RP3: Targeting ATG gene-dependent immunity for novel anti-infective therapeutics

RP3：针对 ATG 基因依赖性免疫的新型抗感染疗法

• 批准号: 10364725
• 负责人: Seungmin Hwang
• 金额: $ 88.28万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-15 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10364725
• 关键词: Adopted; Albers-Schonberg disease; Anti-Infective Agents; Area; Aspergillus; Autophagocytosis; Bacteria; Biochemical; Biopsy; Biotechnology; CRISPR screen; Calicivirus; Cell Differentiation process; Cells; Cellular biology; Chronic Granulomatous Disease; Clinical Trials; Complement; Development; Epidemic; Funding; Genes; Goals; Grant; Human; Immune; Immunity; Infection; Infection Control; Inflammation; Inflammatory; Interferon Type II; Interferons; Laboratory mice; Lead; Malaria; Mediating; Molecular; Molecular Target; Mus; Mycobacterium tuberculosis; Norovirus; Parasites; Pathogenesis; Pharmaceutical Preparations; Pharmacology; Phenotype; Phylogenetic Analysis; Pre-Clinical Model; Process; Property; Proteins; Regulation; Resistance; Role; Salmonella; Screening Result; Tertiary Protein Structure; Therapeutic; Time; Toxic effect; Toxoplasma gondii; Virus; Virus Receptors; activity-based protein profiling; cytokine; design; experimental study; human disease; immunopathology; in vivo; macrophage; microbiome; new therapeutic target; novel; novel therapeutics; pathogen; priority pathogen; programs; receptor; research and development; response; stem cells; therapeutic candidate; tool; whole genome

PROJECT SUMMARY – RP3: The goal of RP3 is to define and pharmacologically target intracellular immune mechanisms utilizing autophagy (Atg) genes (but not degradative autophagy) to control infection and inflammation. We discovered these processes in experiments designed to understand the anti-infective mechanisms of IFNγ, a cytokine essential in both humans and mice for macrophage-mediated resistance to viruses, bacteria, and parasites. IFNγ is used to treat chronic granulomatous disease and osteopetrosis, but other uses of IFNγ have been limited by toxicity, a fact generally attributed to IFNγ inducing the expression of a very large number of pro-inflammatory genes. We theorized that new therapeutics might be generated by defining the effector mechanisms responsible for the potent effects of IFNγ, and then selectively stimulating these mechanisms to generate Atg gene-directed broad-spectrum anti-infectives. This led to the discovery that IFNγ triggers Atg gene-dependent immunity against T. gondii and norovirus (NoV) infection using the same set of Atg genes (Atg5, Atg7, Atg16L1, Atg12), but not Atg14 or degradative autophagy. The fact that Atg gene- dependent mechanism(s) controlled two phylogenetically distinct pathogens suggested to us that we might be able to make compounds that are effective against a broad range of pathogens. During the first CETR funding period we confirmed this prediction by identifying Atg gene-directed broad-spectrum anti-infective compounds with activity against norovirus, salmonella, Mycobacterium tuberculosis and Toxoplasma gondii. This discovery was complemented by discoveries of the role of Atg genes in the regulation of inflammation. RP3 focuses on the Caliciviridae (most importantly NoVs) which are priority pathogens, causing >90% of the epidemic non-bacterial gastroenteritis in the world. We discovered murine NoV (MNoV) and cultured NoVs for the first time, providing a preclinical model in laboratory mice and allowing discovery of Atg gene-dependent immunity. Studies of human NoV (HNoV) infection had been limited until it was recently found that HNoV grows in cells differentiated from biopsy-derived human colonic stem cells. This, together with the discovery that MNoV grows robustly in human cells expressing the proper receptor, and that Atg gene-dependent immunity operates against both T. gondii and MNoV in human cells strongly argues that these mechanisms are evolutionarily conserved and provide the tools to identify compounds relevant to a major human disease via the following aims: Aim 1: Develop Atg gene- directed broad spectrum anti-infective compounds; Aim 2: Define the molecular basis of Atg gene- dependent IFNγ-induced control of norovirus replication, and identify targets and lead compounds that trigger this form of intracellular immunity; Aim 3: Define the molecular basis for the regulation of IFNγ responses by Atg genes and identify lead compounds that up-regulate specific IFNγ effector mechanisms; Aim 4. Collaborate with other CETR projects to identify autophagy- and Atg gene-directed broad-spectrum anti-infective compounds.

项目摘要 – RP3：RP3 的目标是定义和药理学靶向细胞内免疫 利用自噬（Atg）基因（但不是降解自噬）来控制感染和 我们在旨在了解抗感染的实验中发现了这些过程。 IFNγ 的机制，IFNγ 是人类和小鼠巨噬细胞介导的抗性所必需的细胞因子 病毒、细菌和寄生虫 IFNγ 用于治疗慢性肉芽肿病和骨硬化症。 IFNγ的其他用途受到毒性的限制，这一事实通常归因于IFNγ诱导a表达 我们推测，可能会产生大量促炎基因。 定义负责 IFNγ 有效作用的效应机制，然后选择性地刺激 这些机制产生 Atg 基因定向的广谱抗感染药物，这导致了以下发现。 IFNγ 使用同一组触发针对弓形虫和诺如病毒 (NoV) 感染的 Atg 基因依赖性免疫力 Atg 基因（Atg5、Atg7、Atg16L1、Atg12），但不是 Atg14 或降解自噬。 控制两种系统发育上不同的病原体的依赖机制向我们表明，我们可能是 在第一笔 CETR 资助期间，我们能够制造出对多种病原体有效的化合物。 在此期间，我们通过鉴定 Atg 基因定向的广谱抗感染化合物证实了这一预测 这一发现具有对抗诺如病毒、沙门氏菌、结核分枝杆菌和弓形虫的活性。 Atg 基因在炎症调节中的作用的发现补充了 RP3 的作用。 杯状病毒科（最重要的是新病毒）是优先病原体，导致超过 90% 的流行病为非细菌性疾病 我们首次发现了鼠类NoV（MNoV）和培养的NoV，为我们提供了一种新的方法。 实验室小鼠的临床前模型并允许发现人类 Atg 基因依赖性免疫研究。 NoV (HNoV) 感染一直受到限制，直到最近发现 HNoV 在分化的细胞中生长。 活检来源的人类结肠干细胞，以及 MNoV 在人类体内强劲生长的发现。 表达适当受体的细胞，并且 Atg 基因依赖性免疫对弓形虫和 人类细胞中的 MNoV 强烈认为这些机制在进化上是保守的，并提供了工具 通过以下目标鉴定与主要人类疾病相关的化合物： 目标 1：开发 Atg 基因- 定向广谱抗感染化合物；目标 2：确定 Atg 基因的分子基础 依赖 IFNγ 诱导的诺如病毒复制控制，并确定靶标和先导化合物 触发这种形式的细胞内免疫；目标 3：确定 IFNγ 调节的分子基础； Atg 基因的反应并鉴定上调特定 IFNγ 效应子的先导化合物 目标 4. 与其他 CETR 项目合作，识别自噬和 Atg 基因指导的机制； 广谱抗感染化合物。