REGULATION AND FUNCTION OF MAST CELL SECRETED PROTEINASES

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。肥大细胞是组织对先天性和适应性攻击的免疫应答的协调者。肥大细胞的刺激导致多种促炎介质的释放，包括一类分泌的蛋白酶。这些蛋白酶是组织重塑的核心，组织重塑是生理和病理炎症反应的特征。目前建议的长期目标是了解控制肥大细胞促炎反应的信号通路。从这一理解，我们希望确定新的目标，干预炎症性疾病的进展。我们将讨论三个具体目标。 如上所述，肥大细胞响应于各种促炎刺激，包括IgE介导的和先天性免疫激发，以及促分泌素和神经递质。肥大细胞的这些不同的激活机制有一个关键的共同点，都涉及细胞内游离钙水平的持续升高。事实上，在没有其他刺激的情况下，使用离子载体化合物的钙动员足以诱导肥大细胞活化。允许这种持续钙内流的钙通道是肥大细胞反应的治疗调节的重要靶点，但在分子水平上仍不确定。免疫刺激似乎诱导一个高度选择性的，存储操作的钙电导。然而，肥大细胞的几种其他活化刺激物引起NSCC的活化，其可能由TRP家族的成员编码。PI的实验室已经筛选了啮齿动物和人类肥大细胞的TRP通道mRNA和蛋白质的代表。肥大细胞含有TRPV 2蛋白，其未被描述为对脂质配体产生应答。在肥大细胞中检测到TRPV 4信息，而不是蛋白质，并且该通道的多种激活机制可能包括大麻素。我们检测的肥大细胞不含TRPV 1，但表达大麻素敏感的TRPA 1通道。我们现在提出的实验旨在剖析肥大细胞中TRPA 1的调节和功能。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Mast cells are orchestrators of tissue immune responses to innate and adaptive challenge. Stimulation of mast cells results in the release of diverse pro-inflammatory mediators, including a class of secreted proteinases. These proteinases are central to the tissue remodeling that is a feature of physiological, and pathological, inflammatroy responses. The long-term objective of the current proposal is to understand the signaling pathways that control pro-inflammatory responses in mast cells. From this understanding, we hope to identify novel targets for intervention in the progression of inflammatory diseases. We will address three specific aims. As described above, mast cells respond to various pro-inflammatory stimuli, including IgE-mediated and innate immunological challenges, as well as to secretagogues and neurotransmitters. These diverse activation mechanisms for mast cells have one key commonality; all involve the sustained elevation of intracellular free-calcium levels. In fact, calcium mobilization using ionophore compounds is sufficient to induce mast cell activation in the absence of other stimuli. The calcium channels that permit this sustained calcium influx are important targets for therapeutic modulation of mast cell responses, but remain undefined at the molecular level. Immunological stimuli appear to induce a highly selective, store-operated calcium conductance. However, several other activating stimuli for mast cells cause the activation of NSCC, which may be encoded by members of the TRP family. The PI¿s laboratory has screened rodent and human mast cells for the representation of TRP channel mRNA and protein. Mast cells contain TRPV2 protein, which as not been described to respond to lipid ligands. TRPV4 message, but not protein, has been detected in mast cells, and the diverse activation mechanisms for this channel may encompass cannabinoids. The mast cells that we have examined do not contain TRPV1, but do express the cannabinoid-sensitive TRPA1 channel. The experiments that we now propose will aim to dissect the regulation, and function, of TRPA1 in mast cells.