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Nuclease-Resistant Aptamers for Anthrax Opsonization

用于炭疽调理作用的核酸酶抗性适体

基本信息

批准号：
6735824
负责人：
John G Bruno
金额：
$ 9.99万
依托单位：
OPERATIONAL TECHNOLOGIES CORPORATION
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2004-10-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by investigator): The anti-phagocytic poly-D-glutamic acid (PDGA) capsule of Bacillus anthracis is a major virulence factor of inhalation anthrax. Although generation of antibodies to PDGA might be of value in opsonizing vegetative anthrax, such antibodies would require "humanization". A simpler and less expensive approach would be the development of nuclease-resistant ('shielded') DNA aptamers against PDGA with an Fc-TR or C3bR binding domain at the other end of a 'hybrid' aptamer. In Phase I, Operational Technologies (OpTech) in conjunction with the Biochemistry Department of the University of Texas at San Antonio (UTSA), proposes to develop 2'- amino pyrimidine modified shielded DNA aptamers against PDGA, murine Fc-yR, and C3bR by the SELEX process. OpTech will link the aptamers and compare phagocytosis of PDGA-conjugated microbeads by the RAW 264.7 macrophage-like cell line in the presence and absence of shielded aptamers and versus serum opsonized PDGA-conjugated microbeads. Since polyanionic PDGA appears to inhibit phagolysosome formation and killing of ingested bacteria, targeting PDGA with 2'-amino-modified aptamers may serve to partially neutralize the polyanionic charge and enable bacterial killing by the respiratory burst. In Phase II, OpTech will clone and sequence all aptamers, then assess aptamer-opsonization and bacterial killing in RAW 264.7 cells with virulent anthrax at the Southwest Research Foundation. Since anthrax and some other infectious bacteria utilize capsules to evade phagocytosis, any inexpensive reagent that acts as an effective opsonin would be of tremendous value in the medical, veterinary or agricultural communities. If successful, OpTech will possess a pharmaceutical substance of great commercial value in the post-exposure treatment of anthrax. Further development of the concept for shielded aptamers against hyaluronic acid or other capsule materials, for example, would also lead to better treatments for many Streptococcus-related infections. Such shielded aptamers could even be used in inhalers to enhance alveolar macrophage phagocytosis.

描述(由研究人员提供)：炭疽杆菌的抗吞噬多D-谷氨酸(PDGA)胶囊是吸入性炭疽的主要毒力因素。尽管针对PDGA的抗体的产生在对抗植物炭疽病方面可能有价值，但这种抗体需要“人性化”。一种更简单、成本更低的方法是开发抗PDGA的核酸酶抗性(“屏蔽”)DNA适配子，在“杂交”适配子的另一端带有Fc-tr或C3bR结合域。在第一阶段，运营技术公司(Optech)与德克萨斯大学圣安东尼奥分校(UTSA)生物化学系合作，提议通过SELEX过程开发针对PDGA、小鼠Fc-YR和C3bR的2‘-氨基嘧啶修饰屏蔽DNA适配子。Optech将连接适配子，并比较RAW 264.7巨噬细胞样细胞系在存在和不存在屏蔽适配子的情况下对PDGA结合微球的吞噬作用，以及与血清优化的PDGA结合微球的吞噬作用。由于聚阴离子PDGA似乎可以抑制吞噬溶酶体的形成和对吞噬细菌的杀灭，因此用2‘-氨基修饰的适配子靶向PDGA可能有助于部分中和多阴离子电荷，并通过呼吸爆发杀灭细菌。在第二阶段，Optech将克隆所有适配子并对其进行测序，然后在西南研究基金会的RAW 264.7细胞中评估适配子的调理作用和细菌杀灭作用。由于炭疽和其他一些传染性细菌利用胶囊来逃避吞噬作用，任何廉价的调理素试剂在医疗、兽医或农业领域都将具有巨大的价值。如果成功，Optech将拥有一种在暴露后治疗炭疽病方面具有巨大商业价值的药物。例如，进一步发展针对透明质酸或其他胶囊材料的屏蔽适配子的概念也将导致对许多链球菌相关感染的更好治疗。这种屏蔽适配子甚至可以用于吸入器，以增强肺泡巨噬细胞的吞噬功能。