Encapsulated siRNAs for treatment of urological disease

用于治疗泌尿系统疾病的封装 siRNA

• 批准号: 7238990
• 负责人: ROBERT M WEISS
• 金额: $ 20.63万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-07 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7238990
• 关键词: Address; Adverse effects; Animal Disease Models; Animal Model; Animals; Base Pairing; Bladder; Cancer Model; Cancer cell line; Cells; Collaborations; Communicable Diseases; Complex; Disease model; Down-Regulation; Drug Delivery Systems; Encapsulated; Engineering; Extracellular Matrix; Fluorescein; Fluoresceins; Genes; Glycolic-Lactic Acid Polyester; Goals; Half-Life; Human; Hyperreflexia; Immune; Immunotherapy; In Vitro; Individual; Inflammatory; Inflammatory Response; Interstitial Cystitis; Intravesical Instillation; Label; Life; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Measures; Messenger RNA; Microspheres; Modeling; Mus; Nanosphere; Nitrosamines; Oligonucleotides; Overactive Bladder; Parathyroid Hormones; Pathway interactions; Penetration; Polymers; Positioning Attribute; Protein Overexpression; Proteins; Protocols documentation; RNA; RNA Interference; RNA Stability; Rate; Scanning Electron Microscopy; Site; Small RNA; Specificity; Standards of Weights and Measures; System; Techniques; Testing; Therapeutic; Time; Toxic effect; Transcriptional Activation; Transgenic Mice; Transgenic Organisms; Treatment Protocols; Tumor Burden; Up-Regulation; Urologic Diseases; Urothelium; Vascular Endothelial Growth Factors; Viral; bladder transitional cell carcinoma; cancer cell; chemotherapy; controlled release; day; design; drug efficacy; gene therapy; human PTH protein; in vivo; intravesical; mortality; mouse model; polylactic acid-polyglycolic acid copolymer; professor; protein expression; survivin; tool; tumor; uptake

DESCRIPTION (provided by applicant): Intravesical instillation of small interference RNAs (siRNAs) is an exciting new treatment option for urological diseases. siRNA oligonucleotides are an attractive therapeutic option because of their high selectivity and specificity, due to gene selective silencing of target protein expression. siRNAs, however, have a relatively short half-life and thus we will address the technical challenges of stabilization and intravesical delivery of siRNAs. To achieve this goal, we plan to create and test clinically viable, non-viral nanosphere- siRNA complexes that are intravesically instilled for treatment of bladder cancer and other diseases of the urinary tract. As we have: 1) developed transgenic mice that over- express bladder specific human survivin and that is associated with transcriptional activation of a specific gene signature; 2) established a nitrosamine induced murine bladder cancer model in which both survivin and VEGF are upregulated; and 3) developed, in collaboration with Dr.W. Mark Saltzman Professor and Chair, Department of Bio-engineering, poly (lactide-co-glycolide (PLGA) nanospheres for drug delivery, we are in an unique position to determine the effect of intravesical instillation of siRNA- PLGA in mouse disease models which will initially be tested in our transgenic and bladder cancer models. Specifically we plan to: 1) develop and test a siRNA controlled release system to stabilize and deliver siRNA; 2) test in a whole animal system, the ability of intravesically instilled microencapsulated human survivin siRNA to selectively reverse upregulation of transgenically induced bladder specific survivin and the matrix and inflammatory changes associated with survivin upregulation; and 3) test whether the combination of encapsulated survivin siRNA and VEGF siRNA more effectively reduces tumor burden, the time to onset, rate of occurrence and mortality than individual encapsulated siRNAs in a nitrosamine induced bladder cancer model. Thus, the challenges of more effective intravesical instillation protocols for treatment of common urological diseases including bladder cancer, overactive bladder and IC can be addressed using siRNAs encapsulated in microspheres to increase their stability and prolong their efficacy. Standard pharmacologic and chemotherapeutic treatment options for urological diseases including bladder cancer, overactive bladder and interstitial cystitis may cause undesirable side effects or may be ineffective. Small interference RNA (siRNA) can specifically and sensitively degrade RNA messages and thus reduce their target protein levels. In order to exploit the therapeutic potential of these siRNAs, which are short lived and easily degraded, we have designed strategies for stabilization and testing of these siRNAs. Then, these siRNA polymers will be intravesically instilled as a treatment for bladder cancer, using mouse models of bladder cancer. Furthermore, more than one siRNA can be intravesically instilled in order to target different cancer pathways. Thus, we can determine the therapeutic potential of siRNA for treatment of urologic diseases.

描述（由申请人提供）：膀胱内滴注小干扰RNA（siRNA）是泌尿系统疾病的一种令人兴奋的新治疗选择。siRNA寡核苷酸是一种有吸引力的治疗选择，因为它们的高选择性和特异性，由于靶蛋白表达的基因选择性沉默。然而，siRNA具有相对短的半衰期，因此我们将解决siRNA的稳定化和膀胱内递送的技术挑战。为了实现这一目标，我们计划创建和测试临床上可行的非病毒纳米球- siRNA复合物，其被膀胱内滴注用于治疗膀胱癌和其他泌尿道疾病。正如我们所做的：1）开发了过表达膀胱特异性人存活素并且与特异性基因标记的转录激活相关的转基因小鼠; 2）建立了亚硝胺诱导的鼠膀胱癌模型，其中存活素和VEGF均上调;和3）与Dr. W. Mark Saltzman生物工程系教授兼主席，聚（丙交酯-共-乙交酯（PLGA））纳米球用于药物递送，我们处于独特的地位，以确定在小鼠疾病模型中膀胱内滴注siRNA- PLGA的效果，该模型最初将在我们的转基因和膀胱癌模型中进行测试。具体而言，我们计划：2）在整个动物系统中测试膀胱内滴注的微囊化人存活素siRNA选择性逆转转基因诱导的膀胱特异性存活素的上调以及与存活素上调相关的基质和炎性变化的能力;和3）在亚硝胺诱导的膀胱癌模型中测试包封的存活素siRNA和VEGF siRNA的组合是否比单独的包封的siRNA更有效地降低肿瘤负荷、发病时间、发生率和死亡率。因此，用于治疗常见泌尿系统疾病（包括膀胱癌、膀胱过度活动症和IC）的更有效膀胱内滴注方案的挑战可以使用封装在微球中的siRNA来解决，以增加其稳定性并延长其功效。泌尿系统疾病（包括膀胱癌、膀胱过度活动症和间质性膀胱炎）的标准药理学和化学治疗选择可能会导致不良副作用或可能无效。小干扰RNA（siRNA）可以特异性、敏感性地降解RNA信息，从而降低其靶蛋白水平。为了开发这些短寿命且容易降解的siRNA的治疗潜力，我们设计了用于稳定和测试这些siRNA的策略。然后，这些siRNA聚合物将被膀胱内滴注作为膀胱癌的治疗，使用膀胱癌的小鼠模型。此外，可以膀胱内滴注一种以上的siRNA，以靶向不同的癌症途径。因此，我们可以确定siRNA用于治疗泌尿系统疾病的治疗潜力。