CXCL13: a mediator of prostate cancer progression

CXCL13：前列腺癌进展的介质

• 批准号: 9256445
• 负责人: MARCELO G. KAZANIETZ
• 金额: $ 35.61万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9256445
• 关键词: Animals; Antineoplastic Agents; Attention; Automobile Driving; BLR1 gene; Bioinformatics; Breast; CXCL13 gene; Cancer Etiology; Carcinoma; Cell model; Cells; Cessation of life; Collection; Data; Development; Diglycerides; Disease; Disease Progression; Drug Targeting; Engineering; Epithelial; Epithelial Cells; Etiology; Event; Gene Expression Profile; Genes; Growth; Guanine Nucleotide Exchange Factors; Head and Neck Cancer; Human; Human Characteristics; Immunocompetent; Impairment; In Vitro; Individual; Injection of therapeutic agent; Knock-out; Knockout Mice; Laboratories; Lesion; Lung; Lymphocyte Depletion; Maintenance; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Mediator of activation protein; Messenger RNA; Metastatic Prostate Cancer; Microarray Analysis; Modeling; Molecular; Monomeric GTP-Binding Proteins; Mus; Neoplasm Metastasis; Nude Mice; Oncogenic; PIK3CG gene; Pathology; Pathway interactions; Pharmacology; Phenotype; Phosphoric Monoester Hydrolases; Phosphotransferases; Play; Production; Proliferating; Property; Prostate; Prostate Adenocarcinoma; Prostatic Neoplasms; Protein Isoforms; Protein Microarray Assay; Protein Overexpression; RNA Interference; Recurrence; Role; Signal Pathway; Signal Transduction; Site; Specimen; Stimulation of Cell Proliferation; Subfamily lentivirinae; Therapeutic; Transgenic Mice; Transgenic Model; Transgenic Organisms; Tumor Suppressor Proteins; Tumorigenicity; Up-Regulation; autocrine; bone; cancer biomarkers; cancer therapy; cell motility; cell transformation; cellular engineering; chemokine; drug development; grasp; link protein; men; mouse model; novel; overexpression; prognostic; prostate cancer cell; prostate cancer cell line; prostate carcinogenesis; protein kinase C epsilon; public health relevance; rac GTP-Binding Proteins; receptor; small hairpin RNA; tumor; tumor progression; tumorigenesis; tumorigenic

 DESCRIPTION (provided by applicant): Prostate cancer is the second leading cause of cancer-related deaths among men in the US. A large body of evidence links protein kinase C epsilon (PKC) to cancer progression. PKC is markedly up-regulated in epithelial cancers, including prostate cancer, potentially reflecting its involvement in the etiology and progression o the disease. We found that prostate-specific PKC transgenic mice (PB-PKC) develop preneoplastic lesions (PINs), and remarkably, they develop invasive prostatic adenocarcinomas in a Pten haplodeficient background. Understanding the molecular basis of this cooperativity is highly relevant, as loss of the tumor suppressor Pten, a negative regulator of the PI3K pathway, is a common alteration of prostate cancer. We generated a number of cellular models recapitulating PKC overexpression and Pten loss, which display enhanced growth, motility, and invasiveness in vitro, and acquire tumorigenic potential in nude mice. Moreover, gene profiling and bioinformatics analysis revealed that PKC overexpressing/Pten depleted cells display a pattern of gene expression distinctive of human metastatic prostate cancer. We found that the "top hit" in this microarray analysis is the chemokine CXCL13. Notably, PKC cooperates with Pten loss to strongly induce CXCL13 mRNA levels and its release, and RNAi depletion of CXCL13 or its receptor (CXCR5) impairs the ability of PKC overexpressing/Pten-deficient cells to proliferate and migrate. Preliminary data also suggest that PKC and PI3K are downstream effectors of CXCR5, suggesting a positive amplification loop that may act as a vicious cycle. In Specific Aim 1, we will establish the relevance of the CXCL13:CXCR5 axis in prostate tumorigenesis using an inducible silencing approach. A potential contribution of stromal CXCL13 will be examined using immunocompetent mice and depletion of lymphocytes involved in CXCL13 production. In addition, knock-out CXCL13 and CXCR5 mice will be crossed with PB- PKC mice as an approach to assess the requirement of the CXCL13:CXCR5 axis in the development of prostate lesions and the activation of key mitogenic/survival pathways. In Specific Aim 2, we will examine the role of CXCL13:CXCR5 in metastasis using intraprostatic injections and also assess the role of this pathway in metastatic dissemination to bone. Mechanistic studies will be pursued to explore the potential relevance of Rac, a small G-protein, and its exchange factors (Rac-GEFs) in the motile/invasive phenotype driven by the PKC-PI3K-CXCL13-CXCR5 pathway. In Specific Aim 3, we will focus on the mechanisms involved in CXCL13 induction by PKC and PI3K, specifically dissecting the roles of Akt, Erk, Stat3, NF-B and specific p110 PI3Ks in controlling CXCL13 production. We will also investigate PKC as a potential CXCR5 downstream effector. Lastly, in Specific Aim 4 we will establish the relevance of the PKC/Pten-CXCL13 association in human prostate tumors using a large collection of prostate cancer specimens, and assess correlations with disease progression and recurrence. In addition to the mechanistic implications, the proposed studies may have major prognostic and therapeutic impact for personalized prostate cancer treatment.

 描述（由适用提供）：前列腺癌是美国男性与癌症相关死亡的第二大原因。大量证据将蛋白激酶C epsilon（PKC）与癌症的进展联系起来。 PKC在包括前列腺癌在内的上皮癌中明显上调，有可能反映其参与病因学和进展的疾病。我们发现，前列腺特异性PKC转基因小鼠（PB-PKC）产生了肿瘤性病变（PIN），并且非常明显地，它们在PTEN单倍缺陷型背景中发展出侵入性的前列腺腺癌。理解这种协调的分子基础是高度相关的，因为PI3K途径的负调节剂的肿瘤抑制剂PTEN的丧失是前列腺癌的常见改变。我们产生了许多概括PKC过表达和PTEN损失的细胞模型，这些模型在体外表现出增长，运动性和侵入性，并在裸鼠中获得致癌潜力。此外，基因分析和生物信息学分析表明，PKC过表达/PTEN DEPTHD细胞表现出一种与人类转移性前列腺癌不同的基因表达模式。我们发现此微阵列分析中的“最高点击”是趋化因子CXCL13。值得注意的是，PKC会与PTEN损失合作，从而强烈影响CXCL13 mRNA水平及其释放，RNAi对CXCL13或其受体（CXCR5）的定义会损害PKC过表达/PTEN缺乏细胞增殖和迁移的能力。初步数据还表明，PKC和PI3K是CXCR5的下游效应，这表明可能充当恶性循环的阳性扩增环。在特定的目标1中，我们将使用诱导的沉默方法来建立前列腺肿瘤发生中CXCL13：CXCR5轴的相关性。将使用免疫能力的小鼠和CXCL13产生涉及的淋巴细胞的部署来检查基质CXCL13的潜在贡献。此外，将敲除CXCL13和CXCR5小鼠将与PB-PKC小鼠交叉，以评估CXCL13：CXCR5轴在前列腺病变发展中的需求以及关键有丝裂/生存途径的激活。在特定的目标2中，我们将使用手感内注射来检查CXCL13：CXCR5在转移中的作用，并评估该途径在转移到骨骼中的作用。将进行机械研究，以探索RAC，小的G蛋白的潜在相关性及其在PKC-PI3K-CXCL13-CXCR5途径驱动的母亲/侵入性表型中的交换因子（RAC-GEF）。在特定的目标3中，我们将重点关注PKC和PI3K诱导中涉及的机制，特别剖析了AKT，ERK，STAT3，NF-B和特定P110 PI3K在控制CXCL13产量中的作用。我们还将研究PKC作为潜在的CXCR5下游效应子。最后，在特定目标4中，我们将使用大量前列腺癌标本在人类前列腺肿瘤中建立PKC/PTEN-CXCL13关联的相关性，并评估与疾病进展和复发的相关性。除了机械含义外，拟议的研究可能对个性化前列腺癌治疗具有重大的预后和热影响。