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Transcriptomic profiling of hemin-treated bone marrow-derived macrophages in a TGF-β1-induced macrophage–renal tubular epithelial cell co-culture model

ABSTRACT: Kidney fibrosis is characterized by persistent activation of renal tubular epithelial cells and extensive remodeling of the immune microenvironment. Macrophages are highly plastic cells that respond to metabolic and inflammatory signals and may regulate the progression of renal fibrosis. This study investigated the transcriptional changes induced by hemin in bone marrow-derived macrophages in a macrophage–renal proximal tubular epithelial cell co-culture system. Mouse bone marrow-derived macrophages were co-cultured with primary renal proximal tubular epithelial cells using a Transwell system. Renal tubular epithelial cells were exposed to transforming growth factor beta 1 (TGF-β1) to establish a profibrotic microenvironment, with or without hemin treatment. After treatment, macrophages were collected for bulk RNA sequencing. Transcriptomic analysis showed that hemin altered macrophage pathways associated with heme and iron metabolism, antioxidant responses, glutathione metabolism, inflammatory signaling, extracellular matrix regulation, and cellular metabolism. These data provide insight into the macrophage transcriptional reprogramming associated with the protective effects of hemin against renal fibrosis.

ORGANISM(S): Mus musculus

PROVIDER: GSE343718 | GEO | 2026/08/14

REPOSITORIES: GEO

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Project description:We examined, through an oligonucleotide microarray protocol, the renal differential transcriptome profiles of mices submitted to IRI, IPC and Hemin treatment. After identifying the profiles of differentially expressed genes observed for each comparison, we carried out functional enrichment analysis to reveal transcripts putatively involved in potential relevant biological processes and signaling pathways. The most relevant enriched biological processes found in these comparisons were stress, inflammation, apoptosis, pathways in cancer, differentiation, angiogenesis, focal adhesion, ECM-receptor interaction, ion transport, angiogenesis, mitosis and cell cycle, inflammatory response, olfactory transduction and regulation of actin cytoskeleton. In addition, the most important overrepresented pathways were MAPK, ErbB, JAK/STAT, Toll and Nod like receptors, Angiotensin II, Arachidonic acid metabolism, Wnt and coagulation cascade. Also, new insights were gained about the underlying protection mechanisms against renal IRI promoted by IPC and Hemin treatment. Venn diagram analysis allowed us to uncover common and exclusively differentially expressed genes between these two protective maneuvers, underscoring potential common and exclusive biological functions regulated in each case. In summary, IPC exclusively regulated the expression of genes belonging to stress, protein modification and apoptosis, highlighting the role of IPC in controlling exacerbated stress response. Treatment with the Hmox1 inducer Hemin, in turn, exclusively regulated the expression of genes associated with cell differentiation, metabolic pathways, cell cycle, mitosis, development, regulation of actin cytoskeleton and arachidonic acid metabolism, suggesting a pleiotropic effect for Hemin. According to surgery and treatment procedures, mices were divided into five groups: Control, Ischemia-reperfusion injury (IRI), animals pre-treated with Hemin followed by IRI (Hemin + IRI), animals only treated with Hemin (Hemin) and animals pre-conditioned and submitted to IRI (IPC+IRI). To identify the consequences of IRI, Hemin treatment or IPC in gene expression profiles, the following statistical comparisons were made between the groups: IRI vs Control, IPC+IRI vs Control, IPC+IRI vs IRI, IRI+Hemin vs IRI and Hemin vs Control.

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Transcriptomic profiling of hemin-treated bone marrow-derived macrophages in a TGF-β1-induced macrophage–renal tubular epithelial cell co-culture model

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