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Dieguez, L.

Publications and source records attributed to Dieguez, L..

3 recordsLinked to original sources

Engineering a soft tumor microenvironment: fibrin-enriched hydrogel promotes cancer cell invasion in a 3D bioprinted colorectal cancer model

Accurately modeling the tumor microenvironment is crucial for advancing our understanding of colorectal cancer (CRC) progression and therapeutic response. Three-dimensional (3D) hydrogel-based models that mimic the mechanical properties of native tissue serve as a valuable tool for studying tumor-stromal interactions and tumor invasion in vitro. In this work, we developed a 3D bioprinted CRC model by embedding spheroids and human intestinal fibroblasts (HIFs) within the GelMA-PEGDA and GelMA-PEGDA-Fibrin hydrogels to assess the effect of matrix composition and stiffness on spheroid invasiveness. The addition of fibrin resulted in a softer, more viscoelastic hydrogel that promoted fibroblast migration, elongation, and alignment, facilitating more dynamic tumor-stromal interactions. Moreover, both epithelial-like HT29 and mesenchymal-like SW480 spheroids showed more invasive behavior in GelMA-PEGDA-Fibrin hydrogel. This was reflected in distinct phenotypic responses. HT29 spheroids demonstrated greater growth, irregular morphology, and more interaction with elongated fibroblasts, whereas SW480 spheroids exhibited partial dissociation and disruption with a higher number of dispersed cells in GelMA-PEGDA-Fibrin hydrogel. These findings demonstrate the role of matrix softness in promoting the invasiveness of colorectal cancer. Overall, our results highlight the potential of fibrin-enriched soft hydrogel to mimic key features of the tumor microenvironment, offering a powerful tool for studying CRC invasion dynamics and supporting future applications in drug screening and personalized medicine.

bioengineering↗

Single cell-derived spheroids for real-time growth and metabolomic studies in breast cancer

Breast cancer remains a leading cause of cancer-related mortality, with disease progression and metastasis posing significant challenges in treatment. Three-dimensional (3D) cancer models have emerged as valuable tools for studying cancer cell biology in a physiologically relevant microenvironment. Studying the tumour heterogeneity and metabolic adaptations at the single-cell level can be crucial to identify factors driving metastatic progression. Here, we present a novel approach to generate single cell-derived breast cancer spheroids using cell lines (MCF-7 and MCF-10A) within a decellularised adipose tissue extracellular matrix (adECM). Spheroid culture conditions were optimised with integrated plasmonic nanosensors (gold nanostars - GNSs), to enable real-time surface-enhanced Raman scattering (SERS)-based measurements. Our results demonstrated that spheroid growth kinetics and viability in adECM were comparable to commonly used animal-derived matrices, validating its use as a reproducible ECM hydrogel. We further show that the concentration of plasmonic nanosensors used was compatible with cell culture and enabled SERS detection of a model reporter, paving the way for label-free, non-destructive analysis of cancer cell metabolism. This platform offers a promising approach to study cancer progression, including metabolic adaptations, with potential applications in biomarker discovery and preclinical research.

cancer biology↗

Investigating Short-Chain Fatty Acids Effects on Extracellular Vesicles Production in Colorectal Cancer

Colorectal cancer (CRC) is the third most diagnosed and the second leading cause of cancer-related deaths globally, often due to late detection and limited treatment options. Recent studies have linked alterations in gut microbiota to CRC, particularly emphasizing the role of short-chain fatty acids (SCFAs) like acetate, propionate, and butyrate in shaping the tumor microenvironment (TME). SCFAs contribute to CRC pathogenesis by inducing lysosomal membrane permeabilization, cell cycle arrest, and apoptosis in cancer cells. Extracellular vesicles (EVs) are membrane-bound vesicles that facilitate intercellular communication and have gained attention as promising non-invasive biomarkers for cancer diagnosis and treatment monitoring. EVs participate in cellular response mechanisms to external stimuli by transferring proteins, lipids, and nucleic acids between cells, thus modulating target cell behavior and promoting coordinated responses to stress and environmental challenges. This process is essential for cellular adaptation and plays a significant role in pathophysiological processes, including tumor progression and immune modulation, making EVs highly relevant in clinical research. This study examined the impact of SCFAs on EV production and phenotype in CRC cells. The results indicated a notable increase in EV-sized particles following SCFA treatment of colorectal cell lines, particularly in the SW480 CRC cell line. For CRC cell lines, while co-precipitated protein levels remained stable, there was a slight decrease in cellular DNA and an increase in EV-associated DNA. KRAS-mutant SW480 cells exhibited the most pronounced response, emphasizing their heightened sensitivity to SCFA. Notably, microsatellite instability - a key biomarker for immunotherapy in CRC - was detected in both small and large EV populations from BRAF-mutant RKO cells after SCFA treatment, even at low DNA concentrations. These findings underscore the potential of EVs for non-invasive detection of molecular markers, paving the way for further exploration of their role in precision oncology. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=100 SRC="FIGDIR/small/620636v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@b016a4org.highwire.dtl.DTLVardef@9e75a5org.highwire.dtl.DTLVardef@13dccfcorg.highwire.dtl.DTLVardef@5cdf15_HPS_FORMAT_FIGEXP M_FIG Graphical Abstract. Effects of SCFA on EV production and characteristics in CRC cells. Treatment with SCFA led to a significant increase in the number of EV-sized particles, a decrease in cellular DNA and a corresponding increase in EV-DNA. This study also identified MSI in both s-EV and L-EV, even following SCFA treatment and at low DNA concentrations. Created using BioRender. C_FIG

cancer biology↗