SiRNA structural optimization

siRNA结构优化

• 批准号: 7487383
• 负责人: KENNETH A ALEXANDER
• 金额: $ 21.02万
• 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/7487383
• 关键词: Affect; Antiviral Therapy; Binding; Bioavailable; Biological Assay; CCR5 gene; Cells; Cellular Morphology; Chemicals; Class; 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; Transgenic Organisms; 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; size; transmission process; 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）是肛门生殖器和头颈部肿瘤和恶性肿瘤的主要原因。 不幸的是，尚未确定针对HPV感染或HPV诱导的恶性肿瘤的特异性抗病毒疗法。 该U19提案由亚历山大在题为siRNA杀微生物剂的联合提案中作为项目提交。这个目标 一个研究联盟正在开发天然的和化学修饰的siRNA，作为阻断 HIV、HSV 2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 硼烷磷酸骨架短干扰RNA（siRNA）对于有效和持久的干扰作用优于天然siRNA（上级）。 基因沉默，并且抗HPV E6/E7单链硼磷酸骨架siRNA（ssBP-siRNA）可以被 用于有效阻断HPV病毒复制和HPV转化细胞的增殖。通过结合化学 单独显示的增加siRNA沉默效力和持续时间的修饰，我们将创建ssBP-siRNA， 设计用于局部递送，其阻断HPV感染细胞中的附加体复制并诱导不可逆生长 HPV 16转化细胞中的停滞。我们鉴定的抗HPV 16 ssBP-siRNA将在项目3中使用新的 细胞培养和HPV感染的转基因小鼠模型。 项目3还将向Ramratnam和赫罗尔德的项目提供优化的ssBP-siRNA，以允许将ssBP-siRNA与 用于靶向表达猕猴CCR 5、鼠粘着斑激酶和鼠连接蛋白-1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被评估。 在项目3中进行了定量评估。 在这项研究的结论，我们将开发出抗HPV 16 E6/E7 ssBP-siRNA作为有效的局部 用于预防HPV感染和HPV诱导的恶性肿瘤的杀微生物剂。我们还将创造出高效能的 ssBP-siRNA对其他聚生体靶标具有活性，并将这些ssBP-siRNA转发用于微生物测试， 病毒感染的实验室模型。