Expression, function and endogenous modulators of the transient receptor potential (TRP) channels TRPV1, TRPV4, TRPM8 und TRPA1 in human corneal endothelial cells

人角膜内皮细胞瞬时受体电位（TRP）通道TRPV1、TRPV4、TRPM8和TRPA1的表达、功能和内源性调节剂

• 批准号: 288923337
• 负责人: Privatdozent Dr. Stefan Mergler
• 金额: --
• 依托单位: Institut für Anatomie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/288923337?language=en
• 关键词: Expression; function; endogenous; modulators; transient

The human corneal endothelium (HCE) fulfills various required functions mandatory for the maintenance of corneal transparency and normal vision. These functions are dependent on cellular mechanisms that mediate regulation of intracellular calcium levels. Transient receptor potential channels (TRPs) play a substantial role in this process. Recent studies showed that functional TRP channel subtype activity of TRPV1 and TRPM8 modulate apoptosis (calcium-apoptosis link). Therefore, further studies are warranted to determine how modulation of these TRP subtypes contributes to the support of HCE function. In recent studies, the applicants also demonstrated for the first time that HCE cells express other TRP subtypes that include TRPV4 as well as TRPA1. Current investigations confirmed functional expression of TRPM8 and TRPA1 also in primary HCE cell cultures. Furthermore, our results indicate that these TRPs play a role in the temperature-dependent and osmosensitive regulation of intracellular calcium concentration. The aim of this project is to determine the properties of TRPs in regulating corneal endothelial-relevant biological processes and to identify endogenous TRP channel activity modulators, using highly sensitive methods (Ca2+ imaging, patch-clamp, qPCR). In the established in vitro HCEC-12 cell model and fresh and cultured primary HCE cell cultures, we plan to delineate the molecular, functional and pharmacological properties of TRPs as well as their linked signaling pathways, which mediate control of responses that are essential for tissue viability. In addition, we will define mechanisms of functional cross-talk between TRPV1, TRPV4, TRPM8 and TRPA1 and the HCE-relevant membrane cognate receptors for EGF and FGF-2 in the HCE. Furthermore, the impact of long-term exposure to ophthalmic drugs on TRP channel function and regulation and its implication on corneal endothelial function will be investigated. Overall, the results of this study will contribute to a better understanding of HCE biology and hold promise to develop novel strategies for prolonging and improving corneal endothelial cell function before and after keratoplasty and possibly also during long-term exposure to clinically proven effective ophthalmological medications.

人角膜内皮（HCE）具有维持角膜透明性和正常视力所必需的各种功能。这些功能依赖于介导细胞内钙水平调节的细胞机制。瞬时受体电位通道（TRP）在这一过程中发挥了重要作用。最近的研究表明，TRPV 1和TRPM 8的功能性TRP通道亚型活性调节细胞凋亡（钙-细胞凋亡链接）。因此，需要进一步研究以确定这些TRP亚型的调节如何有助于支持HCE功能。在最近的研究中，申请人还首次证明HCE细胞表达其他TRP亚型，包括TRPV 4以及TRPA 1。目前的研究证实了TRPM 8和TRPA 1在原代HCE细胞培养物中也有功能性表达。此外，我们的研究结果表明，这些TRP在细胞内钙浓度的温度依赖性和温度敏感性调节中发挥作用。本项目的目的是确定TRP在调节角膜内皮相关生物过程中的特性，并使用高灵敏度方法（Ca 2+成像，膜片钳，qPCR）鉴定内源性TRP通道活性调节剂。在建立的体外HCEC-12细胞模型和新鲜和培养的原代HCE细胞培养物中，我们计划描述TRP的分子，功能和药理学特性以及它们相关的信号传导通路，其介导对组织活力至关重要的反应的控制。此外，我们将定义TRPV 1，TRPV 4，TRPM 8和TRPA 1和HCE相关的EGF和FGF-2的膜同源受体之间的功能性串扰的机制。此外，将研究长期暴露于眼科药物对TRP通道功能和调节的影响及其对角膜内皮功能的影响。总体而言，本研究的结果将有助于更好地了解HCE生物学，并有望开发新的策略，用于在角膜移植术前后延长和改善角膜内皮细胞功能，也可能在长期暴露于临床证明有效的眼科药物期间。

项目成果

Vascular Endothelial Growth Factor (VEGF) Induced Downstream Responses to Transient Receptor Potential Vanilloid 1 (TRPV1) and 3-Iodothyronamine (3-T1AM) in Human Corneal Keratocytes

• DOI: 10.3389/fendo.2018.00670
• 发表时间: 2018-11
• 期刊: Frontiers in Endocrinology
• 影响因子: 5.2
• 作者: Ersal Türker;F. Garreis;N. Khajavi;P. Reinach;P. Joshi;T. Brockmann;Alexander Lucius;Nina Ljubojevic;Elizabeth Turan;D. Cooper;F. Schick;Rob Reinholz;U. Pleyer;J. Köhrle;S. Mergler