Reactive Oxygen Species (ROS) are tightly regulated in the skin and implicated in both cancer and inflammatory dermatoses, but the role of oxidative stress in cutaneous T-cell lymphoma remains largely unexplored. In this study, we aim to characterize oxidative stress and reduction-oxidation (redox) homeostasis in cutaneous T-cell lymphoma and determine whether increased ROS create a therapeutic vulnerability in malignant T cells and whether they affect neighboring keratinocytes. Lesional mycosis fungoides skin displays increased oxidative DNA and lipid membrane damage and a reduction-oxidation (redox) gene signature distinct from atopic dermatitis, characterized by SOD2 overexpression and PRDX2 repression-a rare redox profile confirmed at the protein level. Primary malignant T cells show widespread cysteine oxidation and constitutively elevated intracellular and mitochondrial ROS. Malignant T cells are selectively killed by 2,3-dimethoxy-1,4-naphthoquinone (and rescued by N-acetyl cysteine), demonstrating that they operate near their maximal antioxidant capacity. Romidepsin-induced apoptosis is similarly N-acetyl cysteine dependent. Supernatants from malignant (but not nonmalignant) T cells induce elevated ROS and recapitulate the SOD2-high/PRDX2-low signature in keratinocytes. This effect is amplified by Staphylococcus aureus activation. Taken together, we find that cutaneous T-cell lymphoma harbors a rare SOD2-high/PRDX2-low redox imbalance and that malignant T cells are sensitive to ROS induction but are able to propagate oxidative stress to keratinocytes.

Metabolic reprogramming is a hallmark of cancer, and the field has predominantly focused on investigating metabolic alterations in tumour cells. However, the relevance, mechanism and consequences of metabolic adaptations in stromal cells remain understudied. Here, we identify aspartoacylase (ASPA) as a metabolic enzyme consistently repressed in tumour stroma and cancer-associated fibroblasts (CAFs). Importantly, we report a reciprocal crosstalk between ASPA and Transforming Growth Factor Beta (TGFβ) signalling that influences fibroblast behaviour. TGFβ suppresses ASPA expression in fibroblasts, whereas ASPA restrains TGFβ-dependent myofibroblast conversion, extracelullar cell matrix (ECM) remodelling, angiogenesis and pro-tumoral macrophage phenotypes. Analyses of human specimens revealed a strong negative prognostic value for ASPA in different tumour types, associated with TGFβ signalling levels and the generation of aggressive pro-tumoral responses. Our findings unveil ASPA expression in fibroblasts as a gatekeeper of TGFβ responses and activation in cancer progression.

Inflammation can affect many diseases. We report here that inflammatory cytokines invoke caspase-8-mediated cleavage of the autophagy adaptor p62/SQSTM1 at aspartic acid 329 in human cells, producing a previously described truncated form, which we term tr-p62. We show that TNF-driven cell death is tr-p62 dependent and that autophagy inhibition promotes death via tr-p62 accumulation. Mechanistically, p62 cleavage is receptor-interacting serine/threonine-protein kinase 1 (RIPK1) dependent, and tr-p62 stabilizes caspase-8 activating complex-IIb. tr-p62-driven cell death downstream of TNF is also RIPK1 and caspase dependent, promoting feedforward caspase-8 activation. p62 cleavage does not, however, affect necroptosis. Surprisingly, this caspase-8 cleavage site in p62 is absent in mice, and introduction of cleavable forms of p62 into mouse cells causes sensitization to TNF-induced death. Moreover, mice with CRISPR-Cas9-generated cleavable p62 exhibit TNF hypersensitivity and intestinal inflammation in vivo. These findings provide significant insights into TNF-induced cell death and introduce a mouse model that may provide better clarity for human-related studies of inflammatory disease.

Abstract Cancer-associated fibroblasts (CAFs) are an abundant cell population of the breast tumor microenvironment (TME). Several CAF phenotypes exist in tumors, being the inflammatory type (iCAF) characterized by the high expression of interleukin 6 (IL-6). Hypoxia promotes the iCAF phenotype and is linked to poor prognosis of breast cancer patients. However, the molecular mechanism driving the iCAF program upon hypoxia exposure remains elusive. We analyzed the changes in the proteome and secretome of patient-derived CAFs exposed to hypoxia and identified a Leucine Rich Repeat Containing Protein (LRRC) as one of the most up-regulated proteins. Further analyses revealed that LRRC in induced by hypoxia at transcriptional level in a HIF1α-dependent manner. Histological analyses show that LRRC is uniquely expressed in CAFs across species, in both human and murine breast tumors. Stromal specificity of LRRC was further confirmed by single-cell RNA sequencing of breast cancer patients. Through loss-of-function approaches in CAFs we uncovered that LRRC is an upstream regulator of IL-6, a key driver of pathological angiogenesis and inflammation in cancer. Notably, LRRC+ CAFs activate STAT signaling in cancer and TME cells. Supporting similar roles in tumors, increased STAT phosphorylation was also observed in tumors of breast cancer patients with high expression of LRRC, in proteomics data from TCGA. Thus, by regulating IL-6, LRRC may function as a driver of pathological angiogenesis and inflammation in breast cancer. Indeed, we observed that LRRC promotes endothelial sprouting angiogenesis and an inflammatory transcriptional program in CAFs and TME cells. Suggesting tumor-promoting roles of LRRC, high levels of LRRC correlate with worsened survival of breast cancer patients in TCGA datasets. To determine the influence of LRRC+ CAFs on the surrounding TME, we are utilizing spatial proteomic analysis of breast cancer patient tissues. Together, our findings position LRRC as a central regulator of inflammatory signaling in breast cancer. By functioning upstream of IL-6 and driving activation of STAT, LRRC emerges as a key molecular nexus linking hypoxia to the iCAF phenotype and its tumor-promoting functions. The discovery of microenvironmental factors that fuel breast cancer inflammation is a sought-after milestone in the field since it may lead to the development of novel therapeutic interventions aimed at disrupting stromal-driven inflammation and tumor progression. Citation Format: Fernanda G. Kugeratski, Lisa Neilson, Adrian Kacperczyk-Perdyan, Juan R. Hernandez-Fernaud, Sergio Lilla, Jakub Mieczkowski, Sara Zanivan. Decoding new mechanisms of stromal-driven inflammation in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 6024.

DNA synthesis inhibition after global translational attenuation via the Unfolded Protein Response requires checkpoint adaptor protein Claspin and activation of checkpoint effector kinase Chk1. Here we show that DNA synthesis is also suppressed and Chk1 activated when ribosomal translation is inhibited by multiple, mechanistically distinct, protein synthesis inhibitors. Inhibition of DNA synthesis is rapid, reversible and not accompanied by detectable DNA damage. Our proteomic and functional characterisation shows that the kinases CK1-alpha and Cdc7 are necessary for transmission of the DNA replication-inhibitory signal in this process, likely by phosphorylating Claspin. Activation of Chk1 during protein synthesis inhibition also requires checkpoint proteins ATR and TopBP1, although activation occurs in the absence of multiple conventional markers of genotoxic stress signalling. We identified the replication and chromatin regulator Rif1 as a component of the DNA replication-inhibitory mechanism, whereas phosphatase PP1, a known functional partner of Rif1, was not required to suppress DNA replication during protein synthesis inhibition. Additionally, protein synthesis inhibition did not alter the phosphorylation of MCM2, a known target of Rif1/PP1, suggesting that Rif1 regulates DNA replication independently of MCM2-7 complex control. These findings elucidate a Claspin-Chk1-Rif1 pathway that links the fundamental processes of protein translation and DNA replication. Characterisation of the response to protein synthesis inhibition identifies a Claspin/Chk1/Rif1 pathway that couples translational attenuation to suppression of DNA replication.

The mitotic spindle is a microtubule-based apparatus that is responsible for accurate segregation of chromosomes into two daughter cells. In this study, we show that the DNA damage kinase Chk1 is required for optimal density and efficient nucleation of spindle microtubules during unperturbed mitosis in vertebrate cells. Chk1 phosphorylates β-tubulin at the identified conserved site threonine-285 (T285) in vitro, and at mitotic centrosomes in prometaphase and metaphase. Impaired β-tubulin-T285 phosphorylation correlates with improper spindles, delayed anaphase onset, erroneous chromosome alignment and segregation, unequal daughter cell-size and reduced cell proliferation. The ATR-interacting protein ATRIP promotes localization of ATR kinase and the mediator protein TopBP1 to mitotic centrosomes; furthermore, interaction of ATRIP with ATR and TopBP1 is required for Chk1 activation and β-tubulin-T285 phosphorylation. These results identify a signaling pathway that promotes spindle maturation and function in human cells, through Chk1-mediated β-tubulin-T285 phosphorylation.

Pancreatic ductal adenocarcinoma (PDAC) has poor prognosis as early-stage asymptomaticity leads to late-stage diagnoses. Strategies to detect PDAC earlier or identify high-risk individuals are therefore paramount. Here, we report results from genetically engineered mice and PDAC patients that identify serum proteins associated with pancreatic intraepithelial neoplasms (PanINs), the most common PDAC precursor, and early-stage PDAC. Initially, we screened previously described PanIN-abundant mice, harbouring pancreatic and duodenal homeobox 1 (Pdx1)-Cre, Lox-STOP-Lox-KrasG12D/+ and floxed alleles of essential autophagy genes autophagy-related 7 (Atg7) or autophagy-related 5 (Atg5). Sera from these mice were assessed by proteomics and hits were compared to those in Lox-STOP-Lox-KrasG12D/+ Lox-STOP-Lox-Trp53R172H/+ Pdx1-Cre (KPC) mice, which closely recapitulate human disease, and early-stage (I-II) PDAC patients. Levels of inter-alpha-trypsin inhibitor heavy chain H3 (ITIH3) were significantly elevated in all three screens, with complement C5, complement factors B and H (CFB/CFH), and monocyte differentiation antigen CD14 increased in KPC mice and PDAC patients; and all were significantly increased co-ordinately in PDAC according to disease stage. Serum levels of C5, CFH and CD14 together constitute a novel panel for identifying PanINs and early-stage PDAC with confidence, and when combined with additional screening, could help increase survival from this dismal disease.

Methylated amino acids accumulate upon the degradation of methylated proteins and are implicated in diverse metabolic and signaling pathways. Disturbed methylated amino acid homeostasis is associated with cardiovascular disease and renal failure. Mitochondria are core processing hubs in conventional amino acid metabolism, but how they interact with methylated amino acids is unclear. Here, we reveal that the orphan mitochondrial solute carrier 25A45 (SLC25A45) is required for the mitochondrial uptake of methylated amino acids. SLC25A45 binds with dimethylarginine and trimethyllysine but has no affinity for unmethylated arginine and lysine. A non-synonymous mutation of human SLC25A45 (R285C) stabilizes the carrier by limiting its proteolytic degradation and associates with altered methylated amino acids in human plasma. Metabolic tracing of trimethyllysine in cancer cells demonstrates that SLC25A45 drives the biosynthesis of the key amino acid derivative, carnitine. SLC25A45 is therefore an essential mediator of compartmentalized methylated amino acid metabolism.

Cancer-associated fibroblasts (CAFs) and immune cells make up two major components of the tumor microenvironment (TME), contributing to an ecosystem that can either support or restrain cancer progression. Metabolism is a key regulator of the TME, providing a means for cells to communicate with and influence each other, modulating tumor progression and anti-tumor immunity. Cells of the TME can metabolically interact directly through metabolite secretion and consumption or by influencing other aspects of the TME that, in turn, stimulate metabolic rewiring in target cells. Recent advances in understanding the subtypes and plasticity of cells in the TME both open up new avenues and create challenges for metabolically targeting the TME to hamper tumor growth and improve response to therapy. This perspective explores ways in which the CAF and immune components of the TME could metabolically influence each other, based on current knowledge of their metabolic states, interactions, and subpopulations.

The vast majority of recurrent somatic mutations arising in tumors affect protein-coding genes in the nuclear genome. Here, through population-scale analysis of 14,106 whole tumor genomes, we report the discovery of highly recurrent mutations affecting both the small (12S, MT-RNR1) and large (16S, MT-RNR2) mitochondrial RNA subunits of the mitochondrial ribosome encoded within mitochondrial DNA (mtDNA). Compared to non-hotspot positions, mitochondrial rRNA hotspots preferentially affected positions under purifying selection in the germline and demonstrated structural clustering within the mitoribosome at mRNA and tRNA interacting positions. Using precision mtDNA base editing, we engineered models of an exemplar MT-RNR1 hotspot mutation, m.1227G>A. Multimodal profiling revealed a heteroplasmy-dependent decrease in mitochondrial function and loss of respiratory chain subunits from a heteroplasmic dosage of ~10%. Mutation of conserved positions in ribosomal RNA that disrupt mitochondrial translation therefore represent a class of functionally dominant, pathogenic mtDNA mutations that are under positive selection in cancer genomes.

All mRNAs require eukaryotic translation initiation factor (eIF) 4A1 for translation through its different functions: loading of the pre-initiation complex onto mRNAs and unwinding of RNA structure. eIF4A1 is the catalytic subunit of the cap binding eIF4F complex and presumed to select the mRNAs for translation through these activities, instructed by signalling pathways driving cell fate. The mechanisms underlying activation of the distinct eIF4A1 functions and establish translational selectivity are unknown. Here, we have unravelled the complexity of mRNA selection by eIF4A1. We have mechanistically characterised the biological and atomic basis of inhibition by gain- and loss-of-function eIF4A1-inhibitors eFT226 and hippuristanol, and used machine learning to model the mRNA features associated with specific inhibition. This uncovered the eIF4A1 function mRNA sequence relationship: 5UTRs containing C CG-rich require efficient mRNA loading by eIF4F which is specifically targeted by hippuristanol, while 5UTRs harbouring alternatives starts sites together with AG-rich motifs utilise eIF4A1 for start site selection, specifically perturbed by eFT226. Our model is validated through a massively parallel reporter assay using 5UTRs from a distinct evolutionary origin. This prompted us to examine the conservation between mRNA sequence and eIF4A1 function, and revealed their co-development. Our findings highlight opportunities for novel therapeutic strategies targeting eIF4A1 and for improved design of mRNA-based therapeutics. ### Competing Interest Statement MB collaborates with Cancer Research Horizons on drug discovery against some of the targets in this paper. JDB is a shared inventor on a provisional patent application filed by Yale University with the US patent office covering the NaPTRAP method and the sequences described here.

Redundancy in the genetic code provides robustness against mutations and errors in translation. It also allows cells to encode additional information within a coding sequence alongside the polypeptide chain itself. Here, we show that genes expressed in different cellular states employ distinct codon usage patterns linked to post-transcriptional regulation of protein expression. During proliferation, gene expression programmes favour codons ending in A or T, while in quiescence, genes are enriched for G or C ending codons. In parallel, the tRNA pool broadly shifts to match the differing codon demands in each state. Further, the majority of tRNA genes up regulated during proliferation are confined to a small, distinct locus on human chromosome 6. Together these findings reveal a striking and previously unreported regulatory programme that exploits redundancy in the genetic code to impose an additional layer of gene expression control, driven by a proliferation-associated tRNA regulatory nexus on chromosome 6.

Integrins are major receptors for secreted extracellular matrix, playing crucial roles in physiological and pathological contexts, such as angiogenesis and cancer. Regulation of the transition between inactive and active conformation is key for integrins to fulfill their functions, and pharmacological control of those dynamics may have therapeutic applications. We create and validate a prototypic luminescent R1 integrin activation sensor (R1IAS) by introducing a split luciferase into an activation reporting site between the RI and the hybrid domains. As a recombinant protein in both solution and living cells, R1IAS accurately reports R1 integrin activation in response to (bio)chemical and physical stimuli. A short interfering RNA (siRNA) high-throughput screening on live R1IAS knockin endothelial cells unveils hitherto unknown regulators of R1 integrin activation, such as R1 integrin inhibitors E3 ligase Pja2 and vascular endothelial growth factor B (VEGF-B). This split-luciferase-based strategy provides an in situ label-free measurement of integrin activation and may be applicable to other R integrins and receptors.

Nucleophosmin (NPM1), a nucleolar protein frequently mutated in hematopoietic malignancies, is overexpressed in several solid tumors with poorly understood functional roles. Here, we demonstrate that Npm1 is upregulated after APC loss in WNT-responsive tissues and supports WNT-driven intestinal and liver tumorigenesis. Mechanistically, NPM1 loss induces ribosome pausing and accumulation at the 5'-end of coding sequences, triggering a protein synthesis stress response and p53 activation, which mediate this antitumorigenic effect. Collectively, our data identify NPM1 as a critical WNT effector that sustains WNT-driven hyperproliferation and tumorigenesis by attenuating the integrated stress response and p53 activation. Notably, NPM1 expression correlates with elevated WNT signaling and proliferation in human colorectal cancer (CRC), while CRCs harboring NPM1 deletions exhibit preferential TP53 inactivation, underscoring the clinical relevance of our findings. Being dispensable for adult epithelial homeostasis, NPM1 represents a promising therapeutic target in p53-proficient WNT-driven tumors, including treatment-refractory KRAS-mutant CRC, and hepatic cancers.

Casitas B-lineage lymphoma (CBL) is an E3 ubiquitin ligase critical for negatively regulating receptor protein tyrosine kinases (RTKs). Deleterious CBL mutants lose E3 activity, but act as adaptors that gain function to cause myeloproliferative neoplasms. Currently, there is no targeted treatment available for patients with CBL mutant-dependent disorders. By combining phage-display technology and structure-based optimization, we discovered CBLock, a nanomolar affinity peptide inhibitor, that binds the substrate-binding site of CBL’s tyrosine kinase binding domain (TKBD). CBLock disrupts the interaction between CBL mutants and RTKs, thereby impairing RTK-mediated priming of adaptor function of CBL mutants and downstream signaling. Notably, CBLock binds TKBD without inducing conformational changes, thereby preserving its ligand-free native conformation. In contrast, when CBL binds RTK substrates, TKBD undergoes a conformational change. Maintaining the native CBL TKBD conformation was crucial for CBLock to inhibit proliferation, induce cell-cycle arrest, and promote apoptosis in leukemia cells harboring CBL mutations. In a mouse xenograft model of acute myeloid leukemia (AML), CBLock reduced tumor burden and improved survival rate. Moreover, CBLock inhibited the proliferation of cells derived from patients with CBL mutations. Therefore, inhibiting CBL TKBD in its native state presents a promising therapeutic opportunity in targeting mutant CBL-dependent leukemia.

Cysteine oxidative modifications are critical signaling events regulating cellular functions, but their low abundance and dynamic nature pose technical challenges. We developed the SICyLIA-TMT workflow, which sequentially labels reduced and reversibly oxidized cysteines with light and heavy iodoacetamide (IAA) within the same sample. The inclusion of tandem mass tags (TMTs) enables simultaneous quantification of oxidative modification dynamics and protein levels across multiple conditions using micrograms of material. To improve the detection of low-abundance oxidized cysteines, a dedicated TMT channel serves as a carrier for heavy IAA-labeled peptides (SICyLIA-cTMT), enhancing quantification and enabling precise stoichiometry calculations. We demonstrate the workflow's applicability to cultured cells and full organs under stress. SICyLIA-cTMT achieves unprecedented depth and accuracy in redox proteome analysis while reducing mass spectrometry time. Combining SICyLIA-TMT with latest mass spectrometry technologies further halves the acquisition time without compromising coverage, improving throughput and enabling comprehensive studies of oxidative signaling.

Cancer-associated fibroblasts (CAFs) are a multifunctional cell population of solid tumors that substantially remodel the tumor microenvironment (TME). The combination of single-cell and spatial technologies with elegant mouse models and analysis of patient samples is enabling unprecedented advances in the characterization of CAF origins, heterogeneity, and functions within the TME. As such, the field is now evolving to delineate tissue-specific subpopulations of CAFs, their markers, and the biological context in which each subset presents with a tumor-promoting or a tumor-restraining function. In this timely review, we discuss recent advances in CAF biology in the context of emerging areas of interest in the field of anticancer therapy: immunotherapy, metabolism, and extracellular vesicles. We also highlight the substantial role of CAFs in modulating the immune microenvironment and the recent advances in targeting CAFs for cancer treatment.

Mitochondrial homeostasis relies on a tight balance between mitochondrial biogenesis and degradation. Although mitophagy is one of the main pathways involved in the clearance of damaged or old mitochondria, its coordination with mitochondrial biogenesis is poorly characterized. Here, by unbiased approaches including last-generation liquid chromatography coupled to mass spectrometry and transcriptomics, we identify the protein phosphatase PP2A-B55α/PPP2R2A as a Parkin-dependent regulator of mitochondrial number. Upon mitochondrial damage, PP2A-B55α determines the amplitude of mitophagy induction and execution by regulating both early and late mitophagy events. A few minutes after the insult, ULK1 is released from the inhibitory regulation of PP2A-B55α, whereas 2 to 4 hours later, PP2A-B55α promotes the nuclear translocation of TFEB, the master regulator of autophagy and lysosome genes, to support mitophagy execution. Moreover, PP2A-B55α controls a transcriptional program of mitochondrial biogenesis by stabilizing the Parkin substrate and PGC-1α inhibitor PARIS. PP2A-B55α targeting rescues neurodegenerative phenotypes in a fly model of Parkinson’s disease, thus suggesting potential therapeutic application.

CYRI proteins promote lamellipodial dynamics by opposing Rac1-mediated activation of the Scar/WAVE complex. This activity also supports resolution of macropinocytic cups, promoting internalisation of surface proteins, including integrins. Here, we show that CYRI-B also promotes focal adhesion maturation and dynamics. Focal adhesions in CYRI-B-depleted cells show accelerated maturation and become excessively large. We probed the composition of these enlarged focal adhesions, using a Bio-ID screen, with paxillin as bait. Our screen revealed changes in adhesion proteins proximal to paxillin suggesting early activation of stress fibre contraction and depletion of the integrin internalisation mediator ERC1. Lack of CYRI-B leads to more stable lamellipodia and accumulation of polymerised actin in stress fibres. This actin acts as a barrier to microtubule targeting for adhesion turnover. Thus, our studies reveal an important connection between lamellipodia dynamics controlled by CYRI-B and microtubule targeting of ERC1 to modulate adhesion maturation and turnover.

Phenotypic switching of vascular smooth muscle cells (VSMCs) from a contractile to a synthetic state contributes to vascular remodelling in hypertension. While oxidative stress and microRNAs (miRNAs) modulate VSMC phenotype, the molecular profile of VSMCs in human hypertension remain undefined. This study aimed to characterize the proteomic, redox, and miRNA signature of VSMCs in human hypertension. VSMCs were isolated from resistance arteries of normotensive (NT) and hypertensive (HT) subjects. Expression of VSMC phenotypic markers and redox-related genes were assessed by immunoblotting and qPCR. Reactive oxygen species (ROS) was measured by Amplex red assay. Global and redox proteomic profiling was performed using tandem mass tag (TMT)-based LC-MS/MS, using isotope-labelled iodoacetamide to quantify cysteine oxidation. miRNA profiling and VSMC secretome were assessed using TaqMan Advanced miRNA arrays and Olink proteomics inflammation panel. In HT, VSMC exhibited reduced expression of contractile proteins (α-SMA, SM22, MYOCD; p<0.05) and increased KLF4 (NT:0.66±0.08, HT:2.83±0.73, p<0.05), indicating de-differentiation. ROS production was increased (NT:9.15±0.66, HT:13.28±1.24), along with Nox1 (NT:1.06±0.18, HT:1.83±0.29) and Nox5 (NT:1.25±0.32, HT:14.03±0.84) mRNA levels (p<0.05). Although Nrf2 mRNA was upregulated (NT:0.89±0.14, HT:1.34±0.07, p<0.05), levels of antioxidant enzymes (SOD1, SOD2, CAT, Trx1) remained unchanged, suggesting impaired redox balance. Proteomic analysis showed upregulation of extracellular matrix (ECM) and inflammatory proteins in HT (fold change>1.5, p<0.05), consistent with a pro-fibrotic/pro-inflammatory phenotype and increased cysteine oxidation, particularly in ECM components (fold change>1.5, p<0.05). miRNA profiling revealed 136 downregulated miRNAs in HT (fold change >1.5), including miR-145-5p, a key regulator of VSMC differentiation. Additionally, four Nox5-targeting miRNAs (miR-505-5p, miR-324-5p, miR-185-5p, miR-491-5p) were downregulated, linking miRNA dysregulation and oxidative stress. Secretome analysis confirmed increased release of pro-inflammatory and pro-fibrotic mediators in HT, including CXCL1/5/6, IL-6/8/24, MCP1–4, IL-18R1, CCL3, and CCL20 (p<0.05). In hypertension, VSMCs exhibit a distinct proteomic, oxidative, and miRNA signatures consistent with a pro-inflammatory, pro-fibrotic phenotype. These findings provide molecular insights into VSMC plasticity and vascular remodelling in human hypertension.