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Pineda, R.

Publications and source records attributed to Pineda, R..

3 recordsLinked to original sources
bioRxiv2025

Single cell transcriptomics in a treatment-segregated cohort exposes a STAT3-regulated therapeutic gap in idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic pulmonary disease of unknown etiology. Since approved IPF drugs only slow disease progression, novel therapeutics are required that improve clinical outcomes. Here we report a single cell lung RNA-Seq and gene regulatory network analysis of the largest IPF cohort assembled to date. Segregating this cohort based on status of treatment with approved first-generation IPF antifibrotics (untreated, nintedanib- and pirfenidone-treated), we describe for the first time the transcriptional landscape of untreated IPF across 40 lung cell types, and the elements of this program that are impacted by these antifibrotics. On average, nearly 60% of the untreated IPF-dysregulated transcriptome is refractory to treatment with these drugs, a transcriptional deficit we refer to as the IPF therapeutic gap. Gene regulatory network analysis indicated a dominant functional footprint for the transcription factor STAT3 in both untreated IPF and the IPF therapeutic gap. Validating our analysis in a translational precision cut lung slice platform that recapitulates IPF explants, pharmacological inhibition of STAT3 reduced the IPF therapeutic gap in numerous lung cell types. Finally, we resolved a STAT3-anchored master regulatory network comprising numerous profibrotic transcription factors in IPF alveolar fibroblasts, a critical fibrotic lineage. Our study represents a comprehensive resource for translational lung fibrosis research and introduces a strategy for drug discovery that is adaptable to human disease more broadly.

cell biology↗
bioRxiv2024

Etrinabdione (VCE-004.8), a B55a activator, promotes angiogenesis and arteriogenesis in critical limb ischemia.

BackgroundVasculogenic therapies explored for the treatment of peripheral artery disease (PAD) have encountered minimal success in clinical trials. Addressing this, B55, an isoform of protein phosphatase 2A (PP2A), emerges as pivotal in vessel remodeling through activation of hypoxia-inducible factor 1 (HIF-1). This study delves into the pharmacological profile of VCE-004.8 (Etrinabdione) and evaluates its efficacy in a preclinical model of critical limb ischemia, with a focus on its potential as a PP2A/B55 activator to induce angiogenesis and arteriogenesis. MethodsVascular endothelial cells were used for in vitro experiments. Aorta ring assay was performed to explore sprouting activity. Matrigel plug-in assay was used to assess the angiogenic potential. Critical limb ischemia (CLI) in mice was induced by double ligation in the femoral arteria. Endothelial vascular and fibrotic biomarkers were studied by immunohistochemistry and qPCR. Arteriogenesis was investigated by microvascular casting and micro-CT. Proteomic analysis in vascular tissues was analyzed by LC-MS/MS. Ex-vivo expression of B55 and biomarkers were investigated in artery samples from PAD patients. ResultsVCE-004.8 exhibited the ability to induce B55 expression and activate the intersecting pathways B55/AMPK/Sirtuin 1/eNOS and B55/PHD2/HIF-1. VCE-004.8 prevented OxLDL and H2O2-induced cytotoxicity, senescence, and inflammation in endothelial cells. Oral VCE-004.8 increased aorta sprouting in vitro and angiogenesis in vivo. In CLI mice VCE-004.8 improved collateral vessel formation and induced endothelial cells proliferation, angiogenic gene expression and prevented fibrosis. The expression of B55, Caveolin 1 and Sirtuin-1 is reduced in arteries from CLI mice and PAD patient, and the expression of these markers was restored in mice treated with VCE-004.8. ConclusionsThe findings presented in this study indicate that Etrinabdione holds promise in mitigating endothelial cell damage and senescence, while concurrently fostering arteriogenesis and angiogenesis. These observations position Etrinabdione as a compelling candidate for the treatment of PAD, and potentially other cardiovascular disorders. Novelty and SignificanceO_ST_ABSWhat Is Known?C_ST_ABSO_LIThe phosphatase PPA2/B55 stabilizes endothelial cells (ECs) in response to cell stress conditions, thereby protecting ECs from apoptosis and promoting angiogenesis. C_LIO_LIEtrinabdione (VCE-004.8) functions as a potent activator of PPA2/B55 inducing PHD2 dephosphorylation at ser125 and fostering HIF activation. C_LIO_LIVCE-004.8 prevents vascular damage in preclinical models of systemic and cardiac fibrosis and alleviates blood-brain barrier disruption in neuroinflammatory conditions. C_LIO_LIVCE-004.8 is also a dual agonist of PPAR{psi} and CB2 receptors and shows antiinflammatory activity. C_LIO_LIOral VCE-004.8 has meet the primary endpoints of safety and tolerability in a Phase IIa clinical trial with systemic sclerosis patients (clinicaltrial.gov: NCT03745001). C_LI What New Information Does This Article Contribute?O_LIEtrinabdione induces HIF-1 expression in endothelial cells through a novel pathway that potentially involves two axes: B55/PHD2 and B55/AMPK/Sirt1 signaling that may converge on HIF stabilization. C_LIO_LIEtrinabdione prevented endothelial cell damage and senescence, while inducing arteriogenesis and angiogenesis in CLI mice. C_LIO_LIIn arteries of patients with PAD and in CLI mouse models, the expression levels of B55, Caveolin 1, and Sirtuin 1 are diminished. However, treatment with Etrinabdione specifically in CLI mice prompts an increase in the levels of these proteins. C_LIO_LIEtrinabdione triggers neovascularization and angiogenesis specifically within hypoxic tissue in a critical ischemia model, with no impact on healthy tissue. C_LI

pharmacology and toxicology↗
bioRxiv2022

Citrus relatives exhibit natural variation in perception and response magnitude to microbial features

Although much is known about the responses of model plants to microbial features, we still lack an understanding of the extent of variation in immune perception across members of a plant family. In this work, we analyzed immune responses in Citrus and wild relatives, surveying 86 Rutaceae genotypes with differing leaf morphologies and disease resistances. We found that responses to microbial features vary both within and between members. Species in two subtribes, the Balsamocitrinae and Clauseninae, can recognize all tested microbial features (flg22, csp22, chitin), including one from Candidatus Liberibacter species (csp22CLas), the bacterium associated with citrus greening disease aka Huanglongbing. We investigated differences at the receptor level for flagellin perception (FLS2 receptor) and chitin perception (LYK5 receptor) in citrus genotypes. We were able to characterize two genetically linked FLS2 homologs from Frost Lisbon lemon (responsive) and Washington navel orange (non-responsive). Surprisingly, FLS2 homologs from responding and non-responding genotypes were expressed in citrus and functional when transferred to a heterologous system. Washington navel orange weakly responds to chitin, but Tango mandarin exhibits a robust response. LYK5 alleles were identical or nearly-identical between the two genotypes and able to complement the Arabidopsis lyk4/lyk5-2 mutant with respect to chitin perception. Collectively, our data indicates that differences in chitin and flg22 perception in these citrus genotypes are not the result of sequence polymorphisms at the receptor level. These findings shed light onto the diversity of perception of microbial features and highlight genotypes capable of recognizing polymorphic pathogen features.

plant biology↗