MSM/WCI Partnership To Investigate Mechanisms of Prostate Cancer (2 Of 2)

描述(申请人提供)：调控昼夜节律性的分子机制由转录翻译反馈环组成，通过该环，Clock1基因产物抑制自己的转录。在实验室啮齿动物模型中，通过大脑损伤、改变光照环境和定时营养限制来实验操纵节律可以影响肿瘤的生长和存活时间。已有研究表明，在确诊的癌症患者中，异常的昼夜节律与癌症发展风险的增加和生存统计数据的减少相关。 利用新开发的生物发光小鼠肿瘤模型，生物发光前列腺癌细胞系， 和最新发展的活体分子成像技术，我们建议用来说明时空 健康前列腺癌和前列腺癌中基因表达昼夜节律的动态变化。我们还将 检查临时限制的营养是否调节前列腺的节律性、肿瘤的发生和 进步。 具体目的1：通过体外和体内生物发光报告基因的追踪，评估前列腺癌是否改变了与健康前列腺组织相比的基因表达的昼夜节律。我们将产生一个发生前列腺癌并表达PER2：荧光素酶融合蛋白的双重转基因小鼠，并研究mPer2基因在前列腺上皮内瘤变(PIN)、前列腺癌、转移灶和健康周围组织中的时间表达谱。同时，我们将开发一种致癌的前列腺癌细胞系，在C57BL/6上是同基因的，它表达昼夜荧光素酶报告基因，以便在体内跟踪麻醉小鼠的肿瘤生长和肿瘤中的分子节律，最终在清醒、行为正常的小鼠中跟踪。 具体目标2：检验对昼夜节律的营养调控不同的假设 影响肿瘤的发展我们将使双转基因小鼠或其具有生物发光肿瘤的WT仔鼠暴露于已知的改变外周器官分子和生理节律的营养操作中：白天限制进食，夜间限制进食，作为对照，限制热量而不携带昼夜节律。肿瘤的发展和进展将通过肿瘤体积的活体成像在小鼠身上进行纵向评估。同时，我们将评估每种昼夜节律操作对前列腺癌不同阶段、健康前列腺癌和皮下生物发光前列腺癌细胞中生物钟基因表达模式的影响。

DESCRIPTION (provided by applicant): The molecular machinery that governs circadian rhythmicity is comprised of transcriptional translational feedback loop whereby clock1 gene products inhibit their own transcription. Experimental manipulation of rhythms by brain lesions altered lighting environments and timed nutritional restriction can affect tumor growth and survival times in laboratory rodent models. It has been shown that aberrant circadian rhythmicity correlates with an increased risk of cancer development and with decreased survival statistics in diagnosed cancer patients. Using newly developed bioluminescent mouse tumor models, bioluminescent prostate cancer cell lines, and newly-developed in vivo molecular imaging techniques, we propose to illustrate the spatiotemporal dynamics of circadian rhythms of gene expression in healthy prostate and prostate cancer. We will also examine whether temporally restricted nutrition regulates prostate rhythmicity, and tumorigenesis and progression. Specific Aim 1: To evaluate whether prostate cancers have altered circadian rhythms of gene expression compared with healthy prostate tissue via in vitro and in vivo tracking of bioluminescent reporter genes. We will generate a dual transgenic mouse that develops prostate cancer and expresses the Per2: luciferase fusion protein and characterize the mPer2 gene temporal expression profile in prostate intraepithelial neoplasia (PIN), prostate adenocarcinoma, and metastatic foci and in healthy surrounding tissue. In parallel, we will develop a tumorigenic prostate cancer cell line, syngeneic on C57BL/6, which expresses circadian luciferase reporter genes to allow for in vivo tracking of tumor growth and molecular rhythms in tumors in anesthetized mice, and eventually in awake, behaving mice. Specific Aim 2: To test the hypothesis that nutritional manipulations of circadian rhythms differentially affect tumor development We will expose the dual transgenic mice or their WT littermates with bioluminescent tumors to nutritional manipulations known to alter molecular and physiological rhythmicity in peripheral organs: daytime restricted feeding, nighttime restricted feeding, and, as a control, caloric restriction without circadian entrainment. Tumor development and progression will be assessed longitudinally in mouse via in vivo imaging of tumor volume. In parallel, we will assess the effects of each circadian manipulation on circadian clock gene expression patterns in various stages of primary prostate cancer, in healthy prostate gland and in subcutaneous bioluminescent prostate adenocarcinoma cells.