Activated cGAS-STING signaling promotes malignancy in metabolic dysfunction-associated fatty liver disease via mitochondrial DNA and immune cell dysfunction

Table 1 Serum biochemistry parameters and hepatocellular carcinoma markers during metabolic dysfunction-associated fatty liver disease progression, mean ± SD

Group	NC (n = 12)	MAFLD (n = 12)	MASH (n = 17)	LC (n = 15)	HCC (n = 10)
ALT (U/L)	11.5 ± 2.0	48.9 ± 12.8c	53.6 ± 10.1c	51.4 ± 16.1c	52.5 ± 18.4c
AST (U/L)	12.3 ± 2.8	42.5 ± 13.6c	45.6 ± 11.2c	53.6 ± 12.4c	56.1 ± 16.8c
TG (mmol/L)	0.58 ± 0.53	1.92 ± 0.36c	1.98 ± 0.58c	1.89 ± 0.68c	1.57 ± 0.85b
Tch (mmol/L)	1.38 ± 0.39	2.92 ± 0.89c	3.97 ± 0.82c	3.69 ± 0.61c	3.62 ± 0.80c
HDL (mmol/L)	2.20 ± 0.42	0.67 ± 0.20c	0.65 ± 0.09c	0.75 ± 0.09c	0.63 ± 0.05c
LDL (mmol/L)	0.31 ± 0.08	1.64 ± 0.54c	1.25 ± 0.1c	0.99 ± 0.25c	0.89 ± 0.68b
AFP (μg/L)	0.85 ± 0.11	1.12 ± 0.31	1.25 ± 0.32a	1.58 ± 0.51b	1.96 ± 0.75c
Wnt3a (μg/L)	1.21 ± 0.23	1.45 ± 1.68	2.25 ± 2.24	6.28 ± 2.12c	9.98 ± 5.62c

^aP < 0.05 vs the normal control (NC) group.

^bP < 0.01 vs the NC group.

^cP < 0.001 vs the NC group.

NC: Normal control; MAFLD: Metabolic dysfunction-associated fatty liver disease; MASH: Metabolic dysfunction-associated steatohepatitis; LC: Liver cirrhosis; HCC: Hepatocellular carcinoma; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; TG: Triglycerides; Tch: Total cholesterol; HDL: High-density lipoprotein; LDL: Low-density lipoprotein; AFP: Alpha-fetoprotein; Wnt3a: Key signaling molecule 3a of the Wnt pathway.

Table 2 Dynamic changes in liver carnitine palmitoyl transferase-II specific concentration and activity, mean ± SD

Group	n	Average AIHC (× 100)	Liver CPT-II (ng/mg protein)	Serum CPT-II (U/L)
NC	12	6.93 ± 1.26	124.18 ± 6.58	5.86 ± 2.51
MAFLD	12	2.98 ± 0.83c	81.24 ± 5.86c	4.12 ± 1.06a
MASH	17	0.95 ± 0.78c	43.68 ± 6.91c	2.61 ± 0.80b
LC	15	0.91 ± 0.48c	40.12 ± 5.58c	2.18 ± 0.57c
HCC	10	0.86 ± 0.72c	36.42 ± 6.88c	1.98 ± 0.47c
F value	-	122.599	431.501	20.299
P value	-	< 0.001	< 0.001	< 0.001

^aP < 0.05 vs the normal control (NC) group.

^bP < 0.01 vs the NC group.

^cP < 0.001 vs the NC group.

CPT-II: Carnitine palmitoyl transferase-II; A_IHC: Average absorbance of liver carnitine palmitoyl transferase-II by immunohistochemistry; NC: Normal control; MAFLD: Metabolic dysfunction-associated fatty liver disease; MASH: Metabolic dysfunction-associated steatohepatitis; LC: Liver cirrhosis; HCC: Hepatocellular carcinoma.

Table 3 Dynamic stimulator of interferon genes immunofluorescence intensity during metabolic dysfunction-associated fatty liver disease malignancy

Group	n	Hepatic STING fluorescence intensity
		-	+	++	+++
NC	12	12	0	0	0
MAFLD	12	6	6	0	0
MASH	17	0	13	4	0
LC	15	0	5	9	1
HCC	10	0	0	1	9

P < 0.001, H = 62.991. NC: Normal control; MAFLD: Metabolic dysfunction-associated fatty liver disease; MASH: Metabolic dysfunction-associated steatohepatitis; LC: Liver cirrhosis; HCC: Hepatocellular carcinoma; STING: Stimulator of interferon genes.

Table 4 Dynamic changes in circulating stimulator of interferon genes levels and inflammatory mediator levels, mean ± SD

Group	n	STING (μg/L)	IFN-I (μg/L)	NF-κB (μg/L)	TNF-α (μg/L)	TGF-β1 (μg/L)
NC	12	5.8 ± 2.8	50.8 ± 3.1	4.3 ± 2.6	85.4 ± 21.8	6.5 ± 2.3
MAFLD	12	11.2 ± 8.2a	121.2 ± 28.6c	11.3 ± 2.8c	158.0 ± 12.8c	9.8 ± 3.2b
MASH	17	26.8 ± 12.9c	195.9 ± 36.5c	17.8 ± 5.4c	228.2±1 0.7c	13.2 ± 6.4b
LC	15	35.5 ± 14.2c	219.2 ±54.8c	31.8 ± 10.4c	301.2 ± 30.8c	15.6 ± 9.8b
HCC	10	108.6 ± 28.1c	328.4 ±89.8c	93.1 ± 25.3c	486.5 ± 61.5c	28.6 ± 21.6c
F value		28.198	93.771	1060.999	280.899	14.602
P value		< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

^aP < 0.05 vs the normal control (NC) group.

^bP < 0.01 vs the NC group.

^cP < 0.001 vs the NC group.

STING: Stimulator of interferon genes; IFN-I: Type-I interferon; NF-κB: Nuclear factor-κB; TNF-α: Tumor necrosis factor-α; TGF-β1: Transforming growth factor-β1; NC: Normal control; MAFLD: Metabolic dysfunction-associated fatty liver disease; MASH: Metabolic dysfunction-associated steatohepatitis; LC: Liver cirrhosis; HCC: Hepatocellular carcinoma.