Neuroendocrine Cancer: Tumor Regulation by PTHrP In Vitro and In Vivo

项目概要/摘要： 我们的总体假设是，PTHrP（甲状旁腺激素相关蛋白）过度表达可能与甲状腺功能低下有关。 前列腺癌的特征在于从这三种肽中产生新的癌症特异性肽。 这种癌蛋白的天然同种型，这些新的肽可以调节肿瘤发生， 前列腺癌的生长和进展，特别是在骨骼中。我们的总体目标是确定 这些肽及其对生长调节和肿瘤进展的作用， 了解这种恶性肿瘤的病理学 具体目标1。为了鉴定和充分表征来自于人的PTHrP肽， 前列腺癌中的天然PTHrP多肽。我们将识别和表征PTHrP 前列腺癌产生的肽。这将通过多方面的办法来实现， 包括（a）免疫化学方法如免疫亲和层析、多位点 免疫测定和免疫组织化学，包括Western和（B）生化程序 基于质谱（MS）。这些方法将通过额外的分子补充。 PTHrP的研究 预期结果总结：我们预期鉴定PTHrP的前列腺特异性肽， 介导对前列腺癌的生长和增殖相关方面的影响。除了PTHrP 经典的肽，我们希望发现新的肽，可以发挥其分子作用，虽然 新的分子途径。 具体目标2。为了确定这些PTHrP加工肽如何调节前列腺癌 骨骼和其他部位的进展。这一目标将在免疫功能低下的 通过肽处理和通过将前列腺癌细胞直接植入骨和其它部位的小鼠 已经被基因工程改造用于PTHrP表达， PTHrP及其肽的调节作用。新的成像程序将用于监测肿瘤 行为这些模型将使我们能够研究PTHrP各自的作用和分子相互作用 介导前列腺癌的体内进展。 预期结果总结：我们期望鉴定PTHrP肽，其调节相应的 在我们的前列腺癌肿瘤动物模型中，骨骼和非骨骼的前列腺癌进展。我们 期望基于目标1的相应体外研究来告知和关注这些体内研究 以及我们在这方面的初步成果。我们希望能找出并阐明- PTHrP及其衍生肽的调节结构域，从而促进PTHrP及其衍生肽的开发， 这些机制用于开发肿瘤的诊断和治疗靶点。

Project Summary/Abstract: It is our overall Hypothesis that PTHrP (parathyroid hormone related protein) overexpression in prostate cancer is characterized by the generation of novel, cancer-specific peptides from the three native isoforms of this oncoprotein and that these novel peptides can regulate tumorigenesis and the growth and progression of prostate cancer, especially in the skeleton. Our overall Aim is to identify these peptides and their effects on growth-regulation and tumor progression in order to fully understand the pathobiology of this malignancy. Specific Aim 1. To identify and fully characterize the PTHrP peptides that are derived from the native PTHrP polypeptides in prostate cancer. We shall identify and characterize the PTHrP peptides produced by prostate cancers. This will be accomplished by a multifaceted approach that includes (a) immunochemical procedures like immunoaffinity chromatography, multi-site immunoassays, and immunohistochemistry, including Westerns and (b) biochemical procedures based on mass spectrometry (MS). These methods will be complemented by additional molecular studies of PTHrP. Summary of Expected Results: We expect to identify prostate-specific peptides of PTHrP that mediate effects on growth- and proliferation-related aspects of prostate cancer. In addition to PTHrP's classical peptides, we expect to discover new peptides that can exert their molecular action though novel molecular pathways. Specific Aim 2. To determine how these PTHrP-processed peptides regulate prostate cancer progression in skeletal and other sites. This aim will be accomplished in immunocompromised mice by peptide treatment and by directly implanting into bone and other sites prostate cancer cells that have been genetically engineered for PTHrP expression to respectively manifest the growth regulatory effects of PTHrP and its peptides. Novel imaging procedures will be used to monitor tumor behavior. These models will allow us to study PTHrP's respective effects and molecular interactions that mediate in vivo progression of prostate cancer. Summary of Expected Results: We expect to identify PTHrP peptides that regulate the respective progression, both skeletal and non-skeletal, of prostate cancer in our animal model for this tumor. We expect to inform and focus these in vivo studies based on the corresponding in vitro studies of Aim 1 as well as our Preliminary Results in this area. We expect to identify and elucidate the growth- regulatory domains of PTHrP and its derived peptides in vivo and to thus facilitate exploitation of these mechanisms for the development of diagnostic and therapeutic targets for the tumor.