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Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes

  • Authors:
    • Xiaona Chen
    • Ziqi Sun
    • Jiabo Chen
    • Jinquan Zhang
    • Zeyu Liu
    • Zhengzheng Yan
    • Quan Li
    • Zhixia Chen
  • View Affiliations / Copyright

    Affiliations: School of Medicine, Southern University of Science and Technology, Shenzhen, Guangdong 518055, P.R. China, Department of Anesthesiology, National Cancer Center/National Clinical Research Center for Cancer/ Cancer Hospital and Shenzhen Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Shenzhen, Guangdong 518116, P.R. China, Department of Anesthesiology, The Tenth Affiliated Hospital, Southern Medical University (Dongguan People's Hospital), Dongguan, Guangdong 523000, P.R. China
    Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 18
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    Published online on: November 12, 2025
       https://doi.org/10.3892/ijmm.2025.5689
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Abstract

Studies have linked the dysregulation of N6‑methyladenosine (m6A) to sepsis‑induced acute kidney injury (SAKI), highlighting the persistent challenge of managing excessive proinflammatory cytokine production and subsequent organ dysfunction. The present study, by analyzing the GSE32707 and GSE69063 datasets, found that fat mass and obesity‑associated protein (FTO) was the sole m6A‑related gene markedly downregulated in the peripheral blood transcriptome of patients with sepsis. It further demonstrated that septic mice subjected to cecal ligation and puncture presented increased m6A modifications and reduced FTO expression in both renal tissues and peritoneal macrophages. The findings revealed that increased levels of FTO was associated with reduced mortality and kidney damage during sepsis and that the upregulation of FTO in lipopolysaccharide‑stimulated macrophages led to decreased production of proinflammatory cytokines. Mechanistically, through multiomic analysis of macrophages, the present study identified a novel mechanism involving matrix metalloproteinase 9 (MMP‑9) as a direct target of FTO, which positively affects its translation efficacy. Furthermore, both in vivo and in vitro data confirmed that reduced MMP‑9 levels exerted adverse effects on mitigating inflammatory responses and alleviating renal injury. Overall, the findings underscored the critical role of the FTO/m6A/MMP‑9 axis in the regulation of proinflammatory secretion and improved our understanding of the transcriptomic landscape during the progression of SAKI, suggesting that targeting the FTO/m6A/MMP‑9 axis may offer therapeutic potential for mitigating renal injury in septic patients.
View Figures

Figure 1

FTO expression is downregulated and
m6A methylation increased in SAKI. (A) Schematic
illustration of the screening process employed to identify key
genes associated with m6A modification in sepsis. (B)
Scatter plot depicting the differential expression of
m6A-related genes, highlighting FTO as the only gene
whose expression was markedly downregulated in the sepsis group
(Sepsis) compared with that in the NC goup in the GSE32707 dataset.
(C) Analysis of the GSE69063 dataset revealed decreased FTO
expression in the transcriptomes of m6A-related proteins
in peripheral blood samples from patients with sepsis (Sepsis)
compared with those from HCs. (D) Representative IHC staining and
(E) quantification of FTO expression in kidney tissues from
CLP-induced septic mice compared with those from sham-operated
controls (n=3; P<0.05). (F) Dot blot analysis revealed a
significant increase in the global m6A modification
level in the kidney tissues of CLP-induced septic mice (n=3;
P<0.05). All the data are presented as the mean±standard error
of the mean. Data (in D and F) were analyzed using an unpaired
two-tailed Student's t-test. *P<0.05. FTO, fat mass
and obesity-associated protein; m6A, N6-methyladenosine;
SAKI, sepsis-induced acute kidney injury; HCs healthy controls; NC,
negative control; IHC, immunohistochemistry; CLP, cecal ligation
and puncture.

Figure 2

FTO modulates macrophage function and
inflammatory responses in SAKI. (A) Western blotting and (B) qPCR
revealed the downregulation of FTO expression in PMs isolated from
septic mice compared with those from sham-operated controls (n=3).
(C) Dot blot analysis revealed a time-dependent increase in
m6A methylation levels in RAW264.7 cells treated with
LPS (1 μg/ml) for 0, 6 and 12 h. (D) qPCR analysis of
m6A-related enzymes in LPS-treated macrophages. (E)
Western blotting and (F) qPCR demonstrated a time-dependent
reduction in FTO protein expression in LPS-treated RAW264.7 cells.
(G) Western blotting confirmed FTO overexpression in oe-FTO
macrophages with or without treatment with LPS for 12 h. (H) CCK-8
and (I) LDH assays revealed that FTO overexpression does not induce
cytotoxicity or affect macrophage proliferation. (J and K)
Phagocytosis assays demonstrated that compared with control
treatment, upregulation of FTO expression did not alter the
phagocytic capacity of macrophages (original magnification, ×63).
(L) ELISA revealed that overexpression of FTO expression markedly
inhibited the secretion of IL-6 and TNF-α. (M) qPCR experiments
confirmed the suppressive effect of FTO on the mRNA expression
levels of IL-6 and TNF-α. (N) Western blot analysis revealed that
FTO attenuated the activation of inflammatory pathways by
inhibiting NF-κB signaling. *P<0.05;
**P<0.01; ***P<0.001;
****P<0.0001; ns, not significant. FTO, fat mass and
obesity-associated protein; SAKI, sepsis-induced acute kidney
injury; qPCR, quantitative PCR; PMs, peritoneal macrophages; LPS,
lipopolysaccharide; m6A, N6-methyladenosine; oe, over
expression; LDH, lactate dehydrogenase; ELISA, enzyme-linked
immunosorbent assay.

Figure 3

MMP-9 is the key target of
FTO-mediated m6A modification. (A) Flowchart of the
multiomics analysis of macrophages. (B) Hierarchical clustering of
differentially expressed genes in macrophages following LPS
treatment. Volcano plot showing (C) downregulated FTO expression
and (D) upregulated MMP-9 expression in LPS-induced macrophages
compared with NC. (E) GO analysis of the DEGs revealed that the
DEGs were enriched in immune response-related terms. (F) GSEA
revealed that key pathways, including the TNF-α signaling and
IL-6/JAK/STAT3 pathways, were markedly associated with the
inflammatory response in LPS-treated macrophages. (G) Motif
analysis of m6A methylomes in macrophages. (H)
Integrative Genomics Viewer image showing that the m6A
peaks of MMP-9 mRNA were near the 3'UTR. (I) mRNA-seq revealed that
DEGs associated with FTO overexpression in LPS-stimulated
macrophages were enriched in cytokine-cytokine receptor interaction
pathways. (J) Volcano plot showing that elevated FTO levels were
associated with increased MMP-9 expression. MMP-9, matrix
metalloproteinase 9; FTO, fat mass and obesity-associated protein;
m6A, N6-methyladenosine; LPS, lipopolysaccharide; NC,
negative control; GO, gene ontology; DEGs, differentially expressed
genes; gene set enrichment analysis; TNF-α, tumor necrosis
factor-α; IL-6, interleukin-6.

Figure 4

FTO upregulates MMP-9 expression in
an m6A-dependent manner in macrophages. (A) MeRIP-qPCR
confirmed the decreased level of m6A modification of
MMP-9 mRNA in LPS-treated macrophages overexpressing FTO. (B) qPCR
and (C) western blot analyses demonstrated that compared with
control macrophage treatment, LPS stimulation increased MMP-9
expression in FTO-overexpressing macrophages. (D)
Immunofluorescence assays confirmed that elevated FTO levels
upregulated MMP-9 protein expression in macrophages following LPS
treatment. (E) Actinomycin D treatment revealed that FTO-mediated
m6A modification did not markedly affect the mRNA
stability of MMP-9 in macrophages treated with LPS for 12 h. (F)
CHX treatment and (G) quantification demonstrated that FTO did not
markedly affect MMP-9 protein stability following LPS treatment.
(H) Puromycin labeling of nascent polypeptides revealed that FTO
enhanced protein synthesis in macrophages treated with LPS for 12
h. (I) overexpression of IGF2BP3 in FTO-overexpressing macrophages
led to the further upregulation of MMP-9. (J) Co-IP experiment
identified the IGF2BP3 did not co-precipitate either with FTO or
MMP9 in macrophages. *P<0.05; **P<0.01;
ns, not significant. FTO, fat mass and obesity-associated protein;
MMP-9, matrix metalloprot einase 9; m6A,
N6-methyladenosine; MeRIP-seq, methylated RNA immunoprecipitation
sequencing; qPCR, quantitative PCR; LPS, lipopolysaccharide; oe,
over expression; IGF2BP3, insulin-like growth factor 2 mRNA-binding
protein 3.

Figure 5

Depletion of MMP-9 increases the
release of proinflammatory factors and exacerbated renal injury in
SAKI model mice. (A) Western blot analysis revealed an increased
abundance of MMP-9 in the kidneys of mice with SAKI (n=3). (B)
Western blot and (C) qPCR analyses revealed an increased abundance
of MMP-9 in CLP-derived PMs (n=3). (D) Western blot analysis
revealed a time-dependent increase in the abundance of the MMP-9
protein in LPS-stimulated macrophages. (E) ELISA confirmed that
MMP-9 depletion markedly increased the production of IL-6 and
TNF-α. (F) qPCR analysis revealed markedly increased expression
levels of IL-6 and TNF-α in MMP-9-depleted oe-FTO macrophages
following LPS treatment. (G and H) ELISA confirmed that MMP-9
depletion markedly increased the production of TNF-α and IL-6 in
MMP-9-depleted oe-FTO macrophages following LPS treatment. (I) The
detection of BUN and Cr in the serum of MMP9−/−
mice revealed that kidney dysfunction was exacerbated 24 h post-CLP
(n=3). (J and K) H&E staining revealed more severe renal injury
in MMP9−/− mice than in wild-type controls
following CLP surgery (n=3, original magnification, ×10). (L) qPCR
analysis revealed that MMP-9 deletion increased the expression
levels of IL-1β and IL-6 in the kidneys of CLP-treated mice (n=3).
(M) IHC staining results showing the expression of F4/80 and
reduced infiltration of F4/80+ macrophages in the kidneys of
MMP9−/− mice after CLP surgery (n=3).
*P<0.05; **P<0.01;
***P<0.001; ****P<0.0001. MMP-9, matrix
metalloproteinase 9; SAKI, sepsis-induced acute kidney injury;
qPCR, quantitative PCR; CLP, cecal ligation and puncture; PMs,
peritoneal macrophages; LPS, lipopolysaccharide; ELISA,
enzyme-linked immunosorbent assay; oe, over expression; BUN, blood
urea nitrogen; Cr, creatinine; CLP, cecal ligation and puncture;
H&E, hematoxylin and eosin; WT, wild-type.

Figure 6

FTO overexpression ameliorated SAKI
and improved survival in CLP mice. (A) Representative IF images
showing successful overexpression of FTO in a murine model and the
expression of F4/80 (n=3). (B) Survival curve using the log-rank
test revealed a significant reduction in sepsis mortality in
FTO-overexpressing mice compared with vector control mice (n=8;
P=0.0022 <0.01). (C and D) H&E staining of kidney sections
revealed that FTO overexpression mitigated CLP-induced renal injury
(n=3). (E) Overexpression of FTO reduced the CLP-induced increase
in serum BUN levels (n=3). (F) ELISAs revealed that FTO
overexpression reduced serum IL-6 and TNF-α production (n=3). (G)
An IHC assay revealed increased MMP-9 abundance in the kidneys of
FTO-overexpressing mice after CLP surgery (n=3, original
magnification, ×20). *P<0.05 was considered to
indicate statistical significance; **P<0.01;
***P<0.001. FTO, fat mass and obesity-associated
protein; SAKI, sepsis-induced acute kidney injury; CLP, cecal
ligation and puncture; IF, immunofluorescence; H&E, hematoxylin
and eosin staining; BUN, blood urea nitrogen; ELISA, enzyme-linked
immunosorbent assay; IHC, immunohistochemistry.

Figure 7

Graphical abstract generated by
Figdraw 2.0 (https://www.figdraw.com/static/index.html#/)
highlights the present study. FTO, fat mass and obesity-associated
protein; AAV, adeno-associated virus; IL-6, interleukin-6; TNF-α,
tumor necrosis factor-α; MMP-9, matrix metalloproteinase 9;
m6A, N6-methyladenosine.
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Copy and paste a formatted citation
Spandidos Publications style
Chen X, Sun Z, Chen J, Zhang J, Liu Z, Yan Z, Li Q and Chen Z: Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes. Int J Mol Med 57: 18, 2026.
APA
Chen, X., Sun, Z., Chen, J., Zhang, J., Liu, Z., Yan, Z. ... Chen, Z. (2026). Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes. International Journal of Molecular Medicine, 57, 18. https://doi.org/10.3892/ijmm.2025.5689
MLA
Chen, X., Sun, Z., Chen, J., Zhang, J., Liu, Z., Yan, Z., Li, Q., Chen, Z."Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes". International Journal of Molecular Medicine 57.1 (2026): 18.
Chicago
Chen, X., Sun, Z., Chen, J., Zhang, J., Liu, Z., Yan, Z., Li, Q., Chen, Z."Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes". International Journal of Molecular Medicine 57, no. 1 (2026): 18. https://doi.org/10.3892/ijmm.2025.5689
Copy and paste a formatted citation
x
Spandidos Publications style
Chen X, Sun Z, Chen J, Zhang J, Liu Z, Yan Z, Li Q and Chen Z: Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes. Int J Mol Med 57: 18, 2026.
APA
Chen, X., Sun, Z., Chen, J., Zhang, J., Liu, Z., Yan, Z. ... Chen, Z. (2026). Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes. International Journal of Molecular Medicine, 57, 18. https://doi.org/10.3892/ijmm.2025.5689
MLA
Chen, X., Sun, Z., Chen, J., Zhang, J., Liu, Z., Yan, Z., Li, Q., Chen, Z."Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes". International Journal of Molecular Medicine 57.1 (2026): 18.
Chicago
Chen, X., Sun, Z., Chen, J., Zhang, J., Liu, Z., Yan, Z., Li, Q., Chen, Z."Elevated FTO alleviates sepsis‑induced acute kidney injury by regulating macrophage inflammatory phenotypes". International Journal of Molecular Medicine 57, no. 1 (2026): 18. https://doi.org/10.3892/ijmm.2025.5689
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