Biomedicinahttps://hdl.handle.net/2072/4787802026-04-08T16:22:02Z2026-04-08T16:22:02ZIdentification of a crosstalk between ClC-1 C-terminal CBS domains and the transmembrane regionGaitán-Peñas, HéctorPérez González, Anna Priscil·laGonzález Subías, MarcZdebik, Anselm A.Gasull Casanova, XavierBuey, Ruben M.Errasti-Murugarren, EkaitzEstévez Povedano, Raúlhttps://hdl.handle.net/2445/2285312026-03-27T14:42:11Z2026-03-26T13:55:39ZIdentification of a crosstalk between ClC-1 C-terminal CBS domains and the transmembrane region
Gaitán-Peñas, Héctor; Pérez González, Anna Priscil·la; González Subías, Marc; Zdebik, Anselm A.; Gasull Casanova, Xavier; Buey, Ruben M.; Errasti-Murugarren, Ekaitz; Estévez Povedano, Raúl
CLC channels and transporters have large C-terminal regions which contain two cystathionine β-synthetase (CBS) domains. It has been hypothesized that conformational changes in these domains upon nucleotide binding modulate the gating of the CLC dimer. It is not clear how rearrangements that occur in the CBS domains are transmitted to the ion pathway, as CBS domains interact with the rest of the channel at multiple locations and some of these sites are not visible in recent solved cryogenic electron microscopy structures or are difficult to model using the AlphaFold server. Using ClC-1 as a model, we started working with a described ClC-1 mutation (H835R) located in the first alpha helix of the CBS2 domain which changes the voltage dependence of gating. We then identified several residues located in the disorganized loop after helix R (R-linker) that revert the phenotype of this mutation. We additionally proved that R-linker's function is connected to the CBS2 domain as current intensity, plasma membrane levels and gating defects of several R-linker variants were corrected by adding the mutation H835R. Furthermore, cross-linking studies using newly developed split-cysless ClC-1 channels containing specific cysteine mutants in the R-linker and the CBS2 domain indicate that these two regions are in close contact. Considering these new results, we propose that conformational changes occurring in the CBS domains could be transmitted to the CLC intracellular chloride binding site by means of its interaction with the R-linker.
2026-03-26T13:55:39ZHuman microglia-like cells differentiated from monocytes with GM-CSF and IL-34 show phagocytosis of α-synuclein aggregates and C/EBPβ-dependent proinflammatory activationLlaves López, AndreaMicoli, EliaBelmonte Mateos, CarlaAguilar, GerardAlba, ClaraMarsal, AnaisPulido Salgado, MartaRabaneda Lombarte, NeusSolà i Subirana, CarmeSerratosa i Serdà, JoanVidal Taboada, José ManuelSaura Martí, Josephttps://hdl.handle.net/2445/2268382026-02-13T08:53:33Z2026-02-12T19:09:22ZHuman microglia-like cells differentiated from monocytes with GM-CSF and IL-34 show phagocytosis of α-synuclein aggregates and C/EBPβ-dependent proinflammatory activation
Llaves López, Andrea; Micoli, Elia; Belmonte Mateos, Carla; Aguilar, Gerard; Alba, Clara; Marsal, Anais; Pulido Salgado, Marta; Rabaneda Lombarte, Neus; Solà i Subirana, Carme; Serratosa i Serdà, Joan; Vidal Taboada, José Manuel; Saura Martí, Josep
Microglia, the main resident immune cells in the central nervous system, are implicated in the pathogenesis of various neurological disorders. Much of our knowledge on microglial biology was obtained using rodent microglial cultures. To understand the role of microglia in human disease, reliable in vitro models of human microglia are necessary. Monocyte-derived microglia-like cells (MDMi) are a promising approach. This study aimed to characterize MDMi cells generated from adult human monocytes using granulocyte–macrophage colony-stimulating factor and interleukin-34. To this end, 49 independent cultures of MDMI were prepared, and various methodological and functional studies were performed. We show that with this protocol, adult human monocytes develop into microglia-like cells, a coating is unnecessary, and high cell density seeding is preferable. When compared to monocytes, MDMi upregulate the expression of many, but not all, microglial markers, indicating that, although these cells display a microglia-like phenotype, they cannot be considered bona fide human microglia. At the functional level, MDMi phagocytose α-synuclein aggregates and responds to lipopolysaccharide (LPS) by nuclear translocation of the transcription factor nuclear factor-kappaB (NFkappaB) and the upregulation of proinflammatory genes. Finally, a long-lasting silencing of the transcription factor CCAAT/enhancer protein β (C/EBPβ) was achieved by small interfering RNA, resulting in the subsequent downregulation of proinflammatory genes. This supports the hypothesis that C/EBPβ plays a key role in proinflammatory gene program activation in human microglia. Altogether, this study sheds new light on the properties of MDMi cells and supports these cells as a promising in vitro model for studying adult human microglia–like cells.
2026-02-12T19:09:22ZAndronet News. Winter 2022/2023Rajpert-De Meyts, Ewahttps://hdl.handle.net/2445/2242322025-11-19T22:14:52Z2025-11-10T12:24:31ZAndronet News. Winter 2022/2023
Rajpert-De Meyts, Ewa
After the Holiday Season at the end of 2022, followed by January
‘hibernation’, we are beginning to work towards the objectives of
our Action
2025-11-10T12:24:31ZIdentification of Dhx15 as a Major Regulator of Liver Development, Regeneration, and Tumor Growth in Zebrafish and MicePortolés, IreneRibera, JordiFernández Galán, EstherLecue Costas, ElenaCasals Mercadal, GregoriMelgar Lesmes, PedroFernández Varo, GuillermoBoix i Ferrero, LoretoSanduzzi Zamparelli, MarcoAishwarya, VeenuReig, MaríaJiménez Povedano, WladimiroMorales Ruiz, Manuelhttps://hdl.handle.net/2445/2227922025-11-19T19:08:04Z2025-08-27T10:41:24ZIdentification of Dhx15 as a Major Regulator of Liver Development, Regeneration, and Tumor Growth in Zebrafish and Mice
Portolés, Irene; Ribera, Jordi; Fernández Galán, Esther; Lecue Costas, Elena; Casals Mercadal, Gregori; Melgar Lesmes, Pedro; Fernández Varo, Guillermo; Boix i Ferrero, Loreto; Sanduzzi Zamparelli, Marco; Aishwarya, Veenu; Reig, María; Jiménez Povedano, Wladimiro; Morales Ruiz, Manuel
RNA helicase DHX15 plays a significant role in vasculature development and lung metastasis in vertebrates. In addition, several studies have demonstrated the overexpression of DHX15 in the context of hepatocellular carcinoma. Therefore, we hypothesized that this helicase may play a significant role in liver regeneration, physiology, and pathology. Dhx15 gene deficiency was generated by CRISPR/Cas9 in zebrafish and by TALEN-RNA in mice. AUM Antisense-Oligonucleotides were used to silence Dhx15 in wild-type mice. The hepatocellular carcinoma tumor induction model was generated by subcutaneous injection of Hepa 1-6 cells. Homozygous Dhx15 gene deficiency was lethal in zebrafish and mouse embryos. Dhx15 gene deficiency impaired liver organogenesis in zebrafish embryos and liver regeneration after partial hepatectomy in mice. Also, heterozygous mice presented decreased number and size of liver metastasis after Hepa 1-6 cells injection compared to wild-type mice. Dhx15 gene silencing with AUM Antisense-Oligonucleotides in wild-type mice resulted in 80% reduced expression in the liver and a significant reduction in other major organs. In addition, Dhx15 gene silencing significantly hindered primary tumor growth in the hepatocellular carcinoma experimental model. Regarding the potential use of DHX15 as a diagnostic marker for liver disease, patients with hepatocellular carcinoma showed increased levels of DHX15 in blood samples compared with subjects without hepatic affectation. In conclusion, Dhx15 is a key regulator of liver physiology and organogenesis, is increased in the blood of cirrhotic and hepatocellular carcinoma patients, and plays a key role in controlling hepatocellular carcinoma tumor growth and expansion in experimental models.
2025-08-27T10:41:24Z