Targeting MT1-MMP to inhibit pathologic inflammation in TB

结核病仍然是世界各地感染致死的主要单一原因；新的治疗方法是 需要改变疫情的形式。一种建议的方法来扩大可用武器库 抗结核药物的目标是宿主因素，以增强细菌杀菌或调节炎症 会导致组织损伤。在感染的背景下，组织损伤通常是基于线性 光谱，“太多”驱动宿主介导的组织破坏，“太少”导致进展 细菌感染。然而，这个线性模型未能捕捉到结核病炎症的复杂性；事实上， 炎症的一些单独成分，包括组织重塑酶，如基质 金属蛋白酶(MMPs)可能在没有促进灭菌的情况下造成破坏。抑制这样的 酶可以在不影响杀菌的情况下改善结果。我们建议采取系统性的 识别和靶向在结核病感染中导致组织破坏的基质酶的方法 有两个首要目标：详细说明个别酶在结核病发病机制中的作用，并开发和 在结核病治疗中测试这些酶的高度特异性抑制剂作为宿主指导的辅助疗法。在……里面 使用空洞性结核病小鼠模型的初步工作，我们进行了感染的序列转录图谱 以确定在感染过程中上调的破坏性基质酶。MT1-MMP脱颖而出 在早期和持续的模式中上调；这种酶以前曾与人类结核病有关。 使用SAGI实验室开发的高度特异的MT1-MMPI抑制剂，我们进行了一项先导性实验 在小鼠空洞模型中确定感染后8周内低水平给药的耐受性 结核病。小鼠对这种抑制剂的耐受性很好，尽管服用这种抑制剂并不是为了达到最大的效果， 组织病理学分析显示病变范围缩小，致密的炎细胞减少。 渗透进来的。在这项拟议的工作中，我们将在这些结果的基础上测试MT1-MMPs在 该模型的发病机制以及抑制对分子、细胞和组织学结果的影响 疾病。在目标1中，我们将优化剂量以在不引起副作用的情况下最大限度地抑制；然后我们将测试 抑制MT1-MMPs对细菌生长和肺组织病理学的影响。在目标2中，我们将使用单细胞 转录图谱用于识别MT1-基质金属蛋白酶产生的细胞驱动因素和测试MT1-基质金属蛋白酶的影响 基质金属蛋白酶抑制炎症细胞向感染肺的募集。然后我们将测试MT1-MMPs的影响 用多重细胞因子图谱和高分辨率显微镜抑制炎症环境。vt.在.的基础上 为了实现这些目标，我们预计已经表征了MT1-基质金属蛋白酶在肿瘤病理进展中的作用。 结核病在小鼠疾病模型中的作用。我们期望这些结果将有助于对基质的作用进行有针对性的研究。 酶在人类结核病发病机制中的作用；此外，我们预计这些结果最终将告知 作为结核病辅助治疗的基质金属蛋白酶抑制剂的研究进展。

Tuberculosis remains the leading single cause of death from infection around the world; new treatments are needed to change the shape of the epidemic. One proposed approach to expanding the available arsenal of anti-TB drugs is targeting host factors to either enhance bacterial sterilization or modulate inflammation that drives tissue damage. Tissue damage in the context of infection is typically conceptualized on a linear spectrum, with “too much” driving host-mediated tissue destruction and “too little” resulting in progression of bacterial infection. However, this linear model fails to capture the complexity of inflammation in TB; in fact, some individual components of inflammation, including tissue remodeling enzymes such as matrix metalloproteases (MMPs), likely contribute to destruction without promoting sterilization. Inhibiting such enzymes could improve outcomes without compromising bacterial killing. We propose to take a systematic approach to identifying and targeting the matrix enzymes that contribute to tissue destruction in TB infection with two overarching goals: detailing the role of individual enzymes in TB pathogenesis and developing and testing highly specific inhibitors of those enzymes as adjunct host-directed therapies in TB treatment. In preliminary work using a murine model of cavitary TB, we performed serial transcriptional profiling of infected lungs to identify the destructive matrix enzymes upregulated during infection. MT1-MMP stood out as upregulated in an early and sustained pattern; this enzyme has previously been associated with human TB. Using a highly specific MT1-MMP inhibitor developed by the Sagi laboratory, we performed a pilot experiment to determine tolerability of low-level dosing over the first 8 weeks post-infection in the murine cavitary model of TB. Mice tolerated the inhibitor well, and although the inhibitor was not dosed for maximum efficacy, histopathologic analysis demonstrated a trend toward decreased lesion size and fewer dense inflammatory cell infiltrates. In this proposed work, we will build upon those results to test both a role for MT1-MMP in pathogenesis in this model and the effect of inhibition on molecular, cellular, and histologic outcomes of disease. In aim 1, we will optimize dosing to maximize inhibition without inducing side-effects; we will then test the effect of MT1-MMP inhibition on bacterial growth and lung histopathology. In aim 2, we will use single-cell transcriptional profiling to both identify cellular drivers of MT1-MMP production and test the impact of MT1- MMP inhibition on inflammatory cell recruitment to the infected lung. We will then test the impact of MT1-MMP inhibition on the inflammatory milieu using multiplexed cytokine profiling and high resolution microscopy. Upon achieving these aims, we anticipate having characterized the role of MT1-MMP in the pathologic progression of TB in a murine model of disease. We anticipate these results will inform targeted studies of the role of matrix enzymes in the pathogenesis of human TB; further, we anticipate these results will ultimately inform the development of MMP inhibitors as adjunctive therapies for TB.