Tumor-targeted silencing of Bcl-2/Bcl-xL by the self-assembled siRNA-nanovectors

通过自组装 siRNA 纳米载体对 Bcl-2/Bcl-xL 进行肿瘤靶向沉默

• 批准号: 7810139
• 负责人: Liang Xu
• 金额: $ 27.38万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-17 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7810139
• 关键词: Antibodies; BCL2 gene; Cell Death; Cell Proliferation; Cells; Data; Drug Delivery Systems; Drug Formulations; European; Functional RNA; Funding; Gene Expression; Genes; Goals; Growth; HMGA2 gene; Human; In Vitro; Legal patent; Ligands; Malignant Neoplasms; Malignant neoplasm of pancreas; Membrane; MicroRNAs; Modeling; Molecular; Molecular Target; Nanostructures; Neoplasm Metastasis; Normal tissue morphology; Nucleotides; Occupations; Oncogenes; Parents; Phase I Clinical Trials; Plasmids; Public Health; RNA Interference; RNA Interference Pathway; Radiation therapy; Recovery; Recurrence; Research; Resistance; Small Interfering RNA; Specificity; Stem cells; Structure; Surface; System; TP53 gene; Testing; Therapeutic; Transferrin; Transferrin Receptor; Tumor Suppressor Proteins; United States National Institutes of Health; Validation; Virion; base; cancer cell; cancer initiation; cancer stem cell; effective therapy; in vivo; knock-down; nanobiotechnology; nanovector; notch protein; novel; pancreatic neoplasm; parent grant; response; restoration; self-renewal; small hairpin RNA; stem cell population; targeted delivery; therapeutic target; tumor; tumor initiation; tumor specificity; zeta potential

DESCRIPTION (provided by applicant): With the support of the current funded R01, we have modified and optimized our nanovectors for tumor targeted delivery of siRNA/shRNA specific to Bcl-2 and Bcl-xL. We found unexpectedly that our nanovectors also targeted to pancreatic cancer stem cells (CSC) and delivered siRNA/shRNA to CSC, inhibited CSC selfrenewal and reduced cancer stem cells in vivo. Based on our exciting new discovery, we propose, in this Competitive Revision project, to expand the scope of the current R01 to further develop the nanovectors for delivery of miRNA-therapeutics targeting cancer stem cells. This is in response to the NOT-OD-09-058: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications. Our long-term goal is to develop nanovector-miRNA as a novel molecular therapy targeting human cancer stem cells. The objective in this two-year project is to explore our above patented nanovector system for targeted delivery of miRNA-based therapeutics to human pancreatic cancer stem cells, and to evaluate the therapeutic potential of the nanovector-delivered tumor suppressor miR-34 in inhibiting CSC. We will test two inter-related basic hypotheses: (1) the self-assembled nanovectors can efficiently deliver miR-34 to the pancreatic cancer stem cells, thus restoring the miR-34 tumor suppressor function in cancer stem cells; (2) miR-34 restoration will inhibit the self-renewal of pancreatic cancer stem cells by simultaneously inhibiting Bcl-2 and Notch, and thus provide a potential novel therapy for pancreatic cancer. To test our hypothesis, we propose to carry out two SPECIFIC AIMS: AIM 1, To prepare and characterize the miRNA-nanovectors targeting CSC in vitro and in vivo; AIM 2, To investigate the anti-tumor activities and the mechanism of action of nanovector-miR-34. Our long-term goal is to develop nanovector-miRNA as a novel molecular therapy targeting human cancer stem cells. Successfully carried out, our studies will provide novel, self-assembled miRNA-nanovectors that can deliver tumor suppressor miR-34 to pancreatic CSC and inhibit their self-renewal and tumor-initiation. The tumor specificity of nanovector-miR-34 is based upon: (1) cancer stem cells have high level of TfR and dysregulation of miR-34-Notch/Bcl-2 for excessive self-renewal; (2) the optimized formulation of nanovectors based on preferred miR-34 delivery to and functional inhibition of CSC; (3) normal stem cells, although also TfR(+), have no dysregulation of Bcl-2/Notch and do not rely on Bcl-2/Notch for survival. The data from the current proposal will provide important impetus to develop the nanovector-miR-34 as a novel and more effective therapy for human pancreatic cancer, potentially by modulating pancreatic cancer stem cells. PUBLIC HEALTH RELAVANCE: Our objective is to explore our patented self-assembled nanovector system for targeted delivery of miRNA-based therapeutics to human pancreatic cancer stem cells, and to evaluate the therapeutic potential of the nanovector-delivered tumor suppressor miR-34 in inhibiting cancer stem cells. Successfully carried out, our studies will provide novel, self-assembled miRNA-nanovectors that can deliver tumor suppressor miR-34 to pancreatic cancer stem cells and inhibit their self-renewal and tumor-initiation. The data from the current proposal will provide important impetus to develop the nanovector-miR-34 as a novel and more effective therapy for human pancreatic cancer by targeting cancer stem cells.

描述（由申请人提供）：在当前资助的 R01 的支持下，我们修改和优化了我们的纳米载体，用于肿瘤靶向递送 Bcl-2 和 Bcl-xL 特异性的 siRNA/shRNA。我们出乎意料地发现，我们的纳米载体还靶向胰腺癌干细胞（CSC），并将siRNA/shRNA递送至CSC，抑制CSC自我更新并减少体内癌症干细胞。基于我们令人兴奋的新发现，我们建议在这个竞争性修订项目中扩大当前 R01 的范围，以进一步开发用于递送靶向癌症干细胞的 miRNA 治疗药物的纳米载体。这是对 NOT-OD-09-058 的回应：NIH 宣布恢复法案基金可用于竞争性修订申请。我们的长期目标是开发纳米载体-miRNA作为针对人类癌症干细胞的新型分子疗法。这个为期两年的项目的目标是探索我们上述专利的纳米载体系统，用于将基于 miRNA 的治疗药物靶向递送至人类胰腺癌干细胞，并评估纳米载体递送的肿瘤抑制因子 miR-34 在抑制 CSC 方面的治疗潜力。我们将测试两个相互关联的基本假设：（1）自组装纳米载体可以有效地将miR-34递送至胰腺癌干细胞，从而恢复癌症干细胞中的miR-34抑癌功能； (2)miR-34恢复将通过同时抑制Bcl-2和Notch来抑制胰腺癌干细胞的自我更新，从而为胰腺癌提供潜在的新疗法。为了检验我们的假设，我们建议实现两个具体目标： 目标 1，在体外和体内制备和表征针对 CSC 的 miRNA 纳米载体；目的2、探讨纳米载体-miR-34的抗肿瘤活性及其作用机制。我们的长期目标是开发纳米载体-miRNA作为针对人类癌症干细胞的新型分子疗法。如果成功进行，我们的研究将提供新型的自组装 miRNA 纳米载体，可以将肿瘤抑制因子 miR-34 传递到胰腺 CSC 并抑制其自我更新和肿瘤起始。 nanovector-miR-34的肿瘤特异性基于：（1）癌症干细胞具有高水平的TfR和miR-34-Notch/Bcl-2的失调以过度自我更新； (2)基于优选的miR-34递送至CSC和功能性抑制的纳米载体的优化配方； (3)正常干细胞虽然也是TfR(+)，但没有Bcl-2/Notch失调，并且不依赖Bcl-2/Notch生存。当前提案的数据将为开发纳米载体-miR-34作为人类胰腺癌的一种新颖且更有效的疗法提供重要动力，可能通过调节胰腺癌干细胞来实现。 公共健康相关性：我们的目标是探索我们获得专利的自组装纳米载体系统，用于将基于 miRNA 的治疗药物靶向递送至人类胰腺癌干细胞，并评估纳米载体递送的肿瘤抑制因子 miR-34 在抑制癌症干细胞方面的治疗潜力。如果成功进行，我们的研究将提供新型的自组装 miRNA 纳米载体，可以将肿瘤抑制因子 miR-34 传递到胰腺癌干细胞并抑制其自我更新和肿瘤启动。当前提案的数据将为开发纳米载体-miR-34作为一种通过靶向癌症干细胞的新型且更有效的人类胰腺癌疗法提供重要动力。