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Updated: May 10, 2026
Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry
Published on: October 18, 2016
Unveiling macrophage dynamics and efferocytosis-related targets in diabetic kidney disease: insights from single-cell
Binshan Zhang1, Yunqi Wu1, Zhongli Wang1
1National Health Commission (NHC) Key Lab of Hormones and Development and Tianjin Key Lab of Metabolic Diseases, Tianjin Medical University Chu Hsien-I Memorial Hospital & Institute of Endocrinology, Tianjin, China.
Background:
Chronic inflammation and immune imbalance mediated by macrophages are considered pivotal in diabetic kidney disease (DKD). The study aims to clarify the macrophage heterogeneity and phenotype dynamics, and pinpoint critical targets within efferocytosis in DKD.
Methods:
Utilizing early human DKD sequencing data, we computed the potential communication between leukocytes and renal intrinsic cells. Subsequently, we scrutinized the single-cell RNA sequencing (scRNA-seq) data from CD45-enriched immune cells, concentrating on the macrophage subsets in DKD. Pseudotime trajectory analysis was conducted to explore cell development. Differential expression genes (DEGs) from macrophage subgroups and bulk RNA-sequencing were used to identify shared hub genes. The NephroseqV5 platform was employed to evaluate the clinical significance, and the expression of key molecules was validated in DKD tissues.
Results:
Macrophage infiltration rose in DKD, causing inflammation through the release of chemokines. As time progressed, the number of resident macrophages substantially dropped, with diminishing M1-like and increasing M2-like phenotypes relative to early stages. Further analysis pointed to the most enrichment of macrophage function is the phagosome. We overlapped the DEGs with efferocytosis-related genes and identified key genes, including CD36, ITGAM, and CX3CR1, which exhibited significant correlations with macrophages and T cells. The Nephroseq database revealed that they are associated with proteinuria and renal function. Consistent with the validation set, in vivo experiments verified elevated expression levels of key molecules.
Conclusions:
In essence, our research elucidated the dynamics in macrophage subtype transitions. It emphasized three pivotal genes as critical modulators of macrophage efferocytosis in DKD, indicating their potential as innovative biomarkers and therapeutic targets.
Insights
Diabetic kidney disease involves shifting macrophage populations and inflammation. Key genes like CD36 regulate efferocytosis, offering potential therapeutic targets for diabetic kidney disease.
Area of Science:
- Immunology
- Nephrology
- Genomics
Background:
- Diabetic kidney disease (DKD) pathogenesis is linked to chronic inflammation and immune cell imbalance, particularly involving macrophages.
- Understanding macrophage heterogeneity and dynamic changes is crucial for DKD progression.
Purpose of the Study:
- To clarify macrophage heterogeneity and phenotype dynamics in DKD.
- To identify critical molecular targets within efferocytosis pathways in DKD.
Main Methods:
- Analysis of human DKD sequencing data to map cell communication.
- Single-cell RNA sequencing (scRNA-seq) of immune cells to focus on macrophage subsets.
- Pseudotime analysis, differential gene expression (DEG) analysis, and clinical validation using NephroseqV5.
Main Results:
- Macrophage infiltration increased in DKD, driving inflammation.
- Phenotypic shifts observed from M1-like to M2-like macrophages with decreasing resident macrophages.
- Identified CD36, ITGAM, and CX3CR1 as key efferocytosis-related genes correlated with disease markers.
Conclusions:
- Elucidated dynamic transitions in macrophage subtypes during DKD.
- Highlighted three pivotal genes (CD36, ITGAM, CX3CR1) as critical modulators of efferocytosis.
- These genes represent potential novel biomarkers and therapeutic targets for DKD.
