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，血管疾病与痴呆密切相关 在一些实施方案中，本发明的组合物用于治疗阿尔茨海默病（AD）和AD相关病症，包括阿尔茨海默病（AD）和AD相关病症，如血管性痴呆（VaD）。的 血管疾病、衰老和痴呆之间的机制联系仍然知之甚少。有 越来越多的证据表明，medin是一种50个氨基酸的肽，是最常见但研究最少的肽之一。 人类淀粉样变性是血管老化病理学的重要驱动因素。Medin在脉管系统中积聚 与衰老有关，并与AD、VaD和主动脉疾病有关。关于药物通过何种机制 诱导细胞和组织损伤，并且目前还不存在medin病理学的动物模型。我们发现， 内皮功能和细胞活力受损，并诱导促炎性激活，部分通过受体 晚期糖基化终产物（Advanced Glycation Endproducts，简称AGEs）。我们的目标是发现medin的生物学途径 使用新的全基因组CRISPR/Cas9敲除遗传筛选，测试F（BA）S肽和 逆转Medin毒性并测试Medin在血管-神经病理学中的体内作用的β-内酰胺酶敲除 通过建立一个内皮细胞过度表达medin的小鼠模型。在目标1中，我们将探索毒性信号 在内皮细胞中使用基于合成致死率的全基因组CRISPR/Cas9筛选的机制。这 将产生一个候选基因/途径的列表，这些基因/途径促进或保护免受Medin毒性的影响，并且可以 成为潜在的药物靶点在目标2中，我们将测试肽F（BA）S是否可以逆转培养的细胞中的medin毒性。 内皮细胞和离体分离的人供体人脑动脉。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，并评估是否将Medin敲除 保护性的在探索性的子目标中，我们将建立一个内皮特异性介导的转基因小鼠模型， 过度表达一旦成功实施，该建议将阐明机制 潜在的medin血管神经病理学，并创建一个有价值的和新的临床前动物模型， 血管老化，可用于识别和测试新的药物靶点。

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.