Investigating the Role of Cell Plasticity in Malignant Transformation

研究细胞可塑性在恶性转化中的作用

• 批准号: 10523137
• 负责人: Andrea Christine Chaikovsky
• 金额: $ 8.92万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10523137
• 关键词: Adenocarcinoma Cell; Bar Codes; Biochemical; Bioinformatics; Biological Assay; CDK4 gene; CRISPR screen; CRISPR/Cas technology; Cancer Etiology; Cancer cell line; Cell Cycle; Cell Cycle Progression; Cell Line; Cells; Cessation of life; Characteristics; Clinic; Complex; Cyclin D1; Cyclin-Dependent Kinase Inhibitor; Cyclins; Diagnosis; Disease; Disease Progression; Drug resistance; G1/S Transition; Gene Targeting; Genes; Genetic study; Genetically Engineered Mouse; Global Change; Goals; In Vitro; Lead; Ligase; Lung Adenocarcinoma; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Molecular; Molecular Genetics; Neurosecretory Systems; Non-Small-Cell Lung Carcinoma; Oncogenic; Pathway interactions; Patient-Focused Outcomes; Patients; Phase; Phosphotransferases; Postdoctoral Fellow; Process; Receptor Protein-Tyrosine Kinases; Relapse; Research; Research Personnel; Research Project Grants; Resistance; Retinoblastoma; Role; Route; Signal Pathway; Stress; Structure; Testing; Time; Tumor Suppressor Proteins; Tyrosine Kinase Inhibitor; Ubiquitination; Work; base; cancer cell; cancer subtypes; cancer type; career; delta protein; driver mutation; experimental study; follow-up; genome-wide; improved; in vivo; in vivo Model; inhibitor; insight; lung cancer cell; lung small cell carcinoma; mouse model; mutant; neuroendocrine cancer; novel; prevent; receptor; resistance mechanism; response; retinoblastoma pathway; retinoblastoma tumor suppressor; small molecule; small molecule inhibitor; success; targeted treatment; therapy resistant; transdifferentiation; tumor; tumor progression; tumorigenesis; ubiquitin-protein ligase

PROJECT SUMMARY As tumors progress, cancer cells acquire characteristics that allow them to adapt to various stresses. In fact, one of the best predictors of patient outcome is disease stage at the time of diagnosis, as advanced tumors are more aggressive and difficult to treat. However, the underlying mechanisms that potentiate increased cell plasticity throughout cancer progression remain poorly understood. The ability of cancer cells to adapt has posed a particular problem for the use of targeted therapies, which are frequently rendered ineffective by the emergence of acquired resistance. The goal of this work is to elucidate molecular mechanisms that regulate the cell cycle and cell fate decisions to influence cancer progression and resistance to targeted therapy. In the F99 phase, I aim to identify novel factors that regulate the retinoblastoma (RB) pathway and influence the cellular response to inhibitors of cyclin-dependent kinases 4 and 6 (CDK4/6). CDK4/6, in complex with Cyclin D, phosphorylate and inactivate the tumor suppressor RB to drive cell cycle progression. Recently developed CDK4/6 inhibitors have shown some promise in the clinic, but every patient given these inhibitors eventually progresses, creating an urgent need to identify mechanisms of resistance. Using an in vitro genome-wide CRISPR/Cas9 screen, I recently identified loss of the E3 ligase adaptor AMBRA1 as a potential mechanism of resistance to CDK4/6 inhibition. Further, AMBRA1 loss increased Cyclin D protein stability. I hypothesize that AMBRA1, with its accompanying E3 ligase complex, targets Cyclin D for degradation, and that AMBRA1 loss could be a mechanism of resistance to CDK4/6 inhibitors in vivo. I will use molecular and biochemical assays to identify the E3 ligase that cooperates with AMBRA1 to target Cyclin D. In addition, I will combine tumor barcoding with multiplexed CRISPR/Cas9-mediated gene targeting in mouse models of non-small cell lung cancer to determine whether loss of AMBRA1 leads to CDK4/6 inhibitor resistance in vivo. In the K00 phase, I aim to elucidate the molecular mechanisms regulating cell identity in lung adenocarcinoma (LUAD). Treatment of LUAD with small molecule inhibitors targeting mutant receptor tyrosine kinases can lead to relapsed tumors that have transdifferentiated into small cell lung cancer, an aggressive neuroendocrine cancer with limited treatment options. However, the mechanism of transdifferentiation is largely unknown. I propose to develop cell line and mouse models of this transdifferentiation process in order to identify factors that regulate LUAD cell identity and ultimately identify means to prevent or reverse transdifferentiation. Together, this body of work will elucidate fundamental principles of acquired resistance and disease progression in lung cancer, which may also be applicable to other cancer types.

项目摘要 随着肿瘤的进展，癌细胞获得了使它们能够适应各种压力的特性。事实上， 患者预后的最佳预测因素之一是诊断时的疾病分期，因为晚期肿瘤 然而，增强细胞增殖的潜在机制， 在癌症进展过程中的可塑性仍然知之甚少。癌细胞的适应能力 这对靶向治疗的使用提出了一个特别的问题，靶向治疗经常由于药物的作用而变得无效。 获得性抵抗的出现。这项工作的目标是阐明调节细胞凋亡的分子机制。 细胞周期和细胞命运决定影响癌症进展和对靶向治疗的抗性。在 F99期，我的目的是确定新的因素，调节视网膜母细胞瘤（RB）的途径和影响， 细胞对细胞周期蛋白依赖性激酶4和6（CDK 4/6）抑制剂的反应。CDK 4/6，与细胞周期蛋白复合 使肿瘤抑制因子RB磷酸化和磷酸化以驱动细胞周期进程。最近开发 CDK 4/6抑制剂在临床上已经显示出一些前景，但最终每个给予这些抑制剂的患者 进展，迫切需要确定耐药机制。使用体外全基因组 在CRISPR/Cas9筛选中，我最近发现E3连接酶接头AMBRA 1的丢失是一种潜在的机制， 对CDK 4/6抑制的抗性。此外，AMBRA 1损失增加了细胞周期蛋白D蛋白的稳定性。我假设 AMBRA 1及其伴随的E3连接酶复合物靶向细胞周期蛋白D进行降解，并且AMBRA 1损失 可能是体内对CDK 4/6抑制剂耐药的机制。我会用分子和生化分析 鉴定与AMBRA 1协同作用靶向细胞周期蛋白D的E3连接酶。另外，我将联合收割机肿瘤 非小细胞肺小鼠模型中多重CRISPR/Cas9介导的基因靶向条形码 癌症，以确定AMBRA 1的缺失是否导致体内CDK 4/6抑制剂抗性。在K 00阶段，我 目的探讨肺腺癌细胞特性的分子调控机制。治疗 LUAD与靶向突变型受体酪氨酸激酶的小分子抑制剂联合使用可导致肿瘤复发 已经转分化成小细胞肺癌，一种侵袭性神经内分泌癌， 治疗方案。然而，转分化的机制在很大程度上尚不清楚。我建议开发细胞 系和小鼠模型，以鉴定调节LUAD细胞的因子 鉴定并最终鉴定防止或逆转转分化的手段。总之，这部分工作将 阐明肺癌获得性耐药和疾病进展的基本原理， 也适用于其他癌症类型。