TIC10 anti-tumor effect through regulation of Foxo3a and TRAIL

项目摘要 TRAIL的效力和安全性促使临床试验将重组蛋白作为一种新的治疗方法 for human人类cancer癌症.虽然TRAIL在杀伤肿瘤细胞方面非常活跃，但重组TRAIL具有药物活性， 限制其疗效的性质，如短血清半衰期，不稳定性，以及不能穿过完整的 血脑屏障为了克服这些限制，我们鉴定了TRAIL基因的小分子诱导剂， TIC 10在稳定性、生物利用度、穿过血脑的能力方面上级重组TRAIL。 障碍、生产成本和活动范围。重要的是，我们的数据显示，TIC 10在 几种侵袭性和耐药的癌症在机制水平上，TIC 10导致有效的抗肿瘤作用， 转录因子Foxo 3a介导的TRAIL诱导作用，Foxo 3a直接调节TRAIL的表达。 TRAIL基因启动子。此外，我们发现TIC 10导致Akt和ERK的双重失活， 从而抑制Fox 3a的组成性磷酸化并增强其向细胞核的转运 并与TRAIL基因启动子结合（艾伦（Allen）等人，科学转化医学（Science Translational Medicine），出版中，2013）。我们 假设TIC 10诱导需要Akt和ERK介导的Foxo 3a的有效抗肿瘤作用 核转位和TRAIL基因的转录激活。为了解决这个假设， 提出以下具体目标：具体目标#1：确定TIC 10诱导的对Foxo 3a表达的影响， 具体目标#2：阐明Akt和ERK激酶在细胞内的作用， TIC 10的作用机制;具体目标#3：确定TRAIL基因的差异调节 FOXO家族成员的转录。这些研究将创造一个全面的分子理解， TIC 10如何利用Foxo 3a来实现其强大的抗肿瘤活性，并有可能隐藏 Foxo 3a活性的新调控机制具有生物学意义。临床前癌症模型 包括原位异种移植物和转基因小鼠沿着与原发性人肿瘤标本， 证实了关于TIC 10和Foxo 3a的功效并验证了机制发现。所有这些 研究将阐明涉及Foxo 3a的关键和新的调控机制， TIC 10产生关于其作用机制以及在结肠背景下的临床样品中的见解 癌症疾病进展。我们的研究将促进一种新型抗癌治疗药物的临床转化 通过临床前实验室研究进一步开发。

Project Summary The potency and safety of TRAIL has prompted clinical trials with the recombinant protein as a novel treatment for human cancer. While TRAIL is very active in killing tumor cells, recombinant TRAIL possesses drug properties that limit its efficacy such as short serum half-life, instability, and the inability to cross the intact blood-brain barrier. To overcome these limitations we identified a small molecule inducer of the TRAIL gene, TIC10 that is superior to recombinant TRAIL in terms of stability, bioavailability, ability to cross the blood-brain barrier, cost of production, and spectrum of activity. Importantly, our data shows that TIC10 is highly active in several aggressive and therapy-resistant cancers. On a mechanistic level, TIC10 causes potent antitumor effects and TRAIL-induction that is mediated by the transcription factor Foxo3a, which directly regulates the TRAIL gene promoter. Furthermore, we found that TIC10 results in the dual inactivation of Akt and ERK, thereby inhibiting their constitutive phosphorylation of Fox3a and potentiating its translocation to the nucleus and binding to the TRAIL gene promoter (Allen et al, Science Translational Medicine, In Press, 2013). We hypothesize that TIC10 induces potent antitumor effects that require Akt- and ERK-mediated Foxo3a nuclear translocation and transcriptional activation of the TRAIL gene. To address the hypothesis we propose the following specific aims: Specific Aim #1: Identify TIC10-induced effects on Foxo3a expression, phosphorylation, and subcellular localization; Specific Aim #2: Elucidate the role of Akt and ERK kinases in the mechanism of action of TIC10; Specific Aim #3: Determine the differential regulation of TRAIL gene transcription by FOXO family members. These studies will create a comprehensive molecular understanding of how TIC10 harnesses Foxo3a to achieve its potent antitumor activity and also has the potential to undercover novel regulatory mechanisms of Foxo3a activity that are biologically significant. Preclinical cancer models including orthotopic xenografts and transgenic mice along with primary human tumor specimens will substantiate the efficacy and validate the mechanistic findings regarding TIC10 and Foxo3a. Together, these studies will elucidate key and novel regulatory mechanisms involving Foxo3a with the first-in-class molecule TIC10 to yield insight regarding its mechanism of action as well as in clinical samples in the context of colon cancer disease progression. Our studies will facilitate the clinical translation of a novel anti-cancer therapeutic agent through further development in preclinical laboratory studies.