Juksar Juri, Mijdam Rachel, Bosman Sabine, van Oudenaarden Alexander, Carlotti Françoise, de Koning Eelco J P
Hubrecht Institute-KNAW (Royal Netherlands Academy of Arts and Sciences), Utrecht, Netherlands.
Department of Internal Medicine, Leiden University Medical Center, Leiden, Netherlands.
Transpl Int. 2025 Apr 1;38:13422. doi: 10.3389/ti.2025.13422. eCollection 2025.
Diabetes mellitus is characterized by the loss of pancreatic insulin-secreting β-cells in the Islets of Langerhans. Understanding the regenerative potential of human islet cells is relevant in the context of putative restoration of islet function after damage and novel islet cell replacement therapies. Adult human pancreatic tissue can be cultured as three-dimensional organoids with the capacity for long-term expansion and the promise of endocrine cell formation. Here, we characterize the endocrine differentiation potential of human adult pancreatic organoids. Because exocrine-to-endocrine differentiation is dependent on the expression of Neurogenin 3 (NEUROG3), we first generated NEUROG3-inducible organoid lines. We show that doxycycline-induced NEUROG3 expression in the organoids leads to the formation of chromogranin A positive (CHGA+) endocrine progenitor cells. The efficiency of this differentiation was improved with the addition of thyroid hormone T3 and the AXL inhibitor R428. Further, compound screening demonstrated that modifying the pivotal embryonic endocrine pancreas signalling pathways driven by Notch, YAP, and EGFR led to increased NEUROG3 expression in organoids. In a similar fashion to embryonic development, adult ductal cells delaminated from the organoids after NEUROG3 induction. Thus, mechanisms in islet (re)generation including the initiation of endocrine differentiation and delamination can be achieved by NEUROG3 induction.
糖尿病的特征是胰岛中分泌胰岛素的胰腺β细胞丢失。了解人类胰岛细胞的再生潜力对于损伤后胰岛功能的假定恢复以及新型胰岛细胞替代疗法具有重要意义。成人胰腺组织可以培养成具有长期扩增能力和形成内分泌细胞前景的三维类器官。在这里，我们描述了成人人类胰腺类器官的内分泌分化潜力。由于外分泌到内分泌的分化依赖于神经生成素3（NEUROG3）的表达，我们首先生成了NEUROG3诱导型类器官系。我们表明，强力霉素诱导类器官中NEUROG3的表达会导致嗜铬粒蛋白A阳性（CHGA+）内分泌祖细胞的形成。添加甲状腺激素T3和AXL抑制剂R428可提高这种分化的效率。此外，化合物筛选表明，改变由Notch、YAP和EGFR驱动的关键胚胎内分泌胰腺信号通路会导致类器官中NEUROG3表达增加。与胚胎发育类似，NEUROG3诱导后，成人导管细胞从类器官中分层。因此，通过NEUROG3诱导可以实现胰岛（再）生成中的机制，包括内分泌分化的启动和分层。
