Adaptive regulation of cancer cell fate following oncogene inhibition

癌基因抑制后癌细胞命运的适应性调节

• 批准号: 8868342
• 负责人: Mohammad Fallahi-Sichani
• 金额: $ 12.02万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-17 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8868342
• 关键词: Accounting; Affect; Antineoplastic Agents; Apoptosis; Apoptotic; Autocrine Communication; Award; Biochemical; Biochemistry; Biological Assay; Biological Markers; Biology; Cell Death; Cell Line; Cell Surface Receptors; Cells; Coculture Techniques; Collaborations; Colon Carcinoma; Combined Modality Therapy; Computer Simulation; Dana-Farber Cancer Institute; Data; Disease; Disease remission; Drug Targeting; Drug resistance; Effectiveness; Engineering; Environment; Feedback; Foundations; Future; Geminin; Genotype; Goals; Gray unit of radiation dose; Growth; Health; Heterogeneity; Immunofluorescence Immunologic; Individual; Inflammatory; JUN gene; Knowledge; Life; Ligands; Link; MAPK8 gene; MEK inhibition; MEKs; Macrophage Activation; Mass Spectrum Analysis; Measurement; Measures; Mediating; Melanoma Cell; Memorial Sloan-Kettering Cancer Center; Mentors; Mentorship; Methods; Microscopy; Modeling; Molecular; Oncogene Proteins; Oncogenes; Oncogenic; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Protein Array; Proteins; Regulation; Research Personnel; Resistance; Ribosomal Protein S6; Role; Signal Transduction; Statistical Models; Systems Biology; Techniques; Testing; Therapeutic; Time; Training; Training Activity; Transcriptional Activation; Tumor Biology; Work; base; cancer cell; cancer pharmacology; cancer type; career; cell fixing; cell killing; cell type; cellular engineering; cellular imaging; design; drug efficacy; drug sensitivity; improved; inhibitor/antagonist; innovation; macrophage; medical schools; melanoma; neoplastic cell; novel; novel strategies; prevent; receptor; resistance mutation; response; senescence; single cell analysis; skills; success; therapeutic effectiveness; therapeutic target; transcription factor; tumor; tumor microenvironment

 DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop a single-cell, and network-level understanding of cell signaling mechanisms involved in adaptive drug responses through the application of engineering and systems biology approaches. I have chosen melanoma and drugs targeting the BRAFV600E oncoprotein, since adaptation is well known to be important in this type of cancer. Treatment of BRAFV600E melanomas with drugs, such as vemurafenib, that inhibit RAF/MEK signaling is effective in the short term, but remission is not durable. Drug resistance is thought to involve short-term adaptive responses that up-regulate compensatory pro-growth and/or anti-apoptotic mechanisms. The discovery and analysis of adaptive responses in melanoma represents a breakthrough in tumor biology and reveals hitherto unsuspected plasticity in signaling biochemistry. However, systematic data comparing BRAFV600E tumor cells is generally lacking and important questions are unanswered. It is not known whether adaptive mechanisms in different cell types are fundamentally similar or they are different from one cell type to the next or even one single cell to the next. Further, it is not clear how different adaptive responses are related to each other, how they are affected by tumor microenvironment, and how they are integrated to determine the fate of an individual cell. Answering these questions is critical for developing single or multi-component biomarkers of drug responsiveness and for designing rational and effective combination therapies to overcome drug adaptation and ultimately drug resistance. Our previous studies show that adaptive responses are diverse across melanomas, involving different combinations of signaling cascades. In particular, we identified an adaptive mechanism involving JNK/c-Jun that diminishes drug efficacy. RAF and JNK inhibitors induce synergistic cell killing in melanoma cells in which c-Jun mediated adaptive response occurs. Single-cell studies show that JNK inhibition enhances suppression of phospho-S6 ribosomal protein, promotes apoptosis in a subset of cells that would otherwise become quiescent and apoptosis-resistant in the presence of vemurafenib alone, and increases drug maximal effect (Emax). This work identified involvement of different pathways in adaptive responses, their diversity with genotype and time, and suggested that it would be critical to examine the diverse phenotypes induced by BRAFV600E inhibition at a single-cell level. Therefore, in this proposal I will couple high-throughput measurement, fixed and live single- cell analysis, and a combination of statistical and mechanistic computational modeling techniques to: (1) identify key molecules (ligands, receptors and transcription factors) linked to JNK/c-Jun mediated adaptive response and crosstalk with other adaptive responses in a set of BRAFV600E melanoma cell lines and primary patient-derived melanoma cells having different genotypes, (2) develop network-level models of adaptive response which discriminate among key adaptive network states observed across different cell types and their association with phenotypic responses and drug sensitivity, (3) assess the diversity and magnitude of adaptive responses across individual cells and determine their association with individual cell phenotypes (proliferation, quiescence, senescence, cell death, etc.), (4) investigate the contribution of other cell types within the tumo microenvironment, in particular tumor-associated macrophages, in drug-induced adaptive and phenotypic responses, and (5) utilize these data to identify mechanism-based biomarkers for different pathway adaptations, use these biomarkers to design and test novel combination therapeutics that take into account malignant cells, the tumor microenvironment, and the dynamics exerted by the treatment itself. The success of these studies is directly linked to the proposed training activities I intend to undertake during the mentored phase of this award. I believe that with my extensive engineering and computational background, being awarded a K99/R00 award will allow me to deepen my understanding of tumor biology (concentrating initially on melanoma) and to obtain advanced training in a highly supportive and innovative training environment of Harvard Medical School. In addition, to support me in my training, and in my transition to the independent phase of my career, I will be benefitting from the mentorship and collaboration with leading experts in the fields I propose studying. This includes my mentor Dr. Peter Sorger (Harvard Medical School), and collaborators Dr. Neal Rosen (Memorial Sloan Kettering Cancer Center), Dr. Nathanael Gray (Dana Farber Cancer Institute), and Dr. Steve Gygi (Harvard Medical School). The skills and knowledge acquired during the mentored phase of this award will be instrumental for the above proposed studies and future studies, and for successfully launching my career as an independent investigator.

 描述（由应用程序提供）：该提案的总体目标是通过应用工程和系统生物学方法来开发单个单元格，网络级别对自适应药物反应涉及的细胞信号传导机制的理解。我选择了针对BRAFV600E癌蛋白的黑色素瘤和药物，因为适应在这种类型的癌症中很重要。用药物（例如vemurafenib）抑制RAF/MEK信号传导的BRAFV600E黑色素瘤在短期内有效，但缓解不持久。耐药性被认为涉及短期自适应反应，这些反应上调了补偿性促进性增长和/或抗凋亡机制。黑色素瘤中适应性反应的发现和分析代表了肿瘤生物学的突破，并揭示了迄今为止信号传导生物化学中未刺激的可塑性。但是，系统数据比较BRAFV600E通常缺乏肿瘤细胞，并且没有解决重要问题。尚不清楚不同细胞类型中的自适应机制在根本上是相似的，还是从一种细胞类型到下一种单元或一个单个细胞到下一个细胞的不同。此外，尚不清楚不同的适应性反应如何相互关联，如何受肿瘤微环境的影响以及如何整合以确定单个细胞的命运。回答这些问题对于开发药物反应性的单一或多组分生物标志物以及设计有效有效的组合疗法以克服药物适应并最终导致药物耐药性至关重要。我们先前的研究表明，梅洛姆斯各地的自适应反应各不相同，涉及信号级联反应的不同组合。特别是，我们确定了一种涉及JNK/C-JUN的自适应机制，可降低药物效率。 RAF和JNK抑制剂诱导黑色素瘤细胞中的协同细胞杀死，其中C-Jun介导的适应性反应发生。单细胞研究表明，JNK抑制抑制磷酸S6核糖体蛋白的抑制作用可促进细胞的一个子集中的凋亡，否则在单独存在vemurafenib的情况下，这些细胞将变得静止且可抗凋亡，并增加药物最大作用（EMAX）。这项工作确定了不同途径参与自适应反应，它们在基因型和时间上的多样性，并建议检查在单细胞水平上抑制BRAFV600E引起的多样性表型至关重要。因此，在这项建议中，我将对高通量测量，固定和实时单细胞分析，以及将统计和机械计算建模技术的组合结合在一起：（1）确定与JNK/C-Jun介导的适应性反应和其他适应性反应的关键分子（配体，受体和转录因子）与其他适应性反应群相关的关键分子（配体，受体和转录因素），以及均与其他适应性反应的旋转群体， melanoma cells having different genotypes, (2) developing network-level models of adaptive response which discriminates among key adaptive network states observed across different cell types and their association with phenotypic responses and drug sensitivity, (3) assess the diversity and magnitude of adaptive responses across individual cells and determine their association with individual cell phenotypes (proliferation, quiescence, senscence, cell death, etc.), (4) investigate the contribution of other cell types within the tumo microenvironment, in particular tumor-associated macrophages, in drug-induced adaptive and phenotypic responses, and (5) utilize these data to identify mechanism-based biomarkers for different pathway adaptations, use these biomarkers to design and test novel combination therapy that take into account that account malignant cells, the tumor microenvironment, and the dynamics executed by the treatment itself.这些研究的成功直接与我打算在此奖项的修订阶段进行的拟议培训活动有关。我相信，借助我广泛的工程和计算背景，获得K99/R00奖项将使我能够加深对肿瘤生物学的理解（最初集中于黑色素瘤），并在哈佛医学院的高度支持和创新的培训环境中获得高级培训。此外，为了在培训中支持我，并在过渡到职业生涯的独立阶段时，我将从我提出的研究领域的主要专家中受益。其中包括我的心理彼得·索格（Peter Sorger）博士（哈佛医学院）和合作者尼尔·罗森（Neal Rosen）博士（纪念斯隆·凯特林（Sloan Kettering）癌症中心），纳塔纳·格雷（Nathanael Gray）博士（达娜·法伯（Dana Farber）癌症研究所）和史蒂夫·吉吉（Steve Gygi）（哈佛医学院）。在该奖项的修订阶段获得的技能和知识将在上述提议的研究和未来的研究中发挥作用，并成功地启动了我作为独立研究者的职业。