Targeting pathologic macrophage activation through inhibition of MyD88 to attenuate laryngotracheal stenosis

通过抑制 MyD88 靶向病理性巨噬细胞激活以减轻喉气管狭窄

• 批准号: 10644555
• 负责人: Kevin Motz
• 金额: $ 19.76万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644555
• 关键词: Adult; Attenuated; Automobile Driving; Biological Response Modifiers; Biopsy; Bypass; CD4 Positive T Lymphocytes; Cells; Collagen; Communication impairment; Critical Illness; Critical Pathways; Data; Development; Disease; Dysphonia; Dyspnea; Excision; Experimental Designs; Fibrosis; Flow Cytometry; Foundations; Gene Expression; Goals; Histologic; Human; Iatrogenesis; Immune; Immune Targeting; Immune response; Immunology; Immunophenotyping; Inflammation; Inflammatory Infiltrate; Injury; Interleukin-1 beta; Intervention; Intratracheal Intubation; Intubation; Investigation; Knowledge; Laboratories; Lamina Propria; Laryngeal Diseases; Larynx; Lead; Life; Macrophage; Macrophage Activation; Mediating; Medical; Mentors; Modeling; Molecular; Mucous Membrane; Mus; Operative Surgical Procedures; Pathogenesis; Pathologic; Pathway interactions; Patients; Pattern; Phenotype; Physiological; Population; Proteins; Regulatory Pathway; Research; Research Training; Resected; Role; S100A8 gene; Secondary to; Serum; Signal Pathway; Signal Transduction; Source; Stenosis; Subglottis structure; TLR4 gene; Therapeutic Intervention; Tissues; Toll-like receptors; Trachea; Tracheostomy procedure; Transgenic Organisms; biomarker development; candidate marker; career; cytokine; experience; genetic regulatory protein; improved; mouse model; pathogen; prevent; protein expression; receptor; single-cell RNA sequencing; skills; targeted treatment; treatment strategy; wound healing

Project Summary: Laryngotracheal Stenosis (LTS) is the pathologic narrowing of the larynx, subglottis, and trachea secondary to mucosal injury from prolonged intubation. This narrowing leads to dyspnea, dysphonia, and can rapidly progress to airway compromise. Therapeutic interventions for the management of LTS are limited to serial dilation, tracheal resection, or permanent tracheostomy which further impairs communication. Medical therapies for LTS are limited by our poor understanding of LTS pathogenesis. Improved understanding of the mechanisms promoting LTS is needed to improve treatment of this debilitating disease. Previous investigation has revealed that an intact immune response is critical to the development of LTS. Characterization of the immune response in LTS has demonstrated increased populations of CD4+ T-cells and macrophages. Preliminary studies in a murine LTS model reveal that depletion of the macrophage population attenuates LTS fibrosis, implicating their pathologic role. However, the local immune mediators and cell signaling pathways promoting pathologic macrophages in LTS are unknown. Macrophage activation is regulated through stimulation of Toll-like receptors (TLRs). TLRs are highly conserved receptors recognizing Pathogen or Damage Associated Molecular Patterns (PAMPs/DAMPs) and lead to downstream activation of regulatory proteins controlling phenotype. Using single cell RNA sequencing the PI has demonstrates increased expression of the TLR4-MyD88 signaling pathway in LTS macrophages. Furthermore, we have identified increased expression of the DAMP S100A8/A9 in LTS tissue. S100A8/A9 is a known activator of TLR4-MyD88 signaling, and worsens fibrosis in our murine LTS model. These findings indicate that TLR4-MyD88 signaling pathways in macrophages may be critical to LTS pathogenesis. However, the relationship between S100A8/A9, TLR4-MyD88 signaling, and pathologic macrophages has not been explored in LTS or other fibrotic diseases, and may represent a critical signaling axis driving pathologic fibrosis. For this study, we will elucidate the signaling networks promoting pathologic macrophages in laryngotracheal stenosis. In Aim 1 we will assess effect of S100A8/A9 on macrophage phenotype and function in a murine LTS model, establish that S100A8/A9s profibrotic effect is mediated by macrophages, and identify the key sources of pathologic S100A8/A9 in human LTS and a murine model. In Aim 2, we will demonstrate the critical role of TLR4-MyD88 signaling in promoting pathologic macrophages in LTS, and elucidate the role of IL1β in promoting LTS fibrosis. Finally, in Aim 3 we will assess S100A8/A9 as a candidate biomarker for the development of laryngotracheal stenosis in patients who have had prolonged intubation. Collectively, this application will lead to an in-depth understanding of the cell signaling networks promoting dysregulated macrophage mediated inflammation and subsequent fibrosis in LTS. The identification of key regulatory pathways promoting pathologic macrophages in LTS will serve as the foundation for targeted treatment strategies that attenuate fibrosis.

项目摘要： 喉气管狭窄（LTS）是喉，sublottis和气管的病理变窄 继发于长时间插管的粘膜损伤。这种变窄会导致呼吸困难，吞咽困难，并且可以 快速进步到气道妥协。 LTS管理的治疗干预措施仅限于 串行词典，气管切除或永久性气管切开术，进一步损害了通信。医疗的 LTS的疗法受到我们对LTS发病机理的不良理解的限制。提高了对 需要促进LTS的机制来改善这种使人衰弱的疾病的治疗。 先前的研究表明，完整的免疫反应对于LT的发展至关重要。 LTS中免疫响应的表征表明，CD4+ T细胞的种群增加了 巨噬细胞。鼠LTS模型中的初步研究表明，巨噬细胞的部署 减弱LTS纤维化，隐含其病理作用。但是，局部免疫介质和细胞信号传导 促进LT中促进病理巨噬细胞的途径尚不清楚。 巨噬细胞激活通过刺激收费受体（TLR）来调节。 TLR高度 保守的受体识别病原体或损害相关的分子模式（PAMP/DAMP）和 导致控制表型的调节蛋白的下游激活。使用单细胞RNA测序 PI表明LTS巨噬细胞中TLR4-MYD88信号通路的表达增加。 此外，我们已经确定了LTS组织中潮湿S100A8/A9的表达增加。 S100A8/A9是 TLR4-MYD88信号传导的已知激活剂，并在我们的鼠LTS模型中恶化纤维化。这些发现 表明巨噬细胞中的TLR4-MYD88信号通路可能对LTS发病机理至关重要。然而， S100A8/A9，TLR4-MYD88信号传导和病理巨噬细胞之间的关系尚未 在LTS或其他纤维化疾病中探索，可能代表关键的信号轴驱动病理纤维化。 在这项研究中，我们将阐明促进喉气管病理巨噬细胞的信号网络 狭窄。在AIM 1中，我们将评估S100A8/A9对鼠LTS中巨噬细胞表型和功能的影响 模型，确定S100A8/A9S纤维化效应是由巨噬细胞介导的，并识别关键来源 人体LTS中的病理S100A8/A9和鼠模型。在AIM 2中，我们将展示 TLR4-MYD88在促进LT中促进病理巨噬细胞的信号传导，并阐明IL1β在促进中的作用 LTS纤维化。最后，在AIM 3中，我们将评估S100A8/A9作为开发的候选生物标志物 插管延长的患者的喉气管狭窄。总的来说，此应用程序将导致 对促进失调的巨噬细胞介导的细胞信号网络的深入了解 LTS的炎症和随后的纤维化。促进病理的关键调节途径的识别 LTS中的巨噬细胞将作为减轻纤维化的靶向治疗策略的基础。