Drug Discovery and Development

核心B：项目总结：该多学科、多站点PPG应用程序的总体目标是构建一个 细胞衰老发现科学的坚实基础，导致治疗策略的管道 减缓或预防与年龄相关的疾病。四个PPG项目中的每一个都关注衰老的作用， 细胞在特定年龄相关的健康结果中的作用：项目1，代谢功能障碍（柯克兰/Tchkonia）;项目2， 骨骼脆弱性（Khosla/Farr）;项目3，血管功能障碍（米勒）;和项目4，骨骼肌损失 功能障碍（LeBrasseur）为了建立因果关系，每个项目将使用遗传学和药理学 去除衰老细胞的方法。重要的是，我们已经确定，senolytics，药物能够 特异性诱导衰老细胞凋亡，具有细胞类型特异性，例如在脂肪细胞中起作用， 不在内皮细胞中，反之亦然。药物发现和开发核心（DDDC）将与 其他三个核心，以协助所有四个项目，确定最有效的senotherapeutics为他们 感兴趣的细胞类型。DDDC已经开发了多种基于原代鼠和人细胞的测定， 筛选和优化敏感化合物和药物组合。senolytics的例子， DDDC鉴定的senomorphics（抑制衰老相关分泌组的药物）包括 BCL-2家族成员的抑制剂，达沙替尼加槲皮素的组合，HSP 90抑制剂， 类黄酮非瑟酮、雷帕霉素、IKK/NF-κB抑制剂、JAK抑制剂、多聚体白藜芦醇和线粒体- 有针对性的自由基清除剂没有一种化合物被证明能有效地清除或 调节所有衰老细胞类型。此外，鉴于迄今为止确定的许多senolytics是FDA 尽管已经批准了抗癌药物，但是它们的衰老清除活性以及它们的细胞类型特异性可能是 被药物化学改良了因此，DDDC的基本作用是开发和实施检测 用于根据需要测量鼠、大鼠和/或人细胞的细胞类型中的sentilant活性， 四个项目。小鼠、大鼠和/或人细胞将通过不同的细胞周期被诱导经历衰老。 机制，包括氧化和遗传毒性应激，复制性衰老和衰老治疗 细胞条件培养基，被认为最适合于感兴趣的细胞类型及其正常生理学， vivo.将开发2D和3D衰老细胞培养测定并用于原代和 二级屏幕。这些试验将用于比较现有的senotherapeutics，筛选新的 化合物，并测试已知的senotherapeutics的类似物以获得改善的活性。此外，DDDC将 提供用于确定药物稳定性和在年轻人中分布的senescent化合物的DMPK分析 和老年小鼠，以协助每个项目确定正确的剂量方案，为他们提出的实验。 最后，核心将验证所鉴定的化合物在体内减少衰老细胞负荷的能力。 使用携带细胞衰老的p16 INKA和p21 Cip 1报告基因的加速老化小鼠模型。

Core B: Project Summary: The overall goal of this multi-disciplinary, multi-site PPG application is to build a firm foundation of discovery science in cellular senescence that leads to a pipeline of therapeutic strategies that slow or prevent age-associated diseases. Each of the four PPG Projects focus on the role of senescent cells in specific age-related health outcomes: Project 1, Metabolic Dysfunction (Kirkland/Tchkonia); Project 2, Skeletal Fragility (Khosla/Farr); Project 3, Vascular Dysfunction (Miller); and Project 4, Skeletal Muscle Loss and Dysfunction (LeBrasseur). To establish cause and effect, each project will use genetic and pharmacologic approaches to ablate senescent cells. Importantly, we have already established that senolytics, drugs able to induce apoptosis specifically of senescent cells, have cell-type specificity, such as working in adipocytes and not in endothelial cell, or vice versa. The Drug Discovery and Development Core (DDDC) will work with the other three cores to assist all four projects with the identification of the most effective senotherapeutics for their cell types of interest. The DDDC has developed multiple assays based on primary murine and human cells for screening and optimizing senotherapeutic compounds and drug combinations. Examples of senolytics and senomorphics (drugs that inhibit the senescence-associated secretome) identified by the DDDC include inhibitors of BCL-2 family members, the combination of dasatinib plus quercetin, HSP90 inhibitors, the flavonoid fisetin, rapamycin, inhibitors of IKK/NF-κB, JAK inhibitors, multimeric resveratrol and mitochondrial- targeted free radical scavengers. No one senotherapeutic compound has proven effective in clearing or modulating all senescent cell types. In addition, given that many of the senolytics identified to date are FDA approved anti-cancer drugs, it is likely that their senolytic activity as well as their cell-type specificity can be improved by medicinal chemistry. Thus, the essential role of the DDDC is to develop and implement assays for measuring senotherapeutic activity in the cells types of murine, rat and/or human cells as needed for the four projects. Mouse, rat and/or human cells will be induced to undergo senescence through different mechanisms, including oxidative and genotoxic stress, replicative senescence and treatment with senescent cell conditioned media, as deemed most appropriate to the cell type of interest and its normal physiology in vivo. 2D and 3D senescent cell culture assays will be developed and utilized for use in both primary and secondary screens. These assays will be used for comparing existing senotherapeutics, screening novel compounds, and testing analogs of known senotherapeutics for improved activity. In addition, the DDDC will provide DMPK analysis of senotherapeutic compounds for determining drug stability and distribution in young and old mice to assist each project in determining the right dosing regimen for their proposed experiments. Finally, the Core will validate the ability of the identified compounds to reduce the senescent cell burden in vivo using mouse models of accelerated aging carrying p16INKA and p21Cip1 reporters of cellular senescence.