Targeting oncogenic pathways for chemoprevention of head and neck cancer by FLLL12

项目总结 通过化学预防延缓或防止癌前病变向浸润性癌的进展 头颈部鳞状细胞癌(SCCHN)是一种严重危害人类健康的疾病。 死亡率。这项研究将调查FLLL12的使用，这是一种结构上与天然 化合物姜黄素，作为化学预防这种致命癌症的新化合物；FLLL12展示了一种 与Janus Kinase(JAK)2结合并抑制STAT3磷酸化的细胞信号转导谱。此外, FLLL12抑制EGFR和AKT转录本，从而抑制EGFR/AKT-mTOR信号转导。这些信号 通路赋予细胞获得无限生长和抵抗细胞死亡的优势-- 致癌的两个特征。一种在浸润性癌前实施的有效化学预防方法 需要发展以减少SCCHN的发生率；然而，目前还没有这样的治疗方案 可用。因此，鉴定有效预防SCCHN癌变的新化合物是必要的。 在这项提案中，我们将开发FLLL12作为SCCHN的化学预防的治疗剂。我们的 初步数据显示，FLLL12的IC50值在高度选择性范围内(&lt；1微米对大多数 对癌前口腔癌细胞系的抑制作用为0.35微米)。药代动力学研究显示 在小鼠身上可以达到药理上相关的浓度。FLLL12也有效地抑制了肿瘤的生长 在SCCHN的异种移植模型中。这项建议将测试FLLL12预防或延缓进展的能力 在致癌物诱导的小鼠口腔癌模型中发现SCCHN癌前病变 细胞信号转导机制(S)对该制剂的作用。我们假设FLLL12调节JAK-STAT3和 EGFR/AKT-mTOR生存通路逆转和/或延缓癌前病变进展为 鳞状细胞癌。提出了三个具体目标。目的1：评估JAK-STAT3通路作为直接信号转导通路 FLLL12的目标。我们将验证FLLL12与JAK2相互作用并抑制JAK-STAT3途径的预测 以调节该化合物的化学预防作用。一种无细胞系统，体外激酶分析，记者 将使用检测和构成活性的STAT3质粒。目标2：界定监管机制 EGFR-AKT通路通过FLLL12。通过启动子缺失突变，我们将鉴定转录因子(S) 抑制EGFR和AKT转录本。目的3：分析FLLL12作为化学预防药物的体内效果 在致癌物诱导的口腔癌变模型中。4NQO诱导的口腔癌小鼠模型将评估 应用FLLL12预防或延缓口腔癌的发生。体内研究的结果将 证实FLLL12在SCCHN中的化学预防作用。重要的是，这个R15奖项将提供一个刺激 为本科生和研究生提供积极参与研究发现的培训机会 提高我们对癌症生物学中涉及的信号转导途径的理解的过程。

PROJECT SUMMARY The delay or prevent the progression of premalignant lesions to invasive cancer by chemoprevention of squamous cell carcinoma of head and neck (SCCHN) is a devastating disease with significant morbidity and mortality. This research will investigate the use of FLLL12, a compound structurally related to the natural compound curcumin, as a novel compound for chemoprevention of this deadly cancer; FLLL12 demonstrates a cell signaling profile of binding to Janus kinase (JAK)2 and inhibition of the phosphorylation of STAT3. In addition, FLLL12 inhibits EGFR and AKT transcripts resulting in inhibition of EGFR/AKT-mTOR signaling. These signaling pathways confer cells with the ability to acquire the advantage of unlimited growth and resistance to cell death - two hallmarks of carcinogenesis. An effective chemoprevention method implemented before an invasive cancer develops is needed to reduce the incidence of SCCHN; however, currently no such treatment regimen is available. Thus, the identification of new compounds effective in preventing SCCHN carcinogenesis is warranted. In this proposal, we will develop FLLL12 as a therapeutic agent for the chemoprevention of SCCHN. Our preliminary data demonstrate that FLLL12 has IC50 values in a highly selective range (<1 µM against most SCCHN cell lines and 0.35 µM against a premalignant oral cancer cell line). Pharmacokinetic studies reveal that a pharmacologically relevant concentration is achievable in mice. FLLL12 also effectively inhibits tumor growth in a xenograft model of SCCHN. This proposal will test the ability of FLLL12 to prevent or delay the progression of premalignant lesions to SCCHN in a carcinogen-induced oral cancer model in a mouse model and uncover the cell signaling mechanism(s) of action for this agent. We hypothesize that FLLL12 regulates JAK-STAT3 and EGFR/AKT-mTOR survival pathways to reverse and/or slow the progression of premalignant lesions to a squamous cell cancer. Three specific aims are proposed. Aim 1: Evaluate the JAK-STAT3 pathway as a direct target of FLLL12. We will test the prediction that FLLL12 interacts with JAK2 and inhibits JAK-STAT3 pathway to mediate the chemoprevention effects of this compound. A cell free system, in vitro kinase assays, reporter assays and a constitutively active STAT3 plasmid will be used. Aim 2: Define the mechanism of regulation of the EGFR-AKT pathway by FLLL12. By employing promoter deletion-mutation, we will identify transcription factor(s) that inhibit EGFR and AKT transcripts. Aim 3: Analyze the in vivo efficacy of FLLL12 as a chemoprevention agent in a carcinogen-induced oral carcinogenesis model. A 4NQO-induced oral cancer mouse model will evaluate the prevention or delay in oral carcinogenesis with administration of FLLL12. The outcome of the in vivo studies will confirm the chemoprevention effects of FLLL12 in SCCHN. Importantly, this R15 award will provide a stimulating training opportunity for undergraduate and graduate students to participate actively in the research and discovery process to improve our understanding of the signal transduction pathways involved in cancer biology.