Stephanou Andreas, Mantri Madhav, Shankaranarayanan Divya, Li Carol, Lagman Mila, Xiang Jenny, Pan Chendong, Sun Yanjie, Muthukumar Thangamani, Machaca Khaled, De Vlaminck Iwijn, Suthanthiran Manikkam
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Ithaca, NY, USA.
Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
bioRxiv. 2025 May 1:2025.04.28.651042. doi: 10.1101/2025.04.28.651042.
Store-operated calcium entry (SOCE) plays a critical role in regulating intracellular calcium signaling and is essential for immune cell functions. SOCE blockade with pyrazole derivative BTP2 has been explored as an anti-inflammatory strategy in preclinical models and Zegocractin (CM4620) is being investigated in Phase 2 clinical trials as an immunoregulatory agent. However, the mode of action and differential effects of SOCE blockade on diverse immune cell types remain largely unknown, limiting the precision of current therapeutic applications. Here, we used multiplexed single-cell RNA sequencing to investigate the effects of two prototypic SOCE blockers, BTP2 and CM4620, on polyclonally-stimulated, normal human peripheral blood mononuclear cells (PBMCs). The data revealed that SOCE blockade suppresses the expression of cytotoxicity-associated genes in CD8 effector T cells and natural killer (NK) cells, restoring them to levels comparable to those in unstimulated cells. Strikingly, SOCE blockade preserved activation-induced expression of anti-inflammatory genes in CD4 regulatory T cells, maintaining their tolerance phenotype even after SOCE blockade. These findings suggest that SOCE blockers modulate immune responses with greater selectivity than conventional immunosuppressants by reducing cytotoxicity while preserving tolerance-associated pathways, highlighting their potential for managing immune-mediated conditions, including organ transplantation.
储存性钙内流（SOCE）在调节细胞内钙信号传导中起关键作用，对免疫细胞功能至关重要。在临床前模型中，已探索使用吡唑衍生物BTP2阻断SOCE作为一种抗炎策略，并且Zegocractin（CM4620）作为一种免疫调节剂正在进行2期临床试验。然而，SOCE阻断对不同免疫细胞类型的作用方式和差异效应在很大程度上仍不清楚，这限制了当前治疗应用的精确性。在此，我们使用多重单细胞RNA测序来研究两种典型的SOCE阻断剂BTP2和CM4620对多克隆刺激的正常人外周血单个核细胞（PBMC）的影响。数据显示，SOCE阻断抑制CD8效应T细胞和自然杀伤（NK）细胞中细胞毒性相关基因的表达，使其恢复到与未刺激细胞相当的水平。令人惊讶的是，SOCE阻断保留了CD4调节性T细胞中抗炎基因的激活诱导表达，即使在SOCE阻断后仍维持其耐受表型。这些发现表明，SOCE阻断剂通过降低细胞毒性同时保留与耐受相关的途径，比传统免疫抑制剂更具选择性地调节免疫反应，突出了它们在管理包括器官移植在内的免疫介导疾病方面的潜力。
