The role of c-Rel in endothelial mechanobiology and atherosclerosis

c-Rel 在内皮力学生物学和动脉粥样硬化中的作用

• 批准号: 1812143
• 金额: --
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1812143
• 关键词: role; Rel; endothelial; mechanobiology; atherosclerosis

Summary - Atherosclerosis develops preferentially at branches and bends of arteries exposed to low shear stress. Evans' group recently demonstrated that the transcription factor c-Rel was expressed preferentially at the low shear site, suggesting a potential role in regulating atherogenesis. The function of c-Rel in endothelial cells (EC) is poorly described but it is known to promote anti-apoptotic signalling in epithelial and other cell types. This Studentship will investigate the function of c-Rel in EC exposed to flow using in vitro systems and murine models. The influence of c-Rel on lesion patterning will be studied by deleting c-Rel from EC in hypercholesterolemic mice. The study brings together Evans' knowledge of endothelial mechanobiology with Oakley's expertise in c-Rel signalling.Hypothesis - It is hypothesised that c-Rel protects against lesion formation by limiting EC apoptosis and dysfunction at low shear sites.Experimental aims1. Does c-Rel reduce injury and dysfucntion of cultured EC exposed to flow? Cultured EC will be exposed to low or high shear using systems established in Evans' lab (parallel plate and orbital flow). The expression of c-Rel will be silenced using siRNA or enhanced by delivery of an expression plasmid containing c-Rel cDNA (available in Oakley's lab). Apoptosis will be assessed by cleavage of Caspase-3 and TUNEL and senescence (known to be induced by low shear) will be studied by measuring SA-b-Gal, p53 and p16. We predict that c-Rel silencing will enhance apoptosis and senescence in sheared EC and overexpression of c-Rel will have the opposite effect.2. Does c-Rel protect against EC dysfucntion at low shear sites in vivo? Wild-type mice will be compared with c-Rel knockout mice (available in Oakley's lab). En face staining for cleaved caspase-3 or TUNEL (apoptosis) and SA-b-Gal/p53/p16 (senescence) will be studied in EC at low (inner curvature of aortic arch) and high (outer curvature) shear sites. It is predicted that c-Rel deletion will enhance EC apoptosis and senescence at low shear sites.3. Does c-Rel regulate the spatial localisation of atherosclerosis? c-Rel knockout mice will be studied. Hypercholesterolemia will be induced using an adenovirus containing PCSK9 (recently established in Sheffield by Evans). After 6-12 weeks, lesion size will be assessed by oil red O staining of the aorta and aortic root. Plaque cellular composition will be studied by immunohistochemistry. It is predicted that c-Rel deletion will alter the patterning of atherosclerosis by enhancing lesion formation at low shear sites.Value -The work will identify c-Rel as a potential therapeutic target in early atherosclerosis.

总结-动脉粥样硬化优先发生在暴露于低剪切应力的动脉分支和弯曲处。Evans的研究小组最近证实，转录因子c-Rel在低剪切位点优先表达，提示其在调节动脉粥样硬化形成中的潜在作用。c-Rel在内皮细胞（EC）中的功能描述不多，但已知其在上皮和其他细胞类型中促进抗凋亡信号传导。本研究将使用体外系统和小鼠模型研究c-Rel在暴露于血流的EC中的功能。将通过在高胆固醇血症小鼠中从EC中删除c-Rel来研究c-Rel对损伤模式的影响。该研究将Evans的内皮机械生物学知识与奥克利在c-Rel信号传导方面的专业知识结合在一起。假设-假设c-Rel通过限制低剪切位点的EC凋亡和功能障碍来防止损伤形成。c-Rel是否能减少暴露于流动的培养EC的损伤和功能障碍？使用Evans实验室建立的系统（平行板和轨道流）将培养的EC暴露于低或高剪切。c-Rel的表达将使用siRNA沉默或通过递送含有c-Rel cDNA的表达质粒（可在奥克利的实验室获得）来增强。将通过半胱天冬酶-3和TUNEL的切割来评估细胞凋亡，并将通过测量SA-b-Gal、p53和p16来研究衰老（已知由低剪切诱导）。我们预测c-Rel基因的沉默将促进剪切内皮细胞的凋亡和衰老，而c-Rel基因的过表达将产生相反的作用. c-Rel是否能在体内保护低剪切位点的EC功能障碍？将野生型小鼠与c-Rel敲除小鼠（可在奥克利实验室获得）进行比较。将在EC中在低（主动脉弓的内曲率）和高（外曲率）剪切位点研究切割的半胱天冬酶-3或TUNEL（凋亡）和SA-b-Gal/p53/p16（衰老）的表面染色。预计c-Rel缺失将增强低剪切位点EC的凋亡和衰老。3. c-Rel是否调节动脉粥样硬化的空间定位？将研究c-Rel敲除小鼠。使用含有PCSK 9的腺病毒（最近由Evans在谢菲尔德建立）诱导高胆固醇血症。6-12周后，将通过主动脉和主动脉根部的油红O染色评估病变大小。将通过免疫组织化学研究斑块细胞组成。据预测，c-Rel缺失将改变动脉粥样硬化的模式，通过增强病变形成在低剪切sites.Value -这项工作将确定c-Rel作为一个潜在的治疗目标，在早期动脉粥样硬化。