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Pavlovic, N.

Publications and source records attributed to Pavlovic, N..

3 recordsLinked to original sources

Contact-dependent regulation of UV-B/C-induced cell fate by neighbouring intact cells

High-energy UV light from the UV-B and UV-C ranges induces severe cellular damage that leads to oxidative stress, cellular senescence, and apoptosis. Most studies of such UV-induced phenotypes have been performed in homogeneous cell cultures where all cells were subjected to comparable levels of damage. However, in physiological conditions, UV exposure generates heterogeneous cell populations in which damaged cells coexist with intact neighbors. How such cellular context influences UV-induced outcomes remains insufficiently understood. Here, we examined the effects of intact neighbouring cells on mammalian cells exposed to combined UV-B and UV-C radiation. We show that the presence of intact cells enhances apoptotic progression and clearance of cells treated with high doses of UV, while having little effect the cells exposed to low and moderate doses. Transcriptomic profiling revealed that UV-treated cells grown in co-culture with intact neighbours exhibit a markedly attenuated transcriptional response to UV exposure, including reduced activation of oxidative stress and reparatory pathways, compared to UV-treated cells grown in monoculture. These effects required direct cell-cell contact and were not mediated by diffusible factors, gap junctions, or tunneling nanotubes. Instead, co-culture conditions were associated with extensive changes in ligand-receptor gene expression profiles, indicating altered intercellular communication in response to UV damage. Our findings demonstrate that UV-induced cellular outcomes are strongly shaped by the surrounding cellular environment and identify a contact-dependent, non-cell-autonomous layer of regulation that influences the resolution of UV-induced damage. These results have implications for understanding tissue-level responses to UV exposure in photodamage, photoaging, and disease contexts.

cell biology↗

A biomimetic liver model recapitulating bio-physical properties and tumour stroma interactions in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a primary liver tumor developing in the wake of chronic liver disease. Chronic liver disease and inflammation leads to a fibrotic environment actively supporting and driving hepatocarcinogenesis. Insight into hepatocarcinogenesis in terms of the interplay between the tumor stroma micro-environment and tumor cells is thus of considerable importance. Three-dimensional (3D) cell culture models are proposed as the missing link between current in vitro 2D cell culture models and in vivo animal models. Our aim was to design a novel 3D biomimetic HCC model with accompanying fibrotic stromal compartment and vasculature. Physiologically relevant hydrogels such as collagen and fibrinogen were incorporated to mimicking the bio-physical properties of the tumor ECM. In our model LX2 and HepG2 cells embedded in a hydrogel matrix were seeded onto the inverted insert membrane of a Transwell system. HUVEC cells were then seeded onto the opposite side of the membrane. Three formulations consisting of ECM-hydrogels embedded with cells were prepared and the bio-physical properties determined by rheology. Cell viability was determined by the AlamarBlue(R) assay over 21-days. The effect of the chemotherapeutic drug doxorubicin was evaluated in both a 2D co-culture and our 3D model for a period of 72h. We show that this model is viable for 25-days and gives rise to metastatic tumor nodules after 17 days in culture. Rheology results show that bio-physical properties of a fibrotic, cirrhotic and HCC liver can be successfully mimicked. Overall, results indicate that this 3D model is more representative of the in vivo situation compared to traditional 2D cultures. Our 3D tumor model showed a decreased response to chemotherapeutics, mimicking drug resistance typically seen in HCC patients. This model could in future provide a valuable new platform to study multifocal HCC or to identify mechanisms that contribute to early stages of metastasis. SUMMARYA protocol for a novel 3D biomimetic HCC model with accompanying fibrotic stromal compartment and vasculature, to study endocrine and paracrine signaling in liver cancer. The model uses physiological relevant hydrogels in ratios mimicking the bio-physical properties of the stromal extracellular matrix, which is an active mediator of cellular interactions, tumor growth and metastasis.

cancer biology↗

Blocking IRE1a-endoribonuclease activity in hepatic stellate cells decreases tumor cell proliferation and metastasis in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a liver tumor that arises in patients with cirrhosis. Hepatic stellate cells are key players in the progression of HCC, as they create a fibrotic micro-environment and produce growth factors and cytokines that enhance tumor cell proliferation and migration. We assessed the role of endoplasmic reticulum (ER) stress in the cross-talk between stellate cells and HCC-cells. Mice with a fibrotic HCC were treated with the IRE1-inhibitor 48C, which reduced tumor burden and collagen deposition. By co-culturing HCC-cells with stellate cells, we found that HCC-cells induce ER-stress in stellate cells, thereby contributing to their activation. Inhibiting IRE1 blocked stellate cell activation, which inhibited tumor cell proliferation and migration in different in vitro 2D and 3D co-cultures. Our results suggest that IRE1 is an important mediator in the communication between stellate cells and cancer cells and components of the ER-stress pathway may be therapeutically relevant for HCC-patients. Impact statementIRE1 is an important mediator in the communication between stellate cells and cancer cells and components of the ER-stress pathway may be therapeutically relevant for liver cancer.

cancer biology↗