Role of CD34+ endothelial cells in in vivo testis formation and in vitro reconstruction of testis

• Project/Area Number: 18K06322
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 44040:Morphology and anatomical structure-related
• Research Institution: Kumamoto Health Science University
• Principal Investigator: Abe Shin-ichi 熊本保健科学大学, 保健科学部, 客員研究員 (90109637)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 精巣形成 / 3次元培養 / organoid / 間質 / CD34 / telocyte / integrin a4b1 / VCAM1 / integrin a4, a9 / 精細管様構造 / 再凝集培養 / integrin / 再凝集 / KSR / ３次元培養 / 再構築 / CD34+内皮細胞 / マウス

Outline of Final Research Achievements

We analyzed the mechanism whereby Leydig cells (LCs) are localized within the re-aggregates of CD34+ cells, which are the main component of the testicular interstitium, in a 3-D re-aggregate culture in which successful reconstruction of testicular structures had been established. While LCs have adult type (ALCs) and fetal type (FLCs), most of the LCs localized within the CD34+ cell-re-aggregates were ALCs. Live cell imaging showed that CD34+ cells on the dish displayed chemotactic behavior toward ALC-re-aggregates, and the behavior was inhibited by VCAM1 antibody. Combined with expression studies by immunostaining, these results indicate that VCAM1-a4b1 integrin interaction between CD34+ cells (telocytes) and ALCs plays an important role in the formation, maintenance and regeneration of the structure of testicular interstitium. The testicular organoid model that we established will contribute to the analysis of the mechanism of spermatogenesis and also to medical application.

Academic Significance and Societal Importance of the Research Achievements

近年、精巣を含め哺乳類のいろいろな器官の間質にCD34+PDGFRa+ telocytesが存在し、周りの細胞とcommunicationを取っているという形態学的報告がある。再凝集培養系を用いて、精巣の間質様構造の再構築にCD34+細胞とadult LC間のVCAM1-a4b1 integrin相互作用が重要な役割を果たしていることを示唆した今回の結果は、生体におけるLCの分化・再生におけるtelocytesの機能的役割を示唆する先駆的な報告と言えるだろう。また、我々が確立した精巣organoidが精巣再構築の解析に有用なツールとなることが証明されたことから、今後医学的な応用も期待される。

Research Products

• [Int'l Joint Research] ZunYi Medical University(中国)
• [Int'l Joint Research] Monash University/Monash Biomedicine Discovery Institute/Hudson Institute of Medical Research(オーストラリア)
• [Int'l Joint Research] Cardiff University(英国)
• [Journal Article] CD34 is expressed in endothelial cells in embryonic testes and is additionally expressed in non-endothelial cells in postnatal mouse testes (2022)
  • Author(s): Abe K, Kameyama H, Abe S-I
  • Journal Title: Zoological Science Volume: in press
  • Peer Reviewed
• [Journal Article] VCAM1-a4b1 integrin interaction mediates interstitial tissue reconstruction in 3-D re-aggregate culture of dissociated prepubertal mouse testicular cells (2021)
  • Author(s): Abe K, Kon S, Kameyama H, Zhang J, Morohashi K, Shimamura K, Abe S-I
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 18332-18332
  • DOI: 10.1038/s41598-021-97729-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mesenchymal Niche-Derived Neuregulin-1 Drives Intestinal Stem Cell Proliferation and Regeneration of Damaged Epithelium. (2020)
  • Author(s): Jarde, T., Chan, W.H., Rossello, F.J., Kahlon, T.K., Theocharous, M., Arackal, T.K., Flores, T., Giraud, M., Richards, E., Chan, E., Kerr, G., Engel, R.M., Prasko, M., Donoghue, J.F., Abe, S-I., Phesse, T.J., Nefzger, C.M., McMurrick, P.J., Powell, D.R., Daly R.J., Polo, J.M., Abud, H.E.
  • Journal Title: Cell Stem Cell Volume: 27 Issue: 4 Pages: 646-662
  • DOI: 10.1016/j.stem.2020.06.021
  • Peer Reviewed / Int'l Joint Research
• [Presentation] ３次元培養によるマウス精細管様構造再構築におけるPDGFとNeurotrophin signalingの役割について (2019)
  • Author(s): 安部眞一、安部和子、亀山広喜
  • Organizer: 第90回日本動物学会大会（大阪）
• [Presentation] ３次元培養によるマウス精細管様構造再構築におけるPDGFとNeurotrophin signalingの役割について (2018)
  • Author(s): 安部眞一、安部和子
  • Organizer: 日本動物学会