawano N, Motomura K, Kuroda A, Sekiguchi K, Miyado M, Woojin K, Miyamoto Y, Hanai M, Iwai M, Yamada M, Hamatani T, Saito T, Saito H, Tanaka M, Umezawa A, Miyado K.
  • Journal Title: Genes Cells. Volume: 23(10) Issue: 10 Pages: 904-914
  • DOI: 10.1111/gtc.12639
  • Peer Reviewed / Open Access
• [Journal Article] Chemotactic behavior of egg mitochondria in response to sperm fusion in mice. (2018)
  • Author(s): Iwai M, Harada Y, Miyabayashi R, Kang W, Nakamura A, Kawano N, Miyamoto Y, Yamada M, Hamatani T, Miyado M, Yoshida K, Saito H, Tanaka M, Umezawa A, Miyado K.
  • Journal Title: Heliyon Volume: 4(11)
  • Peer Reviewed
• [Journal Article] Ubiquitin-activating enzyme E1 inhibitor PYR-41 retards sperm enlargement after fusion to the egg. (2018)
  • Author(s): Yoshida K, Kang W, Nakamura A, Kawano N, Hanai M, Miyado M, Miyamoto Y, Iwai M, Hamatani T, Saito H, Miyado K and Umezawa A.
  • Journal Title: Reprod Toxicol. Volume: 76 Pages: 71-77
  • DOI: 10.1016/j.reprotox.2018.01.001
  • Peer Reviewed
• [Journal Article] Clinical efficacy of a combination of Percoll continuous density gradient and swim-up techniques for semen processing in HIV-1 serodiscordant couples. (2017)
  • Author(s): Inoue O, Kuji N, Ito H, Yamada M, Hamatani T, Oyadomari A, Kato S, Hanabusa H, Isaka K, Tanaka M.
  • Journal Title: Asian J Androl. Volume: 19 Issue: 2 Pages: 208-213
  • DOI: 10.4103/1008-682x.173442
  • Peer Reviewed / Open Access
• [Journal Article] 不妊治療における慢性炎症と着床不全 (2017)
  • Author(s): 吉井 紀子, 浜谷 敏生．産婦人科領域と炎症（青木大輔編著）
  • Journal Title: 別冊Bio Clinica:慢性炎症と疾患 Volume: 6(4) Pages: 78-83
• [Journal Article] Factors affecting pregnancy outcomes in young women treated with fertility-preserving therapy for well-differentiated endometrial cancer or atypical endometrial hyperplasia. (2016)
  • Author(s): Inoue O, Hamatani T, Susumu N, Yamagami W, Ogawa S, Takemoto T, Hirasawa A, Banno K, Kuji N, Tanaka M, Aoki D.
  • Journal Title: Reprod Biol Endocrinol Volume: 14(1) Issue: 1 Pages: 2-2
  • DOI: 10.1186/s12958-015-0136-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 子宮内膜上皮の膜タンパク質CD9は細胞内局在・細胞外分泌によって子宮機能を制御す (2019)
  • Author(s): 祝井麻希，浜谷敏生，佐々木拓幸，上條慎太郎，小川誠司，山田満稔，宮戸健二，田中守
  • Organizer: 第64回日本生殖医学会学術講演・総会
• [Presentation] Zfp371 is a novel DNA repair gene responsible for genome stability during mitosis (2019)
  • Author(s): Seiji Ogawa, Mitsutoshi Yamada, Toshio Hamatani, Hidenori Akutsu, Akihiro Umezawa, Mamoru Tanaka, Daisuke Aoki
  • Organizer: The 70th Annual Congress of the Japan Society of Obstetrics and Gynecology
• [Presentation] 高用量黄体ホルモン（MPA）療法後の内膜病理検査結果と妊娠予後の検討 (2019)
  • Author(s): 上條慎太郎，浜谷敏生，祝井麻希，山田満稔，田中守．
  • Organizer: 第64回日本生殖医学会学術講演・総会
• [Presentation] 栄養外胚葉の推定細胞数に基づく胚盤胞形態の定量的評価と継続妊娠率の関連 (2019)
  • Author(s): 宇津野 宏樹, 石丸 智子, 松本 美保, 渡邉 久美, 上條 慎太郎, 浜谷 敏生, 田中 守
  • Organizer: 第64回日本生殖医学会学術講演・総会
• [Presentation] 再生医療と生殖医療の接点 ZGA遺伝子はゲノムの安定性を介して,初期発生に寄与する (2019)
  • Author(s): 小川 誠司, 山田 満稔, 浜谷 敏生．
  • Organizer: 第64回日本生殖医学会学術講演・総会
• [Presentation] Zfp371 is a novel DNA repair gene responsible for genome stability during mitosis. (2018)
  • Author(s): Seiji Ogawa, Mitsutoshi Yamada, Toshio Hamatani, Hidenori Akutsu, Akihiro Umezawa, Mamoru Tanaka, Daisuke Aoki.
  • Organizer: The 70th Annual Congress of the Japan Society of Obstetrics and Gynecology, SENDAI INTERNATIONAL CENTER, May 12, 2018.
  • Int'l Joint Research
• [Presentation] 胚性遺伝子KzpiはDifferentially methylated regionsにおけるメチル化を保護して胚発生に寄与する． (2017)
  • Author(s): 上條 慎太郎, 浜谷 敏生, 山田 満稔, 小川 誠司, 阿久津 英憲, 秦 健一郎, 田中 守, 青木 大輔
  • Organizer: 第69回日本産科婦人科学会学術講演会
• [Presentation] 卵子加齢機構の探索と臨床への示唆．シンポジウム「卵子の成熟と老化 －最新知見－ (2017)
  • Author(s): 浜谷敏生
  • Organizer: 第35回日本受精着床学会総会・学術講演会
• [Presentation] 胚培養液について考える (2016)
  • Author(s): 浜谷敏生
  • Organizer: 第22回勝川ART研究会
  • Place of Presentation: ホテルプラザ勝川（愛知県春日井市
  • Invited
• [Presentation] 次世代培養液を考える (2016)
  • Author(s): 浜谷敏生
  • Organizer: 第54回東北生殖医学会総会・学術講演会
  • Place of Presentation: 岩手医科大学創立60周年記念館（盛岡市）
  • Invited