Biology subjects

Guirola, O.

Publications and source records attributed to Guirola, O..

4 recordsLinked to original sources

bioRxiv2026

Integrated Analysis of HeberFERON-Driven Comparative Proteomic regulation in Glioblastoma Cells U-87MG

Glioblastoma is a very aggressive brain tumor with few therapeutics options. Type I and II Interferons (IFNs) co-formulation HeberFERON has been used in cancer treatment, with promising results in high grade brain tumors. High throughput techniques in easy-to-handle models have been important to interrogate biomolecules changes, describe mechanisms and find pharmacodynamic biomarkers. This study aims to elucidate the effect of HeberFERON over the cell proteome in comparison to its individual IFNs components. Proteomic changes with HeberFERON in the glioblastoma-derived cell line U-87MG, in comparison with individual IFN-2b and IFN-{gamma}, were studied using a nanoLC instrument EasyLC coupled to Velos Pro mass spectrometer; Maxquant and Perseus were also used. Several enrichment tools, networking analysis and canSAR for drug targets were employed. Translation, RNA processing, mitotic cell cycle, cytoskeleton and chromosome organization, apoptosis, autophagy, DNA repair are enriched to limit cellular growing together with changes in immune response components, supporting HeberFERON as a multitarget treatment. This co-formulation is distinguished at modulating RNA splicing with SMN complex, cytoskeleton organization and microtubule-based movement, nuclear envelope breakdown, DNA conformational changes, and oxidative phosphorylation, with a better drawing of effects over a variety of systems inside the tumoral cell. Together with previous microarray experiment, informative genes and proteins as pharmacodynamic biomarkers for antiproliferative effects showed up (ex. STAT1/2, CENPE, ATRIP, MAP1B, LIMA1, VCP, several ribosomal, spliceosome and proteasomal complexes proteins). This study complements transcriptomic and phosphoproteomic previous experiments in this model and underscore HeberFERON as a glioblastoma therapeutic.

cancer biology↗

bioRxiv2022

Proteomics and phosphoproteomics profiling of the co-formulation of type I and II interferons, HeberFERON, in the glioblastoma-derived cell line U-87 MG

HeberFERON is a co-formulation of Interferon (IFN)-2b and IFN-{gamma} in synergic proportions, with a demonstrated effect on skin cancer and other solid tumors. It has antiproliferative effects over glioblastoma multiform (GBM) clones and cell lines in culture, including U-87 MG. Omics studies in U-87 MG showed distinctive expression patterns compared to individual IFNs. Kinase signaling pathways dysregulation can also contribute to HeberFERON effects. Here, we report the first label-free quantitative proteomic and phosphoproteomic analyses to evaluate changes induced by HeberFERON after 72h incubation of U-87 MG cell line. LC-MS/MS analysis identified 7627 proteins with a fold change >2 (p<0.05); 122 and 211 were down- and up-regulated by HeberFERON, respectively. We identified 23549 peptides (5692 proteins) and 8900 phosphopeptides, 412 of these phosphopeptides (359 proteins) were differentially modified with fold change >2 (p<0.05). Proteomic enrichment analysis showed IFN signaling and its control, together to direct and indirect antiviral mechanisms were the main modulated processes. Enrichment analysis of phosphoproteome pointed to the cell cycle, cytoskeleton organization, translation and RNA splicing, autophagy, and DNA repair as biological processes represented. There is a high interconnection of phosphoproteins in a molecular network, where mTOR occupies a centric hub. HeberFERON regulates many phosphosites newly reported or with no clear association to kinases. Of interest is phosphosites increasing phosphorylation were mainly modified by CDK and ERK kinases, thus new cascades regulations can be determining the antiproliferation outcome. Our results contribute to a better mechanistic understanding of HeberFERON in the context of GBM. Significance of the StudyHeberFERON is a co-formulation of IFN-2b and -{gamma} in synergic proportion, registered for skin basal cell carcinoma treatment, also demonstrating clinical effect over solid tumors, including GBM. GBM is a very lethal tumor, protected by the blood-brain barrier (BBB), highly mutated in proliferative signaling pathways with little treatment success. Interferons have been widely used in cancer; they pass BBB and act at JAK/STAT, PI3K/AKT/mTOR, and MAPKs cascades. We observed antiproliferative effects over GBM clones and cell lines in culture. U-87 MG is used as a model to understand the HeberFERON mechanism of action in GBM. We completed the first proteomic and label-free quantitative phosphoproteomic analysis after incubation of U-87 MG cell line with HeberFERON for 72h. The main contribution of this article is the description of phosphosites regulated in proteins participating in cell cycle, cytoskeleton organization, translation, autophagy, and DNA repair in a highly interconnected molecular network, where mTOR occupies a centric hub. Together with reported phosphosites, we described new ones and others with no associated kinases. Increased phosphorylation is mainly accounted by CDK and ERK kinases pointing to possibly new cascades regulations. This knowledge will contribute to the functional understanding of HeberFERON in GBM joined to general regulatory mechanisms in cancer cells.

cancer biology↗

bioRxiv2021

Phosphoproteomic Landscape of AML Cells Treated with the ATP-Competitive CK2 Inhibitor CX-4945

Casein kinase 2 (CK2) regulates a plethora of proteins with pivotal roles in solid and hematological neoplasia. Particularly, in acute myeloid leukemia (AML) CK2 has been pointed as an attractive therapeutic target and prognostic marker. Here, we explored the impact of CK2 inhibition over the phosphoproteome of two cell lines representing major AML subtypes. Quantitative phosphoproteomic analysis was conducted to evaluate changes in phosphorylation levels after incubation with the ATP-competitive CK2 inhibitor CX-4945. Functional enrichment, network analysis, and database mining were performed to identify biological processes, signaling pathways, and CK2 substrates that are responsive to CX-4945. A total of 273 and 1310 phosphopeptides were found differentially modulated in HL-60 and OCI-AML3 cells, respectively. Despite regulated phosphopeptides belong to proteins involved in multiple biological processes and signaling pathways, most of these perturbations can be explain by direct CK2 inhibition rather than off-target effects. Furthermore, CK2 substrates regulated by CX-4945 are mainly related to mRNA processing, translation, DNA repair, and cell cycle. Overall, we evidenced that CK2 inhibitor CX-4945 impinge on mediators of signaling pathways and biological processes essential for primary AML cells survival and chemosensitivity, reinforcing the rationale behind the pharmacologic blockade of protein kinase CK2 for AML targeted therapy.

cancer biology↗

bioRxiv2020

The low-density lipoprotein receptor-related protein-1 is essential for Dengue virus infection

Dengue virus (DENV) causes the most prevalent and rapidly spreading arboviral disease of humans. It enters human cells by receptor-mediated endocytosis. Numerous cell surface proteins have been proposed as DENV entry factors. Among these, the phosphatidylserine receptor TIM-1 is the only one known to mediate virus internalization. However, several cellular models lacking TIM-1 are permissive to DENV infection, suggesting that other receptors exist. Here we show that the Low-density lipoprotein receptor-related protein-1 (LRP1) binds DENV virions by interacting with the DIII of the viral envelope glycoprotein. DENV infection is effectively inhibited by the purified receptor at 5x10-8 mol/L and the interaction of the envelope protein with LRP1 is also blocked by a natural ligand of LRP1. Depletion of LRP1 causes 100-fold lower production of infectious virus than controls. Our results indicate that LRP1 is another DENV receptor thus, becoming an attractive target to evaluate for the development of effective antiviral drugs against DENV. Author summaryDengue virus (DENV) is a complex of four related viruses, recognized as serotypes, designated as DENV1-4. Any of the four DENV serotypes can cause a self-limited disease of mild flu-like symptoms known as dengue or its life threatening form, severe dengue, with hemorrhagic manifestations, organ impairment and shock. This disease is widely spread in tropical and sub-tropical areas worldwide, where the incidence of severe dengue has been increasing steadily. So far, efforts that target components of the viral replication machinery in order to develop a specific antiviral drug for dengue disease patients have failed. Thus, identifying the cell surface receptors used by DENV to enter host cells would provide a new molecular target to develop inhibitory drugs. Here, we evaluate the Low density lipoprotein receptor-related protein-1 (LRP1) as a putative DENV receptor. We present evidence demonstrating that LRP1 binds DENV through the viral envelope protein. We show that the production of infective virus is impaired on cells lacking LRP1, and that purified LRP1 is a potent blocker of DENV infection. These results are consistent with LRP1 playing an important role on DENV entry, making this receptor a molecule of interest on the investigation for medical treatments of dengue/severe dengue disease.

biochemistry↗