SiRNA structural optimization

siRNA结构优化

• 批准号: 7906869
• 负责人: KENNETH A ALEXANDER
• 金额: $ 22.22万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7906869
• 关键词: Affect; Antiviral Therapy; Binding; Bioavailable; Biological Assay; CCR5 gene; Cell Culture Techniques; Cells; Cellular Morphology; Chemicals; Clinical; Complex; Cultured Cells; Drug Formulations; Episome; Fluorescence Polarization; Focal Adhesion Kinase 1; Gene Silencing; Genes; Genome; Goals; Growth; HIV; Head and Neck Neoplasms; Human Herpesvirus 2; Human Papillomavirus; Human papillomavirus 16; Hydrophobicity; Infection; Interferons; Kinetics; Laboratories; Local Microbicides; Macaca; Macaca mulatta; Maintenance; Malignant Neoplasms; Measures; Modeling; Modification; Mucous Membrane; Mus; NIH Program Announcements; PVRL1; Play; Prevention; Property; Proteins; Pyrimidine; Pyrimidines; RNA Interference; RNA-Induced Silencing Complex; Research; Research Personnel; Ribonucleosides; Role; Simplexvirus; Small Interfering RNA; Structure; Testing; Tissues; Toxic effect; Transgenic Mice; Vagina; Vertebral column; Viral; Viral Genome; Virus Diseases; Work; cell growth; cell transformation; cytotoxic; cytotoxicity; design; enhanced green fluorescent protein; human DICER1 protein; keratinocyte; locked nucleic acid; melting; microbicide; mouse model; mucosal uptake; novel; nuclease; phosphodiester; pre-clinical; programs; protein expression; response; riboside; siRNA Microbicides; transmission process; vaginal microbicide; virus episome maintenance

Human papillomaviruses (HPVs) are a major cause of anogenital and head and neck neoplasms and malignancies. Unfortunately, no specific antiviral therapies for HPV infection or HPV-induced malignancies have been identified. This U19 proposal is submitted as project by Alexander in a consortium proposal entitled siRNA Microbicides. The goal of this research consortium is to develop native and chemically modified siRNAs as effective means for blocking transmission of HIV, HSV2 and HPV. In project by Alexander, we hypothesize that chemically modified single-stranded boranophosphate-backbone short interfering RNAs (siRNAs) are superior to native siRNAs for potent and durable gene silencing, and that anti-HPV E6/E7 single-stranded boranophosphate-backbone siRNAs (ssBP-siRNAs) can be used to effectively block HPV viral replication and proliferation of HPV-transformed cells. By combining chemical modifications shown individually to increase siRNA silencing potency and duration, we will create ssBP-siRNAs, designed for topical delivery, that block episome replication in HPV-infected cells and induce irreversible growth arrest in HPV16-transformed cells. Anti-HPV16 ssBP-siRNAs identified by us will be tested in project 3 using novel cell culture and transgenic mouse models of HPV infection. Project 3 will also supply optimized ssBP-siRNAs to projects by Ramratnam and Herold to allow comparison of ssBP-siRNAs with conventional siRNAs for targeting expression of macaque CCR5, murine focal adhesion kinase, and murine nectin-1. To allow immediate work optimizing ssBP-siRNA delivery to vaginal mucosa, we will supply existing highly active ssBP-siRNAs to Core B (Formulations). The vaginal mucosal uptake and activity of formulated ssBP-siRNAs will be assessed quantitatively in project 3. At the conclusion of this study, we will have developed anti-HPV16 E6/E7 ssBP-siRNAs as effective topical microbicides for prevention of HPV infection and HPV-induced malignancies. We will also have created high potency ssBP-siRNAs active against other consortium targets, and forwarded these ssBP-siRNAs for microbicidal testing in laboratory models of viral infection.

人乳头瘤病毒 (HPV) 是肛门生殖器、头颈肿瘤和恶性肿瘤的主要原因。 不幸的是，尚未发现针对 HPV 感染或 HPV 诱发的恶性肿瘤的特异性抗病毒疗法。 该 U19 提案是 Alexander 在题为 siRNA Microbicides 的联盟提案中作为项目提交的。此举的目标 研究联盟将开发天然的和化学修饰的 siRNA 作为阻断的有效手段 HIV、HSV2 和 HPV 的传播。在亚历山大的项目中，我们假设化学修饰的单链 硼磷酸盐主链短干扰 RNA (siRNA) 优于天然 siRNA，具有有效和持久的功效 基因沉默，抗 HPV E6/E7 单链磷酸硼骨架 siRNA (ssBP-siRNA) 可以 用于有效阻断HPV病毒复制和HPV转化细胞的增殖。通过化学结合 单独显示的修饰可增加 siRNA 沉默效力和持续时间，我们将创建 ssBP-siRNA， 专为局部给药而设计，可阻断 HPV 感染细胞中的游离体复制并诱导不可逆生长 HPV16 转化细胞中的停滞。我们鉴定出的抗 HPV16 ssBP-siRNA 将在项目 3 中使用新颖的方法进行测试 HPV 感染的细胞培养和转基因小鼠模型。 项目 3 还将向 Ramratnam 和 Herold 的项目提供优化的 ssBP-siRNA，以便将 ssBP-siRNA 与 用于靶向猕猴 CCR5、鼠粘着斑激酶和鼠 nectin-1 表达的常规 siRNA。 为了立即优化 ssBP-siRNA 向阴道粘膜的递送，我们将提供现有的高活性 ssBP-siRNA 至核心 B（制剂）。配制的 ssBP-siRNA 的阴道粘膜摄取和活性将是 在项目3中进行定量评估。 在本研究结束时，我们将开发出抗 HPV16 E6/E7 ssBP-siRNA 作为有效的局部用药 用于预防 HPV 感染和 HPV 诱发的恶性肿瘤的杀微生物剂。我们也将创造出高效力 ssBP-siRNA 对其他联合体靶点具有活性，并将这些 ssBP-siRNA 转发至 病毒感染的实验室模型。