Targeting PDE4D as a candidate oncogene in prostate cancer

将 PDE4D 作为前列腺癌的候选癌基因

• 批准号: 7913935
• 负责人: Kimberly Dawn petry Hammer
• 金额: $ 4.58万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-24 至 2012-03-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7913935
• 关键词: Alternative Splicing; Candidate Disease Gene; Cell Proliferation; Clinical Trials; Complementary DNA; Data; Development; Epithelial Cells; Exons; Future; Growth; Health; Histology; Human; Immigration; Injection of therapeutic agent; Laboratories; Malignant neoplasm of prostate; Modeling; Mus; Mutagenesis; Mutate; Nuclear; Nude Mice; Oncogenes; PC3 cell line; Pharmaceutical Preparations; Phenotype; Pre-Clinical Model; Prostate; Prostate Cancer therapy; Prostatic; Prostatic Epithelium; Protein Isoforms; Rattus; Research Proposals; Role; Safety; Sampling; Signal Pathway; Site; Technology; Testing; Tissues; Transgenic Mice; Transgenic Organisms; Tumor Suppressor Proteins; Up-Regulation; Xenograft procedure; efficacy testing; inhibitor/antagonist; novel; phosphoric diester hydrolase; prevent; probasin; promoter; protein expression; public health relevance; small hairpin RNA; small molecule

DESCRIPTION (provided by applicant): This proposal will investigate candidate oncogene PDE4D in prostate cancer and explore the potential of a small molecule PDE4D inhibitor, NVP-ABE171, as an anti-prostate cancer agent. Our laboratory used a transposon-mutagenesis strategy to identify common transposon insertion sites that were recurrently mutated in mice with phenotypic changes resembling prostate cancer. One novel candidate gene that was identified was a phosphodiesterase called PDE4D. Preliminary data showed that PDE4D protein expression was increased in human prostate cancer cell lines and in human prostate cancer tissue. Finally, shRNA knockdown of exons common to all PDE4D isoforms reduced prostate cell proliferation and migration in prostate cancer cell lines and in xenograft-bearing mice. Analysis of alternative splicing of PDE4D identified one isoform of PDE4D, called PDE4D5v2, that was the predominant isoform found in human prostate cancer samples. This data suggested that PDE4D5v2 is a potential prostate cancer oncogene. The hypothesis of this proposal is that the up-regulation of isoform-specific PDE4D is critical for prostate cancer development, and the PDE4D inhibitor NVP-ABE171 will decrease the growth and/or progression of prostate cancer in pre- clinical models. In Aim 1, isoform specific knockdown of PDE4D5v2 with shRNA technology will be used to assess the impact of PDE4D5v2 on proliferation and signaling pathways important in prostate cancer. In Aim 2, the over-expression of PDE4D5v2 in human prostate cancer will be modeled in mice using a transgenic approach to drive expression of PDE4D5v2 specifically in prostatic epithelial cells. A cDNA for human PDE4D5v2 will be cloned into an expression construct that places PDE4D expression under the control of the ARR2PB version of the rat probasin promoter. The expression construct will be used to create transgenic mice with over-expression of PDE4D5v2 in the prostatic epithelium using pro-nuclear injection. Changes in the prostates of the ARR2PB-PDE4D5v2 transgenic mice that resemble features of human prostate cancer will be identified using histology and immunohistochemical approaches. In Aim 3, the PDE4D specific inhibitor, NVP- ABE171, will be tested for the ability to slow prostate cancer growth and/or progression first in prostate cancer cell lines and in xenografts of prostate cancer cell lines in nude mice, and finally for its ability to prevent and/or reverse the prostatic phenotypes caused by PDE4D5v2 over-expression in transgenic mice and in mice with prostate specific deletion of the tumor suppressor Pten. This research proposal is applicable to human health because it will define the role of a novel prostate cancer oncogene in the development of prostate cancer and will assess the efficacy of NVP-ABE171 as an anti-prostate cancer agent. The potential of NVP-ABE171 to be used in future clinical trials for prostate cancer is significant if the data can demonstrate efficacy in the pre-clinical models that I have proposed to study. PUBLIC HEALTH RELEVANCE: This proposal will explore a novel prostate cancer oncogene and will test the efficacy of a small molecule inhibitor of the novel oncogene. This inhibitor can be taken orally and extensive safety data is available for this class of drugs, making it an ideal potential anti-prostate cancer therapy. The potential of this inhibitor to be used in future clinical trials for prostate cancer is significant if the data can demonstrate efficacy in the pre- clinical models of prostate cancer that I have proposed to study.

描述（由申请方提供）：本提案将研究前列腺癌中的候选癌基因PDE 4D，并探索小分子PDE 4D抑制剂NVP-ABE 171作为抗前列腺癌药物的潜力。我们的实验室使用转座子诱变策略来识别常见的转座子插入位点，这些位点在具有类似前列腺癌的表型变化的小鼠中反复突变。一个新的候选基因被确定是磷酸二酯酶称为PDE 4D。初步数据显示，PDE 4D蛋白表达在人前列腺癌细胞系和人前列腺癌组织中增加。最后，在前列腺癌细胞系和异种移植小鼠中，所有PDE 4D亚型共有的外显子的shRNA敲低减少了前列腺细胞增殖和迁移。PDE 4D的选择性剪接分析鉴定了PDE 4D的一种亚型，称为PDE 4D 5v 2，这是在人前列腺癌样品中发现的主要亚型。这些数据表明，PDE 4D 5v 2是一个潜在的前列腺癌癌基因。该提议的假设是，亚型特异性PDE 4D的上调对于前列腺癌的发展至关重要，并且PDE 4D抑制剂NVP-ABE 171将在临床前模型中降低前列腺癌的生长和/或进展。在目标1中，将使用shRNA技术对PDE 4D 5v 2的同种型特异性敲低来评估PDE 4D 5v 2对前列腺癌中重要的增殖和信号传导途径的影响。在目的2中，将使用转基因方法在小鼠中对人前列腺癌中PDE 4D 5v 2的过表达进行建模，以驱动PDE 4D 5v 2在前列腺上皮细胞中的特异性表达。将人PDE 4D 5v 2的cDNA克隆到表达构建体中，所述表达构建体将PDE 4D表达置于大鼠probasin启动子的ARR 2 PB形式的控制下。表达构建体将用于使用前核注射产生在前列腺上皮中过表达PDE 4D 5v 2的转基因小鼠。ARR 2 PB-PDE 4D 5v 2转基因小鼠前列腺中类似于人前列腺癌特征的变化将使用组织学和免疫组织化学方法进行鉴定。在目标3中，将首先在前列腺癌细胞系和裸鼠中的前列腺癌细胞系的异种移植物中测试PDE 4D特异性抑制剂NVP-ABE 171减缓前列腺癌生长和/或进展的能力，最后，其预防和/或逆转由PDE 4D 5v 2过度表达引起的前列腺表型的能力，在转基因小鼠和前列腺特异性缺失肿瘤抑制因子Pten的小鼠中的表达。这项研究计划适用于人类健康，因为它将确定一种新的前列腺癌癌基因在前列腺癌发展中的作用，并将评估NVP-ABE 171作为抗前列腺癌药物的疗效。NVP-ABE 171用于未来前列腺癌临床试验的潜力是显著的，如果数据可以在我提议研究的临床前模型中证明疗效。 公共卫生关系：这项提案将探索一种新的前列腺癌癌基因，并将测试新癌基因的小分子抑制剂的疗效。这种抑制剂可以口服，并且这类药物有广泛的安全性数据，使其成为理想的潜在抗前列腺癌疗法。如果数据能够证明在我提议研究的前列腺癌临床前模型中的疗效，这种抑制剂在未来前列腺癌临床试验中使用的潜力是显著的。