2023 Fiscal Year Final Research Report

Development of regenerative therapy for Hirschsprung's disease by extracellular matrix molecules activation and selective neural crest cell transplantation

Research Project

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Project/Area Number	21K08650
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Review Section	Basic Section 55010:General surgery and pediatric surgery-related
Research Institution	Juntendo University
Principal Investigator	Tanaka Nana 順天堂大学, 医学部, 先任准教授 (50530656)
Co-Investigator(Kenkyū-buntansha)	浦尾正彦順天堂大学, 医学部, 教授 (00213504) 宮原克順天堂大学, 大学院医学研究科, 技術員 (00420844) 藤原なほ順天堂大学, 医学部, 准教授 (20589543)
Project Period (FY)	2021-04-01 – 2024-03-31
Keywords	ヒルシュスプルング病 / 腸管神経系 / 腸管神経堤細胞 / 細胞治療
Outline of Final Research Achievements	ENCCs were isolated from Sox10 Venus transgenic mouse gut on embryonic day (E) 18.5 and neurospheres (NS) were generated. Then, NS were transplanted into aganglionic KO and wildtype gut that had been transected just distal to the ENCC wavefront (KO-wf, WT), and into distal KO gut transected at a site equivalent to WT (KO-d) on E12.5. ENCC differentiation was evaluated using whole-mount immunohistochemistry with Tuj-1 and GFAP antibodies. The transplanted ENCCs migrated to form the myenteric and submucosal plexus in all groups. The ratio of the area of Tuj-1 positive cells:GFAP-positive cells in migrated cells in recipient gut was significantly lower in KO-d compared to KO-wf and WT, while there was no significant difference between KO-wf and WT. This suggests that neuronal versus glial differentiation was decreased in KO-d compared to that in KO-wf and WT. Our study showed the differences in ENCC differentiation depending on the site of transplantation.
Free Research Field	小児外科
Academic Significance and Societal Importance of the Research Achievements	ヒルシュスプルング病（H病）では、口側から肛門側へ発達する腸管神経堤細胞（ENCC）が途中で停止することより無神経節腸管が生じるとされている。しかし、無神経節性腸管の腸内微小環境が、正常腸管と、またＨ病近位の神経節節腸管の腸内微小環境とどのように異なるかはいまだ不明である。本研究で我々は、無神経節腸管の異なる部位にENCCを移植することにより、移植部位によるENCCの分化の違いを初めて明らかにした。H病に対する細胞治療をさらに発展させるためには、レシピエントの腸内環境が移植ENCCに与える影響を考慮することが重要である。