RP2: Targeting genes and pathways for autophagy-dependent inhibition of bacterial infection

项目摘要-RP 2：新兴耐药病原体的发展速度已经超过了药物发现的速度，这构成了 开发安全有效药物的重大挑战。我们的战略方针 通过整合人类遗传学、功能基因组学、新型动物模型和 创新化学具体来说，RP 2旨在开发宿主导向的治疗方法，利用先天性 通过诱导自噬治疗细菌病原体的细胞内防御机制 包括S.鼠伤寒沙门氏菌属单核细胞增多症、S.金黄色葡萄球菌、其多重耐药菌株，以及另外的 RP 1、RP 3和RP 4的优先病原体。在上一个CETR资助期间，我们（1）完成了3个 小分子筛选，以确定自噬依赖的抗感染分子，（2）完成了几个 CRISPR筛选以识别控制选择性自噬和溶酶体稳态的新靶标，以及（3） 利用人类遗传学和功能基因组学来确定治疗诱导的新靶点， 抗菌自噬总的来说，这些研究推进了我们开发自噬导向的 通过从表型筛选中产生新的先导化合物和从功能筛选中产生精确靶点， 基因组学此外，我们的工作已经发现了新的调控机制，控制自噬和翻译， 这些发现为自噬疗法确定了新的切入点。我们与RP 1、RP 3的合作 和RP 4导致了小分子的发现，这些小分子可以增强细胞内对多种疾病的天然防御， 病原微生物，包括M. tuberculosis，S. flexneri，S.鼠伤寒，虫媒病毒，诺如病毒，和 T.刚地。此外，我们还与诺华公司合作，推进针对新靶点的先导化合物， 以促进国家自主研发的研究。在这个CETR提案中，我们将推进这些自噬依赖的抗- 感染性分子使用创新的化学（RP 5）和验证新的治疗目标，从功能 基因组学和人类遗传学研究。我们建议利用以前CETR计划的发现， 目标1：药物化学从三个独立的自噬筛选中获得初步筛选结果 (LC3 puncta、NDP 52-沙门氏菌共定位和GPR 65激动剂）以导致候选自噬依赖性 用于体外和体内疗效研究的广谱抗感染药;目标2：靶向策略的开发 通过TFEB诱导自噬，TFEB是自噬和溶酶体生物发生的主要转录调节因子 目的3：验证从功能基因组学和人类遗传学研究中鉴定的新基因， 目标4：验证产生的新型抗感染候选药物 通过RP 1-RP 5作为抗细菌自噬的诱导剂，使用体外和体内模型。我们的CETR团队 和行业合作伙伴处于独特的地位，能够迅速推进针对新兴病原体的新疗法， 传染病

PROJECT SUMMARY – RP2: Emerging drug-resistant pathogens have outpaced drug discovery, which poses significant challenges for the development of safe and efficacious drugs. Our strategic approach addresses challenges in drug discovery by integrating human genetics, functional genomics, novel animal models, and innovative chemistry. Specifically, RP2 aims to develop host-directed therapeutics that harness innate intracellular defense mechanisms through induction of autophagy for the treatment of bacterial pathogens including S. Typhimurium, L. monocytogenes, S. aureus, multiple-drug-resistant strains thereof, and additional priority pathogens with RP1, RP3, and RP4. During the previous CETR funding period we have (1) completed 3 small molecule screens to identify autophagy-dependent anti-infective molecules, (2) completed several CRISPR screens to identify new targets controlling selective autophagy and lysosome homeostasis, and (3) leveraged human genetics and functional genomics to identify novel targets for therapeutic induction of antibacterial autophagy. Collectively, these studies advanced our objective of developing autophagy-directed therapeutics by generating novel lead compounds from phenotypic screens and precision targets from functional genomics. Moreover, our work has uncovered novel regulatory mechanisms governing autophagy and translated these discoveries to identify new points of entry for autophagy therapeutics. Our collaboration with RP1, RP3 and RP4 led to the discovery of small molecules that augment innate intracellular defense against diverse pathogenic microorganisms, including M. tuberculosis, S. flexneri, S. Typhimurium, arboviruses, norovirus, and T. gondii. In addition, we have partnered with Novartis to advance lead compounds directed at novel targets and to facilitate IND-enabling studies. In this CETR proposal, we will advance these autophagy-dependent anti- infective molecules using innovative chemistry (RP5) and validate new therapeutic targets from functional genomic and human genetic studies. We propose to leverage discoveries from the previous CETR Program to advance: Aim 1: medicinal chemistry to progress primary screen hits from three independent autophagy screens (LC3 puncta, NDP52-Salmonella co-localization and GPR65 agonist) to lead candidate autophagy-dependent broad-spectrum anti-infectives for in vitro and in vivo efficacy studies; Aim 2: development of targeting strategies to induce autophagy through TFEB, a master transcriptional regulator of autophagy and lysosome biogenesis genes; Aim 3: validation of novel genes identified from functional genomic and human genetic studies as therapeutic targets for antibacterial autophagy; and Aim 4: validation of novel anti-infective candidates generated by RP1-RP5 as inducers of anti-bacterial autophagy using in vitro and in vivo models. Together, our CETR team and industry partners are uniquely positioned to rapidly advance new treatments for emerging pathogens and infectious diseases.