PDF(Pubmed)
Abstract：
UNASSIGNED: Clinical trials for reducing fibrosis in steatotic liver disease (SLD) have targeted macrophages with variable results. We evaluated intrahepatic macrophages in patients with SLD to determine if activity scores or fibrosis stages influenced phenotypes and expression of druggable targets, such as CCR2 and galectin-3.
UNASSIGNED: Liver biopsies from controls or patients with minimal or advanced fibrosis were subject to gene expression analysis using nCounter to determine differences in macrophage-related genes (n = 30). To investigate variability among individual patients, we compared additional biopsies by staining them with multiplex antibody panels (CD68/CD14/CD16/CD163/Mac387 or CD163/CCR2/galectin-3/Mac387) followed by spectral imaging and spatial analysis. Algorithms that utilize deep learning/artificial intelligence were applied to create cell cluster plots, phenotype profile maps, and to determine levels of protein expression (n = 34).
UNASSIGNED: Several genes known to be pro-fibrotic (e.g. CD206, TREM2, CD163, and ARG1) showed either no significant differences or significantly decreased with advanced fibrosis. Although marked variability in gene expression was observed in individual patients with cirrhosis, several druggable targets and their ligands (e.g. CCR2, CCR5, CCL2, CCL5, and LGALS3) were significantly increased when compared to patients with minimal fibrosis. Antibody panels identified populations that were significantly increased (e.g. Mac387+), decreased (e.g. CD14+), or enriched (e.g. interactions of Mac387) in patients that had progression of disease or advanced fibrosis. Despite heterogeneity in patients with SLD, several macrophage phenotypes and druggable targets showed a positive correlation with increasing NAFLD activity scores and fibrosis stages.
UNASSIGNED: Patients with SLD have markedly varied macrophage- and druggable target-related gene and protein expression in their livers. Several patients had relatively high expression, while others were like controls. Overall, patients with more advanced disease had significantly higher expression of CCR2 and galectin-3 at both the gene and protein levels.
UNASSIGNED: Appreciating individual differences within the hepatic microenvironment of patients with SLD may be paramount to developing effective treatments. These results may explain why such a small percentage of patients have responded to macrophage-targeting therapies and provide additional support for precision medicine-guided treatment of chronic liver diseases.
UNASSIGNED: Liver biopsies from controls or patients with minimal or advanced fibrosis were subject to gene expression analysis using nCounter to determine differences in macrophage-related genes (n = 30). To investigate variability among individual patients, we compared additional biopsies by staining them with multiplex antibody panels (CD68/CD14/CD16/CD163/Mac387 or CD163/CCR2/galectin-3/Mac387) followed by spectral imaging and spatial analysis. Algorithms that utilize deep learning/artificial intelligence were applied to create cell cluster plots, phenotype profile maps, and to determine levels of protein expression (n = 34).
UNASSIGNED: Several genes known to be pro-fibrotic (e.g. CD206, TREM2, CD163, and ARG1) showed either no significant differences or significantly decreased with advanced fibrosis. Although marked variability in gene expression was observed in individual patients with cirrhosis, several druggable targets and their ligands (e.g. CCR2, CCR5, CCL2, CCL5, and LGALS3) were significantly increased when compared to patients with minimal fibrosis. Antibody panels identified populations that were significantly increased (e.g. Mac387+), decreased (e.g. CD14+), or enriched (e.g. interactions of Mac387) in patients that had progression of disease or advanced fibrosis. Despite heterogeneity in patients with SLD, several macrophage phenotypes and druggable targets showed a positive correlation with increasing NAFLD activity scores and fibrosis stages.
UNASSIGNED: Patients with SLD have markedly varied macrophage- and druggable target-related gene and protein expression in their livers. Several patients had relatively high expression, while others were like controls. Overall, patients with more advanced disease had significantly higher expression of CCR2 and galectin-3 at both the gene and protein levels.
UNASSIGNED: Appreciating individual differences within the hepatic microenvironment of patients with SLD may be paramount to developing effective treatments. These results may explain why such a small percentage of patients have responded to macrophage-targeting therapies and provide additional support for precision medicine-guided treatment of chronic liver diseases.
摘要：
■减少脂肪变性肝病（SLD）纤维化的临床试验已靶向巨噬细胞，结果可变。我们评估了SLD患者的肝内巨噬细胞，以确定活动评分或纤维化阶段是否影响表型和可成药靶标的表达，如CCR2和半乳糖凝集素-3。
■使用nCounter进行基因表达分析，以确定巨噬细胞相关基因的差异（n=30）。为了调查个体患者之间的变异性，我们通过多重抗体组(CD68/CD14/CD16/CD163/Mac387或CD163/CCR2/galectin-3/Mac387)染色,然后进行光谱成像和空间分析,比较了其他活检.利用深度学习/人工智能的算法被应用于创建细胞簇图，表型图谱，并测定蛋白质表达水平(n=34)。
几种已知是促纤维化的基因(例如CD206、TREM2、CD163和ARG1)在晚期纤维化中没有显著差异或显著降低。虽然在个别肝硬化患者中观察到基因表达的显著变异性，与纤维化程度最小的患者相比,一些可药物治疗的靶点及其配体(如CCR2,CCR5,CCL2,CCL5和LGALS3)显著增加.抗体小组确定了显著增加的群体(例如Mac387+)，减少(例如CD14+)，或富集（例如，Mac387的相互作用）在疾病进展或晚期纤维化的患者中。尽管SLD患者存在异质性，几种巨噬细胞表型和可含药靶点与NAFLD活性评分和纤维化分期的增加呈正相关.
■患有SLD的患者在其肝脏中具有显著变化的巨噬细胞和可药用靶相关基因和蛋白质表达。几个患者有相对较高的表达，而其他人就像控制一样。总的来说,疾病更晚期的患者在基因和蛋白质水平上CCR2和半乳糖凝集素-3的表达均显著较高.
■了解SLD患者肝微环境中的个体差异对于开发有效的治疗方法至关重要。这些结果可以解释为什么如此小比例的患者对巨噬细胞靶向治疗有反应，并为精准医学指导治疗慢性肝病提供额外的支持。
■使用nCounter进行基因表达分析，以确定巨噬细胞相关基因的差异（n=30）。为了调查个体患者之间的变异性，我们通过多重抗体组(CD68/CD14/CD16/CD163/Mac387或CD163/CCR2/galectin-3/Mac387)染色,然后进行光谱成像和空间分析,比较了其他活检.利用深度学习/人工智能的算法被应用于创建细胞簇图，表型图谱，并测定蛋白质表达水平(n=34)。
几种已知是促纤维化的基因(例如CD206、TREM2、CD163和ARG1)在晚期纤维化中没有显著差异或显著降低。虽然在个别肝硬化患者中观察到基因表达的显著变异性，与纤维化程度最小的患者相比,一些可药物治疗的靶点及其配体(如CCR2,CCR5,CCL2,CCL5和LGALS3)显著增加.抗体小组确定了显著增加的群体(例如Mac387+)，减少(例如CD14+)，或富集（例如，Mac387的相互作用）在疾病进展或晚期纤维化的患者中。尽管SLD患者存在异质性，几种巨噬细胞表型和可含药靶点与NAFLD活性评分和纤维化分期的增加呈正相关.
■患有SLD的患者在其肝脏中具有显著变化的巨噬细胞和可药用靶相关基因和蛋白质表达。几个患者有相对较高的表达，而其他人就像控制一样。总的来说,疾病更晚期的患者在基因和蛋白质水平上CCR2和半乳糖凝集素-3的表达均显著较高.
■了解SLD患者肝微环境中的个体差异对于开发有效的治疗方法至关重要。这些结果可以解释为什么如此小比例的患者对巨噬细胞靶向治疗有反应，并为精准医学指导治疗慢性肝病提供额外的支持。
