New Approaches for Development of PcP Therapy

描述（由申请人提供）：卡氏肺囊虫（PcP）仍然是免疫功能低下宿主（包括艾滋病患者）感染的重要原因。最广泛使用的治疗方法是甲氧苄啶-磺胺甲恶唑（TMP-SMX）。最近，DHPS基因座突变的发展引起了人们的关注，DHPS基因座介导对SMX的敏感性。因此，Dr. Cushion在更好地理解作为潜在治疗靶点的固醇和线粒体代谢途径的基础上，提出了开发肺囊虫病治疗的新策略。他们建议首先使用体外生存能力测定确定针对Pc的甾醇生物合成和线粒体途径的酶促步骤的有效化合物。在第二个目标下，他们将通过使用宏阵列分析基因表达来评估这些抑制剂在每种途径上的潜在作用机制。在第三个目标中，他们将试图通过构建抑制剂等线形图来确定这些途径内部和之间的甾醇和线粒体抑制剂的协同组合。最后，他们将选择最有效的药物组合，并再次使用macroarray方法评估其对基因表达的影响。

DESCRIPTION (provided by applicant): Pneumocystis carinii (PcP) remains an important cause of infection in immunocompromised hosts including patients with AIDS. The most widely utilized therapy is trimethoprim-sulfamethoxazole (TMP-SMX). Recent concerns have arisen over development of mutations to the DHPS locus, which mediates sensitivity to SMX. Accordingly, Dr. Cushion proposes new strategies to develop treatments for Pneumocystis based upon a better understanding of the sterol and mitochondrial metabolism pathways as potential targets for therapy. They propose to first identify efficacious compounds which target enzymatic steps in the sterol biosynthetic and mitochondrial pathways of Pc using in vitro viability assay. Under the second aim, they will assess the potential mechanisms of action of these inhibitors on each pathway by analysis of gene expression using macroarrays. In the third aim, they will attempt to identify synergistic combinations of the sterol ad mitochondrial inhibitors within and between these pathways by construction of inhibitor isobolograms. Finally, they will select the most efficacious combinations of agents and evaluate their effects on gene expression again using the macroarray approach.