Understanding vascular aging-related dementia through medin signaling

摘要 年龄是心脑血管疾病和痴呆症最重要的危险因素。 流行病学、临床前和临床数据表明，血管疾病与痴呆症密切相关 疾病，包括阿尔茨海默病(AD)和与AD相关的疾病，如血管性痴呆(VAD)。这个 血管疾病、衰老和痴呆症之间的机制联系仍然知之甚少。的确有 越来越多的证据表明，麦迪因，一种由50个氨基酸组成的多肽，形成了最常见但研究最少的多肽之一 人类淀粉样变性是血管衰老病理的重要驱动因素。麦迪素在血管系统中积聚 随着年龄的增长，并与AD、VAD和主动脉疾病有关。对Medin的作用机制知之甚少 诱导细胞和组织损伤，目前尚无Medin病理学的动物模型。我们向梅林展示了 部分通过受体损伤血管内皮细胞功能和细胞活性并诱导促炎活性 用于晚期糖基化终末产物(RAGE)。我们的目标是发现梅毒的生物学途径 用新的全基因组CRISPR/CAS9基因敲除筛选、检测F(BA)S肽和 RAGE基因敲除逆转Medin毒性并测试Medin在血管神经病理中的体内作用 通过建立内皮细胞过表达Medin的小鼠模型。在目标1中，我们将探索有毒信号 在内皮细胞中使用基于合成致死性的全基因组CRISPR/Cas9筛选的机制。这 将生成促进或保护免受梅丁毒性的候选基因/途径的列表，并可能 成为潜在的毒品目标。在目标2中，我们将测试F(BA)S多肽是否能逆转培养的Medin毒性 分离的人供体脑动脉内皮细胞和体外培养。在目标3中，我们将在体内测试 Medin在衰老诱导的血管和认知功能障碍中的作用以及评估RAGE基因敲除是否会 防护性的。在一个探索性的子目标中，我们将创建一种内皮特异性药物的转基因小鼠模型 过度表达。一旦成功实施，该提案将阐明这些机制 基础Medin血管神经病理学和创建一种有价值的新的临床前动物模型 血管老化，可用于识别和测试新药靶点。

Abstract Age is the most important risk factor for cardio-cerebrovascular diseases and dementia disorders. Epidemiologic, preclinical and clinical data show that vascular disease is strongly associated with dementia disorders, including Alzheimer's disease (AD) and AD-related disorders such as vascular dementia (VaD). The mechanistic links among vascular disease, aging and dementia remain poorly understood. There is growing evidence that medin, a 50-amino acid peptide that forms one of the most common yet least studied human amyloidoses, is an important driver of vascular aging pathologies. Medin accumulates in the vasculature with aging and is implicated in AD, VaD and aortic disease. Little is known as to the mechanisms by which medin induces cell and tissue injury, and no animal model of medin pathology currently exists. We showed that medin impaired endothelial function and cell viability and induced pro-inflammatory activation, in part through receptor for advanced glycation endproducts (RAGE). Our goals are to discover biological pathways of medin toxicity using novel genome-wide CRISPR/Cas9 knockout genetic screening, test F(BA)S peptide and RAGE knockout to reverse medin toxicity and test the in vivo role of medin in vasculo-neuropathology by creating a mouse model with endothelial overexpression of medin. In Aim 1, we will probe toxic signaling mechanisms using a synthetic lethality-based genome-wide CRISPR/Cas9 screening in endothelial cells. This will generate a list of candidate genes/pathways that facilitate or protect against medin toxicity and could be potential drug targets. In Aim 2, we will test if peptide F(BA)S can reverse medin toxicity in cultured endothelial cells and ex vivo in isolated human donor human cerebral arteries. In Aim 3, we will test in vivo the role of medin in aging-induced vascular and cognitive dysfunction and assess whether RAGE knockout will be protective. In an exploratory subaim, we will create a transgenic mouse model of endothelium-specific medin overexpression. Once successfully implemented, the proposal will shed light on the mechanisms underlying medin vasculo-neuropathology and create a valuable and novel preclinical animal model of vascular aging that can be used to identify and test new drug targets.