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.

项目概要 随着肿瘤的进展，癌细胞获得了能够适应各种压力的特征。实际上， 患者结果的最佳预测因素之一是诊断时的疾病阶段，因为晚期肿瘤是 更具侵略性且难以治疗。然而，增强细胞增加的潜在机制 整个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 给靶向治疗的使用带来了一个特殊的问题，这些治疗经常因 获得性抵抗的出现。这项工作的目标是阐明调节的分子机制 细胞周期和细胞命运决定影响癌症进展和对靶向治疗的抵抗力。在 F99 阶段，我的目标是识别调节视网膜母细胞瘤 (RB) 通路并影响视网膜母细胞瘤 (RB) 通路的新因子。 细胞对细胞周期蛋白依赖性激酶 4 和 6 (CDK4/6) 抑制剂的反应。 CDK4/6，与 Cyclin 复合 D、磷酸化并灭活肿瘤抑制因子RB以驱动细胞周期进展。最近开发的 CDK4/6 抑制剂在临床上显示出一些希望，但每个患者最终都会接受这些抑制剂 进展，迫切需要确定耐药机制。使用体外全基因组 CRISPR/Cas9 筛选，我最近发现 E3 连接酶适配器 AMBRA1 的丢失是一个潜在的机制 对 CDK4/6 抑制的抵抗。此外，AMBRA1 缺失增加了 Cyclin D 蛋白的稳定性。我假设 AMBRA1 及其伴随的 E3 连接酶复合物以 Cyclin D 为目标进行降解，并且 AMBRA1 丢失 可能是体内对 CDK4/6 抑制剂产生耐药性的机制。我将使用分子和生化检测 鉴定与AMBRA1配合靶向Cyclin D的E3连接酶。另外，我会结合肿瘤 在非小细胞肺小鼠模型中使用多重 CRISPR/Cas9 介导的基因靶向进行条形码 癌症以确定 AMBRA1 缺失是否会导致体内 CDK4/6 抑制剂耐药性。在K00阶段，我 旨在阐明调节肺腺癌（LUAD）细胞身份的分子机制。治疗 LUAD与针对突变受体酪氨酸激酶的小分子抑制剂可导致肿瘤复发 已转分化为小细胞肺癌，这是一种侵袭性神经内分泌癌，其作用有限 治疗方案。然而，转分化的机制很大程度上未知。我建议开发细胞 这种转分化过程的细胞系和小鼠模型，以确定调节 LUAD 细胞的因子 身份并最终确定阻止或逆转转分化的方法。总的来说，这一系列工作将 阐明肺癌获得性耐药和疾病进展的基本原理，这可能 也适用于其他癌症类型。