Degroote Laure, Martens Pieter-Jan, Viaene Marijke, Heremans Yves, Leuckx Gunter, Geukens Nick, De Leu Nico, Staels Willem, Mathieu Chantal, Gysemans Conny
Leuven Diabetes Lab, Clinical and Experimental Endocrinology (CEE), CHROMETA, KU Leuven, Leuven, Belgium.
Genetics, Reproduction, and Development (GRAD), Beta Cell Neogenesis (BENE) Research Unit, Vrije Universiteit Brussel (VUB), Brussels, Belgium.
Diabetologia. 2025 Jul 12. doi: 10.1007/s00125-025-06490-8.
AIMS/HYPOTHESIS: Verapamil, a calcium channel blocker, and low doses of anti-thymocyte globulin (ATG) have individually shown efficacy in preserving beta cell function in people with recent-onset symptomatic type 1 diabetes (stage 3). We hypothesised that combining interventions with complementary modes of action and different targets would increase their efficacy in arresting beta cell demise and promoting disease recovery.
Continuous administration of verapamil via drinking water, combined with a short course of low-dose rabbit-anti-mouse ATG (mATG), was studied in female recent-onset diabetic NOD mice for its potential to induce disease remission and mechanism of action.
Verapamil stably reversed diabetes in 3 out of 15 mice (20%) by day 56 after therapy start. Low-dose mATG reversed diabetes in 7 out of 18 mice (39%) by day 7 after therapy start, yet the effect waned to 3 out of 18 mice (17%) by day 56. The combination of verapamil with mATG induced durable diabetes reversal in 9 out of 20 mice (45%) by day 56, which was associated with preserved beta cell function, higher pancreatic insulin content and increased total beta cell volume with decreased severe insulitis. mATG, both alone and in combination, induced a temporary depletion of lymphocytes in peripheral blood on day 3 after therapy start, which largely recovered by day 14, when naive cells had shifted to a memory phenotype in both CD4 and CD8 T cells. Only in combination-treated mice was a higher CD4 regulatory T cell to CD8 effector memory T cell ratio observed in the pancreatic draining lymph nodes. The expression of the glucose-induced gene encoding thioredoxin-interacting protein (Txnip), a key regulator of beta cell apoptosis and dysfunction, was reduced in pancreatic beta cells in reversed mice, irrespective of whether they received verapamil or not.
CONCLUSIONS/INTERPRETATION: The combination of verapamil and low-dose mATG outperformed monotherapy in reversing recent-onset type 1 diabetes in NOD mice. This approach targets both the beta cell and immune axes, suggesting a promising strategy for disease reversal in human type 1 diabetes.
目的/假设：维拉帕米是一种钙通道阻滞剂，低剂量抗胸腺细胞球蛋白（ATG）单独使用时，已显示出对近期出现症状的1型糖尿病患者（3期）β细胞功能的保护作用。我们推测，将具有互补作用方式和不同靶点的干预措施联合使用，可提高其阻止β细胞死亡和促进疾病恢复的效果。
在近期发病的雌性糖尿病NOD小鼠中，研究通过饮水持续给予维拉帕米并联合短疗程低剂量兔抗小鼠ATG（mATG）诱导疾病缓解的潜力及其作用机制。
治疗开始后第56天，15只小鼠中有3只（20%）通过维拉帕米治疗实现了糖尿病的稳定逆转。治疗开始后第7天，低剂量mATG使18只小鼠中的7只（39%）糖尿病得到逆转，但到第56天时，这一效果降至18只小鼠中的3只（17%）。维拉帕米与mATG联合使用，在治疗开始后第56天，使20只小鼠中的9只（45%）实现了持久的糖尿病逆转，这与β细胞功能的保留、胰腺胰岛素含量增加、总β细胞体积增大以及严重胰岛炎减轻有关。单独使用mATG及其联合使用时，在治疗开始后第3天均导致外周血淋巴细胞暂时减少，到第14天时基本恢复，此时CD4和CD8 T细胞中的幼稚细胞已转变为记忆表型。仅在联合治疗的小鼠胰腺引流淋巴结中观察到较高的CD4调节性T细胞与CD8效应记忆性T细胞比例。无论是否接受维拉帕米治疗，逆转的小鼠胰腺β细胞中编码硫氧还蛋白相互作用蛋白（Txnip）的葡萄糖诱导基因的表达均降低，Txnip是β细胞凋亡和功能障碍的关键调节因子。
结论/解读：在逆转NOD小鼠近期发病的1型糖尿病方面，维拉帕米与低剂量mATG联合使用优于单一疗法。这种方法同时针对β细胞和免疫轴，提示这是一种在人类1型糖尿病疾病逆转方面有前景的策略。
