Research on the effects of infection and inflammation on embryo development

• Project/Area Number: 20H03654
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Osama Woman's and Children's Hospital
• Principal Investigator: Yanagihara Itaru 地方独立行政法人大阪府立病院機構大阪母子医療センター(研究所), 免疫部門, 部長 (60314415)
• Co-Investigator(Kenkyū-buntansha): 西海 史子 地方独立行政法人大阪府立病院機構大阪母子医療センター(研究所), 免疫部門, 流動研究員 (60599596) ; 呉 恒寧 地方独立行政法人大阪府立病院機構大阪母子医療センター(研究所), 免疫部門, 研究技術員 (80648139)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2022: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2021: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2020: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: ウレアプラズマ / 生殖細胞 / 不妊 / ES細胞 / 生殖始原細胞様細胞 / 感染 / 卵母細胞 / 流早産原因細菌 / 病原性解析 / 細胞毒性因子 / マウス卵母細胞 / 発生 / 始原生殖細胞様細胞 / 初期胚発生 / 核酸増幅 / 感染性流早産

Outline of Research at the Start

受精後、胚はゲノムにおけるメチル化の大部分を消去し、その後メチル化状態を再構築する。一連のエピジェネティックな調節は胚発生に必須であり、この過程の異常は不妊や流産を始め先天性インプリント異常症、小児癌の発症にもつながる。感染による炎症は、癌の発症と関連してメチル化異常を起こすが、発生段階におけるメチル化異常症との関連における分子メカニズムは不明である。周産期特有の感染や炎症への曝露がマウス胚における脱メチル化やメチル化によるリプログラミングにどのように影響するのか、ES/iPS細胞からのマウス卵母細胞再構成系を用いて検証し、DNAのメチル化を指標にエピジェネティックな病態への関与を解明する。

Outline of Final Research Achievements

Infertility affects about 15% of couples of reproductive age. Infection of mouse sperm with Ureaplasma inhibited motility in a bacterial lode-dependent manner, and in vitro fertilization using infected mouse sperm allowed Ureaplasma to invade into mouse fertilized eggs and impede their development. We also identified a novel Ureaplasma membrane-disrupting molecule that impaired vacuolar transport in a yeast screen. This factor, named Ureaplasma parvum vacuolating factor (UpVF), evaded ER stress in cervical cancer cells and acted as a cancer enhancer. We also provided epidemiological evidence that Ureaplasma infection in utero is associated with long-term respiratory prognosis in infants with extremely low birth weight.

Academic Significance and Societal Importance of the Research Achievements

我々はこれまで、ウレアプラズマが胎盤における羊膜絨毛膜炎を引き起こし、感染性流早産の最大の起因微生物であることを明らかにした。マウス精子の運動性を菌量依存的に低下させるところや、感染精子を用いたIVFで受精卵の発生が阻害されることを明らかにし、不妊症との関連を科学的に証明することができた。また、後方視的な解析で、胎内ウレアプラズマ感染が早産児の長期的な児の呼吸器予後（小気道閉塞）に関わっていることを世界で初めて明らかにした。超低出生体重児においては、長期的な呼吸器のフォローが必要である。

Research Products

• [Int'l Joint Research] J Craig Venter Institute(米国)
• [Int'l Joint Research] University of the Philippines(フィリピン)
• [Int'l Joint Research] J Craig Venter Institute(米国)
• [Int'l Joint Research] フィリピン大学(フィリピン)
• [Journal Article] Diagnostic accuracy of amniotic fluid interleukin-6 for fetal inflammatory response syndrome. (2023)
  • Author(s): Ohkuma K, Ono T, Oshima Y, So K, Tsumura K, Yamasaki F, Nakura Y, Yanagihara I, Nomiyama M, Yokoyama M.
  • Journal Title: J Obstet Gynaecol Res. Volume: in press Issue: 6 Pages: 1498-1505
  • DOI: 10.1111/jog.15630
  • Peer Reviewed
• [Journal Article] Relationship of maternal inflammatory response and fetal inflammatory response to duration and intensity of intra-amniotic infection and inflammation (2023)
  • Author(s): Gondo K, Yamasaki F, Nomiyama M, Hisamoto N, Yamashita N, Nakagawa T, Ikeda M, Tsuda S, Ishimatsu M, Oshima Y, Ono T, Kozuma Y, Nakura Y, Yanagihara I, Tsumura K
  • Journal Title: Placenta Volume: 137 Pages: 23-30
  • DOI: 10.1016/j.placenta.2023.04.003
  • Peer Reviewed
• [Journal Article] Genetic Characterization of Ovarian Tumor Tissues from Patients with Epithelial Ovarian Cancer in a Philippine Tertiary Hospital: A Descriptive Study (2023)
  • Author(s): Lintao RCV, Padua AJP, Nakura Y, Llamas-Clark EF, Yanagihara I.
  • Journal Title: Asian Pac J Cancer Prev . Volume: 24 Issue: 4 Pages: 1187-1197
  • DOI: 10.31557/apjcp.2023.24.4.1187
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Estimated time to emergence of secondary intra-amniotic infection or inflammation since the onset of the preterm premature rupture of membranes (2022)
  • Author(s): Tsuda S, Shinagawa T, Tsumura K, So K, Yamasaki F, Kawaguchi A, Nakura Y, Yanagihara I, Nomiyama M, Yokoyama M.
  • Journal Title: Taiwan J Obstet Gynecol Volume: 61（4） Issue: 4 Pages: 634-640
  • DOI: 10.1016/j.tjog.2022.01.002
  • Peer Reviewed / Open Access
• [Journal Article] A fatal case of hemophagocytic lymphohistiocytosis associated with gestational psittacosis without symptoms of pneumonia (2022)
  • Author(s): Yoshimura Michinobu、Shimizu Kanako、Nakura Yukiko、Kawahara Kazumi、Katano Harutaka、Motooka Daisuke、Takeuchi Makoto、Nagamune Kisaburo、Imamura Yoshiaki、Nakamura Shota、Yasukawa Kiyoshi、Hasegawa Hideki、Yoshida Yoshio、Yanagihara Itaru
  • Journal Title: Journal of Obstetrics and Gynaecology Research Volume: 48 Issue: 12 Pages: 3325-3330
  • DOI: 10.1111/jog.15429
  • Peer Reviewed / Open Access
• [Journal Article] Co-infection of human papillomavirus and other sexually transmitted bacteria in cervical cancer patients in the Philippines (2022)
  • Author(s): Tantengco OAG, Nakura Y, Yoshimura M, Nishiumi F, Llamas-Clark EF, Yanagihara I.
  • Journal Title: Gynecol Oncol Rep Volume: 40 Pages: 100943-100943
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Urine Desmosine as a Novel Biomarker for Bronchopulmonary Dysplasia and Postprematurity Respiratory Disease in Extremely Preterm or Low Birth Weight Infants (2022)
  • Author(s): Hirata K, Nishikawa M, Nozaki M, Kitajima H, Yanagihara I, Wada K, Fujimura M.
  • Journal Title: Am J Perinatol Volume: in press Issue: S 01 Pages: e1030-e1036
  • DOI: 10.1055/a-1979-8501
  • Peer Reviewed
• [Journal Article] Intrauterine Ureaplasma is associated with small airway obstruction in extremely preterm infants (2022)
  • Author(s): Kitajima H, Fujimura M, Takeuchi M, Kawamoto Y, Sumi K, Matsunami K, Shiraishi J, Hirano S, Nakura Y, Yanagihara I.
  • Journal Title: Ped Pulmonol Volume: 57(11) Issue: 11 Pages: 1-11
  • DOI: 10.1002/ppul.26098
  • Peer Reviewed / Open Access
• [Journal Article] A case of a preschool child with a successful kidney transplant following the long-term administration of antibiotics to treat peritoneal dialysis-related ESI/peritonitis by Mycobacterium abscessus (2022)
  • Author(s): Nakano S, Yamamura-Miyazaki N, Michigami T, Yazawa K, Yanagihara I, Yamamoto K.
  • Journal Title: CEN Case Rep Volume: 11(4) Issue: 4 Pages: 408-411
  • DOI: 10.1007/s13730-022-00689-z
  • Peer Reviewed
• [Journal Article] Blockade of endoplasmic reticulum stress‐induced cell death by Ureaplasma parvum vacuolating factor (2021)
  • Author(s): Nishiumi Fumiko、Kawai Yasuhiro、Nakura Yukiko、Yoshimura Michinobu、Wu Heng Ning、Hamaguchi Mitsuhide、Kakizawa Shigeyuki、Suzuki Yo、Glass John I.、Yanagihara Itaru
  • Journal Title: Cellular Microbiology Volume: 23 Issue: 12 Pages: 1-19
  • DOI: 10.1111/cmi.13392
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Development of an efficient one-step real-time reverse transcription polymerase chain reaction method for severe acute respiratory syndrome-coronavirus-2 detection (2021)
  • Author(s): Nakura Yukiko、Wu Heng Ning、Okamoto Yuya、Takeuchi Muneyuki、Suzuki Koichiro、Tamura Yoshitaka、Oba Yuichiro、Nishiumi Fumiko、Hatori Nobuaki、Fujiwara Shinsuke、Yasukawa Kiyoshi、Ida Shinobu、Yanagihara Itaru
  • Journal Title: PLOS ONE Volume: 16 Issue: 6 Pages: e0252789-e0252789
  • DOI: 10.1371/journal.pone.0252789
  • NAID: 120007041647
  • Peer Reviewed / Open Access
• [Journal Article] Validation of the loop-mediated isothermal amplification method for rapid and sensitive detection of Ureaplasma species in respiratory tracts of preterm infants (2021)
  • Author(s): Mikami Yuta、Fuwa Kazumasa、Arima Eriko、Suda Yasuo、Yanagihara Itaru、Ibara Satoshi
  • Journal Title: PLOS ONE Volume: 16 Issue: 3 Pages: e0247618-e0247618
  • DOI: 10.1371/journal.pone.0247618
  • Peer Reviewed
• [Journal Article] リアルタイムPCR・デジタルPCRの臨床診断への応用 (2021)
  • Author(s): 柳原格
  • Journal Title: 臨床検査 Volume: 45 Pages: 1142-1147
• [Journal Article] Clostridium tertium Peritonitis and Bacteremia in a Neonate With Congenital Intestinal Atresia: A Case Report (2021)
  • Author(s): Morikawa K, Nozaki M, Hirata K, Abe T, Yanagihara I, Wada K
  • Journal Title: The Pediatric Infectious Disease Journal Volume: 40(2) Issue: 2 Pages: 159-161
  • DOI: 10.1097/inf.0000000000002920
  • Peer Reviewed
• [Journal Article] The ability of Ureaplasma parvum to invade mouse sperm, fertilize eggs through infected sperm, impair mouse sperm function, and embryo development (2020)
  • Author(s): Ito K, Akai K, Nishiumi F, Nakura Y, Wu HN, Kurata T, Onodera A, Kawai Y, Kajiyama S, Yanagihara I
  • Journal Title: F&S Science Volume: 2(1) Issue: 1 Pages: 13-23
  • DOI: 10.1016/j.xfss.2020.12.003
  • Peer Reviewed
• [Presentation] ヒト流早産原因細菌ウレアプラズマの宿主細胞への影響 (2022)
  • Author(s): 柳原格、西海史子
  • Organizer: 近畿腸管微生物研究会
  • Invited
• [Presentation] 新型コロナウイルスの核酸増幅法と変異型ゲノム解析 (2022)
  • Author(s): 柳原格
  • Organizer: 第235回 大阪小児科学会
  • Invited
• [Presentation] 耐熱性DNAポリメラーゼの鎖置換活性に関与する領域の比較解析 (2022)
  • Author(s): 井上瑛介、柳原格、保川清、藤原伸介、福田青郎
  • Organizer: 第95回日本生化学会
• [Presentation] 病理解剖により診断に至った妊娠オウム病の1例 (2022)
  • Author(s): 佐藤由美子、井関明子、杉井裕和、阿萬紫、柳原格、若狭朋子
  • Organizer: 第68回日本病理学会
• [Presentation] 合成ゲノム生物のもたらすサイエンスと安全性 (2022)
  • Author(s): 柳原格、高屋明子
  • Organizer: 第96回日本細菌学会シンポジウム（S９）
  • Invited
• [Presentation] ウレアプラズマの病原性解析 (2021)
  • Author(s): 柳原格
  • Organizer: 第48回 日本マイコプラズマ学会学術集会
  • Invited
• [Presentation] U. parvum感染による細胞障害と新規遺伝子の機能解析 (2021)
  • Author(s): 西海史子, 河合泰弘、名倉由起子、吉村芳修、呉恒寧、柳原格
  • Organizer: 第48回 日本マイコプラズマ学会学術集会
• [Presentation] Ureaplasma parvumのマウス精子からの除去 (2021)
  • Author(s): 伊東和俊、西海史子、名倉由起子、呉恒寧、柳原格
  • Organizer: 第48回 日本マイコプラズマ学会学術集会
• [Presentation] Cell in cell observed in Syn3.0 and reconstruction of attachment ability and pathogenicity to HeLa cells (2021)
  • Author(s): Shigeyuki Kakizawa, Itaru Yanagihara
  • Organizer: Minimal Cell Workshop
  • Int'l Joint Research / Invited
• [Presentation] 高い正電荷数を有する分岐鎖ポリアミンを用いた抗菌剤の開発 (2021)
  • Author(s): 廣田裕介、福田青郎、山内光陽、増尾貞弘、柳原格、林翰佳、藤原伸介
  • Organizer: 第73回日本生物工学会大会
• [Presentation] RNA検出を目指したThermus thermophilus由来DNAポリメラーゼの改変 (2021)
  • Author(s): 安藤友理、保川清、柳原格、福田青郎、藤原伸介
  • Organizer: 第73回日本生物工学会大会
• [Presentation] 新規鎖置換型DNAポリメラーゼ生産菌の探索と酵素活性の比較 (2021)
  • Author(s): 井上瑛介、福田青郎、保川清、柳原格、藤原伸介
  • Organizer: 第73回日本生物工学会大会
• [Presentation] 新規開発one-step real-time RT-PCR試薬の性能評価 (2021)
  • Author(s): 羽場 暢晃、鈴木孝一朗、高橋美恵、名倉由起子、保川清、藤原伸介、柳原格
  • Organizer: 第40回日本衛生検査所協会近畿支部学術研究発表会
• [Presentation] Development and optimization of reaction condition of recombinase polymerase amplification to detect SARS-CoV-2 DNA and RNA using a statistical method. (2021)
  • Author(s): Kevin Maafu Juma, Teisuke Takita, Kenji Ito, Masaya Yamagata, Shihomi Akagi, Emi Arikawa, Kenji Kojima, Manish Biyani, Shinsuke Fujiwara, Yukiko Nakura, Itaru Yanagihara, Kiyoshi Yasukawa
  • Organizer: 日本生物高分子学会2021年度大会
• [Presentation] Identification of novel endonucleases from Ureaplasma parvum SV3F4 strain (2021)
  • Author(s): Yuya Fujisawa, Heng Ning Wu, Tetsuya Matsukawa, Itaru Yanagihara, Shin’ichiro Kajiyama
  • Organizer: International Chemical Congress of Pacifichem 2021
  • Int'l Joint Research
• [Presentation] Molecular Diagnostic Tests of COVID-19 (2021)
  • Author(s): Itaru Yanagihara, Yoshitaka Tamura, Yuichiro Oba, Muneyuki Takeuchi, Shinobu Ida, Yukiko Nakura, Koichiro Suzuk2, Shinsuke Fujiwara, Kiyoshi Yasukawa
  • Organizer: 日本農芸化学会2021年度大会
  • Int'l Joint Research / Invited
• [Presentation] U. parvum感染による細胞障害性と新規遺伝子の機能解析 (2020)
  • Author(s): 西海 史子、河合 泰弘、名倉 由起子、吉村 芳修、呉 恒寧、柳原 格
  • Organizer: 第47回日本マイコプラズマ学会学術集会
• [Presentation] Ureaplasma parvum感染のマウス精子のDNAと細胞膜への影響 (2020)
  • Author(s): 伊東 和俊、西海 史子、名倉 由起子、呉 恒寧、柳原 格
  • Organizer: 第47回日本マイコプラズマ学会学術集会
• [Presentation] 当部門で同定した泌尿生殖器マイコプラズマの菌種について (2020)
  • Author(s): 名倉 由起子、柳原 格
  • Organizer: 第47回日本マイコプラズマ学会学術集会
• [Remarks] 大阪母子医療センター研究所免疫部門
  • URL: https://www.wch.opho.jp/research/developmental
• [Remarks] 大阪母子医療センター研究所免疫部門
  • URL: https://www.wch.opho.jp/research/developmental/index.html
• [Patent(Industrial Property Rights)] ワンステップRT-PCR反応用組成物／キット (2021)
  • Inventor(s): 柳原格、名倉由起子他
  • Industrial Property Rights Holder: 柳原格、名倉由起子他
  • Industrial Property Rights Type: 特許
  • Filing Date: 2021