Reverse Engineering of Cell Senescence

细胞衰老的逆向工程

• 批准号: 10573323
• 负责人: MARC Wallace KIRSCHNER
• 金额: $ 69.39万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-15 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10573323
• 关键词: Address; Affect; Age; Aging; Animals; Biochemical; Biological Assay; Cell Aging; Cell Culture Techniques; Cell Cycle; Cell Cycle Arrest; Cell Line; Cell Senescence Induction; Cell Size; Cell Survival; Cells; Cellular biology; Cessation of life; Clustered Regularly Interspaced Short Palindromic Repeats; Critical Pathways; Cultured Cells; DNA Damage; Dasatinib; Dependence; Development; Disease; Dryness; Event; Exposure to; FRAP1 gene; Genes; Goals; Growth; Growth Factor; Hepatocyte; Heterogeneity; Hormones; Human; Hydrogen Peroxide; Hypertrophy; Inflammatory; Insulin; Ionizing radiation; Kinetics; Laboratories; Learning; Life; Lipids; Liver; Logic; Machine Learning; Mass Spectrum Analysis; Measures; Methods; Microscopic; Microscopy; Molecular; Mus; Mutagenesis; Normal Cell; Normal tissue morphology; Oncogene Activation; Pathology; Pathway interactions; Pattern; Pharmaceutical Preparations; Pharmacologic Substance; Pharmacology; Phenotype; Phosphoproteins; Phosphorylated Peptide; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Process; Proteins; Proteomics; Radiation; Reactive Oxygen Species; Research Personnel; Reverse engineering; Role; Signal Transduction; Sirolimus; Site; Small Interfering RNA; Stress; Swelling; Symptoms; Testing; Time; Tissues; cell growth; cell type; chemokine; experimental study; healthspan; insight; interest; kinase inhibitor; knock-down; mTOR inhibition; new technology; novel therapeutics; pharmacologic; phosphoproteomics; prevent; programs; proteostasis; response; senescence; stressor; tool

Abstract Cellular senescence is not just a symptom of aging, but a contributor to aging pathologies. Recent experiments in the mouse show that elimination of senescent cells (a.k.a. senolysis) can reverse several features of the aging process and extend life- and health-span. This has stimulated interest in cell senescence and in finding ways to suppress it that can be easily ported from mouse to human and made even more effective. This implies looking for such drugs as an early senolytic candidate – a kinase inhibitor dasatinib, whose mode of action is unclear. Taking senolytic therapies into human will also require a better understanding of the cell type specific progression towards senescence and diverse sub-types of senescence. In addition to aging per se cells respond to insults such as DNA damage, cell cycle arrest or oncogene activation by expressing aging phenotypes, such as hypertrophy, cell cycle arrest markers, and by secreting growth factors and chemokines, which are thought to produce degenerative responses in nearby cells. Hypertrophy may be one of the most pervasive senescent cell responses but has been hard to study, as until recently we have not had accurate enough means to measure cell mass and cell size, protein and lipid content. Yet, hypertrophy is of special interest because it is deeply connected to cell growth, which is normally under strict control in normal cells. We propose to study how cells, prompted by stressors like radiation or drugs or simply by age, become hypertrophic and how they come to express senescence markers. We have three goals: 1) to trace the progression of cell senescence in molecular terms by quantitative mass spectrometry and phospho-mass spectrometry, and in physiological terms by Raman microscopy, 2) to identify protein circuits responsible for cell senescence, and 3) to find drugs that will prevent, reverse, or eliminate senescent cells. As our tools of perturbation, we have chosen kinase inhibitors. Employing a machine learning approach, we will probe senescent cell development and senescent cell viability via a small set of well-characterized poly-specific kinase inhibitors; then regress a phenotype, such as senescence- specific signaling and senescent cell formation or death, to the activity of key kinases and their downstream circuits. Kinases implicated that way will be independently validated using siRNA knock downs and CRISPR. Additionally, extensive phospho-proteomic profiling will identify the phosphorylation sites on key proteins that most contribute to the phenotypes of senescence and which could be druggable by other means. Taken together, these methods will produce insight into the mode of action of already identified senolytic drugs and suggest new targets and new drugs for seno-therapies. This combination of pharmacology, machine learning, quantitative proteomics and new forms of microscopy can provide fresh insights into aging and suggest potential therapies for aging-related disease.

抽象的 细胞感应不仅是衰老的症状，而且是导致衰老病变的症状。最近的 小鼠中的实验表明，消除感觉细胞（又称鼻溶）可以逆转几个 衰老过程的特征并扩展寿命和健康。这激发了对细胞感应的兴趣 并找到抑制它可以轻松从鼠标移植到人的方法 有效的。这意味着寻找像早期塞溶剂候选药物的药物 - 激酶抑制剂dasatinib， 其作用方式不清楚。将塞溶性疗法带入人也将需要更好 了解细胞类型的特异性进展，向感应和潜水员的sectim型进展。 除了衰老本质上，细胞还会对DNA损伤，细胞周期停滞或癌基因做出反应 通过表达衰老表型（例如肥大，细胞周期停止标记）和分泌来激活 生长因子和趋化因子被认为会在附近的细胞中产生退化反应。 肥大可能是最普遍的感觉细胞反应之一，但很难研究，直到 最近，我们没有足够准确的方法来测量细胞质量和细胞大小，蛋白质和脂质 内容。然而，肥大是特别感兴趣的，因为它与细胞生长密切相关，这就是 通常在正常细胞中严格控制。我们建议研究细胞的压力源之类的细胞如何 辐射或药物或仅仅按年龄按年龄变成肥厚症，以及它们如何表达感应标记。 我们有三个目标：1）通过定量质量以分子术语跟踪细胞传感的进展 光谱法和磷酸质量光谱法，并以拉曼显微镜为物理，2） 确定负责细胞感应的蛋白质回路，3）找到可以防止，逆转或 消除感觉细胞。作为扰动工具，我们选择了激酶抑制剂。雇用 机器学习方法，我们将通过小的探测细胞的发育和感官的细胞活力 一组良好的多特异性激酶抑制剂；然后回归表型，例如衰老 特定的信号传导和感觉细胞形成或死亡，对关键激酶的活性及其下游 界。这种方式实施的激酶将使用sirna stock downs和crispr独立验证。 此外，广泛的磷酸蛋白质分析将识别关键蛋白上的磷酸化位点 大多数有助于感应的表型，并且可以通过其他方式吸毒。拍摄 这些方法将共同洞悉已经鉴定出的鼻溶剂的作用方式 并提出新的靶标和新药物的鼻孔。药理学，机器的结合 学习，定量蛋白质组学和新形式的显微镜可以为衰老和 提示与衰老相关疾病的潜在疗法。