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Background <p>This study aimed to identify a key target gene in proximal tubule cells (PTCs) of sepsis-induced acute kidney injury (S-AKI) and elucidate the underlying mechanisms.</p> Methods <p>We screened and analyzed GEO datasets and identified a key gene, GATM, in S-AKI. An S-AKI mouse model was established via intraperitoneal injection of lipopolysaccharide (LPS), and HK-2 cells were used for in vitro experiments. The role of GATM was evaluated using adeno-associated virus (AAV)-mediated overexpression in mice and plasmid-mediated overexpression in HK-2 cells. To identify the downstream target genes of GATM, transcriptome sequencing was conducted. Pathological evaluation was performed using hematoxylin–eosin (HE) and periodic acid–Schiff (PAS) staining. Protein levels were determined by Western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF) assays; Apoptosis was evaluated by TUNEL staining; Mitochondrial morphology and function were assessed by transmission electron microscopy (TEM), JC-1 and MitoSOX assays. Lactate concentration and cellular ATP levels were measured.</p> Results <p>Through analysis of four datasets (GSE151658, GSE247727, GSE220812, and GSE139061), GATM was identified as a key gene in PTCs during S-AKI. GATM expression was downregulated in both S-AKI mice and LPS-stimulated HK-2 cells. In vivo, GATM overexpression improved renal function, alleviated tubular damage, decreased the expression of KIM-1, IL-6, Caspase-3, and 4-HNE, and reduced mitochondrial injury. In vitro, HK-2 cell viability was enhanced, TUNEL-positive cells were reduced, and damaged mitochondria were decreased. Transcriptome sequencing revealed that the PDK4-mediated glycolysis pathway was a downstream target of GATM. GATM overexpression downregulated PDK4 expression, reduced glycolytic enzyme levels (p-PDHA, HK2, LDHA, GLUT1) and lactate, and increased ATP production. However, PDK4 overexpression in HK-2 cells abolished the protective effects of GATM, enhanced glycolysis, increased lactate levels, and reduced ATP production.</p> Conclusion <p>GATM plays a protective role in S-AKI by inhibiting PDK4-mediated aerobic glycolysis, enhancing ATP production, and restoring energy metabolism and mitochondrial function in PTCs.</p>

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GATM alleviates sepsis-induced acute kidney injury via PDK4-mediated glycolytic reprogramming in renal tubular epithelial cells

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年5月9日

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  • Ruhao Yang,
  • Niandan Hu,
  • Hairui Chen,
  • Wenqiang Li,
  • Ting Zheng

摘要

Background

This study aimed to identify a key target gene in proximal tubule cells (PTCs) of sepsis-induced acute kidney injury (S-AKI) and elucidate the underlying mechanisms.

Methods

We screened and analyzed GEO datasets and identified a key gene, GATM, in S-AKI. An S-AKI mouse model was established via intraperitoneal injection of lipopolysaccharide (LPS), and HK-2 cells were used for in vitro experiments. The role of GATM was evaluated using adeno-associated virus (AAV)-mediated overexpression in mice and plasmid-mediated overexpression in HK-2 cells. To identify the downstream target genes of GATM, transcriptome sequencing was conducted. Pathological evaluation was performed using hematoxylin–eosin (HE) and periodic acid–Schiff (PAS) staining. Protein levels were determined by Western blotting (WB), immunohistochemistry (IHC), and immunofluorescence (IF) assays; Apoptosis was evaluated by TUNEL staining; Mitochondrial morphology and function were assessed by transmission electron microscopy (TEM), JC-1 and MitoSOX assays. Lactate concentration and cellular ATP levels were measured.

Results

Through analysis of four datasets (GSE151658, GSE247727, GSE220812, and GSE139061), GATM was identified as a key gene in PTCs during S-AKI. GATM expression was downregulated in both S-AKI mice and LPS-stimulated HK-2 cells. In vivo, GATM overexpression improved renal function, alleviated tubular damage, decreased the expression of KIM-1, IL-6, Caspase-3, and 4-HNE, and reduced mitochondrial injury. In vitro, HK-2 cell viability was enhanced, TUNEL-positive cells were reduced, and damaged mitochondria were decreased. Transcriptome sequencing revealed that the PDK4-mediated glycolysis pathway was a downstream target of GATM. GATM overexpression downregulated PDK4 expression, reduced glycolytic enzyme levels (p-PDHA, HK2, LDHA, GLUT1) and lactate, and increased ATP production. However, PDK4 overexpression in HK-2 cells abolished the protective effects of GATM, enhanced glycolysis, increased lactate levels, and reduced ATP production.

Conclusion

GATM plays a protective role in S-AKI by inhibiting PDK4-mediated aerobic glycolysis, enhancing ATP production, and restoring energy metabolism and mitochondrial function in PTCs.