Revised: February 25, 2026
Accepted: April 28, 2026
Published online: June 27, 2026
Processing time: 219 Days and 20.3 Hours
Primary biliary cholangitis (PBC) is an autoimmune liver disease involving dysre
To investigate whether microRNAs (miRNA) modulate NK cell activation and granzyme B secretion in PBC via the nuclear factor of activated T cell (NFATC) pathway, focusing on the regulatory role of miR-137-3p.
Peripheral blood samples from 168 patients with PBC and 74 healthy controls were analyzed. NK cells were isolated, and miRNA expression was quantified via quantitative polymerase chain reaction. Flow cytometry assessed cluster of differentiation (CD) molecule expression, whereas dual luciferase assays validated miR-137-3p binding to the NFATC1 gene.
There were significantly lower Ct values (indicating higher expression) for miR-137-3p, miR-124-3p, and miR-506-3p in NK cells from patients with PBC vs cells from healthy controls (all P < 0.05). The miR-137-3p directly bound NFATC1, enhancing granzyme B secretion (348.59 ± 7.47 pg/mL vs 373.92 ± 15.50 pg/mL; P < 0.05) and increasing the number of CD16+ NK cells (73.2% vs 46.8; P < 0.05) compared to controls.
Exosomal miR-137-3p promotes NK cell activation in PBC by targeting NFATC1, driving CD16 expression and granzyme B secretion. These miRNAs, particularly miR-137-3p, may serve as novel diagnostic biomarkers or therapeutic targets for PBC.
Core Tip: This study identifies elevated exosomal microRNA-137-3p (miR-137-3p) in primary biliary cholangitis (PBC) plasma as a key driver of natural killer cell hyperactivation. Mechanistically, miR-137-3p directly targets and inhibits nuclear factor of activated T cell 1, leading to upregulation of the activation marker cluster of differentiation 16 and enhanced secretion of the cytotoxic molecule granzyme B. These findings define miR-137-3p as a novel diagnostic biomarker and suggest that targeting the miR-137-3p/nuclear factor of activated T cell 1 axis could be a promising therapeutic strategy for PBC by alleviating natural killer cell-mediated bile duct injury.
- Citation: Yang YF, Liu QX, Su WH, Zang B, Liu BQ, Zhao CY, Han YB, Liu LW, Leung PSC, Liu B. Plasma exosomal miR-137 activates natural killer cells in primary biliary cholangitis by targeting NFATC1. World J Hepatol 2026; 18(6): 116738
- URL: https://www.wjgnet.com/1948-5182/full/v18/i6/116738.htm
- DOI: https://dx.doi.org/10.4254/wjh.116738
Primary biliary cholangitis (PBC) is a female-predominant chronic autoimmune liver disease characterized by cholestasis, a high titer of anti-mitochondrial antibodies (AMAs), lymphocyte infiltration in the portal vein area, and destruction of small bile duct epithelial cells, leading to intrahepatic cholestasis and ultimately, liver cirrhosis[1,2]. PBC is caused by a combination of genetic, inflammatory, and environmental factors[3].
Natural killer (NK) cells represent > 40% of the intrahepatic lymphocytes and play an important role in the progression of PBC due to their adaptive immune characteristics[4,5]. Among patients with PBC, the number of NK cells increases and accumulates around the affected small bile ducts, inducing apoptosis of biliary epithelial cells (BECs). Immunogenetic studies have shown that polymorphisms in the nuclear factor of activated T cells (NFAT) gene family and interleukin-12/signal transducer and activator of transcription 4 pathways are associated with PBC[6]. NFATC1, also called NFATC or NFAT2, is a gene that induces immune responses and regulates cells by binding to the cluster of differentiation 16 (CD16) molecule on the surface of NK cells. As early as 1995, Aramburu et al[7] demonstrated that NFAT pre-existing (NFATC2/NFAT1) regulates the transcription of several cytokines and mediates CD16-induced activation of cytokine genes in human NK cells. A recent study showed that blocking the NFAT pathway had a relieving effect on intrahepatic cholestasis[8]. NFAT signaling is involved in the regulating development of the immune system and adaptive immune responses[9]. In NK cells, lactic acid or hydrochloric acid inhibits upregulation of NFAT activation, and data suggest that inhibition of NFAT is the main reason for the inhibitory effect of NK cell activation[10-12]. Studies have also indicated the significant role of NFATC1 in regulating immune tolerance and autoimmunity[13,14]. Modulating NFATC1 gene expression, which regulates the NFAT pathway, is pivotal for NK cell regulation.
The microRNAs (miRNAs) are short noncoding, single-stranded RNAs involved in RNA silencing and post-transcriptional regulation of gene expression. Various miRNAs have been implicated in the development of PBC[15,16]. From its intracellular origin, miRNA can be secreted in cell-derived extracellular vesicles/exosomes and serve as personalized signatures reflecting the disease status or perform cell-to-cell communication[17,18]. There is growing evidence that miRNAs can be used as novel biomarkers for diagnosis and monitoring of the treatment of diseases such as cancer, neurodegenerative diseases, heart disease, and infections[19-22].
Differentially expressed miRNAs in the serum and peripheral blood mononuclear cells have been reported in patients with PBC, indicating the potential application of profile analysis in the diagnosis and evaluation of treatment efficacy in PBC[23]. Compared with healthy controls (HCs), the levels of miR-21 and miR-210 in the liver tissue of patients with PBC were elevated, and 35 miRNAs were found to be differentially expressed in patients with end-stage PBC compared with HCs. Another study confirmed that upregulation of miR-122a and miR-26a, and downregulation of miR-328 and miR-299-5p in the liver tissue of patients with PBC were closely related to the pathogenesis of PBC, including biliary inflammation, apoptosis, and reactive oxygen species[20,23,24]. More than 400 miRNAs and 300 miRNAs have been found in human and mouse NK cells, respectively; some of which play important roles in the development and maturation of NK cells. Killer immunoglobulin-like receptors (KIR), killer lectin-like receptors (KLR), and natural killer group 2D (NKG2D) are common genes related to NK cell activation, but no miRNA correlation has been found in the gene prediction process[25].
In this study, we examined NFAT signaling and the role of miRNA in NK cell activation in the pathogenesis of PBC.
There were 168 inpatients with PBC and outpatients at Affiliated Hospital of Qingdao University (Qingdao, Shandong Province, China), and 74 HCs from a health screening center. Whole blood and plasma were collected from patients with PBC and HCs, and the clinical information of all enrolled individuals was recorded. All patients met the European Association for the Study of the Liver clinical practice guidelines for the diagnosis of PBC[26]. The inclusion criteria were: (1) Evidence of serum biochemical indicators, such as elevated alkaline phosphatase or gamma glutamyl transferase without a clear cause; (2) Positive serum antimitochondrial antibodies (AMA), including the AMA-M2 subtype, or positive anti-gp210/sp100 antibodies; and (3) Nonsuppurative or destructive cholangitis in liver tissue, without lobular bile duct damage. If two of the three tests were positive and the patient did not have hepatitis virus or human immunodeficiency virus infections, autoimmune liver diseases or fatty liver disease, they were excluded. Samples were obtained with written consent of all participants. The study protocol was conducted in accordance with the Declaration of Helsinki[27]. This study was approved by the Ethics Committee of Affiliated Hospital of Qingdao University (No. QYFY
Exosomes were obtained from plasma samples from 32 patients with PBC and 29 HCs by serial differential centrifugation, as previously described[28]. Plasma (1.5 mL) was centrifuged at 300 g for 10 minutes, and the supernatant was collected and centrifuged again at 2000 g for 10 minutes and 10000 g for 30 minutes. Thereafter, the supernatant was subjected to ultracentrifugation at 100000 g for 70 minutes at 4 °C to generate a crude exosome precipitate (containing a small amount of tramp protein). The pellet was resuspended in 200 mL phosphate-buffered saline (PBS) and centrifuged again at 100000 g for 70 minutes at 4 °C to obtain pure exosomes.
The human miRNAs associated with NFATC1 were predicted by using TargetScane, miRDB, miRactDB, and mitarBase. The miRNAs associated with KIR, KLR, NKG2D, and NFATC1, namely miR-137-3p, miR-124-3p, miR-506-3p, miR-331-3p, miR-338-3p, miR-193a-3p, miR-485-5p, miR-24-3p, miR-143-3p, and miR-218-3p, were selected. Figure 1 shows the predicted miRNA overlap of the four sites, and Table 1 shows the screened miRNA sequences.
| MiRNA | Sequences |
| U6 | 5’-CTCGCTTCGGCAGCACA-3’ |
| Hsa-miR-124-3p | 5’-UAAGGCACGCGGUGAAUGCCAA-3’ |
| Hsa-miR-506-3p | 5’-GCCACCACCATCAGCCATAC-3’ |
| Hsa-miR-137-3p | 5’-UUAUUGCUUAAGAAUACGCGUAG-3’ |
| Hsa-miR-331-3p | 5’-GCCCCUGGGCCUAUCCUAGAA-3’ |
| Hsa-miR-338-3p | 5’-UCCAGCAUCAGUGAUUUUGUUG-3’ |
| Hsa-miR-193a-3p | 5’-AACUGGCCUACAAAGUCCCAGU-3’ |
| Hsa-miR-485-5p | 5’-AGAGGCUGGCCGUGAUGAAUUC-3’ |
| Hsa-miR-24-3p | 5’-UGGCUCAGUUCAGCAGGAACAG-3’ |
| Hsa-miR-143-3p | 5’-UGAGAUGAAGCACUGUAGCUC-3’ |
| Hsa-miR-218-1-3p | 5’-AUGGUUCCGUCAAGCACCAUGG-3’ |
Total RNA from NK cells and exosomes was extracted using AG RNAex Pro Reagent (No. AG21101; AccurateBio, Hunan Province, China). RNA was reverse transcribed with primer initiation according to the instructions of the miRNA cDNA First Strand Synthesis Kit, followed by quantitative polymerase chain reaction (qPCR) using the SYBR Green Pro Taq HS Kit and the ABI PRISM 7500 FAST Real Time PCR System. The relative expression of the target genes was analyzed using the 2-ΔΔCt method with U6 as an internal reference.
A fragment (5’-CTCGCTTCGGCAGCACA-3’) of the NFATC1 3’-untranslated region (UTR) was synthesized and introduced into the psiCHECK-2 vector (Promega, Madison, WI, United States). Mutation sites were designed on the wild-type (WT) gene for NFATC1 with complementary sequences to the seed sequence. Subsequently, luciferase reporter plasmids of NFATC1 3’ UTR WT and NFATC1 3’ UTR mutant were obtained. Plasmids and miR-137 were co-transfected with miR-137 mimic and mock-negative control (NC), respectively, into 4 × 105 HEK-293T cells (a commonly used human embryonic kidney cell line for molecular biology research, favored for its high transfection efficiency and protein expression). Cells were lysed, and luciferase activity was measured using the Dual Luciferase Reporter Kit (No. 11401ES60; Yeasen Biotechnology, Shanghai, China) 48 hours after transfection.
Peripheral blood was collected using EDTA-containing anticoagulation tubes. Peripheral blood monocytes were obtained by Ficoll Hypaque centrifugation. The buffy coat layer containing the lymphocytes was harvested. A sample was taken, stained with an equal volume of 0.4% Trypan blue, and the number of monocytes was measured using a hemocytometer (8 × 106 to 8 × 107). Lymphocytes were incubated with Biotin-Antibody and MicroBead Cocktail (No. 130-092-661; Miltenyi Biotec, Germany) and further purified using the LS column (miltenyibiotec.com).
NK cells were seeded in culture solution (No. CM-0533; PricellaBio, Wuhan, Hubei Province, China) in 24-well plates and incubated at 37 °C in 5% CO2, until confluence reached 60%-80%. Cells were transfected with miRNA mimic miR10000430 and miR20000435 (RiboBio, Guangzhou, Guangdong Province, China). The transfection efficiency was verified by qPCR.
Sandwich enzyme-linked immunosorbent assay was performed on 96-well plates to detect perforin (No. E-EL-H1123; Elabscience, Houston, TX, United States), granzyme B (No. E-EL-H1617; Elabscience), tumor necrosis factor alpha (TNF-α) (No. E-EL-H0109; Elabscience), and interferon gamma (IFN-γ) (No. E-EL-H0108; Elabscience) in NK cell cultures. Standards were diluted in sample dilution buffer to generate a standard curve. The blocking buffer was removed and the standards/samples were loaded in duplicate in the wells. Plates were incubated at 37 °C for 2 hours and washed five times. To detect captured antigen, plates were incubated with biotinylated secondary antibody for 1 hour at 37 °C and washed five times. Bound antibody was detected by incubating with anti-biotin-horseradish peroxidase at 37 °C for 30 minutes. After washing, the plates were developed for 10 minutes in the dark at room temperature and the reaction was stopped using the termination solution.
Total exosome protein was extracted using high performance RIPA lysis buffer. Protein concentration was determined using a BCA kit (No. E112-01/02; Vazyme, Nanjing, China). Proteins (5 mL) were resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and electrotransferred onto polyvinylidene fluoride membranes. The membranes were blocked in 5% bovine serum albumin for 1 hour at room temperature and probed overnight at 4 °C with the following: (1) Rabbit antibodies to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (No. GB15004; Servicebio, Hubei, China); (2) CD9 (No. GB115697; Servicebio); (3) CD81 (No. GB111073; Servicebio); and (4) tumor susceptibility gene 101 (TSG101) (No. GB15619; Servicebio). The membranes were washed three times (5 minutes each) with Tris-buffered saline/Tween-20 and incubated with horseradish peroxidase-labeled goat anti-rabbit immunoglobulin G for
Flow cytometry was used to determine the purity (> 95%) and number of NK cells (4 × 104 to 4 × 106). NK cells were isolated from 15 independent donors per group (PBC and HC). Flow cytometry analysis was performed with technical duplicates for each sample. NK cells were blocked in 4% paraformaldehyde (No. 60536ES; Yeasen) and incubated with anti-CD3/fluorescein isothiocyanate (No. E-AB-F1013C; Elabscience), anti-CD56/APC (No. E-AB-F1239E; Elabscience), and anti-CD16/PE (No. E-AB-F1236D; Elabscience) at 4 °C for 30 minutes. PBS was added to wash away unbound antibodies; the cells were centrifuged to obtain stained cells and resuspended in 300 μL PBS. Cells were subjected to flow cytometry and analyzed using FlowJo software (Ashland, OR, United States).
Non-normally distributed data were described using medians and quartiles, and normally distributed data were described using the mean ± SD. The Kolmogorov-Smirnov test was used for one-way non-normal distributions. All data are expressed as the mean ± SD of at least two biological replicate series. Differences between two groups were analyzed using SPSS 22.0 (IBM SPSS Statistics, Armonk, NY, United States) and plotted using Graphpad Prism (GraphPad Software, Inc., San Diego, CA, United States). P < 0.05 was considered statistically significant.
RESULTS
Evaluation of the isolated exosomes by transmission electron microscopy (Figure 2A) showed spherical vesicles in all samples with diameters of 80-160 nm measured by nanoparticle tracking analysis (Figure 2B). Three commonly used exosomal protein markers, namely CD9, CD81 and TSG101, were examined by western blotting. With the same amount of protein loading in each lane, all three exosomal protein markers were highly enriched in isolated exosomes (Figure 2C). The miRNA in plasma exosomes isolated from 32 patients with PBC and 45 HCs were quantified by qPCR to verify whether there were differences in miRNA in plasma exosomes. Expression of miR-137-3p (P = 0.0020), miR-124-3p (P = 0.0077), and miR-506-3p (P = 0.0086), miR-338-3p (P = 0.0053), and miR-24-3p (P = 0.0037) was significantly altered in exosomes derived from the plasma of patients with PBC compared to those from HCs (Figure 3).
Ten human miRNAs associated with the NFATC1 gene were screened by gene prediction, and tested by qPCR in NK cells. Patients with PBC (n = 37) and HCs (n = 37) were grouped according to sex and similar age. The miR-137 (P = 0.0053), miR-124 (P = 0.0034), and miR-506 (P = 0.0010) differed significantly between patients with PBC and HCs by qPCR. Ct values of miR-137-3p (11.33 ± 4.40 vs 11.85 ± 4.40; P = 0.0350), miR-124-3p (8.61 ± 2.22 vs 8.94 ± 2.99; P = 0.0369) and miR-506-3p 7.18 ± 1.89 vs 8.29 ± 1.46; P = 0.0010) were higher in patients with PBC than in HCs, whereas other miRNAs did not differ significantly between the two groups (P > 0.05). This indicated that the difference in the expression of these three miRNAs by NK cells in patients with PBC and HCs was significant (Figure 4).
To determine whether any differential miRNAs were associated with NFATC1, we predicted the binding sites of hsa-miR-137-3p, hsa-miR-124-3p, and hsa-miR-506-3p to NFATC1 followed by a dual luciferase assay in HEK-293T cells. HEK-293T cells carrying either WT and 3’ UTR mutant NFATC1 in luciferase reporting plasmid were transfected with miRNA mimic or miR-137-3p NC. The fluorescence intensity by firefly luciferase was measured and corrected by sea kidney luciferase. The fluorescence intensity of miR-137-3p bound to NFATC1 WT was less than that of miR-137-3p NC bound to NFATC1 WT (P < 0.05), while miR-124-3p and miR-506-3p had no significant effect on WT, indicating that miR-137-3p bound to NFATC1 (Figure 5).
Overexpression plasmid mimics of miR-137-3p, miR-124-3p, and miR-506-3p were constructed and transfected into NK cells from 64 patients with PBC. The levels of perforin, granzyme B, TNF-α, and IFN-γ in the culture medium were determined after transfection. Overexpression of miR-137- 3p increased granzyme B secretion by NK cells compared to miR-137-3p-NC (348.27 [217.62-354.75] pg/mL vs 336.47 [214.38–344.33] pg/mL; P = 0.035), while there was no significant effect on perforin, TNF-α, or IFN-γ (P > 0.05). Overexpression of miR-137-3p significantly enhanced granzyme B secretion by NK cells compared to the NC group (P = 0.036). By contrast, overexpression of miR-124-3p or miR-506-3p did not significantly alter the secretion of perforin, granzyme B, TNF-α, or IFN-γ (all P > 0.05; Table 2, Figure 6).
| Item | Mimic | Negative control | P value | Analysis of variance |
| Perforin (pg/mL) | ||||
| MiR-124 | 1133.07 (260.53, 3044.59) | 1097.67 (210.53, 3101.26) | 0.820 | |
| MiR-137 | 1016.45 (497.13, 1263.06) | 930.28 (480.20, 1097.03) | 0.754 | 0.503 |
| MiR-506 | 3125.92 (2368.10, 4089.81) | 4890.15 (4289.15, 5913.06) | 0.065 | |
| Granzyme B (pg/mL) | ||||
| MiR-124 | 341.14 (113.09, 350.63) | 340.93 (140.95, 350.32) | 0.164 | |
| MiR-137 | 347.55 (206.95, 357.04) | 336.47 (196.57, 342.71) | 0.036 | P = 0.037 |
| MiR-506 | 206.945 (197.61, 224.89) | 216.195 (199.47, 245.05) | 0.579 | |
| Interferon gamma (pg/mL) | ||||
| MiR-124 | 13.61 (10.29, 14.97) | 13.29 (12.28,14.10) | 0.833 | |
| MiR-137 | 13.58 (12.97, 17.45) | 13.64 (12.86, 14.03) | 0.825 | 0.819 |
| MiR-506 | 15.405 (13.64, 16.43) | 16.265 (1578, 16.43) | 0.833 | |
| Tumor necrosis factor alpha (pg/mL) | ||||
| MiR-124 | 9.52 (7.09, 11.87) | 7.39 (5.28, 8.99) | 0.208 | |
| MiR-137 | 9.41 (4.36, 10.00) | 10.35 (8.89, 10.72) | 0.171 | 0.856 |
| MiR-506 | 10.33 (9.41, 10.82) | 10.49 (10.07, 10.73) | 0.753 | |
NFATC1 bound to CD molecules on the surface of NK cells. miR-137 bound to NFATC1. To determine whether miR-137 had any effect on expression of other surface markers, NK cells were transfected with a miR-137 mimic overexpression vector. CD16 and CD56 on the surface of NK cells were measured using flow cytometry. Compared with the miR-137-3p NC group, the proportion of CD3-CD16+ NK cells was significantly increased after overexpression of miR-137-3p (73.2% vs 46.8%; P < 0.05). Conversely, the proportion of CD3-CD56+ NK cells was decreased upon miR-137-3p overexpression. This indicated that the binding of miR-137-3p to NFATC1 reduced the binding of CD16 molecules on the surface of NK cells, and the free state of NK cells was reduced. We also found that overexpression of miR-137 and NFATC1 reduced the binding of CD16 molecules on the surface of NK cells and increased CD16+ on the surface of NK cells. Overexpression of miR-137 reduced CD3-CD56+ NK cells (Figure 7).
The identification of pathogenic mechanisms in PBC remains a critical research gap. Our study provides novel evidence that miR-137 from exosomes increases the proportion of CD16+ NK cells by targeting NFATC1, thereby promoting secretion of granzyme B. Transfection with miR-137 mimics enhanced granzyme B secretion in NK cells. Mechanistically, miR-137 directly binds to NFATC1 mRNA, upregulating CD16 surface expression on NK cells (Figure 8). This finding positions miR-137 as a key regulator of the NFAT-CD16 axis, highlighting its potential role in PBC immunopathology. The increase of CD16+ NK cells may activate downstream effects (such as granzyme B) through Fc gamma receptor III (FcγRIII) signaling, but the specific signaling axis needs verification[29,30].
While our study definitively establishes that the analyzed exosomes were isolated from the total plasma pool, a pertinent question arises regarding their cellular origins. The significant upregulation of specific miRNAs, particularly miR-137-3p, which has established roles in NK cell function, invites the hypothesis that a fraction of these plasma exosomes may be partially derived from NK cells. This is a plausible speculation, as activated immune cells, including NK cells, are known to actively release exosomes that reflect their functional state and molecular cargo. The observed functional outcomes – specifically, the enhancement of NK cell cytotoxicity following exposure to these PBC plasma exosomes – could be consistent with a model where exosomes originating from activated NK cells contribute to a positive feedback loop, further amplifying the hyperactivation of NK cells in a paracrine manner. However, it is crucial to emphasize that the plasma exosome population is heterogeneous, and contributions from other cell types implicated in PBC pathogenesis, such as BECs or T cells, cannot be ruled out. Future studies using techniques such as immunoprecipitation of exosomes using cell-specific surface markers (e.g., anti-CD56 for NK cells) are necessary to unequivocally delineate the precise cellular sources of these pathogenic exosomes and their relative contributions to disease progression.
Ultimately, the pathological hallmark of PBC is the destruction of small bile ducts mediated by apoptosis of BECs. Our findings, which demonstrate that exosomal miR-137-3p enhances NK cell cytotoxicity via the NFATC1/CD16/granzyme B axis, provide a potential mechanistic link between circulating factors and intrahepatic damage. We hypothesize that the hyperactivated NK cells, upon infiltration into the liver, may directly contribute to BEC injury through release of granzyme B and other cytotoxic molecules. Future studies investigating the homing capability of these activated NK cells to the bile ducts and their direct cytotoxic effects on BECs in vitro and in vivo will be crucial to validate this proposed pathway.
Comparative analysis revealed distinct miRNA expression profiles between NK cells and circulating exosomes. While miR-124 demonstrated differential expression in exosomes from both patients with PBC and HCs, its overexpression via mimics failed to alter NK cell activation markers (CD16/CD56) in our experimental system. This contrasts with reports linking hypoxic miR-124 silencing to NFATC1-mediated cartilage formation and MALAT1/miR-124/NFATC1 signaling in osteoclastogenesis[31], suggesting tissue-specific regulatory roles. miR-506 showed significant upregulation in PBC NK cells, corroborating prior studies implicating miR-506 in cholangiocyte dysfunction and immune activation[21,32]. These divergent behaviors imply alternative regulatory pathways for miR-124/miR-506 in NK cells, warranting further investigation.
The CD16 (FcγRIII) receptor, a 50–70-kDa low-affinity immunoglobulin G receptor, exists as two isoforms: (1) Transmembrane FcγRIIIA on NK cells/macrophages; and (2) Glycosylphosphatidylinositol-anchored FcγRIIIB on neutrophils[30]. Our data extend current understanding by demonstrating that NFATC1-mediated CD16 upregulation enhances NK cell cytotoxicity through granzyme B mobilization. This aligns with the established role of CD3ζ chain phosphorylation in NFAT pathway activation[8,33]. Crucially, NFATC1 autoregulation in peripheral lymphocytes[34-36] may explain sustained CD16 expression post-activation, creating a feed-forward loop in PBC pathogenesis.
NK cell functional impairment in PBC involves complex interactions between apoptotic (FasL/Fas) and cytotoxic (perforin/granzyme) mechanisms, with calcium signaling acting as a critical convergence point[37]. Our findings complement this by revealing miR-137 as an upstream regulator of granzyme B release via CD16. The observed synergy between IFN-γ and CD16 in promoting granzyme B secretion[38] suggests potential therapeutic combinations for PBC management.
The overexpression of miR-137-3p, miR-124-3p, and miR-506-3p in the NK cells of patients with PBC (vs HCs) positions these miRNAs as potential diagnostic biomarkers. In particular, the regulatory effect of miR-137 on the NFAT-CD16 axis presents a promising therapeutic target. However, the focus of the current study on PBC-derived NK cells limits comparative analyses with HCs. Future investigations should incorporate HC-derived cells and explore synergistic effects with existing therapies.
This work establishes miR-137 as a novel modulator of NK cell cytotoxicity in PBC through NFATC1-mediated CD16 regulation. While miR-124 and miR-506 exhibit disease-associated expression patterns, their functional roles require further elucidation. Pharmacological inhibition of miR-137 may represent a viable strategy to attenuate NK cell hyperactivation and improve clinical outcomes in PBC.
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