The analysis of roles of NLRP7 in molar pregnancies using NLRP7-deleted HTR-8/SVneo cells.

• Project/Area Number: 16K11120
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Obstetrics and gynecology
• Research Institution: Kio University
• Principal Investigator: Maehara Kayoko 畿央大学, 健康科学部, 教授 (80421311)
• Co-Investigator(Kenkyū-buntansha): 祐實 泰子 畿央大学, 健康科学部, 講師 (80425454)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 反復胞状奇胎 / NLRP7遺伝子 / ゲノム編集 / 遺伝子 / 細胞・組織 / 産科学

Outline of Final Research Achievements

Hydatidiform moles (HMs) are abnormal pregnancies with trophoblastic hyperplasia. Recent studies have shown that mutations of NLRP7 gene are associated with recurrent hydatidiform moles (RHMs), although the mechanism underlying molar pregnancies affected by the mutations of NLRP7 remains unknown. To investigate the effects of NLRP7 on cell growth in trophoblasts, we deleted NLRP7 from HTR-8/SVneo cells, a trophoblast cell line, by the CRISPR/Cas9 system. To minimize the off-target effect, D10A mutant nickase version of Cas9 (Cas9D10A) combined with guide RNA (gRNA) pairs having high sequence-specificity evaluated by our developing tool (fcrisprt) was applied. Of 50 clones screened, NLRP7 was successfully disrupted in two clones; clone 1 and clone 2 obtained biallelic 20 bp-deletion and 2 bp-insertion within exon 4 of NLRP7 gene, respectively.

Academic Significance and Societal Importance of the Research Achievements

遺伝子の機能を解析には、細胞やモデル生物（個体）を利用する方法があり、マウスなどのげっ歯類が利用されている。しかしNLPR7遺伝子はげっ歯類に保存されておらず、ノックアウトマウスを用いた解析などができない。近年開発されたゲノム編集を利用することで、NLRP7遺伝子の破壊をヒトの細胞で行い、疾患モデル細胞を作成できた。反復胞状奇胎の病態形成を研究するための有用なツールになると考える。

Research Products

• [Journal Article] Elucidation of the developmental mechanism of ovarian mature cystic teratomas using B allele-frequency plots of single nucleotide polymorphism array data (2018)
  • Author(s): Usui Hirokazu、Nakabayashi Kazuhiko、Kaku Hiroshi、Maehara Kayoko、Hata Kenichiro、Shozu Makio
  • Journal Title: Genes, Chromosomes and Cancer Volume: 57 Issue: 8 Pages: 409-419
  • DOI: 10.1002/gcc.1
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A versatile technology for targeted genome editing has an impact on our lives. (2017)
  • Author(s): 前原 佳代子
  • Journal Title: 畿央大学紀要 Volume: 14 Issue: 1 Pages: 1-8
  • DOI: 10.24482/00000009
  • NAID: 120006323665
  • URL: https://kio.repo.nii.ac.jp/records/18
  • Year and Date: 2017-06-30
  • Peer Reviewed / Open Access
• [Journal Article] Novel Nonsense Mutation in the NLRP7 Gene Associated with Recurrent Hydatidiform Mole. (2015)
  • Author(s): Ito Y, Maehara K, Kaneki E, Matsuoka K, Sugahara N, Miyata T, Kamura H, Yamaguchi Y, Kono A, Nakabayashi K, Migita O, Higashimoto K, Soejima H, Okamoto A, Nakamura H, Kimura T, Wake N, Taniguchi T, Hata K.
  • Journal Title: Gynecol Obstet Invest Volume: Epub ahead of print Issue: 4 Pages: 1-6
  • DOI: 10.1159/000441780
  • Peer Reviewed / Int'l Joint Research
• [Presentation] ヒト早老症モデル細胞の開発 (2018)
  • Author(s): 前原佳代子
  • Organizer: 第36回日本ヒト細胞学会学術集会
  • Invited
• [Book] 学生と考える生命倫理［第2版］ (2018)
  • Author(s): 金子 章道、金内 雅夫、河野 由美、島 恒生、前原 佳代子、堀江 尚子、藤澤 弘枝、福本 貴彦、山本 隆、森岡 周、西井 康惠、訓覇 秋麿、細越 寛樹、小野 尚香、安井 義和
  • Total Pages: 232
  • Publisher: ナカニシヤ出版
  • ISBN: 9784779512216
• [Remarks] 畿央大学教員紹介データベース 前原佳代子
  • URL: http://webinfo.kio.ac.jp/kio1/view2.asp?msg_no=275
• [Remarks] 畿央大学ホームページ
  • URL: http://www.kio.ac.jp/