Biology subjects

Unciti-Broceta, A.

Publications and source records attributed to Unciti-Broceta, A..

5 recordsLinked to original sources

bioRxiv2026

Molidustat Targets a Synthetic Lethal Vulnerability in APC-Mutant Colorectal Cancer through GSTP1 and PHD2 Co-Inhibition

Mutations in the adenomatous polyposis coli (APC) gene are a defining feature of colorectal cancer (CRC) and impose metabolic and stress-adaptation requirements that may create exploitable vulnerabilities. Prolyl hydroxylase domain (PHD) inhibitors have been explored as therapeutic agents in CRC, however, their mechanisms of action and off-target effects remain elusive. Serendipitously, we found that Molidustat, a PHD2 inhibitor, induced cell death in APC mutant CRC cells. Ablation of PHD2 alone did not affect cell viability, suggesting an off-target mechanism. Using thermal proteome profiling and chemical proteomics, we identify glutathione S-transferase P1 (GSTP1) as a previously unrecognised off-target of Molidustat and demonstrate direct inhibition of its enzymatic activity. Genetic ablation of PHD2 alone did not phenocopy the cytotoxic effects of Molidustat, whereas combined loss of PHD2 and GSTP1 induced synergistic proteomic changes associated with cell-cycle suppression and apoptotic signalling. Integrated proteomic and metabolomic analyses further revealed energetic and metabolic perturbations specific to simultaneous GSTP1 and PHD2 loss. Consistent with these findings, APC-mutant colonic organoids displayed selective sensitivity to Molidustat that was not reproduced by hydroxylase inhibition alone, supporting a synthetic lethal interaction between GSTP1 and PHD2 in APC-mutant contexts. Together, these results identify a functional interaction between GSTP1 and PHD2 in a subset of colorectal cancer and suggest that off-target engagement of GSTP1 contributes to the anti-tumour activity of Molidustat.

cancer biology↗

bioRxiv2025

The c-Src inhibitor eCF506 diminishes opioid tolerance creating bias against β-arrestin2 recruitment

Opioids reduce severe pain, but persistent use is compromised by tolerance, attenuated by either {beta}-arrestin2 depletion, prompting development of biased opioids limited by partial efficacy, or c-Src kinase inhibitors, potentially acting through off-target effects. We tested eCF506, a conformationally selective c-Src inhibitor, on morphine antinociception and examined its effect on receptor signaling and {beta}-arrestin2 recruitment. Oral eCF506 inhibited morphine tolerance in C57BL/6J mice. Exposure of PathHunter CHO cells to eCF506 did not affect inhibition of cAMP accumulation by the agonist, DAMGO, but reduced {beta}-arrestin2 recruitment. This effect, mimicked by targeted degradation of c-Src, occurred through inhibition of c-Src catalytic function as evidenced by its diminution by the catalytically inactive Src250-536(K298M) construct. This mutant also restricted the effect of c-Src inhibitors on {beta}-arrestin2 recruitment. eCF506 additionally increased surface expression of receptors and limited their internalization by endomorphin-2 but did not alter DAMGO-evoked GRK-mediated receptor phosphorylation. These findings suggest that eCF506 prolongs opioid antinociception by inducing signalling bias, diminishing {beta}-arrestin2-mediated receptor regulation.

neuroscience↗

bioRxiv2025

Tissue scarring provides a biomechanical framework to promote mammalian bile duct regeneration through the activation of integrin-SRC/FAK signalling.

Following chronic injury, the adult mammalian bile duct regenerates by forming new branches, essentially replumbing the ductular system to overcome blockages and breaks. To regenerate effectively, biliary epithelial cells (BECs) receive a range of pro-mitogenic signals from myofibroblasts, which concurrently deposit a collagen-rich scar around the duct as it regrows. Despite epithelial regeneration and scarring occurring side-by-side, whether the deposition of scar tissue regulates ductular regeneration per se remains unclear. By inducing ductular fibrosis and regeneration in vivo, we show that the formation of collagen-I-rich scars around regenerating ducts changes the local biomechanical properties of these tissues, promoting the growth of ducts. Critically, this changing structural landscape is perceived by a spatially restricted population of biliary epithelial cells which forms a "leading-tip" of integrin-2-high cells. This leading-tip undergoes partial-EMT-type reprogramming, allowing it to become migratory and coordinate ductular regeneration. We show that this process is directly driven through an integrin-2-SRC/FAK signalling axis; thereby connecting epithelial regeneration directly to the changing fibrotic environment in chronic ductular disease. HighlightsO_LIChronic liver disease results in the formation of stiff, collagen scars around ducts. C_LIO_LINew ducts acquire high levels of integrin-2 which is spatially localised to a "leading-tip", which loses epithelial features. C_LIO_LIIntegrin-2{beta}1-SRC/FAK signalling regulates ductular migration by linking changes in the bio-structural composition of the liver with ductular cells. C_LI

cell biology↗

bioRxiv2025

Identification of drug candidates against glioblastoma with machine learning and high-throughput screening of heterogeneous cellular models

Glioblastoma multiforme (GBM) is an aggressive primary brain tumour that presents significant treatment challenges due to its complex pathology and heterogeneity. The lack of validated molecular targets is a major obstacle for discovering new therapeutic candidates, with no new effective GBM therapies delivered to patients in over two decades. Here, we report the identification of compounds that target the GBM stem cell survival phenotype. Our approach employs machine learning (ML) predictors of cell survival trained on high-throughput, image-based, phenotypic screening data for 3,561 compounds, at multiple concentrations, across a panel of six heterogeneous, patient-derived, GBM stem cell lines. We computationally screened more than 12,000 compounds spanning various chemical classes. Experimental validation of ML-identified candidates across the GBM stem cell lines led to the identification of three compounds with activity against the GBM phenotype. Notably, one of our validated hits, the Hsp90 inhibitor XL888, displayed targeted elimination of all six GBM stem cell lines with IC50 in the nanomolar range. The other two compounds, which displayed broad activity across multiple GBM cell lines with distinct cell line sensitivities, offer routes for future personalised medicine campaigns. Our work demonstrates the use of phenotypic screening in tandem with ML can effectively identify therapeutic leads for personalised treatments in highly heterogeneous indications with few known molecular targets.

cancer biology↗

bioRxiv2023

Single-cell morphological tracking of liver cell states to identify small-molecule modulators of liver differentiation

Alternative therapeutic strategies are urgently required to treat liver disease, which is responsible for 2 million deaths anually. By combining Cell Painting, a morphological profiling assay that captures diverse cellular states, with the bi-potent HepaRG(R) liver progenitor cell line, we have developed a high-throughput, single-cell technique, to track liver cell fate and map small-molecule induced changes using a morphological atlas of bi-lineage liver cell differentiation. To our knowledge this is the first-time single-cell trajectory inference has been applied to image-based Cell Painting data and leveraged for drug screening. The overarching goal of this new method is to aid research into understanding liver cell regeneration mechanisms and facilitate the development of cell-based and small-molecule therapies. Using this approach, we have identified a class of small-molecule SRC family kinase inhibitors that promote differentiation of HepaRG(R) single-cells towards the hepatocyte-like lineage and promotes differentiation of primary human hepatic progenitor cells towards a hepatocyte-like phenotype in vitro.

cell biology↗