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: 9892870
• 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/9892870
• 关键词: 3-Dimensional; Alcohol consumption; Algorithms; Animal Model; Apoptosis; Area; Bioinformatics; Biological; 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; Oncology; 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; Smoker; Solid Neoplasm; Sum; Technology; Testing; Therapeutic; Tobacco use; Toxic effect; Translational Research; Treatment Efficacy; Tumor-Derived; Validation; Veterans; Xenograft Model; Xenograft procedure; aggressive therapy; base; cancer cell; cancer therapy; career development; cell injury; combinatorial; cost; data registry; differential expression; disorder control; druggable target; effective therapy; experimental study; extracellular; genomic aberrations; head and neck cancer patient; improved; in vivo; inhibitor/antagonist; innovation; interest; monolayer; mouse model; neoplasm registry; new therapeutic target; novel; pre-clinical; protein expression; radiation resistance; research and development; resistance mechanism; response; skills; synergism; targeted treatment; therapy resistant; three dimensional cell culture; tobacco products; translational scientist; 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) 是全球第六大常见恶性肿瘤，已确诊两次 退伍军人中经常出现这种情况。放射治疗（RT）是癌症的重要组成部分 治疗;然而，其功效受到放射抗性的限制，有时癌症 返回治疗区域内。对放疗的抵抗至少部分是由适应性介导的 治疗引起的信号事件。然而，我们还不了解所使用的途径 细胞逃避 RT 造成的细胞损伤。长期目标是更好地理解， 并随后针对癌细胞用来逃避癌细胞的抵抗机制 辐射引起的死亡。组合适应性反应疗法（CART）代表了一种新颖的 平台允许快速、系统地识别治疗组合， 克服治疗耐药性并导致综合致死率。 CART 利用 反相蛋白质微阵列分析 (RPPA)，提供高通量、灵敏且 定量方法分析差异蛋白表达以确定目标 组合疗法。 CART 方法对 RT 的适用性此前尚未被证实 调查了。这个职业发展应用程序的中心目标是发展我自己 作为具有 HNC 专业知识的独立转化科学家，并成为该领域的领导者 转化肿瘤学领域，实施 CART 方法以提高疗效 3D 培养和异种移植模型中的 RT。拟议研究的理由是合理的 将新发现的全身治疗方法（例如谷氨酰胺酶抑制剂 CB-839）与 放疗将产生最大疗效，同时最大限度地减少潜在毒性。强者引导 初步数据表明谷氨酰胺酶在适应性放疗抵抗中发挥作用，我们将 追求三个具体目标：首先（SA1），我们将确定 RT 与 谷氨酰胺酶抑制剂 (CB-839) 导致有氧呼吸减少和细胞死亡增加 2D 和 3D 文化。为了实现这一目标，上调有氧呼吸途径，包括 由谷氨酰胺酶催化，将选择性地单独靶向或与 3D 中的 RT 组合 增殖和凋亡标志物分析。海马技术将用于评估 有氧呼吸。其次（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或与 RT 结合进行测试 衍生的异种移植动物模型。最后 (SA3)，我们将确定其他新颖的 HNC 信号传导 使用 RPPA 的 RT 显着改变的途径。为了这个目的，球体生长自 口腔肿瘤衍生细胞系将在 3D 培养中生长并进行非致死 RT RPPA 评估蛋白质水平的剂量，以确定候选目标蛋白质 与 RT 差异表达。这种创新方法采用了尖端、高通量、 灵敏的定量方法（RPPA）来识别全新的治疗靶点 在蛋白质水平上与 RT 结合使用。拟议的研究意义重大，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。总之，该提案概述了 复杂、合理、快速的方法来识别和测试新的治疗靶点 这对于与头颈癌作斗争的退伍军人来说是不成比例的好处。