POST-ENDOCYTOTIC INFLAMMATORY SIGNALING AFTER TRAUMA

创伤性损伤包括休克、组织损伤、骨折和输血，释放循环因子(如肿瘤坏死因子α、IL-1和IPS)，促进全身炎症反应。这些药物立即激活循环中的白细胞、内皮细胞和血管细胞上的信号受体，从而启动或改变这些细胞类型及其随后的反应。由配体激活的受体从质膜表面传递信号的初始阶段，是由第二信使、离子通量和蛋白质磷酸化在几分钟内介导的。激活的受体招募接头蛋白，通常与钙离子和局部膜成分的变化相协调。一系列组装事件形成聚集的信号复合体，建立多结构域蛋白支架，同时进行内吞作用和 激活激酶模块，如MAPK和IKK。这些模块导致转录组的许多持续变化，从而影响随后几小时或几天的所有细胞反应，包括细胞周期或细胞凋亡。在白细胞中，MAPK模块调节促炎作用：超氧化物的分泌，中性粒细胞的脱颗粒，细胞因子和趋化因子的合成和释放。在血管内皮细胞和平滑肌细胞中，核因子-kB和AP-1上调白细胞黏附蛋白的表达，促进炎症和渗漏。 细胞生物学家已经知道，笼蛋白介导的内吞作用(CME)可以被 高张力和伯胺阻断(通过影响内体组装或扰乱其成熟)。在几个转化的细胞中，干扰CME可以阻止生物活性介质(如儿茶酚、脂类和肿瘤坏死因子α)传递MAPK或NF-kB信号。然而，目前尚不清楚CME处理的不同阶段是如何通过哺乳动物，特别是人类细胞中内化的受体来调节激酶信号的。此外，在原代细胞中的结果是否反映了动物模型中的炎症信号？值得注意的是，复苏方面的进展表明，高张力在预防促炎启动方面具有额外的优势。另外，一些伯胺和微管效应器似乎在炎症控制方面很有希望。在这项建议中，我们评估抑制内吞作用是否会影响一组排列的罪魁祸首和细胞的内吞后信号。如果是这样的话，这些机制能否提出改进或替代方案？

Traumatic injury involving shock, tissue injury, fracture and transfusion, releases circulating factors (such as TNFalpha, IL-1 and IPS) that promote systemic hyper-inflammation. These agents immediately activate signaling receptors on circulating leukocytes, endothelium and vascular cells, thereby priming or altering these cell-types, and their subsequent responses. The initial phase of signaling by ligand activated receptor from the plasma membrane surface, is mediated by second messengers, ion-fluxes, and protein phosphorylation occurring within minutes. Activated receptors recruit adaptor proteins often coordinated with change in Ca++ and local membrane composition. A series of assembly events forms clustered signaling complexes that build-up multi-domain protein scaffolds which simultaneously conduct endocytosis and activate kinase modules such as MAPKs and IKK. These modules cause many sustained changes to the transcriptome, thereby affecting all subsequent cellular responses for hours or days, including cell-cycling or apoptosis. In leukocytes, MAPK modules regulate pro-inflammatory actions: superoxide secretion, and degranulation in neutrophils, synthesis and release of cytokines and chemokines. In vascular endothelial and smooth muscle cells, NF-kB and AP-1 upregulate expression of leuko-adhesive proteins, promoting inflammation and leak. Cell biologists have known that that clathrin-mediated endocytosis (CME) can be blocked by hypertonicity and primary amines block (by affecting endosome assembly or disrupting its maturation). In several transformed cells, interfering with CME prevents MAPK or NF-kB signaling by bioactive mediators (such as catechols, lipids and TNFalpha). However, it is unclear how the different stages of CME processing regulates kinase signaling by internalized receptors in mammalian, especially human, cells. Further, would the results in primary cells reflect inflammatory signaling in animal models? Remarkably, advances in resuscitation suggest that hypertonicity has additional advantages for preventing pro-inflammatory priming. Separately, some primary amines and microtubule effectors appear promising for inflammation control. In this proposal we assess whether suppressing endocytosis affects postendocytotic signaling for a permuted list of culpable agents and cells. If so, could the mechanisms suggest improvements or alternatives?