Differential expressions of exosomal miRNAs in patients with chronic heart failure and hyperuricemia: diagnostic values of miR-27a-5p and miR-139-3p

doi:10.12122/j.issn.1673-4254.2025.01.06

Abstract

Abstract:

Objective To analyze the differentially expressed exosomal miRNAs in patients with chronic heart failure (CHF) complicated by hyperuricemia (HUA) and explore their potential as novel diagnostic molecular markers and their target genes. Methods This study was conducted among 30 CHF patients with HUA (observation group) and 30 healthy volunteers (control group) enrolled between September, 2020 and September, 2023. Peripheral blood samples were collected from 6 CHF patients with HUA for analyzing exosomal miRNAs by high-throughput sequencing, and the results were validated in the remaining 24 patients using qRT-PCR. GO and KEGG enrichment analyses were performed to predict the the target genes of the identified differential miRNAs. We also validated the differentially expressed miRNAs by animal experiment. Results A total of 42 differentially expressed exosomal miRNAs were detected in observation group by high-throughput sequencing; among them, miR-27a-5p was significantly upregulated (P=0.000179), and miR-139-3p was significantly downregulated (P=0.000058). In the 24 patients with both CHF and PUA, qRT-PCR validated significant upregulation of miR-27a-5p (P=0.004) and downregulation of miR-139-3p (P=0.005) in serum exosomes. When combined, miR-27a-5p and miR-139-3p had a maximum area under the curve (AUC) of 0.899 (95% CI: 0812-0.987) for predicting CHF complicated by HUA. GO and KEGG enrichment analyses suggested that the differential expressions of miR-27a-5p and miR-139-3p was associated with the activation of the AMPK-mTOR signaling pathway to activate the autophagic response. We obtained the same conclusion from animal experiment. Conclusion Upregulated exosomal miR-27a-5p combined with downregulated exosomal miR-139-3p expression can serve as a novel molecular marker for diagnosis of CHF complicated by HUA, and their differential expression may promote autophagy in cardiomyocytes by activating the AMPK-mTOR signaling pathway.

Key words: chronic heart failure, hyperuricemia, exosomes, microRNAs, AMP-activated protein kinases, TOR serine-threonine kinases, autophagy

Zhiliang CHEN, Yonggang YANG, Xia HUANG, Yan CHENG, Yuan QU, Qiqi HENG, Yujia FU, Kewei LI, Ning GU. Differential expressions of exosomal miRNAs in patients with chronic heart failure and hyperuricemia: diagnostic values of miR-27a-5p and miR-139-3p[J]. Journal of Southern Medical University, 2025, 45(1): 43-51.

Figures/Tables 13

References 36

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Target genes	Primer	Sequences	Annealing temp (℃)	Product size (bp)	Notes
hsa-miR-27a-5p	hsa-miR-27a-5p-RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACTGCTCA	-	66	Stem ring primers
	hsa-miR-27a-5p F	CCAGCGTGAGGGCTTAGC	60		Forward primers
	microRNA_uniRev	CAGTGCAGGGTCCGAGGTAT	60		Reverse primers
hsa-miR-139-3p	hsa-miR-139-3p-RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACACTCCA	-	67	Stem ring primers
	hsa-miR-139-3p F	CCAGCGTGTGGAGACGC	60		Forward primers
	microRNA_uniRev	CAGTGCAGGGTCCGAGGTAT	60		Reverse primers
hsa-U6	hsa-U6 snRNA For	CTCGCTTCGGCAGCACATA	60	85	Forward primers
hsa-U6	hsa-U6 snRNA Rev	CGAATTTGCGTGTCATCCT	60	85	Reverse primers

Target genes	Primer	Sequences	Annealing temp (℃)	Product size (bp)	Notes
hsa-miR-27a-5p	hsa-miR-27a-5p-RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACTGCTCA	-	66	Stem ring primers
	hsa-miR-27a-5p F	CCAGCGTGAGGGCTTAGC	60		Forward primers
	microRNA_uniRev	CAGTGCAGGGTCCGAGGTAT	60		Reverse primers
hsa-miR-139-3p	hsa-miR-139-3p-RT	GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACACTCCA	-	67	Stem ring primers
	hsa-miR-139-3p F	CCAGCGTGTGGAGACGC	60		Forward primers
	microRNA_uniRev	CAGTGCAGGGTCCGAGGTAT	60		Reverse primers
hsa-U6	hsa-U6 snRNA For	CTCGCTTCGGCAGCACATA	60	85	Forward primers
hsa-U6	hsa-U6 snRNA Rev	CGAATTTGCGTGTCATCCT	60	85	Reverse primers

Characteristic	Observation group	Control group	P
Gender (%)			0.417
Male	60	70
Female	40	30
Age (year)	67.03±18.10	63.07±13.08	0.335
SUA (μmol/L)	528.36±83.48	314.74±58.76	<0.001
LVEF (%)	38.81±9.43	65.30±2.28	<0.001
ALT (U/L)	30.80±23.04	25.73±13.66	0.305
AST (U/L)	28.20±16.53	21.20±9.95	0.051
SCr (μmol/L)	96.10±22.06	65.80±13.67	<0.001
BUN (mmol/L)	7.19±1.79	5.27±1.30	<0.001
TC (mmol/L)	4.44±1.20	4.94±0.90	0.067

Characteristic	Observation group	Control group	P
Gender (%)			0.417
Male	60	70
Female	40	30
Age (year)	67.03±18.10	63.07±13.08	0.335
SUA (μmol/L)	528.36±83.48	314.74±58.76	<0.001
LVEF (%)	38.81±9.43	65.30±2.28	<0.001
ALT (U/L)	30.80±23.04	25.73±13.66	0.305
AST (U/L)	28.20±16.53	21.20±9.95	0.051
SCr (μmol/L)	96.10±22.06	65.80±13.67	<0.001
BUN (mmol/L)	7.19±1.79	5.27±1.30	<0.001
TC (mmol/L)	4.44±1.20	4.94±0.90	0.067

Upregulated miRNAs	BaseMean	Log FC	P
MiR-27a-5p	4.740923	5.8306	0.000179
MiR-193b-3p	64.58635	2.2848	0.000523
MiR-210-5p	4.917714	5.8752	0.000554
MiR-508-5p	4.249289	1.9014	0.002869