Cell Biology, Cancer Research, Biomaterials, Transplantation
108
Scopus Publications
Scopus Publications
Modeling hepatocellular carcinoma and its microenvironment on a chip Orsola Mocellin, Stéphane Treillard, Abbie Robinson, Aleksandra Olczyk, Thomas Olivier, et al. Cell Death Discovery, 2026 Hepatocellular carcinoma (HCC) is the most common type of liver cancer. Its incidence is increasing and is closely related to advanced liver disease. Interactions in the HCC microenvironment between tumor cells and the associated stroma actively regulate tumor initiation, progression, metastasis, and therapy response. Effective drug development increasingly requires advanced models that can be utilized in the earliest stages of compound and target discovery. Here we report a phenotypic screen on an advanced HCC patient-derived chip (PDChip) model. The vascularized HCC PDChip models include relevant cellular players of the HCC microenvironment. We assessed the effect of 28 treatment conditions on a panel of 8 primary HCC tumors and 2 cell lines. Approximately 1200 HCC PDchips were grown under perfusion flow, exposed to treatments, and subsequently assessed for viability, tumor-associated vasculature responses and chemokine and cytokine changes. Although the SoC therapeutics sorafenib and lenvatinib reduced culture viability and produced profound changes in the organization of the vascular beds, they did not affect the tumor cell population in these cultures. Atorvastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor, reduced PDChips viability but did not affect vascular bed organization. Sorafenib, lenvatinib and atorvastatin also affected chemokine and cytokine release. Tocilizumab, galunisertib, and vactosertib decreased the level of IL6, a relevant prognostic marker for HCC, while IL6 was increased by halofuginone. In conclusion, HCC PDChip models enabled a detailed evaluation of drug-induced responses in the tumor and associated microenvironment, highlighting their importance in preclinical research for understanding diseases and developing new drugs.
Beyond Decellularization: Remnant Mitochondrial DNA Can Act as Hidden Damage-Associated Molecular Pattern Elena V. A. van Hengel, Kuan Liu, Henk P. Roest, Jorke Willemse, Kimberley Ober-Vliegen, et al. Bioengineering, 2026 Tissue decellularization aims to obtain bioscaffolds for regenerative applications by removing all cellular components while preserving the extracellular matrix (ECM) architecture. Although decellularization removes the majority of linear nuclear DNA (nDNA), residual amounts remain detectable. However, the fate of circular mitochondrial DNA (mtDNA) after decellularization has not yet been reported. Cell death or injury can cause the release of mtDNA, which is resistant to breakdown by exonucleases. Extracellular mtDNA acts as a damage-associated molecular pattern (DAMP) that can trigger immune responses. The aim of this study is to assess the presence of residual mtDNA in the liver, bile duct, and vascular scaffolds after decellularization and whether this causes inflammatory responses in macrophages. Decellularized tissues showed a marked reduction in total DNA content well below the threshold of 50 ng/mg tissue. However, in liver and vascular scaffolds, a relative increase in the mtDNA:nDNA ratio was detected in the remnant DNA fraction. Residual mtDNA in bioscaffolds acted as DAMPs causing macrophage activation, as shown by increased cell proliferation and cytokine production. Strategies to further reduce remnant mtDNA were tested. We found that treatment with the endonuclease enzyme HpaII was effective in degrading residual mtDNA. Importantly, mtDNA removal resulted in a significantly reduced macrophage activation. In conclusion, our study shows that mtDNA is relatively resistant to the decellularization procedure and can act as a DAMP in bioscaffolds. This underscores the importance of removing mtDNA from decellularized bioscaffolds to improve the immunocompatibility for biomedical applications.
Removal of Porcine Endogenous Retroviruses in Decellularized Liver Bioscaffolds Elena V. A. van Hengel, Dubravka Drabek, Henk. P. Roest, Jorke Willemse, Lieve J. Reniers, et al. Xenotransplantation, 2025 Tissue engineering using decellularized liver scaffolds presents a promising approach in regenerative medicine, offering a potential alternative to donor organ transplantation. The use of human livers as a bioscaffold is restricted by their limited availability and quality. Porcine livers offer an alternative due to their anatomical and physiological similarities to human livers. However, applying porcine‐derived biomaterials in a clinical setting poses a risk of pathogen transmission, which is a noteworthy concern. Porcine endogenous retroviruses (PERVs), which are integrated into the genome of all pig breeds, are of particular concern, as subclasses PERV‐A and PERV‐B have shown to infect human cells in vitro. It is therefore essential to effectively remove all PERVs when manufacturing porcine scaffolds. In this study, we assessed the presence of PERV‐specific DNA, RNA, and protein in decellularized porcine livers. Our findings prove that genetic and protein PERV material was effectively removed from porcine livers during our decellularization procedure. This finding substantiates the potential of using decellularized scaffolds of porcine origin for clinical applications without risk of PERV transmission.
Apoptosis regulators of the Bcl-2 family play a key role in chemoresistance of cholangiocarcinoma organoids Wunan Mi, Gilles S. van Tienderen, Shaojun Shi, Amy Broeders, Kathryn Monfils, et al. International Journal of Cancer, 2025 Cholangiocarcinoma (CCA) is a rare but devastating liver cancer which is commonly diagnosed at a late stage and often resistant to chemotherapy. Bcl‐2 family members, which control apoptotic cell death, are known to be involved in the chemoresistance of some cancer types. This study investigated the role of Bcl‐2 family members in the chemoresistance of cholangiocarcinoma organoids (CCAOs) in both undifferentiated and matured branching phenotypes (BRCCAOs). Patient‐derived CCAOs and BRCCAOs were cultured to assess chemoresistance to an FDA‐approved anticancer drug panel by testing cell viability using ATP quantification and apoptotic cell death by cleaved caspase 3 staining. More specifically, sensitivity to the first‐line drug gemcitabine was tested in combination with Bcl‐2 family inhibitors or activators. We found that in gemcitabine‐resistant CCAOs, inhibition of Bcl‐xl could overcome gemcitabine resistance and induce apoptotic cell death. Although inhibition of Mcl‐1 or activation of Bax induced spontaneous cell death, this could not overcome gemcitabine resistance. The BRCCAOs, which mimic tumor architecture better than CCAOs, show broader chemoresistance to anticancer drugs. Of note, in the resistant BRCCAOs, Bcl‐xl inhibition could restore gemcitabine sensitivity. In conclusion, this study shows that targeting Bcl‐xl can overcome chemoresistance to gemcitabine in CCA organoids. CCAOs and BRCCAOs provide good preclinical models for testing new drug combinations and assessing personalized therapeutic approaches.
Optimizing the Infusion Route of Human Bone Marrow Mesenchymal Stromal Cells to Mitigate Liver Ischemia–Reperfusion Injury in a Porcine Model Stefan H. Luijmes, Job P. van Kooten, Henk P. Roest, Jubi de Haan, Michail Doukas, et al. Cells, 2025 Mesenchymal stromal cells (MSC) have been shown to mitigate IRI through their anti-inflammatory and immune-modulating capacities. This study aims to demonstrate the feasibility, safety, and effectiveness of hepatic administration of bone marrow-derived (BM)-MSCs in a large pig model relevant to human anatomy. After complete vascular exclusion for 45 min, 3 × 106 human BM-MSCs/kg body weight were infused via the portal vein or hepatic artery. BM-MSC infusion did not cause obstruction of hepatic or pulmonary blood flow within 6 h after infusion. Cells were effectively retained in the liver, being undetectable in peripheral blood, lung, and spleen samples. Human B2M expression, as a marker for BM-MSC presence, was significantly higher for the left liver lobe in arterial infusion compared to portal infusion. In liver samples with high BM-MSC levels, we identified the prevention of up- or downregulation of some genes related to inflammation and energy metabolism that was present in non-treated control samples, indicating biological effects within 6 h of infusion. We conclude that hepatic BM-MSC infusion is feasible and safe, with the hepatic artery serving as the optimal administration route for homogenous distribution. These findings pave the way for clinical studies on MSC infusion in IRI, either in situ in liver conditions or ex situ during machine perfusion.
Hemopoëtische stamcellen (HSC) uit navelstrengbloed; bevorderen van uitgroei in SCID-muizen door middel van macrofagendepletie Tijdschrift Voor Kindergeneeskunde, 1996
