Elucidating the mode of action of the abietanes to develop potential therapeutic agents_supplement

项目摘要 真核细胞起始因子（eIFs）的过度表达在多种人类疾病中均有报道 包括乳腺癌（BC）。我们的目标是开发基于松香烷天然产物的工具化合物 选择性地攻击eIF大约15-20%的乳腺癌病例的特征是三阴性乳腺癌。 癌症（TNBC），一种缺乏有效靶向治疗方式的亚型，1并且经常受到 因此，有必要发现新的化学物质，可以选择性地 针对TNBC，以降低与这种疾病相关的死亡率。 我们已经表明，松香烷天然产物衍生物（SJ 38）显着降低蛋白质合成 和靶向TNBC细胞模型中的eIF 2/eIF 4信号传导轴。我们发现a）SJ 38选择性抑制 低微摩尔浓度范围（<10μM）的癌细胞活力，同时不影响非癌性乳腺上皮细胞 B）SJ 38显著降低参与蛋白质表达的调节因子的表达， c）在细胞培养物中用氨基酸进行稳定同位素标记（SILAC）蛋白质组学提供的基因 本体论分析SJ 38主要靶向蛋白质合成和免疫途径分析表明， SJ 38主要通过eIF 2和eIF 4/p70 S6 K信号通路发挥其抗癌作用。 我们的长期目标是阐明SJ 38在TNBC中的分子机制，重点是翻译控制。我们 一项研究解决了知识上的差距，以发现新的治疗药物和生物靶点，以潜在地治疗 TNBC。我们假设SJ 38通过以下途径减少乳腺癌进展并使细胞对治疗敏感： 翻译控制我们提出了以下具体目标：1）开发一种新的合成策略，以获得一个 一系列不同的松香烷衍生物，以促进结构活性关系（SAR）和靶点接合 问题研究2)确定所产生的化合物作为蛋白质合成调节剂的生物学性质， 它们在TNBC细胞模型中抑制增殖和迁移的能力。3)确定 松香烷靶向癌症细胞模型。 影响：拟议研究的结果将为制定有针对性的 成功治疗人类疾病包括癌症所需的治疗策略。

PROJECT ABSTRACT Over expression of eukaryotic initiation factors (eIFs) has been reported in various human diseases including breast cancer (BC). Our objective is to develop tool compounds based on the abietane natural products to selectively target eIFs. Approximately 15-20% breast cancer cases are characterized as triple negative breast cancer (TNBC), a subtype that lacks effective targeted therapy modalities,1 and is often plagued with dysfunctional eIFs expression.2 Therefore, there is a need to discover new chemical matter that could selectively targeted TNBC to reduce mortality rates associated with this disease. We have shown that abietane natural product derivatives (SJ38) significantly reduce protein synthesis and target eIF2/eIF4 signaling axis in TNBC cellular models. We discovered that a) SJ38 selectively inhibits cancer cell viability at the low micromolar range (<10μM) while not affecting non-cancerous mammary epithelial cell viability or signaling, b) SJ38 significantly reduces the expression of regulatory factors involved in protein synthesis, and c) Stable isotope labeling with amino acids in cell culture (SILAC) proteomics provided Gene Ontology analysis that SJ38 mainly targeted protein synthesis and Ingenuity Pathway Analysis suggested that SJ38 exerts its anticancer effects primarily through the eIF2, and eIF4/p70S6K signaling pathway. Our long-term goal is to elucidate SJ38 molecular mechanism in TNBC focusing on translational control. Our study addresses a gap in knowledge to discover new therapeutic agents and biological targets to potentially treat TNBC. We hypothesize that SJ38 reduces breast cancer progression and sensitizes cells to therapy via translational control. We propose the following specific aims: 1) Develop a novel synthetic strategy to access a series of diverse abietane derivatives to facilitate structure activity relationship (SAR), and target engagement studies. 2) Determine the biological properties of the generated compounds as protein synthesis modulators and their capability to inhibit proliferation and migration in TNBC cellular models. 3) Identify the mechanism by which the abietanes target cancer cellular models. IMPACT: Results from the proposed studies will provide critical knowledge towards the development of targeted therapeutic strategies needed for the successful treatment of human diseases including cancer.