Tumor targeted RNAi by novel nanovectors for molecular therapy of prostate cancer

新型纳米载体肿瘤靶向RNAi用于前列腺癌的分子治疗

• 批准号: 7238432
• 负责人: Liang Xu
• 金额: $ 9.73万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7238432
• 关键词: Animal Model; Antibodies; Antisense Oligonucleotides; Apoptosis; Apoptotic; BCL-Xs protein; Biological; Cell Death; Charge; Clinical Research; Development; Double-Stranded RNA; Drug Formulations; Drug resistance; Gene Delivery; Gene Expression; Gene Silencing; Gene Targeting; Genes; Goals; Human; In Vitro; Legal patent; Ligands; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Membrane; Molecular; Molecular Target; Nanostructures; Neoplasm Metastasis; Normal Cell; Normal tissue morphology; Nude Mice; Oncogenes; Phase I Clinical Trials; Pilot Projects; Plasmids; Prostate; Prostate Cancer therapy; Prostatic Neoplasms; Protein Overexpression; Proteins; RNA; RNA Interference; Radiation therapy; Resistance; Small Interfering RNA; Specificity; Structure; Surface; System; Testing; Therapeutic; Transferrin; Transferrin Receptor; Treatment Efficacy; United States Food and Drug Administration; Validation; Viral; Virion; Xenograft Model; anticancer research; base; cancer cell; cancer therapy; concept; design; gene therapy; human disease; improved; in vivo; knock-down; nanoparticle; nanovector; novel; novel therapeutics; receptor; size; small hairpin RNA; success; targeted delivery; therapy resistant; tool; tumor; tumor initiation; tumor progression

DESCRIPTION (provided by applicant): The potent, sequence-specific gene silencing by small interfering RNA (siRNA) has become a powerful tool in cancer research and holds significant potential as novel molecular therapy for cancer. However, delivering the siRNA-based therapeutics efficiently and specifically to prostate cancer and its metastases remains a great challenge. We have developed a tumor-specific, ligand-targeting, self-assembled DNA-nanovector system for prostate cancer gene delivery. The nanovectors show promising efficiency and specificity in targeted delivery of various genes and anti-sense oligonucleotides to human prostate cancer, with limited effect on normal tissues (US Patent No. 6,749,863). This nanovector system is now in Phase I clinical trial for non-viral p53 gene therapy. The objective of this pilot project proposal is to explore the feasibility of using our patented nanovector system for tumor-targeted delivery of siRNA-based therapeutics for human prostate cancer. In our preliminary studies, we have designed siRNAs for human Bcl-2 and Bcl-xL that can potently knock-down Bcl-2/Bcl-xL up to 95%, leading to extensive cancer cell death (US Patent pending). We propose to use Bcl-2/Bcl-xL siRNA to test two inter-related hypotheses: (1) the self-assembled nanovectors can selectively deliver siRNA to prostate cancer and efficiently silence the target Bcl-2/Bcl-xL; (2); Knock-down of the anti-apoptotic Bcl-2/Bcl-xL will induce apoptosis in human prostate cancer cells that depend on Bcl-2/Bcl-xL for survival, thus overcome resistance and restore sensitivity of prostate cancer cells to chemo/radiotherapy. Our long-term goal is to develop the tumor-targeting siRNA-nanovectors as novel molecular therapy for prostate cancer. To test our hypothesis, we propose to carry out two SPECIFIC AIMS: AIM 1: To prepare and optimize the nanovectors for efficient RNA interference for human prostate cancer in vitro and in vivo; AIM 2: To investigate anti-tumor activities and target validation of Bcl-2/Bcl-xL siRNA-nanovectors in nude mouse xenograft models of human prostate cancer with high levels of Bcl-2/Bcl-xL. Anti-apoptotic proteins Bcl-2 and Bcl-xL are overexpressed in most of prostate cancer and contribute to prostate tumor initiation, progression and resistance to therapy. Molecular modulation of Bcl-2/Bcl-xL represents a promising strategy for overcoming the resistance to apoptosis induced by current cancer therapy. Combining siRNA-based molecular therapy with conventional therapy would improve the efficacy and overcome the resistance to current cancer treatment, especially for tumor metastasis, in which Bcl-2/Bcl-xL protein is overexpressed and for which conventional therapy is not very effective. Successfully carried out, our study will provide proof-of-concept that siRNA can be delivered by the self-assembled nanovectors for tumor-targeted RNA interference of the genes critical for prostate cancer progression and resistance. Combining tumor-targeted RNA interference of Bcl-2/Bcl-xL with conventional therapy would improve the efficacy and overcome the drug resistance to current cancer treatment, especially for metastasis, in which Bcl- 2/Bcl-xL protein is overexpressed and for which conventional therapy is not very effective. Successfully carried out, our studies will provide proof-of-concept that siRNA can be delivered by the self-assembled nanovectors for tumor-targeted silencing of the genes critical for prostate tumor progression and resistance. The success of tumor-targeted RNA interference by nanovectors in vivo will have a major impact on the development of siRNA-based novel therapeutics for various molecular targets of human prostate cancer.

描述（由申请人提供）：小型干扰RNA（siRNA）的有效的，序列特异性的基因沉默已成为癌症研究中的强大工具，并具有作为癌症分子疗法的新型潜力。但是，有效地，专门针对前列腺癌及其转移的疗法提供了基于siRNA的治疗剂仍然是一个巨大的挑战。我们已经开发了一种用于前列腺癌基因递送的肿瘤特异性，靶向的，自组装的DNA-Nanovector系统。纳米分离器在针对性递送各种基因和反义寡核苷酸到人前列腺癌的靶向递送方面表现出有希望的效率和特异性，对正常组织的影响有限（美国6,749,863）。现在，该纳米动物系统正在进行I期非病毒p53基因治疗的I期临床试验。该试点项目提案的目的是探索使用我们专利的纳米植物系统以针对人类前列腺癌的基于siRNA的疗法进行肿瘤递送的可行性。在我们的初步研究中，我们为人类BCL-2和BCL-XL设计了siRNA，这些siRNA可以有效敲低BCL-2/BCL-XL高达95％，从而导致广泛的癌细胞死亡（美国专利待处理）。我们建议使用Bcl-2/bcl-XL siRNA检验两个相互关联的假设：（1）自组装的纳米旋转器可以选择性地输送siRNA以使前列腺癌并有效地使靶标Bcl-2/bcl-XL有效地静音； （2）;抗凋亡BCL-2/BCL-XL的敲除将诱导依赖Bcl-2/bcl-XL的人类前列腺癌细胞中的凋亡，从而克服耐药性并恢复前列腺癌细胞对化学/放射治疗的敏感性。我们的长期目标是开发靶向肿瘤的siRNA-NANONOVECTOR作为前列腺癌的新分子疗法。为了检验我们的假设，我们建议执行两个具体的目的：目标1：准备和优化纳米分离器，以有效地在体外和体内对人前列腺癌的有效干扰；目的2：研究抗肿瘤活性和靶向验证Bcl-2/bcl-XL siRNA-NANANONONOVETOR中的裸小鼠异种移植物模型，具有高水平的Bcl-2/bcl-XL。抗凋亡蛋白Bcl-2和Bcl-XL在大多数前列腺癌中过表达，并有助于前列腺肿瘤的启动，进展和对治疗的抗性。 Bcl-2/bcl-XL的分子调节是克服当前癌症治疗引起的对凋亡的抵抗力的有前途的策略。将基于siRNA的分子疗法与常规疗法相结合将提高疗效并克服对当前癌症治疗的抗药性，尤其是对于肿瘤转移，在这种肿瘤转移中，Bcl-2/bcl-XL蛋白过表达，并且常规治疗不是很有效。成功进行的研究将提供概念证明，即siRNA可以由自组装的纳米分离器传递，以实现针对前列腺癌进展和耐药性至关重要的肿瘤靶向的RNA干扰。将Bcl-2/BCl-XL与常规疗法相结合，将肿瘤靶向的RNA干扰结合在一起，将提高功效并克服对当前癌症治疗的耐药性，尤其是对于转移的耐药性，在这种转移中，BCL-2/BCl-XL蛋白过表达了，并且对于哪种常规疗法不太有效。成功进行的研究将提供概念证明，即siRNA可以由自组装的纳米分离器传递，以实现对前列腺肿瘤进展和耐药性至关重要的基因的沉默。纳米分离器在体内受到肿瘤靶向的RNA干扰的成功将对基于siRNA的新型疗法的发展产生重大影响，用于人类前列腺癌的各种分子靶标。