Translational Research Career Development: Overcoming Resistance to Radiotherapy

转化研究职业发展：克服放射治疗的耐药性

• 批准号: 10454199
• 负责人: Vinita Takiar
• 金额: --
• 依托单位: CINCINNATI VA MEDICAL CENTER RESEARCH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454199
• 关键词: 3-Dimensional; Alcohol consumption; Algorithms; Animal Model; Apoptosis; Area; Bioinformatics; Biological Assay; Cancer cell line; Caring; Case Study; Cell Death; Cell Line; Cell Respiration; Cell physiology; Cells; Cessation of life; Chemotherapy and/or radiation; Clinical Data; Clinical Trials; Combined Modality Therapy; Data; Development; Diagnosis; Dose; Enzymes; Event; Foundations; Funding; Future; General Population; Genus Hippocampus; Glutaminase; Goals; Head and Neck Cancer; Immunofluorescence Immunologic; Immunohistochemistry; In Vitro; Incidence; Ionizing radiation; Laboratories; Leadership; Malignant Neoplasms; Mediating; Mediator of activation protein; Mentors; Metabolic; Methods; Midwestern United States; Mission; Modality; Modeling; Molecular; Mouth Neoplasms; Outcome; Pathway interactions; Patients; Pharmacology; Phase; Phase I Clinical Trials; Phosphoproteins; Play; Population; Predictive Value; Protein Analysis; Protein Microarray Assay; Proteins; Proteome; Publishing; Quality of life; Radiation; Radiation Dose Unit; Radiation therapy; Recurrence; Research; Resistance; Risk Factors; Role; Sampling; Signal Pathway; Signal Transduction; Smoke; Solid Neoplasm; Sum; Technology; Testing; Therapeutic; Tobacco use; Toxic effect; Translational Research; Treatment Efficacy; Tumor-Derived; Validation; Veterans; Xenograft Model; aggressive therapy; base; cancer cell; cancer therapy; career development; cell injury; combinatorial; cost; data registry; differential expression; disorder control; druggable target; effective therapy; efficacy validation; experimental study; extracellular; former smoker; genomic aberrations; head and neck cancer patient; improved; in vivo; inhibitor; innovation; interest; monolayer; mouse model; neoplasm registry; new therapeutic target; novel; patient derived xenograft model; pre-clinical; protein expression; radiation resistance; research and development; resistance mechanism; response; skills; synergism; targeted treatment; therapy resistant; three dimensional cell culture; translational oncology; translational scientist; treatment response; tumor

Head and neck cancer (HNC) is the sixth most common malignancy worldwide, diagnosed twice as frequently in veterans. Radiation therapy (RT) is an important component of cancer treatment; however, its efficacy is limited by radioresistance, with the cancer sometimes returning within the treated area. Resistance to RT is, at least in part, mediated by adaptive signaling events induced by treatment. However, we do not yet understand the pathways used by cells to evade the cellular damage caused by RT. The long-term goal is to better understand, and subsequently target, the mechanisms of resistance that cancer cells use to evade a radiation-induced death. Combinatorial adaptive response therapy (CART) represents a novel platform that allows for the rapid and systematic identification of treatment combinations that overcome therapeutic resistance and result in synthetic lethality. CART takes advantage of Reverse Phase Protein Microarray Analysis (RPPA), providing a high throughput, sensitive, and quantitative approach to analyze differential protein expression to identify targets for combinatorial therapy. The applicability of the CART approach to RT has not previously been investigated. The central objective of this career development application is to develop myself as an independent translational scientist with expertise in HNC, and to become a leader in the field of translational oncology, with implementation of a CART approach to increase the efficacy of RT in 3D culture and xenograft models. The rationale for the proposed research is rationally combining newly identified systemic treatments, such as the glutaminase inhibitor, CB-839, with RT will result in maximal efficacy while minimizing potential toxicities. Guided by strong preliminary data implicating glutaminase as playing a role in adaptive resistance to RT, we will pursue three specific aims: First (SA1), we will determine whether the combination of RT with a glutaminase inhibitor (CB-839) results in decreased aerobic respiration and increased cell death in 2D and 3D culture. To pursue this, upregulated aerobic respiration pathways, including those catalyzed by glutaminase, will be selectively targeted alone or in combination with RT in 3D with analysis of proliferation and apoptosis markers. Seahorse technology will be used to assess aerobic respiration. Second (SA2), we will validate the efficacy of glutaminase inhibition by testing it both alone and in combination with RT in preclinical heterotopic cell line and patient- derived xenograft animal models. Finally (SA3), we will identify other novel HNC signaling pathways that are significantly altered by RT using RPPA. For this aim, spheroids grown from oral cavity tumor derived cell lines will be grown in 3D culture and subjected to non-lethal RT doses with protein levels assessed by RPPA to identify candidate target proteins that are differentially expressed with RT. This innovative approach uses a cutting-edge, high-throughput, sensitive, and quantitative method (RPPA) to identify entirely novel therapeutic targets to be used in combination with RT, at the protein level. The proposed research is significant because it is testing a rationally selected treatment (CB-839) to increase the efficacy of RT. These experiments will lay the groundwork for future clinical trials and are expected to identify additional unknown, yet effective treatment combinations. In sum, this proposal outlines a sophisticated, rational, and rapid approach to identifying and testing novel therapeutic targets which would be of disproportionate benefit to veterans battling head and neck cancer.

头颈癌（HNC）是全球第六大最常见的恶性肿瘤，被诊断出两次 在退伍军人中经常。放射治疗（RT）是癌症的重要组成部分 治疗;但是，它的功效受放射性的限制，有时会导致癌症 在处理区域内返回。对RT的抗性至少部分是由自适应介导的 治疗引起的信号事件。但是，我们还不了解所使用的途径 通过细胞逃避由RT造成的细胞损伤。长期目标是更好地理解， 随后，癌细胞用来逃避A的抗药性机制 辐射引起的死亡。组合自适应反应疗法（CART）代表一种新颖 平台可以快速，系统地识别治疗组合 克服治疗性抗性并导致合成致死性。购物车利用 反相蛋白微阵列分析（RPPA），提供高通量，敏感和敏感 定量方法分析差异蛋白表达以鉴定靶标 组合治疗。购物车方法对RT的适用性以前尚未 调查。这项职业发展应用程序的核心目标是发展自己 作为具有HNC专业知识的独立翻译科学家，并成为领导者 翻译肿瘤学领域，采用推车方法来提高功效 3D培养和异种移植模型中的RT。拟议研究的理由是合理的 将新鉴定的全身治疗方法（例如谷氨酰胺酶抑制剂CB-839）结合在一起 RT将导致最大功效，同时最大程度地减少潜在毒性。在强者的指导下 初步数据暗示谷氨酰胺酶在自适应抗RT中发挥作用，我们将 追求三个具体目标：第一个（SA1），我们将确定RT与 谷氨酰胺酶抑制剂（CB-839）导致有氧呼吸减少并增加细胞死亡 在2D和3D文化中。为了追求这一点，上调有氧呼吸途径，包括 由谷氨酰胺酶催化，将被选择性地靶向单独或与3D中的RT结合 分析增殖和凋亡标记。海马技术将用于评估 有氧呼吸。第二（SA2），我们将验证谷氨酰胺酶抑制的功效 在临床前异位细胞系和患者中单独测试并与RT结合使用RT 衍生的异种移植动物模型。最后（SA3），我们将确定其他新型的HNC信号传导 使用RPPA通过RT显着改变的途径。为此，球体从 口腔肿瘤衍生的细胞系将在3D培养物中生长，并进行非致命的RT 由RPPA评估的蛋白质水平的剂量以鉴定候选靶蛋白是 用RT差异表达。这种创新的方法使用了尖端，高通量的 敏感和定量方法（RPPA）确定完全新颖的治疗靶标为 在蛋白质水平上与RT结合使用。拟议的研究很重要，因为 它正在测试合理选择的治疗方法（CB-839）以提高RT的功效。这些 实验将为以后的临床试验奠定基础，并有望确定 其他未知但有效的治疗组合。总而言之，该提议概述了 识别和测试新型治疗靶标的精致，理性和快速的方法 这对与头颈癌作斗争的退伍军人将是不成比例的好处。