Immortalization of normal human neudoendocrine cells

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K22670
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 55:Surgery of the organs maintaining homeostasis and related fields
• Research Institution: National Cancer Center Japan
• Principal Investigator: Kiyono Tohru 国立研究開発法人国立がん研究センター, 先端医療開発センター, プロジェクトリーダー (10186356)
• Project Period (FY): 2019-06-28 – 2022-03-31
• Keywords: 不死化 / 神経内分泌細胞

Outline of Final Research Achievements

Among normal human cells, neuroendocrine cells are most difficult to cultivate and immortalize, and thus hormone replacement therapy by using hormone-producing cell transplantation has not been available. In this study, normal human pancreatic islet cells and hormone-producing pituitary adenoma cells have been cultivated and immortalized by transducing mutant CDK4, cyclin D1 and TERT. For the future hormone-producing cell transplant therapy, we planned to immortalize normal ACTH-producing cells. So far, three iPS cell clones transduced with a PiggyBAC vector conditionally expressing mutant CDK4 and cyclin D1 were established, and now are subjected to differentiation into pituitary organoid. Mutant CDK4 and cyclin D1 as well as mCheery are designed to be induced by doxycyclin, and their induction was confirmed upon dox treatment. A clone from which ACTH secretion was enhanced upon doxycyclin administration was obtained. Now we are trying to immortalize them by TERT transduction.

Free Research Field

ウイルス腫瘍学

Academic Significance and Societal Importance of the Research Achievements

膵島や下垂体などのペプチドホルモンを産生する神経内分泌系細胞を単離培養できれば、細胞移植療法による補充治療に直結する。しかし神経内分泌細胞の単離、培養、増幅は難しく、機能性神経内分泌細胞株はほとんどない。膵島細胞や、下垂体腺腫細胞に変異CDK4, cyclin D1, TERTの3遺伝子の導入により不死化することには成功したが、ホルモン産生能が低下していた。そこで、より実用化を促進するためiPS細胞から分化したホルモン産生細胞を増幅不死化する方法を考案した。iPS細胞から分化誘導した神経内分泌細胞の増幅・不死化法を確立できれば細胞移植療法による補充治療への実用化を促進することができる。