Cavity and Granuloma Oriented Inflammation and Tissue Pharmacokinetics in Pulmonary Tuberculosis (COOK TB)

项目摘要 结核病（TB）是全球传染病死亡的第二大原因， 2020.肺结核的一个特点是倾向于在约30-85%的患者中形成空洞性病变。腔 为结核分枝杆菌（Mtb）复制提供理想的环境， 抗生素的渗透，并可能导致不可逆转的肺损伤。重要的是，它们与贫穷有关， 临床结果包括获得性耐药性和治疗失败。空洞是由发展而来的 坏死性肺肉芽肿;然而，其形成的机制尚不清楚。改进 了解驱动组织坏死的炎症反应和抗生素的能力， 需要坏死性肉芽肿内的治疗浓度来1）鉴定宿主定向的靶点， HDT可以限制组织损伤和2）优化抗生素方案。利用创新 成像和空间多组学的方法，我们将绘制转录途径的分布， 与人类坏死性肉芽肿和空洞有关的生物介质以及新实施的抗结核药物 总体目标是为改善结核病治疗提供关键的新数据 这项研究的长期目标是提供数据，以指导开发一种串联治疗药物。 根据抗结核药物在所有患者体内达到杀菌浓度的能力， 病变区域结合宿主靶向治疗以限制病理性炎症。具体目标是 建议是（1）鉴定与每个组织区域相关的宿主代谢和脂质表型， 人类坏死性肺肉芽肿;（2）利用空间转录组学和靶向成像来识别 与坏死性肉芽肿中的组织坏死相关的病理程序;以及（3）利用靶部位 药代动力学（PK）和PK建模，以加强对新实施的抗结核药物的理解。的 该项目的目标将通过招募一个独特的肺结核患者队列来实现， 手术切除的肺部病变的后续研究。采用科学方法进行 我们的目标包括使用增强的MALDI-2质谱成像（MSI），激光捕获 显微切割（LCM），以分离高分辨率代谢组学，脂质组学和 药物浓度测定，以及新的空间转录组学和先进的数据分析方法， 空间分辨的多组学数据集和目标位点PK数据模型。 该提案将直接涉及结核病研究议程中的关键优先事项，包括实现更好的 理解M的决定因素。肉芽肿中的结核病控制以及抗结核药物如何定位和 渗透进肉芽肿和空洞建议工作的具体目标是确定宿主炎症 可用于宿主定向治疗的途径，并定义关键的组织穿透特性， 用于耐药结核病的药物，以优化临床试验检测和治疗的药物方案设计。

PROJECT SUMMARY Tuberculosis (TB) is the 2nd leading cause of infectious disease mortality worldwide with ~1.5 million deaths in 2020. A hallmark of pulmonary TB is the propensity to form cavitary lesions in ~30-85% of patients. Cavities provide an ideal environment for Mycobacterium tuberculosis (Mtb) replication, are associated with decreased penetration of antibiotics, and can lead to irreversible lung damage. Importantly, they are associated with poor clinical outcomes including acquired drug resistance and treatment failure. Cavities develop from progression of necrotic lung granulomas; however, mechanisms underlying their formation are not clear. Improved understanding of the inflammatory responses that drive tissue necrosis and the ability of antibiotics to achieve therapeutic concentrations within necrotic granulomas are needed to 1) identify targets for host directed therapies (HDT) that can limit tissue damage and 2) to optimize antibiotic regimens. Utilizing innovative methods in imaging and spatial multiomics, we will map the distribution of transcriptional pathways and biomediators associated with human necrotic granulomas and cavities and of newly implemented anti-TB drugs in such lesions with an overall goal of providing critical new data to improve TB treatment The long term objective of this research is to provide data to guide development of a tandem therapeutic approach of optimizing anti-TB drug regimens based on their ability to reach bactericidal concentrations in all lesion areas combined with host-targeted therapies to limit pathologic inflammation. The specific aims of this proposal are to (1) identify the host metabolic and lipid phenotypes associated with each tissue region of human necrotic lung granulomas; (2) utilize spatial transcriptomics and targeted imaging to identify pathological programs associated with tissue necrosis in necrotic granulomas; and (3) utilize target site pharmacokinetics (PK) and PK modeling to enhance understanding of newly implemented anti-TB drugs. The aims of this project will be achieved by enrolling a unique cohort of patients with pulmonary TB undergoing adjunctive surgery and subsequent study of their resected lung lesions. Scientific methods employed to carry out our aims include the use of enhanced MALDI-2 mass spectrometry imaging (MSI), laser capture microdissection (LCM) to isolate targeted granulomas regions for high-resolution metabolomics, lipidomics and drug concentration assays, and novel spatial transcriptomics and advanced data analytic methods to integrate spatially resolved multi-omics data sets and model target site PK data. This proposal will directly address key priories in the TB research agenda including attaining a better understanding of the determinants of M. tuberculosis control in granulomas and how anti-TB drugs localize and penetrate into granulomas and cavities. Specific goals of the proposed work are to identify host inflammatory pathways that can be exploited for host-directed therapy and to define tissue penetrating properties of key drugs used for drug-resistant TB to optimize drug regimen design for clinical trial testing and treatment.