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，诊断过两次 在退伍军人中也是如此。放射治疗(RT)是癌症的重要组成部分 治疗；然而，它的疗效受到辐射抵抗的限制，有时会出现癌症 在治疗区域内返回。对RT的抗性至少在一定程度上是由适应性 治疗引起的信号事件。然而，我们还不了解所使用的路径 通过细胞来逃避RT引起的细胞损伤。长期目标是更好地理解， 并随后针对癌细胞用来逃避 辐射导致的死亡。组合适应性反应疗法(CART)代表了一种新的 允许快速和系统地识别治疗组合的平台 克服治疗阻力，导致合成致命性。Cart利用了 反相蛋白质微阵列分析(RPPA)，提供高通量、灵敏和 定量分析差异蛋白表达以确定靶点 综合疗法。CART方法对RT的适用性以前从未出现过 调查过了。这份职业发展申请的中心目标是发展我自己 作为一名独立的翻译科学家，在HNC方面具有专业知识，并成为 翻译肿瘤学领域，实施CART方法以提高疗效 RT在3D培养和异种移植模型中的表达。建议进行研究的理由是合理的。 将新发现的系统性治疗，如谷氨酰胺酶抑制剂CB-839，与 RT将产生最大的疗效，同时将潜在的毒性降至最低。以Strong为指导 初步数据表明谷氨酰胺酶在适应性RT抵抗中发挥作用，我们将 追求三个具体目标：第一，(SA1)，我们将确定RT与A 谷氨酰胺酶抑制剂(CB-839)导致有氧呼吸减少和细胞死亡增加 在2D和3D文化中。为了实现这一点，上调了有氧呼吸途径，包括 由谷氨酰胺酶催化，将在3D中选择性地单独靶向或与RT结合 细胞增殖和凋亡标记物分析。将使用海马技术来评估 有氧呼吸。第二(SA2)，我们将通过以下方式验证谷氨酰胺酶抑制的有效性 在临床前异位细胞系和患者中单独和联合应用RT进行测试。 衍生异种移植动物模型。最后(SA3)，我们将识别其他新的HNC信号 使用RPPA的RT显著改变的通路。为了达到这个目的，球体从 口腔肿瘤细胞系将在3D培养中生长并接受非致死性RT RPPA评估的蛋白质水平的剂量以确定符合以下条件的候选目标蛋白质 与RT差异表达。这一创新方法使用尖端、高吞吐量、 灵敏、定量的方法(RPPA)确定全新的治疗靶点 与RT结合使用，在蛋白质水平上。这项拟议的研究具有重要意义，因为 它正在测试一种合理选择的治疗方法(CB-839)，以提高RT的疗效。这些 实验将为未来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，这项提案概述了 识别和测试新的治疗靶点的复杂、合理和快速的方法 这对与头颈部癌症作斗争的退伍军人来说将是不成比例的好处。