Identification and Functional Analysis of Circular RNAs in Ankylosing
Spondylitis Patient Peripheral Blood Mononuclear Cells
Abstract
Objectives:To analyze the function and roles of circRNAs in the
peripheral blood mononuclear cells (PBMCs) of ankylosing spondylitis
(AS) patients. Methods: The expression of AS-related circRNAs were
detected by high-throughput RNA-sequencing within the PBMCs of 5 AS
cases and healthy controls. After profiling circRNA expression in these
samples, differentially expressed circRNAs (DECs) were identified, and a
qPCR-based validation approach was used to confirm the differential
expression of six of these DECs in patient samples. Spearman’s
correlation tests and ROC curve analyses were further used to assess the
relationship between disease severity and the expression of DECs of
interest in AS patients, after which a putative circRNA-microRNAs
(miRNAs) interaction network was constructed leading to the detection of
six validated DECs with competing endogenous RNA (ceRNA) functionality.
Besides, cell experiments were also performed to investigate the
potential mechanism of key circRNA in AS. Results: 10,441 circRNAs were
identified in these 10 PBMC samples, with 131 total DECs including 89
and 42 that were up- and down-regulated, respectively. In qPCR
validation assays, patterns of hsa_circ_0000702, hsa_circ_0006209,
hsa_circ_0047920, hsa_circ_0001543, hsa_circ_0072697 and
hsa_circ_0005076 were confirmed to align well with RNA-seq results. In
addition, the expression levels of hsa_circ_0006209,
hsa_circ_0047920, and hsa_circ_0072697 were detected to be
positively correlated with disease severity. ROC curve analyses
suggested that hsa_circ_0072697 may offer value as a diagnostic
biomarker for AS. Cell experiments indicated that hsa_circ_0072697
could suppress the progression of AS by targeting NF-κB pathway
Conclusions: The identification of six circRNAs with putative ceRNA
functionality in AS patients highlights potential molecular mechanisms
governing this debilitating disease. However, further research will be
necessary to formally confirm the roles of these DECs and their target
miRNAs in AS. Further, hsa_circ_0072697 has a good diagnostic value in
AS patients, and it could suppress the progression of AS by targeting
NF-κB pathwa