C. elegans Model for Neurodegenerative Diseases of Aging

A.总结 衰老是退行性疾病的主要危险因素，并与累积的蛋白质损伤和细胞凋亡相关。 细胞功能下降。我们以前的工作表明，聚集倾向蛋白的表达 in C.神经退行性疾病的elegans模型启动了一系列蛋白质损伤， 其他亚稳态蛋白质的错误折叠。使用计算方法，我们表明， 聚合不是随机的，而是有共同的序列元素，预测其内在的 亚稳定性和聚集倾向。此外，各种神经元和非神经元细胞中的错误折叠和聚集 在整个生命周期中，组织不是零星的，而是在C. elegans 成年期与可诱导细胞应激反应的稳健性的急剧下降相一致。在这 MERIT扩展应用中，我们提出来检验假设，即蛋白质组的组成。和 在老化过程中可溶性和不溶性物质平衡的改变被急性和慢性 蛋白毒性应激，并通过选择性诱导蛋白稳态网络和寿命途径来保护。这 将使用两个互补的模型系统进行演示：使用C的有机体方法。优雅的， 提供了一个精确的衰老模型和测试控制蛋白质稳态和寿命的遗传途径的能力， .和成人原代细胞和诱导性多能细胞，以评估衰老蛋白质组的变化是否 并且控制这些过程的机制是保守的。目的：（1）评价特异性 以及在急性和慢性蛋白毒性应激和衰老中蛋白质组聚集的选择性。我们将 在C的多个点进行蛋白质组学分析。elegans成年和衰老，并量化可溶性 和聚集的部分在动物攻击的急性（热休克）和慢性（表达 聚谷氨酰胺。ApoA和tau）蛋白毒性应激。转变为聚集状态的蛋白质将通过以下方法进行验证： 产生相应系列的转基因蛋白-GFP报告细胞系，并用于评估折叠 在活体动物的不同区室和组织中的转变。人的比较蛋白质组学分析 来自不同年龄段供体的原代细胞将揭示是否存在相同的蛋白质或 途径在溶解度和聚集方面经历类似的变化。蛋白质组学数据将与 相应的一组RNA-seq数据，并用于开发模型，以确定蛋白质组学风险，失败，或 保护作用可以在组织水平上进行评估，以建立对蛋白质组范围的生物理解。 网络. (2)确定蛋白质组的稳定性是否可以通过调节蛋白质组的结构来选择性改变。 蛋白质稳态网络和寿命途径。我们将决定后果的可溶性和 增强或抑制PN不同臂的聚集蛋白质组。例如通过组成性活化 或抑制热休克反应和细胞器未折叠蛋白反应。同样， 通过激活或抑制调节的寿命途径， 通过热量限制、生殖系干细胞信号传导、胰岛素信号传导和热休克反应，以及（3） 建立应激和衰老蛋白质组的多维系统网络图。发展 使用目标1和目标2的数据绘制的蛋白质组动态随时间变化的可视化图像。连同 关于蛋白质组和PN表达的档案数据。这张图将被用来预测蛋白质， 这些区域或组织受到衰老和压力的影响，并受到伴侣蛋白网络和细胞膜的保护。 PN。

A. Summary Aging is a major risk factor for degenerative diseases, and associated with cumulative protein damage and the decline of cellular function. Our previous work demonstrated that the expression of aggregation-prone proteins in C. elegans models of neurodegenerative disease initiates a cascade of protein damage that results in misfolding of other metastable proteins. Using a computation approach, we showed that proteins at-risk for aggregation are not randonri, but rather have in common sequence elements that predict their intrinsic metastability and tendency to aggregate. Moreover, misfolding and aggregation in various neuronal and nonneuronal tissues is not sporadic throughout lifespan, but rather occurs at a much earlier point in C. elegans adulthood coincident with a dramatic decline in the robustness of inducible cell stress responses. In this MERIT extension application, we propose to test the hypothesis that the composition of the proteome. and alterations in the balance of soluble and insoluble species during aging are accelerated by acute and chronic proteotoxic stress and protected by selective induction of proteostasis network and lifespan pathways. This will be demonstrated using two complementary model systems: an organismal approach using C. elegans that affords a precise model for aging and the ability to test genetic pathways that control proteostasis and lifespan, .and adult human primary cells and inducible pluripotent cells to assess whether changes in the aging proteome and mechanisms that control these processes are conserved. The Aims are: (1) Assessing the specificity and selectivity of proteome aggregation in acute and chronic proteotoxic stress and aging. We will perform a proteomic analysis at multiple points of C. elegans adulthood and aging, and quantify the soluble and aggregated fractions in animals challenged by acute (heat shock) and chronic (expression of polyglutamine. Aß, and tau) proteotoxic stress. Proteins that shift to the aggregated state will be validated by generating a corresponding series of transgenic protein-GFP reporter lines and used to assess folding transitions in different compartments and tissues in living animals. Comparative proteomic analysis of human primary cells from donors over a wide range of chronological age will reveal whether the same proteins or pathways undergo similar changes in solubility and aggregation. The proteomic data will be integrated with a corresponding set of RNA-seq data and used to develop models to establish whether proteomic risk, failure, or protection can be assessed at the tissue-level to establish an organismal understanding of proteome-wide networks. (2) Establishing whether proteome stability can be selectively altered by regulation of the proteostasis networif and lifespan pathways. We will determine the consequences to the soluble and aggregated proteome of enhancing or inhibiting different arms of the PN. for example by constitutive activation or inhibition of the heat shock response and the organellar unfolded protein responses. Likewise, are the consequences to the proteome the same or distinct, by activating or inhibiting the lifespan pathways regulated by caloric restriction, germline stem cell signaling, insulin-signaling, and the heat shock response, and '(3) Developing a multi-dimensional systems network map of the stressed and aging proteome. To develop a visual image of proteome dynamics over chronological age using data from Aims 1 and 2. together with archival data on expression of the proteome and the PN. This map will be used to predict the proteins, compartments, or tissues that are affected by aging and stress, and protected by chaperone networks and the PN.