Biology subjects

Ruiz-Ortega, M.

Publications and source records attributed to Ruiz-Ortega, M..

6 recordsLinked to original sources

bioRxiv2026

Early Lipid Remodeling During Ischemia Reperfusion Injury Is Associated with Damage- and Protection-Related Lipid Signatures in Acute Kidney Injury.

Acute kidney injury (AKI) secondary to kidney ischemia-reperfusion (IRI) is driven by profound metabolic perturbations that shape oxidative stress, inflammation, and cell death responses. Despite growing evidence of lipid dysregulation in AKI, its biological and mechanistic relevance remains unclear. To address this gap, we performed comprehensive untargeted kidney lipidomics using LC-QTOF-MS to delineate dynamic lipid changes during the acute phase of murine kidney IRI. Integration of lipidomic signatures with kidney gene-expression profiling and curated datamining revealed early activation of lipid pathways associated with injury responses. IRI induced marked lipidomic reprogramming, characterized by a marked accumulation of glycerolipids, including triacylglycerols (TGs) species detected exclusively after injury, together with increased levels of sphingolipids (ceramides, sphingomyelins, and hexosylceramides). Cardiolipins and most glycerophospholipids declined sharply following IRI, whereas specific phosphatidylcholines (PCs) exhibited opposite regulation, consistent with dynamic membrane remodeling. Datamining linked TG accumulation and altered transcriptional regulation of PC metabolism to repair-prone type 1 injured proximal tubular cells, highlighting its role in early tubular injury. Correlation analyses revealed strong associations between glycerolipids/sphingolipids and markers of renal dysfunction and inflammation, identifying TG 54:9 as a candidate for injury biomarker. Conversely, PE 40:6, PI 38:6, together with several lysophosphatidylcholines and ether-linked phospholipids, correlated positively with nephroprotective and antioxidant markers. Together, these patterns delineate two major lipid modules: a damage-associated module and a nephroprotection-associated module. These findings establish lipid remodeling as a central yet underexplored determinant of AKI pathogenesis and underscore lipidomics as a powerful discovery tool for identifying novel biomarkers and mechanistic targets in kidney disease.

pathology↗

bioRxiv2025

Multi-omics integrative approach of senescent endothelial cells and derived extracellular vesicles in a replicative senescence model

Aging is a major unmodifiable risk factor for cardiovascular disease (CVD). Replicative endothelial senescence (RES), characterized by permanent cell-cycle arrest and a senescence-associated secretory phenotype (SASP), is a hallmark of vascular aging. However, the molecular mechanisms underlying RES, particularly the role of extracellular vesicles (EVs), remain poorly understood. To this aim, an integrated multi-omics approach (proteomics, mRNA, and miRNA profiling) was applied to characterize senescent and early-passage human umbilical vein endothelial cells (HUVECs) and their secreted EVs. Senescent HUVECs and EVs displayed a canonical senescence profile, including cell-cycle arrest, DDR activation, NF-{kappa}B signaling, and SASP induction, alongside marked suppression of RNA metabolism, ribosome biogenesis, and DNA repair. Multi-omics integration has linked endothelial senescence to vascular aging, maladaptive angiogenesis, and ECM remodeling, which are key processes in CVD development. Moreover, senescent EVs propagate senescence by exporting fewer reparative and antioxidant factors while carrying pro-fibrotic, adipogenic, and angiogenic mediators. Multi-omics profiling revealed consistent transcript-protein changes in senescent cells and EVs, defining a robust molecular signature of RES, including endothelial cell-specific markers and post-transcriptional regulators (lncRNAs and miRNAs). Among these, miR-22-3p and miR-126-5p emerged as key modulators in both cells and EVs, with miR-22-3p demonstrating compartment-specific regulation. Integration further uncovered a coordinated regulatory network involving miRNAs (miR-23, miR-335-3p, miR-29 family, miR-590-3p, and miR-126-5p) that were inversely correlated with transcription factors (e.g., REST, KLFs, and ZNFs) and downstream targets, collectively driving endothelial senescence through impaired PI3K-Akt signaling. Although further validation is needed, these findings provide new insights into the molecular mechanisms of RES and open perspectives for modulating secondary senescence and age-related vascular diseases.

cell biology↗

bioRxiv2024

Targeting STING in experimental peritoneal damage: a novel approach in peritoneal dialysis therapy

Peritoneal dialysis (PD) is a widely used kidney replacement therapy for end-stage kidney disease (ESKD) patients. However, long-term exposure to PD fluids (PDF) can lead to peritoneal membrane (PM) damage, causing ultrafiltration failure and thus PD discontinuation. Investigating the molecular mechanisms underlying this damage is crucial for identifying new therapeutic targets to mitigate peritoneal deterioration in PD patients. Therefore, in this work we study the role of STING in peritoneal inflammation and fibrosis. To this aim, we performed different preclinical mouse models of peritoneal inflammation, fibrosis, and adhesions. In a chlorhexidine gluconate (CHX)-induced inflammation model, we found changes in the peritoneal transcriptomic profile, and cytosolic DNA-sensing signaling was one of the most enriched KEGG pathways. STING, as a conspicuous member of this pathway, was upregulated in CHX-and PDF-exposed mice, and in peritoneal biopsies from PD patients. STING genetic deficiency diminished peritoneal inflammation, by downregulating inflammatory gene expression, preventing NF-{kappa}B pathway activation, and decreasing cell infiltration, in early (10 days) and advanced (30 days) stages of the CHX model. STING absence also decreased PM thickness and fibrosis in the advanced CHX model, reduced adhesion scores in a post-surgical intra-abdominal adhesion model, and decreased inflammation in an S. epidermidis-induced peritonitis model. Furthermore, pharmacological inhibition of STING with C-176 decreased inflammation and macrophage-mediated mesothelial-to-mesenchymal transition in cultured mesothelial cells, and reduced CHX-induced PM thickness and inflammation in mice. Altogether, these findings highlight STING as a key mediator of peritoneal damage and suggest it may be a novel therapeutic target for preventing PD-associated peritoneal deterioration.

pathology↗

bioRxiv2023

Ccn2 deficiency causes smooth muscle cell de-differentiation and severe atherosclerosis in hyperlipidemic mice

Cellular communication network factor 2 (CCN2/CTGF) is a matricellular protein with an established role in fibrotic diseases and cancers, and therapies targeting CCN2 is currently in Phase II and III clinical trials for idiopathic pulmonary fibrosis, pancreatic cancer and Duchenne Muscular Dystrophy. Recent studies have highlighed a protective role of CCN2 in aortic aneurysm disease, but its role in atherosclerosis remains to be investigated. We identified arteries as having the highest relative expression of CCN2 across 54 human tissues. In aortas, CCN2 was among the highest expressed genes, and in situ hybridization of human internal thoracic arteries revealed vascular smooth muscle cells (SMCs) as its principal source. Hypothesizing a role for CCN2 in SMC phenotype maintenance and athero-protection, we investigated inducible Ccn2 knockout (Ccn2{Delta}/{Delta}) mice in normo- and hyper-lipidemic settings. Induction of hyperlipidemia by single intravenous injection of 1{middle dot}1011 viral genomes of rAAV8-D377Y-mPcsk9 combined with 24 weeks of western type diet resulted in severe enlargement (3-5-fold increase of relative aorta mass compared to wildtype littermates, p < 0.0001) and whitening of Ccn2{Delta}/{Delta} aortas. Oil Red O-staining of en face prepared thoracic aortas showed a marked increase in atherosclerosis in Ccn2{Delta}/{Delta} mice as compared to wildtype littermates (75% vs. 10% Oil Red O-positive aortic area, p < 0.0001). Transcriptomic profiling of cultivated SMCs derived from aortas of normolipidemic mice showed signatures of dedifferentiation (reduced expression of e.g. Myocd, Acta2 and Myh11) and modulation toward a synthetic, pro-inflammatory phenotype of Ccn2{Delta}/{Delta} SMCs. These effects were verified in vivo and in CCN2-silenced human aortic SMCs. Taken together, we find that CCN2 plays a critical athero-protective role in artery tissues, likely through maintaining SMCs in a differentiated, contractile phenotype.

pathology↗

bioRxiv2020

Renal tubule Cpt1a overexpression mitigates kidney fibrosis by restoring mitochondrial homeostasis

Chronic kidney disease (CKD) remains a major epidemiological, clinical and biomedical challenge. During CKD, renal tubular epithelial cells (TECs) suffer a persistent inflammatory and profibrotic response. Fatty acid oxidation (FAO), the main source of energy for TECs, is reduced in kidney fibrosis and contributes to its pathogenesis. To determine if FAO gain-of-function (FAO-GOF) could protect from fibrosis, we generated a conditional transgenic mouse model with overexpression of the fatty acid shuttling enzyme carnitine palmitoyl-transferase 1 A (CPT1A) in TECs. Cpt1a knock-in (CPT1A KI) mice subjected to three different models of renal fibrosis (unilateral ureteral obstruction, folic acid nephropathy-FAN and adenine induced nephrotoxicity) exhibited decreased expression of fibrotic markers, a blunted pro-inflammatory response and reduced epithelial cell damage and macrophage influx. Protection from fibrosis was also observed when Cpt1a overexpression was induced after FAN. FAO-GOF restituted oxidative metabolism and mitochondrial number and enhanced bioenergetics increasing palmitate oxidation and ATP levels, changes also recapitulated in TECs exposed to profibrotic stimuli. Studies in patients evidenced decreased CPT1 levels and increased accumulation of short and middle chain acyl-carnitines, reflecting impaired FAO in human CKD. We propose that strategies based on FAO-GOF may constitute powerful alternatives to combat fibrosis inherent to CKD. Graphical abstractOverexpression of CPT1A impedes tubular epithelial renal cell dedifferentiation and a pro-fibrotic phenotype by restoring mitochondrial metabolic bioenergetics. O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY

biochemistry↗

bioRxiv2019

MiR-9-5p protects from kidney fibrosis by metabolic reprogramming

MicroRNAs (miRNAs) regulate gene expression post-transcriptionally and control biological processes, including fibrogenesis. Kidney fibrosis remains a clinical challenge and miRNAs may represent a valid therapeutic avenue. We show that miR-9-5p protected from renal fibrosis in the mouse model of unilateral ureteral obstruction (UUO). This was reflected in reduced expression of pro-fibrotic markers, decreased number of infiltrating monocytes/macrophages and diminished tubular epithelial cell injury and transforming growth factor-beta 1 (TGF-{beta}1)-dependent de-differentiation in human kidney proximal tubular (HKC-8) cells. RNA sequencing (RNA-Seq) studies in the UUO model revealed that this protection was mediated by a global shift in the expression profile of genes related to key metabolic pathways, including mitochondrial dysfunction, oxidative phosphorylation (OXPHOS), fatty acid oxidation (FAO) and glycolysis, preventing their UUO-dependent down-regulation. This effect was mirrored by a prevention in the TGF-{beta}1-induced bioenergetics changes in HKC-8 cells. The expression of the FAO-related axis peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1)-peroxisome proliferator-activated receptor alpha (PPAR) was reduced by UUO, although preserved by the administration of miR-9-5p. We found that in mice null for the mitochondrial master regulator PGC-1, miR-9-5p was unable to promote a protective effect in the UUO model. We propose that miR-9-5p elicits a protective response to chronic kidney injury and renal fibrosis by inducing reprogramming of the metabolic derangement and mitochondrial dysfunction affecting tubular epithelial cells.

molecular biology↗