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：
7361525
负责人：
Long-Cheng Li
金额：
$ 20.82万
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 2009-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7361525
关键词：
Animal Model Apoptosis Biological Assay Body Weight CD44 gene CDKN1A 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 Prostate Prostatic Neoplasms Proteins RNA 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 Week Western Blotting Work Xenograft procedure base cancer cell caspase-7 cell growth concept design gene function gene therapy in vivo male men migration neoplastic cell novel therapeutics oncoprotein p21 p27 Cell Cycle Protein p27 Enzyme Inhibitor promoter size 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。该提案的长期目标是为前列腺癌开发基于RNA的小型治疗策略。小型双链RNA（DSRNA）已被广泛用作小型干扰RNA（siRNA）通过靶向mRNA序列（一种称为RNA干扰（RNAI）的现象）来敲除基因表达。最近，我们和其他人确定了一种新的RNA介导的基因调节的新机制，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，这一现象称为“ RNA诱导的基因激活”（RNAA）（RNAA）（RNAA）（RNAA）和DSRNA分子“小型活性RNAS”（SARNAS）。我们证明RNAA在基因激活中有效，其作用是持久的，这表明RNAA具有有希望的治疗作用。我们假设SARNA分子在前列腺癌中靶向激活肿瘤抑制基因（TSG）的体外和体内具有抗肿瘤活性。该假设将以两个具体的目的进行检验。特定目标＃1。确定用于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。要靶向激活的基因包括（1）负细胞周期调节剂，例如p16，p21，p27和p53，（2）抑制肿瘤细胞浸润和转移的基因，包括e-钙粘着蛋白，KAI1和CD44，以及（3）包括Par-4和Caspase-4和Caspase-7。候选SARNA靶向基因启动子将被转染到人前列腺癌细胞系LNCAP和PC3中，然后将在mRNA和蛋白质水平上评估靶基因的表达。功能性SARNA将进一步评估其对细胞周期分布，细胞凋亡，克隆性，侵袭和迁移的抗增长活性和影响。特定目标＃2。评估SARNA在异种移植人类前列腺肿瘤模型中的抗肿瘤活性。异种移植前列腺肿瘤将使用人类前列腺癌PC-3细胞在无胸腺小鼠中建立。具有已建立肿瘤的小鼠将通过肿瘤内注射用功能性SARNA治疗。 SARNA处理的小鼠的肿瘤生长将与用对照肉干治疗的小鼠进行比较。这些研究构成了我们先前工作的逻辑发展。该提案的成功完成将为RNAA的进一步临床试验提供理由。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。迫切需要为前列腺癌发展新的治疗策略。该项目将研究使用小RNA分子的新鉴定的基因治疗方法的治疗用途。