Interaction of cannabidiol (CBD) with targeted inhibitors of essential cancer signaling pathways

项目摘要/摘要 被诊断为晚期癌症的患者通常需要全身治疗，其中包括化疗， 靶向治疗和免疫治疗。这项建议将解决影响临床结果的主要问题； 哪位患者对哪种治疗有反应的问题，以及与哪些因素相关的问题 固有的或后天的抵抗力。尽管内在耐药有时可以明确地归因于肿瘤特异性 例如特定的体细胞突变等因素，在许多情况下，耐药的潜在基础是模糊的。这 一项提案将解决新的假设，即使用大麻二酚(CBD)提供了一种被低估的来源 对临床上重要的癌症治疗的反应的变异。越来越多的人使用CBD在 癌症治疗是为了减轻癌症疼痛，甚至是因为他们认为治疗癌症有好处。在 过去，由于问题的存在，对CBD和其他大麻衍生植物化学物质的研究相对有限 社会耻辱和法律限制；然而，越来越多的研究产生了令人信服的 有证据表明，CBD影响某些类型肿瘤的生长，并与细胞毒性化疗药物相互作用 调节它们的活动。一些研究表明，CBD细胞受体与 作为重要治疗靶点的EGFR、SRC和其他信号蛋白的信号转导，意味着CBD可能 影响针对这些蛋白质的药物的反应。然而，任何癌症类型的已发表研究都没有。 系统地探讨了CBD与肿瘤治疗中常用的靶向信号通路的相互作用。 在试点研究中，我们一直在研究CBD与175种激酶靶向抑制剂和 限制癌细胞生长的额外对照化合物。这已经确定了几个具体的CBD- 抑制剂的相互作用，这在两个被测试的细胞模型之间有所不同，但在EGFR- 依赖细胞模型。鉴于检测到CBD和EGFR信令之间存在多个连接， 这项建议的目的是评估CBD对靶向通路活性的影响程度 两种EGFR依赖肿瘤类型(头颈部鳞状细胞癌(HNSCC)和 结直肠癌(CRC)，并表征CBD-抑制物相互作用的潜在机制。在目标1中， 我们将在额外的HNSCC和CRC细胞模型中筛选靶向抑制物库，以建立一致的 相互作用模式，使用CellTiterBlue和核计数作为初步筛选分析。积极的点击率将是 进一步探讨了克隆生长和3D球体生长分析，并定义了相互作用的剂量范围。 反相蛋白质阵列(RPPA)分析将用于建立特别受影响的信号通路 通过CBD-抑制剂的相互作用。在目标2中，对于选定的特定蛋白靶向抑制物，我们将定义 在体内异种移植和患者来源的异种移植(PDX)模型中，CBD-抑制剂的相互作用证实了CBD- 在体外定义的依赖信号变化与体内相关，并使用大数据的生物信息学分析 了解CBD受体表达、预后和药物反应之间的关系的资源。

PROJECT SUMMARY/ABSTRACT Patients diagnosed with advanced cancers typically required systemic therapies, which include chemotherapy, targeted therapy, and immunotherapy. This proposal will address major issues affecting clinical outcomes; the question of which patient will respond to which therapy, and the question of what factors are associated with intrinsic or acquired resistance. Although intrinsic resistance can sometimes be clearly assigned to tumor-specific factors, such as specific somatic mutations, in many cases the underlying basis for resistance is obscure. This proposal will address the novel hypothesis that use of cannabidiol (CBD) provides an under-appreciated source of variation in response to clinically important cancer therapies. A growing number of individuals use CBD during cancer treatment to alleviate cancer pain, or even because of perceived benefits for treating their cancers. In the past, there has been relatively limited study of CBD and other cannabis-derived phytochemicals due to issues of social stigma and legal restrictions; however, a growing number of studies have generated convincing evidence that CBD affects the growth of some types of tumors and interacts with cytotoxic chemotherapies to modulate their activity. Some studies have demonstrated interaction between CBD cellular receptors and signaling by EGFR, SRC, and other signaling proteins that are important therapeutic targets, implying CBD might affect response to drugs targeting those proteins. However, no published studies in any cancer type have systematically probed the interaction of CBD with the signaling pathways commonly targeted in cancer therapy. In pilot studies, we have been investigating the interaction of CBD with 175 kinase-targeted inhibitors and additional control compounds in restricting the growth of cancer cells. This has identified several specific CBD- inhibitor interactions, which differed between two cell models tested, but were more notable in an EGFR- dependent cell model. Given multiple connections identified between CBD and EGFR signaling detected, the objective of this proposal is to evaluate the degree to which CBD influences the activity of pathway-targeted inhibitors in two EGFR-dependent tumor types (head and neck squamous cell carcinoma (HNSCC), and colorectal cancer (CRC)), and to characterize the underlying mechanisms of CBD-inhibitor interaction. In Aim 1, we will screen the targeted inhibitor library in additional HNSCC and CRC cell models to establish consistent patterns of interaction, using CellTiterBlue and nuclear count as initial screening assays. Positive hits will be further explored using clonogenic and 3D spheroid growth analysis, and dose range of interaction defined. Reverse-phase protein array (RPPA) analysis will be used to establish signaling pathways specifically affected by CBD-inhibitor interaction. In Aim 2, for selected specific protein-targeted inhibitors of interest, we will define the CBD-inhibitor interaction in xenograft and patient-derived xenograft (PDX) models in vivo, confirm CBD- dependent signaling changes defined in vitro pertain in vivo, and use bioinformatic analysis of large data resources to understand the relationship between CBD receptor expression, prognosis, and drug response.