预测自噬相关基因并揭示肝缺血再灌注损伤中肝内皮细胞的异质性。

BACKGROUND: Hepatic Ischemia-Reperfusion Injury (HIRI) is a major complication in liver transplants and surgeries, significantly affecting postoperative outcomes. The role of mitophagy, essential for removing dysfunctional mitochondria and maintaining cellular balance, remains unclear in HIRI. METHODS: To unravel the role of mitophagy-related genes (MRGs) in HIRI, we assembled a comprehensive dataset comprising 44 HIRI samples alongside 44 normal control samples from the Gene Expression Omnibus (GEO) database for this analysis. Using Random Forests and Support Vector Machines - Recursive Feature Elimination (SVM-RFE), we pinpointed eight pivotal genes and developed a logistic regression model based on these findings. Further, we employed consensus cluster analysis for classifying HIRI patients according to their MRG expression profiles and conducted weighted gene co-expression network analysis (WGCNA) to identify clusters of genes that exhibit high correlation within different modules. Additionally, we conducted single-cell RNA sequencing data analysis to explore insights into the behavior of MRGs within the HIRI. RESULTS: We identified eight key genes (FUNDC1, VDAC1, MFN2, PINK1, CSNK2A2, ULK1, UBC, MAP1LC3B) with distinct expressions between HIRI and controls, confirmed by PCR validation. Our diagnostic model, based on these genes, accurately predicted HIRI outcomes. Analysis revealed a strong positive correlation of these genes with monocytic lineage and a negative correlation with B and T cells. HIRI patients were divided into three subclusters based on MRG profiles, with WGCNA uncovering highly correlated gene modules. Single-cell analysis identified two types of endothelial cells with different MRG scores, indicating their varied roles in HIRI. CONCLUSIONS: Our study highlights the critical role of MRGs in HIRI and the heterogeneity of endothelial cells. We identified the macrophage migration inhibitory factor (MIF) and cGAS-STING (GAS) pathways as regulators of mitophagy's impact on HIRI. These findings advance our understanding of mitophagy in HIRI and set the stage for future research and therapeutic developments.

背景：肝缺血再灌注损伤（HIRI）是肝移植和手术中的主要并发症，显著影响术后结果。线粒体自噬在 HIRI 中的作用对于去除功能失调的线粒体和维持细胞平衡至关重要，但目前尚不清楚。

方法：为了揭示与线粒体自噬相关的基因（MRGs）在 HIRI 中的作用，我们从基因表达综合数据库（GEO）中收集了 44 个 HIRI 样本和 44 个正常对照样本，进行了全面的数据集组装。使用随机森林和支持向量机-递归特征消除（SVM-RFE），我们确定了 8 个关键基因，并基于这些发现开发了一个逻辑回归模型。此外，我们根据这些基因的表达谱对 HIRI 患者进行了共识聚类分析，并进行了加权基因共表达网络分析（WGCNA），以识别不同模块内高相关性的基因簇。此外，我们进行了单细胞 RNA 测序数据分析，以探索 HIRI 中 MRGs 行为的见解。

结果：我们确定了 8 个关键基因（FUNDC1、VDAC1、MFN2、PINK1、CSNK2A2、ULK1、UBC、MAP1LC3B），它们在 HIRI 和对照组之间的表达不同，通过 PCR 验证得到了确认。我们基于这些基因的诊断模型准确预测了 HIRI 结果。分析表明，这些基因与单核细胞谱系呈强正相关，与 B 和 T 细胞呈负相关。根据 MRG 谱，HIRI 患者被分为 3 个亚群，WGCNA 揭示了高度相关的基因模块。单细胞分析确定了两种具有不同 MRG 评分的内皮细胞类型，表明它们在 HIRI 中具有不同的作用。

结论：我们的研究强调了 MRGs 在 HIRI 中的关键作用和内皮细胞的异质性。我们确定了巨噬细胞移动抑制因子（MIF）和 cGAS-STING（GAS）途径作为调节线粒体自噬对 HIRI 影响的调节剂。这些发现提高了我们对 HIRI 中线粒体自噬的理解，并为未来的研究和治疗发展奠定了基础。