Dataset Information

PAD2-Mediated Histone Citrullination Drives Tumor Progression by Enhancing Cell Proliferation and Modifying the Microenvironment in Pancreatic Cancer

ABSTRACT: Histone citrullination is catalyzed by peptidyl arginine deiminases (PAD) that play a role in gene regulation. However, the clinical significance and molecular mechanisms of histone citrullination and PADs in pancreatic ductal adenocarcinoma (PDAC) remain unclear. This study aimed to investigate the role and potential molecular mechanisms of PADs in PDAC. Histone citrullination was upregulated and strongly associated with PAD2 nuclear expression in human PDAC samples. PAD2 nuclear expression correlated with aggressive tumor characteristics and poor prognosis. PAD2 overexpression increased PDAC cell proliferation, whereas its knockdown had the opposite effect in vitro. PAD2 bound to the promoter regions of PRUNE1 and E2F1, resulting in the activation of their mRNA expression via upregulation of histone citrullination and chromatin accessibility at their promoter regions. PAD2 overexpression enhanced tumorigenicity, and increased PRUNE1 expression and M2 tumor-associated macrophage (M2 TAM) infiltration in subcutaneous and orthotopic PDAC models. In vitro assay showed that macrophages cultured in a conditioned medium from PAD2-expressing PDAC cells exhibited an increase of M2 TAM-related markers in a PRUNE1-dependent manner. Treatment with the PAD2 inhibitors Cl-amidine and AFM-30a selectively impaired the growth and tumorigenicity of PAD2-expressing PDAC cells in vitro and in vivo. Furthermore, the triple-high expression of nuclear PAD2, PRUNE1, and the M2 TAM marker CD206 may serve as independent adverse prognostic factors for human PDAC. Conclusively, PAD2-mediated histone citrullination drives PDAC progression by epigenetically regulating downstream target genes and influencing the tumor microenvironment. The PAD2-PRUNE1-M2 TAM axis may be a promising therapeutic target and prognostic indicator for PDAC.

ORGANISM(S): Homo sapiens

PROVIDER: GSE281497 | GEO | 2025/06/30

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Abstract: Peptidylarginine deiminases (PADs or PADIs) catalyze the conversion of positively charged arginine to neutral citrulline, which alters target protein structure and function. It is well established that a gonadotropin releasing hormone agonist (GnRHa) stimulates PAD2-catalyzed histone citrullination to epigenetically regulate gonadotropin gene expression in the gonadotrope derived LBT2 cell line. However, PADs are also found in the cytoplasm. Given this, we used mass spectrometry to identify additional non-histone proteins that are citrullinated following GnRHa stimulation and characterize the temporal dynamics of this modification. Our results show that actin and tubulin are rapidly citrullinated, which led us to hypothesize that GnRHa might induce their citrullination to modulate cytoskeletal dynamics and architecture. Data shows that 10 nM GnRHa induces citrullination of beta-actin with maximal levels occurring at 10 minutes. The level of beta-actin citrullination is reduced in the presence of the pan-PAD inhibitor bi-phenyl-benzimidazole-Cl-amidine (BB-ClA), which also prevents GnRHa induced actin reorganization in dispersed mouse gonadotrope cells. GnRHa induces citrullination of beta-tubulin with elevated levels occurring at 30 minutes; similarly to beta-actin, this response is attenuated in the presence of BB-ClA. To examine the functional consequence of beta-tubulin citrullination, we utilized fluorescently tagged end binding protein 1 (EB1-GFP) to track the growing plus end of microtubules (MT) in real time in transfected LBT2 cells. Time-lapse confocal microscopy of EB1-GFP reveals that MT average lifetime increases following 30 minutes of GnRHa treatment, but this increase is attenuated by PAD inhibition. Taken together, our data suggest that GnRHa induced citrullination alters actin reorganization and MT lifetime in gonadotrope cells. Keywords: gonadotrope, peptidylarginine deiminase, citrullination, cytoskeleton, beta tubulin, beta actin, microtubules

Project description:Purpose: Corneal endothelial cells (CECs), located in the innermost layer of cornea, are crucial for maintaining its transparency. Peptidylarginine deiminase 2 (PAD2) is an enzyme responsible for catalyzing the post-translational modification of arginine into citrulline, a process known as citrullination. In this study, we investigated the effect of AMF30a, PAD2 inhibitor, on survival and function of CECs. Methods: Cultured human CECs (hCECs) were treated with AMF30a, followed by treatments with or without AMF30a under transforming growth factor-beta (TGF-β). Cells were cultured from donor corneas and analyzed for viability (CCK-8), proliferation (BrdU assay), cytotoxicity (LDH assay), and oxidative stress (DCF-DA). Adhesion and morphology were evaluated via crystal violet staining and imaging software. Western blot and immunofluorescence identified protein expression and localization. RNA sequencing revealed differentially expressed genes, with functional enrichment via GO analysis. Results: AMF30a enhanced cell viability, proliferation, and adhesion while reducing cytotoxicity and oxidative stress. Morphological analysis revealed reduced cell size and elongation factor, indicating structural changes. AMF30a modulated the HIPPO signaling pathway by increasing YAP phosphorylation and reducing its nuclear translocation, while downregulating ERK1/2 activation. Transcriptome analysis identified differentially expressed genes (DEGs) associated with AMF30a treatment, highlighting pathways related to GPCR signaling and protein targeting. Additionally, AMF30a counteracted TGF-β-induced effects, including increased oxidative stress, citrullination, and ROCK/HIPPO signaling activation, thereby promoting cell cycle progression and reducing senescence. Conclusion: PAD2-mediated citrullination is essential for TGF-β/ROCK/HIPPO signaling pathway. AMF30a promoted the proliferation and protected against TGF-β-induced senescence in hCECs, suggesting that PAD2 plays a significant role in the survival and function of hCECs. Thus, AMF30a may be a promising therapeutic strategy for hCEC diseases.

Dataset Information

PAD2-Mediated Histone Citrullination Drives Tumor Progression by Enhancing Cell Proliferation and Modifying the Microenvironment in Pancreatic Cancer

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets