Biology subjects

Heindryckx, F.

Publications and source records attributed to Heindryckx, F..

7 recordsLinked to original sources

bioRxiv2025

Irinotecan induces intestinal atrophy and compromises duodenal motility and mucosal permeability: in vivo and in vitro evaluations

PurposeIntestinal toxicity is a significant adverse effect of chemotherapy, with no effective treatments currently available. This study aimed to evaluate the effects of irinotecan (IRT) on; (i) duodenal motility and mucosal barrier function, (ii) histopathological changes along the small intestine and colon, and (iii) faecal water content, to better understand IRT-induced intestinal toxicity, and to support future treatment strategies to minimize these effects. MethodsSprague Dawley rats were anaesthetized with thiobarbiturate, and a 30-mm long segment of the proximal duodenum with an intact blood supply was perfused in situ. The effects of IRT on duodenal motility and blood-to-lumen clearance of 3H-mannitol were investigated. In separate experiments, intestinal toxicity was induced in rats by intravenous (i.v.) dosing with IRT. After 72 h post IRT administration, the rats were sacrificed and the small intestine and the colon were excised for histological analysis. Fecal water contents and change in body weight were also measured. In addition, effects of IRT on cell viability were evaluated via Caco-2 cell culture, IRT treatment, and resazurin reduction assays. ResultIntravenous dosing of IRT (60 mg/kg) significantly induced robust and sustained increase in both duodenal motility (+277%) and in mucosal paracellular permeability (+85%). Moreover, IRT in doses of 180 mg/kg and 200 mg/kg (i.p.), induced similar reduction in villus length evaluated 72 h post administration; in duodenum (-52% and -45%, respectively) and jejunum (-37% and -33%), but was without effects in ileum. IRT induced increases in jejunal crypt depth (+48% and +41%), but did not significantly influence the crypt depth in duodenum, ileum or colon. IRT induced increases in fecal water content by +19% and +16%, respectively, and caused significant body weight reductions by 10% and 13%, respectively. IRT exposing of Caco-2 cells resulted in a significant decrease in viability, and a resulting IC50 value of 1289 M. ConclusionsOur novel results show that IRT highly increases duodenal motility and impair mucosal barrier function. In addition, IRT induces diarrhoea, and demonstrates histopathological in the proximal small intestine. These findings contribute to the basic physiological understanding, and provide potentially new innovations for treatment of IRT-induced intestinal toxicity.

cancer biology↗

bioRxiv2025

A comparative study of the chemotoxic effects of idarubicin and doxorubicin in a chemically-induced in vivo mouse model and in vitro models for hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a significant clinical challenge, with limited therapeutic options. Anthracyclines such as doxorubicin (DOX) and idarubicin (IDA) are commonly used in cancer treatment but have shown variable efficacy and side effects in HCC. This study aimed to compare the cytotoxic effects of DOX and IDA on HCC in both in vivo and in vitro models, assess their impact on the tumor and its microenvironment, as well as identify potential adverse effects. In vivo, both DOX and IDA treatments led to a significant reduction in body weight and spleen-to-body weight ratio, with IDA showing a more pronounced effect. However, neither treatment significantly affected intestinal morphology, compared to untreated mice with HCC. None of the treatments had a significant impact on macroscopic or microscopic tumor burden. Notably, DOX and IDA treatments resulted in a significant reduction in collagen deposition and liver fibrosis. DOX reduced hepatic stellate cell (HSC) activation, despite having no significant impact on the expression levels of fibrotic markers TGF-{beta} and CTGF. In contrast, IDA not only increased HSC activation but also upregulated TGF-{beta} and CTGF expression in both tumor and peritumoral tissues. Molecular analysis further revealed that DOX and IDA treatments increased mRNA levels of ER-stress and proliferation markers in non-tumor tissues, with significant findings within the PERK pathway. IDA, in particular, induced higher ATF4 expression in hepatocytes and enhanced macrophage recruitment in tissue sections. While DOX and IDA exhibit limited effectiveness in reducing HCC tumor burden in vivo, the in vitro analyses showed that DOX and IDA demonstrated strong, concentration-dependent cytotoxicity, significantly reducing cell viability in all tested HCC cell lines. Increasing complexity of the in vitro models, by culturing in 3D and adding HSCs, decreased the sensitivity to the anthracyclines. This, along with the effects on liver fibrosis, stellate cell activation and inflammation seen in vivo, may be the result of the significant contribution of the tumor microenvironment in mediating drug response. The differential expression of ER-stress and proliferation markers, particularly in the PERK pathway, further highlights the complexity of the tumor microenvironments influence on treatment outcomes. More research into these molecular responses and underlying mechanisms is needed to provide insights into improving therapeutic strategies for HCC.

cancer biology↗

bioRxiv2025

CombiCTx: Screening diffusion gradients of anti-cancer drug combinations

The reduced effectiveness of chemotherapy in many patients highlights the need for novel drug combinations and optimal ratios that target multiple survival mechanisms, which tumors may engage to confer drug resistance. Dynamic conditions within the tumor microenvironment shape cell behavior and influence the response to anti-cancer drugs, varying by cell type and local context. Accordingly, assays that identify effective concentrations and drug interactions (additive, synergistic, or antagonistic) in a relevant tumor tissue model are required to discover new combination treatments. To address this need for combinatorial chemotherapeutic (CTx) screening assays, we reconfigured CombiANT, a device for testing antibiotic interactions, and present a new assay called CombiCTx. The assay uses a device with three reservoirs containing gels loaded with chemotherapeutics or other anti-cancer drugs. The drug-loaded device is inverted and placed in a standard culture dish containing cancer cells, and both are then enclosed in gel. As drugs diffuse from the reservoirs, cancer cells are exposed to overlapping dynamic gradients of anti-cancer drugs, which can interact in various ways. We imaged doxorubicin diffusion in the assay using timelapse microscopy and employed the apoptotic agent staurosporine as a model drug, and established an imaging protocol for quantifying MDA-MB-231 breast cancer cell apoptosis along drug diffusion gradients. Finally, we tested navitoclax and gemcitabine to demonstrate the capacity of CombiCTx to evaluate combined cytotoxic effects while accounting for drug diffusion. Evaluating drug combinations with an assay that accounts for drug properties that influence diffusion in a complex tumor matrix may provide clinically relevant information.

cancer biology↗

bioRxiv2021

Anthracyclins increase free PUFAs and etherPEs with PUFAs as potential hallmarks of lipid peroxidation and ferroptosis

Metabolic and personalized interventions in cancer treatment require a better under-standing of the relationship between the induction of cell death and metabolism. Consequently, we treated three primary liver cancer cell lines with two anthracyclins (doxorubicin and idarubin) and studied the changes of the lipidome. We found that both anthracyclins in the three cell lines increased the levels of polyunsaturated fatty acids (PUFAs) and alkylacylglycerophosphoethano-lamines (etherPEs) with PUFAs. As PUFAs and alkylacylglycerophospholipids with PUFAs are fundamental in lipid peroxidation during ferroptotic cell death, our results suggests supplementa-tion with PUFAs and/or etherPEs with PUFAs as a potential general adjuvant of anthracyclins. In contrast, neither the markers of de novo lipogenesis nor cholesterol lipids presented the same trend in all cell lines and treatments. In agreement with previous research, this suggests that modulation of the metabolism of cholesterol could be considered a specific adjuvant of anthracyclins depend-ing on the type of tumor and the individual. Finally, we discuss the changes in the lipidome in re-lation to the endoplasmic reticulum stress and the sensitivity to anthracyclins of the different cells. In conclusion, our results suggest that the modulation of different lipid metabolic pathways may be considered for generalized and personalized metabochemotherapies.

cancer biology↗

bioRxiv2020

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↗

bioRxiv2019

Fibrin fragment E potentiates TGF-β-induced myofibroblast activation and recruitment

BackgroundFibrin is an essential constituent of the coagulation cascade, and the formation of hemostatic fibrin clots is central to wound healing. Fibrin clots are over time degraded into fibrin degradation products as the injured tissue is replaced by granulation tissue.\n\nObjectivesstudy the role of the fibrin degradation product fragment E (FnE) in fibroblast activation and migration.\n\nMethodsRat kidney fibroblasts (NRK-49F), human fetal lung fibroblasts (HFL1) and immortalized human foreskin fibroblasts (BJ) were exposed to FnE and TGF-{beta}. Fibroblast activation was measured via quantitative RT-PCR and western blot for -SMA and Collagen I1. A microfluidic chemotaxis assay was used to study directional migration. To investigate if FnE interacts with integrins, direct binding experiments were performed using surface plasmon resonance biosensor technology. Infusion pumps releasing FnE were implanted subcutaneously in mice and profibrotic effects of FnE were analyzed using immunohistochemical staining of the wound area.\n\nResultsWe present evidence that FnE is a chemoattractant for fibroblasts and that FnE can potentiate TGF-{beta}-induced myofibroblast formation. FnE forms a stable complex with V{beta}3 integrin, and the integrin {beta}3 subunit is required both for FnE-induced fibroblast migration and for potentiation of TGF-{beta}-induced myofibroblast formation. Finally, subcutaneous infusion of FnE in mice results in a fibrotic response in the hypodermis. These results support a model where FnE released from clots in wounded tissue promote wound healing and fibrosis by both recruitment and activation of fibroblasts. Fibrin fragment E could thus represent a therapeutic target for treatment of pathological fibrosis.\n\nEssentials- Fibrin is an essential constituent of the coagulation cascade, and fibrin clotting is central to wound healing.\n- Fibrin clots are over time degraded into fibrin degradation products, including fibrin fragment E (FnE).\n- FnE is a chemoattractant for fibroblasts and can potentiate TGF-{beta}-induced myofibroblast formation.\n- FnE released from clots in wounded tissue could promote wound healing and fibrosis.

cell biology↗

bioRxiv2019

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↗