Role of Tumor Stem Cells Recurrent Prostate Cancer

项目总结(见说明)： 癌症的进展从起始的癌症干细胞(CSC)到临床意义，并最终 治疗失败后的复发，是一种不断演变的相互作用的顶峰 在CSC和“干细胞利基”之间。这一提议的假设是，雄激素剥夺 干扰前列腺内皮细胞在调节前列腺雄激素环境中的关键作用 为了验证这一假说，在组织微环境和干细胞“生态位”的调节，促进/加速去势复发性前列腺癌的出现方面，提出了以下具体目标。 目的1研究内皮细胞小窝在循环雄激素跨过内皮细胞屏障、内吞循环雄激素导致内皮细胞稳态和信号转导相关基因的AR反式激活以及通过细胞表面受体介导的循环雄激素“非基因组信号传递”中的作用。目的2将确定雄激素剥夺诱导的前列腺内皮细胞杀伤是否导致前列腺微血管内皮细胞的短暂或不可逆的扰动，导致组织雄激素环境的失调(用LC/MS/MS表征)，以及产生/揭开独特的、有针对性的“表位”或“血管地址”(用噬菌体多肽展示技术表征)。目的3确定雄激素剥夺对前列腺癌干细胞与成体干细胞(ASCs)差异表达的基因和基因途径的影响，确定前列腺癌ASCs/CSCs的细胞表面“表位”指纹，并确定干细胞“生态位”的扰动是否导致前列腺癌CSCs中HNF-4N转录网络的表观遗传调控失调。在AIMS 1-3中产生的数据和试剂将支持AIM 4的研究，这些研究侧重于开发利用雄激素剥夺的新方法，以实现前列腺癌的特异性治疗，证实：雄激素剥夺使ASC/CSC暴露于靶向治疗；循环中雄激素的跨细胞作用可以被抑制，而不会扰乱内皮细胞的动态平衡；内皮细胞向ASCs/CSCs发出的信号可以被重新编程，以诱导干细胞离开“利基”并致力于分化。

PROJECT SUMMARY (See instmctions): The progression of a cancer from the initiating cancer stem cell (CSC) to clinical significance, and ultimately to recurrence after treatment failure, is the culmination of a continuously evolving reciprocal interaction between the CSC and the "stem cell niche". The hypothesis of this proposal is that androgen deprivation perturbs the critical role of the prostate endothelial cell in regulation of the androgenic milieu of the prostate tissue microenvironment, and in regulation ofthe stem cell "niche", facilitating/accelerating the emergence of castration-recurrent prostate cancer To validate the hypothesis, the following Specific Aims are proposed. Aim 1 will characterize the role of endothelial cell caveolae in transcytosis of circulating androgens across the endothelial cell barrier, in endocytosis of circulating androgens resulting in AR-transactivation of genes associated with endothelial cell homeostasis and signaling, and in "non-genomic signaling" mediated through cell surface receptors for circulating androgens. Aim 2 will determine if androgen-deprivation induced killing of prostate endothelial cells results in transient, or irreversible, perturbation of the endothelial cells of the prostate microvasculature resulting in dysregulation of the tissue androgenic milieu (characterized using LC/MS/MS) and in creation/unmasking of unique, targetable "epitopes" or "vascular addresses" (characterized using phage peptide-display technology). Aim 3 will identify the effect of androgen deprivation on genes and gene pathways that are differentially expressed in prostate CSCs compared to adult stem cells (ASCs), identify the cell surface "epitope" fingerprint of prostate ASCs/CSCs, and determine if perturbation of the stem cell "niche" results in epigenetically-modulated dysregulation of the HNF-4n transcription network in prostate CSCs. The data and reagents generated in Aims 1-3 will support the studies of Aim 4 focused on development of novel approaches for utilizing androgen-deprivation to enable prostate-specific therapies by validating that: androgen deprivation exposes the ASC/CSC to targeted therapy; transcytosis of circulating androgens can be inhibited without perturbation of endothelial cell homeostasis; and endothelial cell initiated signaling to ASCs/CSCs can be reprogrammed to induce the stem cell to exit the "niche" and commit to differentiation.