Targeted-delivery of small interference RNA against anthrax

针对炭疽病的小干扰 RNA 的靶向递送

• 批准号: 9089923
• 负责人: MINGTAO ZENG
• 金额: $ 22.95万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-15 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9089923
• 关键词: A/J Mouse; Aluminum Hydroxide; Amino Acids; Animals; Anthrax Vaccines; Anthrax disease; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antigens; Antimicrobial Resistance; Arginine; Bacillus (bacterium); Bacillus anthracis; Bacillus anthracis spore; Bacteria; Bacterial Infections; Binding; Bioterrorism; Biothrax; Blood Circulation; Blood capillaries; Cardiac Myocytes; Cell Survival; Cells; Cessation of life; Charge; Chimeric Proteins; Cleaved cell; Communicable Diseases; Complement; Complex; Containment; Cytosol; Development; Disease; Edema; Endocytosis; Exposure to; Formulation; Future; Gram-Positive Bacteria; Health; Hepatocyte; Human; Immune; Immune system; Immunity; Immunization; Immunization Schedule; Infection; Influenza; Kinetics; Label; Licensing; Lung; Mass Immunization; Modeling; Morphogenesis; Mus; Names; Patients; Peptides; Property; Prophylactic treatment; Proteins; Public Health; RNA Binding; RNA Interference; Receptor Cell; Reporter; Reporting; Reproduction spores; Research; Safety; Shock; Small Interfering RNA; Smooth Muscle; System; Technology; Therapeutic; Toxic effect; Toxin; Travel; Vaccination; Vaccines; Virulence; Virulence Factors; animal tissue; anthrax lethal factor; anthrax toxin; anthrax toxin receptors; antigen binding; biothreat; capillary; combat; cytotoxicity; design; edema factor; immunogenicity; improved; lymph nodes; macrophage; mouse model; novel; pathogen; prevent; prophylactic; protective efficacy; reaction rate; receptor binding; respiratory; success; targeted delivery; targeted treatment; tumor endothelial marker 8; vaccine candidate; weapons

 DESCRIPTION: Targeted-delivery of small interference RNA against anthrax. Anthrax is a serious disease caused by Bacillus afgfnthracis, a bacterium that forms spores. Anthrax most commonly occurs in wild and domestic mammalian species; but can also occur in humans when they are exposed to infected animals or animal tissues, or when anthrax spores are dispersed as a bioterrorist weapon. The complicated immunization schedule with the licensed vaccine BioThrax calls for a new and easily administered anthrax vaccine. Since anthrax is a disease that rarely occurs naturally in humans, it is more realistic to develop a post exposure prophylaxis or therapy instead of mass immunization, as with the current vaccine. Following exposure, macrophages ingest anthrax spores and travel to the lymph node where these spores germinate. The B. anthracis bacteria are then released into the bloodstream and produce toxins that are key factors in the virulence of disease: protective antigen (PA), edema factor (EF), and lethal factor (LF). PA is the receptor binding toxin component that attaches to either of two host cell receptors: anthrax toxin receptor 1 (ANTXR1 or tumor endothelial marker 8/TEM8) and anthrax toxin receptor 2 (ANTXR2 or capillary morphogenesis protein 2/CMG2). After binding, PA is cleaved and the receptor-bound portions form a heptameric pore that binds EF or LF. The toxin complexes are endocytosed and delivered into the cytosol. The activities of LeTx and EdTx result in malfunction of the immune system, edema, shock, and death. Our preliminary study has shown that inhibition of ANTXR expression by RNA interference (RNAi) technology using specific anti-ANTXR small interference RNA (siRNA) could prevent cytotoxicity of anthrax toxins. We hypothesize that a detoxified anthrax toxin could be used as a delivery vehicle for anti-ANTXR siRNA. In order to evaluate this hypothesis, we propose the following three specific aims: Specific Aim 1: To generate detoxified anthrax toxins for ANTXR-targeted siRNA delivery. Specific Aim 2: To assess the inhibitory effect on anthrax toxin induced cytotoxicity in ANTXR-silenced cells. Specific Aim 3: To evaluate efficacy of the anti-ANTXR siRNA treatment in a mouse model of anthrax. We anticipate that the proposed host-targeted treatment strategy will prevent severe illness and death in patients exposed to both wild type and even antibiotic-resistant B. anthracis spores by natural infection or a bioterrorist attack. Furthermore, this technology can be developed as a platform to treat other antimicrobial-resistant pathogens that employ pore-forming toxins as virulence factors.

 描述：针对炭疽的小干扰RNA的靶向分娩。炭疽病是由Afgfnthracis芽孢杆菌引起的严重疾病，一种形成孢子的细菌。炭疽最常见发生在野生和家庭哺乳动物中。但是，当人类暴露于感染的动物或动物组织时，或者将炭疽孢子作为生物恐怖主义武器分散时，也可能发生在人类中。持牌疫苗Biothrax的复杂免疫时间表要求采用一种新的且易于施用的炭疽疫苗。由于炭疽是一种很少在人类中自然发生的疾病，因此在预防后进行暴露后更现实 或治疗而不是质量免疫，例如当前的疫苗。暴露后，巨噬细胞摄入炭疽孢子并传播到这些孢子发芽的淋巴结。然后，炭疽芽孢杆菌细菌被释放到血液中，并产生作为病毒关键因素的毒素：受保护的抗原（PA），水肿因子（EF）和致死因子（LF）。 PA是连接到两个宿主细胞受体中的任何一个受体结合毒素成分：炭疽毒素受体1（ANTXR1或肿瘤内皮标记物8/TEM8）和炭疽毒素受体2（ANTXR2或毛细血管形态发生蛋白2/CMG2）。结合后，将PA裂解，并结合受体结合的部分结合EF或LF的肝孔。毒素络合物被内吞并输送到细胞质中。 LETX和EDTX的活动导致免疫系统，水肿，冲击和死亡的故障。我们的初步研究表明，使用特定的抗ANTXR小型干扰RNA（siRNA）对RNA干扰（RNAI）技术抑制ANTXR表达可以防止炭疽毒素的细胞毒性。我们假设一个排毒的炭疽毒素可以用作抗抗Antxr siRNA的递送工具。为了评估这一假设，我们提出以下三个特定目的：特定目的1：生成排毒的炭疽毒素以进行ANTXR靶向的siRNA递送。具体目的2：评估对ANTXR脱毛细胞中炭疽毒素诱导的细胞毒性的抑制作用。特定目的3：评估在炭疽小鼠模型中抗ANTXR siRNA处理的有效性。我们预计，拟议的以宿主为目标的治疗策略将防止因自然感染或生物恐怖主义攻击而暴露于野生型甚至抗生素的鼻孢子孢子的患者严重疾病和死亡。此外，这项技术可以作为一种平台来处理其他抗菌病原体，这些病原体将员工形成毛孔的毒素作为病毒因素。