Stress-activated Protein Kinases and Chemotherapy

描述（由申请人提供）：抗癌药物与其主要靶点的相互作用仅代表药物作用机制的第一步。如果要在提高这些药物的有效性方面取得进展，那么随后诱导的凋亡信号机制的知识是至关重要的。我们研究的总体目标是通过了解这些凋亡信号通路的性质和调节来改善癌症化疗。具体来说，我们有兴趣在定义的作用，应激激活，c-Jun氨基末端蛋白激酶（JNK）的抗有丝分裂抗癌药物，特别是长春碱的作用机制。在具体目标1中，我们将特异性抑制JNK信号传导至c-Jun/AP-1，并检查AP-1抑制对长春碱诱导的细胞凋亡和基因表达的影响，以鉴定推定的AP-1靶基因。具体目标2将使用分子和细胞方法的组合来测试作为长春碱诱导的细胞凋亡的中间体的相关AP-1靶基因。完成这一目标将提供实验验证的候选基因的作用，并更好地了解长春碱激活的JNK/AP-1的凋亡信号转导机制。Specific Aim 3将利用染色质免疫沉淀来证明c-Jun募集到相关AP-1靶基因的调控区域，并鉴定可能与c-Jun合作的新型调控伙伴。Specific Aim 4将利用ec-jun-/-永生化成纤维细胞作为独特的模型系统，进一步探索c-Jun在细胞对长春碱的反应中的作用。这项研究将提供JNKJc-Jun在长春碱诱导的细胞凋亡中的作用的分子见解，并有助于我们对这一作用机制和相关药物的基本理解。反过来，这项研究可能会提出新的方法来改变细胞死亡的阈值，使这些药物更有效，减少毒性和提高特异性。

DESCRIPTION (provided by applicant): The interaction of an anticancer agent with its primary target represents only the first step in the drug's mechanism of action. Knowledge of the apoptotic signaling mechanisms subsequently induced is of critical importance if advances are to be made in improving the effectiveness of these agents. The overall goal of our research is to improve cancer chemotherapy by understanding the nature and regulation of these apoptotic signaling pathways. Specifically, we are interested in defining the role of the stress-activated, c-Jun NH2-terminal protein kinase (JNK) in the mechanism of action of antimitotic anticancer drugs, particularly vinblastine. In Specific Aim 1 we will specifically inhibit JNK signaling to c-Jun/AP-1 and examine the effects of AP-1 inhibition on vinblastine-induced apoptosis and on gene expression in order to identify putative AP-1 target genes. Specific Aim 2 will test the relevant AP-1 target genes as intermediates in vinblastine-induced apoptosis, using a combination of molecular and cellular approaches. Completion of this aim will provide experimental verification of the role of the candidate genes identified, and a better understanding of the mechanisms of apoptotic signaling by vinblastine-activated JNK/AP-1. Specific Aim 3 will utilize chromatin immunoprecipitation to demonstrate recruitment of c-Jun to the regulatory regions of the relevant AP-1 target genes, and to identify novel regulatory partners that may cooperate with c-Jun. Specific Aim 4 will utilizec-jun-/- immortalized fibroblasts as a unique model system to further explore the role of c-Jun in the cellular response to vinblastine. This research will provide molecular insight into the role of JNKJc-Jun in vinblastine-induced apoptosis, and contribute to our basic understanding of the mechanism of action of this and related agents. In turn, this research may suggest novel approaches to alter the threshold for cell death to make these drugs more effective, lessening toxicity and improving specificity.