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Small RNA targeted gene activation for the treatment of prostate cancer

小RNA靶向基因激活治疗前列腺癌

基本信息

批准号：
7541428
负责人：
Long-Cheng Li
金额：
$ 17.38万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-01-01 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7541428
关键词：
Animal Model Apoptosis Biological Assay Body Weight CD44 gene Cancer Etiology Categories Cell Cycle Cell Proliferation Cell division Cells Cessation of life Clinical Trials Complex Disease Double-Stranded RNA E-Cadherin Enzyme-Linked Immunosorbent Assay Epigenetic Process Exhibits Flow Cytometry Gene Activation Gene Expression Gene Expression Regulation Gene Targeting Genes Growth Guide RNA Human In Vitro Injection of therapeutic agent KAI1 gene LNCaP Malignant Neoplasms Malignant neoplasm of prostate Mediating Messenger RNA Methods Migration Assay Modeling Molecular Target Mus Neoplasm Metastasis Nude Mice Oncogenes PAWR gene PC3 cell line Phenotype Promoter Regions Prostatic Neoplasms Proteins RNA Interference Reporting Research Reverse Transcriptase Polymerase Chain Reaction Role Sarna Second Primary Cancers Small Interfering RNA Small RNA Specificity TP53 gene TdT-Mediated dUTP Nick End Labeling Assay Technology Testing Therapeutic Therapeutic Uses Time Tumor Burden Tumor Cell Invasion Tumor Suppressor Genes Western Blotting Work Xenograft procedure base cancer cell caspase-7 cell growth design gene function gene therapy in vitro activity in vivo male men migration neoplastic cell novel therapeutics promoter small molecule tool tumor tumor growth

项目摘要

DESCRIPTION (provided by applicant): In the U.S., prostate cancer is the most common male cancer and the second leading cause of cancer deaths in men. The long-term objective of this proposal is to develop small RNA-based therapeutic strategies for prostate cancer. Small double-stranded RNA (dsRNA) has been widely used as small interfering RNA (siRNA) to knockdown gene expression by targeting mRNA sequences, a phenomenon known as RNA interference (RNAi). Recently we and others identified a new mechanism of small RNA-mediated gene regulation in which dsRNA molecules targeting gene promoter regions induce sequence-specific transcriptional gene activation, a phenomenon termed "RNA-induced gene activation" (RNAa) and the dsRNA molecules "small activating RNAs" (saRNAs). We demonstrated that RNAa is potent in gene activation and its effects are long-lasting, suggesting a promising therapeutic role for RNAa. We hypothesize that saRNA molecules have anti-tumor activity in vitro and in vivo via targeted activation of tumor suppressor genes (TSGs) in prostate cancer. This hypothesis will be tested in two specific aims. Specific Aim #1. Identify functional saRNAs for TSG activation and evaluate their anti-growth effects in cultured human prostate cancer cells. Genes to be targeted for activation include (1) negative cell cycle regulators such as p16, p21, p27 and p53, (2) genes that inhibit tumor cell invasion and metastasis including E-cadherin, KAI1 and CD44, and (3) pro-apoptosis genes including Par-4 and caspase-7. Candidate saRNAs targeting gene promoters will be transfected into human prostate cancer cell lines LNCaP and PC3, and the expression of targeted genes will then be evaluated at both mRNA and protein levels. Functional saRNAs will be further assessed for their anti-growth activity and effects on cell cycle distribution, apoptosis, clonogenicity, invasion and migration. Specific Aim #2. Evaluate saRNA's anti-tumor activity in xenograft human prostate tumor models. Xenograft prostate tumors will be established in athymic mice using human prostate cancer PC-3 cells. Mice with established tumors will be treated with functional saRNAs by intratumoral injections. Tumor growth in saRNA-treated mice will be compared with mice treated with control saRNAs. These studies constitute a logical progression of our previous work. Successful completion of this proposal will provide rationale for further clinical trials of RNAa. In the U.S., prostate cancer is the most common male cancer and the second leading cause of cancer deaths in men. There is an urgent need to develop novel therapeutic strategies for prostate cancer. This project will investigate the therapeutic use of a newly identified method of gene therapy using small RNA molecules.

描述（由申请人提供）：在美国，前列腺癌是最常见的男性癌症，也是男性癌症死亡的第二大原因。本提案的长期目标是开发基于小rna的前列腺癌治疗策略。小双链RNA （dsRNA）被广泛用作小干扰RNA (siRNA)，通过靶向mRNA序列来敲低基因表达，这种现象被称为RNA干扰（RNAi）。最近，我们和其他人发现了一种新的小rna介导的基因调控机制，其中dsRNA分子靶向基因启动子区域诱导序列特异性转录基因激活，这种现象称为“rna诱导基因激活”（RNAa）， dsRNA分子称为“小激活rna”（saRNAs）。我们证明了RNAa在基因激活中是有效的，其作用是持久的，这表明RNAa在治疗中具有很好的作用。我们假设saRNA分子在体外和体内通过靶向激活前列腺癌的肿瘤抑制基因（TSGs）而具有抗肿瘤活性。这一假设将在两个具体目标中得到检验。明确目标#1。鉴定TSG激活的功能性sarna并评估其在培养的人前列腺癌细胞中的抗生长作用。被激活的目标基因包括(1)细胞周期负调控基因，如p16、p21、p27和p53；(2)抑制肿瘤细胞侵袭和转移的基因，如E-cadherin、KAI1和CD44；(3)促凋亡基因，如Par-4和caspase-7。将靶向基因启动子的候选saRNAs转染到人前列腺癌细胞LNCaP和PC3中，然后在mRNA和蛋白水平上评估靶向基因的表达。功能性sarna将进一步评估其抗生长活性以及对细胞周期分布、凋亡、克隆原性、侵袭和迁移的影响。具体目标2。评价saRNA在异种移植人前列腺肿瘤模型中的抗肿瘤活性。将利用人前列腺癌PC-3细胞在胸腺小鼠体内建立异种前列腺肿瘤。已形成肿瘤的小鼠将通过瘤内注射功能性sarna进行治疗。用sarna处理的小鼠的肿瘤生长将与用对照sarna处理的小鼠进行比较。这些研究构成了我们以前工作的合乎逻辑的进展。该建议的成功完成将为RNAa的进一步临床试验提供理论依据。在美国，前列腺癌是最常见的男性癌症，也是男性癌症死亡的第二大原因。目前迫切需要开发新的前列腺癌治疗策略。该项目将研究一种新发现的使用小RNA分子的基因治疗方法的治疗用途。