Suppression of BCR-ABL by siRNA-loaded Nanoparticles for the Treatment of CML

负载 siRNA 的纳米颗粒抑制 BCR-ABL 治疗 CML

• 批准号: 8704716
• 负责人: Kseniya E Gavrilov
• 金额: $ 4.08万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8704716
• 关键词: ABL1 gene; Abnormal Myeloid Cell; Appearance; Artificial nanoparticles; Biological Models; Cardiovascular system; Cell Communication; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cells; Chromosomes, Human, Pair 9; Chronic Myeloid Leukemia; Dasatinib; Devices; Disease; Disease remission; Effectiveness; Encapsulated; Engineering; Gene Silencing; Generations; Gleevec; Glycolates; Goals; Hematological Disease; Hematopoietic; Human; Imatinib; Imatinib mesylate; In Vitro; K-562; K562 Cells; Kinetics; Knowledge; Lead; Leukemic Cell; Leukocytes; Ligands; Methods; Modification; Morphology; Mus; Myeloid Leukemia; Oncogene Proteins; Particle Size; Patients; Philadelphia Chromosome; Protein Tyrosine Kinase; Proteins; Quality of life; RNA; RNA Interference; Research; Resistance; Site; Small Interfering RNA; Specificity; Surface; System; Techniques; Technology; Testing; Therapeutic; Thermodynamics; Transcript; Tyrosine Kinase Inhibitor; Variant; base; bcr-abl Fusion Proteins; cell growth; controlled release; design; disease characteristic; fluid flow; improved; in vivo; inhibitor/antagonist; innovation; knock-down; leukemia; mouse model; nanoparticle; novel therapeutics; public health relevance; targeted delivery; uptake

DESCRIPTION (provided by applicant): Chronic Myeloid Leukemia (CML) is a disorder of hematopoietic cells wherein the appearance of the Philadelphia chromosome (a translocation of chromosomes 9 and 22) results in expression of the fusion oncoprotein BCR-ABL, whose disregulated tyrosine kinase activity causes the abnormal myeloid cell growth characteristic of the disease. In the last 20 years, substantial advances have been made in targeting BCR-ABL therapeutically to treat CML; current therapy for the disease involves daily administration of the tyrosine kinase inhibitor Imatinib Mesylate (Gleevec) and/or several second-generation inhibitors (Dasatinib, Nilotinib). However, increasing resistance to the current line of ATP-competitive inhibitors of BCR-ABL warrants discovery of novel therapeutics that can target all BCR-ABL proteins with high specificity. Recently the potential of using fusion junction-targeted small interfering RNA (siRNA) to silence BCR-ABL has been demonstrated in vitro and ex vivo. This technology could be used to design a powerful line of therapeutics that suppresses all variants of activated Abl, even imatinib/dasatinib resistant forms. But advancement of this approach toward gene silencing in vivo requires developing methods for optimizing junction-targeted siRNA efficiency, specificity, and delivery. Aim 1 of this proposed research is to design a potent and specific junction-targeted small silencing RNA for suppression of the predominant transcript variant of BCR-ABL (b3a2) and to apply techniques to load this construct into polymeric nanoparticles for delivery. In Aim 2, strategies for enhancing delivery of polymeric nanoparticles to leukemic cells via surface modification will be explored using a micro-scale fluid flow device that mimics nanoparticle-cell interactions expected in the circulatory system. After optimization of the small silencing RNA and its delivery vehicle, Aim 3 seeks to assess the efficacy of the engineered therapeutic in a humanized mouse model of leukemia. Through these aims, significant progress will be made toward the discovery and optimization of an RNAi based therapeutic for more broad, but equally potent, treatment of CML.

描述（由申请人提供）：慢性髓性白血病（CML）是一种造血细胞疾病，其中费城染色体的出现（9号和22号染色体的易位）导致融合癌蛋白BCR-ABL的表达，其酪氨酸激酶活性失调导致该疾病的异常髓性细胞生长特征。在过去的20年中，在靶向BCR-ABL治疗CML方面取得了实质性进展;目前对该疾病的治疗包括每日给予酪氨酸激酶抑制剂甲磺酸伊马替尼（格列卫）和/或几种第二代抑制剂（达沙替尼，尼洛替尼）。然而，对目前BCR-ABL的ATP竞争性抑制剂的耐药性不断增加，需要发现能够以高特异性靶向所有BCR-ABL蛋白的新疗法。最近，使用融合连接靶向的小干扰RNA（siRNA）沉默BCR-ABL的潜力已在体外和离体实验中得到证实。该技术可用于设计一种强大的治疗方法，抑制所有活化Abl的变体，甚至是伊马替尼/达沙替尼耐药形式。但是，这种方法在体内基因沉默的进步需要开发优化连接靶向siRNA效率，特异性和交付的方法。这项研究的目的1是设计一种有效的和特异性的连接靶向的小沉默RNA，用于抑制BCR-ABL（b3 a2）的主要转录变体，并应用技术将该构建体加载到聚合物纳米颗粒中进行递送。在目标2中，将使用微尺度流体探索通过表面修饰增强聚合物纳米颗粒向白血病细胞递送的策略 模拟循环系统中预期的纳米颗粒-细胞相互作用的流动装置。在优化小沉默RNA及其递送载体后，Aim 3试图评估工程化治疗剂在白血病人源化小鼠模型中的功效。通过这些目标，将在发现和优化基于RNAi的治疗方法方面取得重大进展，用于更广泛但同样有效的CML治疗。