New Approaches for Development of PcP Therapy

描述(申请人提供)：卡氏肺孢子虫(PCP)仍然是包括艾滋病患者在内的免疫功能低下的宿主感染的重要原因。应用最广泛的治疗方法是甲氧苄氨嘧啶-磺胺甲恶唑(TMP-SMX)。最近出现了对DHPS基因突变的关注，DHPS基因突变介导了对SMX的敏感性。因此，垫博士提出了新的策略，以更好地了解作为潜在治疗靶点的类固醇和线粒体代谢途径，来开发肺孢子虫的治疗方法。他们建议首先使用体外活性测试来鉴定针对Pc生物合成和线粒体途径中的酶步骤的有效化合物。在第二个目标下，他们将通过使用大分子阵列分析基因表达来评估这些抑制剂在每条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物同功图来确定这些途径内和这些途径之间的类固醇和线粒体抑制物的协同组合。最后，他们将选择最有效的药物组合，并使用大阵列方法再次评估它们对基因表达的影响。

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.