Krivić Denis, Panasiuk Olga, Pilic Johannes, Malli Roland, Bacsa Bernadett, Ćurčić Sanja, Groschner Klaus
Division of Medical Physics and Biophysics, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.
Division of Molecular Biology and Biochemistry, Gottfried Schatz Research Center, Medical University of Graz, Graz, Austria.
Front Immunol. 2025 Jul 24;16:1595036. doi: 10.3389/fimmu.2025.1595036. eCollection 2025.
Photopharmacology has recently emerged as a strategy for high-precision modulation of immune functions. Here we explored efficiency and specificity of interventions based on light-induced TRPC6 activation in the RBL-2H3 mast cell model.
Expression of TRPC6 fusion constructs in RBL-2H3 allowed for generation of temporally well-defined, cytosolic Ca transients in response to photoisomerization of the TRPC6 actuator OptoBI-1. These Ca signals originated exclusively from Ca entry across the plasma membrane. Transient TRPC6 activation in response to UV pulses of 1s duration (3 mW/cm) just exceeded the detection threshold for monitoring of Ca signals within the TRPC6-jRGECO1a nano/microdomain. Activation of TRPC6-jRGECO1a by single, 1s UV light pulses was sufficient to trigger maximal cytosolic to nuclear translocation of NFATc1 (NFAT2) equivalent to the level generated by ionomycin (1 µM)-induced Ca entry. TRPC6 photopharmacology enabled control over NFATc1 nuclear translocation devoid of any detectable degranulation responses.
We report here the exceptionally efficient and specific modulation of mast cell activity by TRPC6 photopharmacology.
光药理学最近已成为一种高精度调节免疫功能的策略。在此，我们在RBL-2H3肥大细胞模型中探索了基于光诱导TRPC6激活的干预措施的效率和特异性。
RBL-2H3中TRPC6融合构建体的表达使得能够响应TRPC6激活剂OptoBI-1的光异构化产生时间上定义明确的胞质钙瞬变。这些钙信号仅源于跨质膜的钙内流。响应持续时间为1秒（3 mW/cm）的紫外线脉冲的瞬时TRPC6激活刚好超过了监测TRPC6-jRGECO1a纳米/微域内钙信号的检测阈值。单次1秒紫外线脉冲激活TRPC6-jRGECO就足以触发NFATc1（NFAT2）从胞质到细胞核的最大转位，其水平相当于离子霉素（1 µM）诱导的钙内流所产生的水平。TRPC6光药理学能够控制NFATc1的核转位，而不会产生任何可检测到的脱颗粒反应。
我们在此报告了通过TRPC6光药理学对肥大细胞活性进行的异常高效且特异性的调节。
