Perdrieau, H. | Search bioRxiv

bioRxiv2026

Oncolytic measles virus reprograms the tumor microenvironment in a vascularized mesothelioma-on-chip model

Pleural mesothelioma (PM) is a rare, aggressive cancer primarily caused by asbestos exposure and remains resistant to conventional chemotherapy. Although dual immune checkpoint inhibition (anti-PD-1/anti-CTLA-4) is now approved as first-line therapy, clinical benefit is limited to a small subset of patients, necessitating the need for alternative strategies. Oncolytic viruses (OVs) represent a promising approach as they selectively infect and lyse tumor cells while reprogramming the immunosuppressive tumor microenvironment (TME) into an immunostimulatory state. In PM, we previously showed that the attenuated Schwarz strain of measles virus (MV) oncolytic activity is mainly dependent on alterations in the type I interferon (IFN-I) pathway, rendering tumor cells sensitive to infection. Recently, we showed that monocytes/macrophages exposed to MV produce IFN-I, which protects PM cells via paracrine IFNAR signaling. This underscores the necessity of modeling the TME to accurately evaluate OV efficacy. Conventional rodent models are non-permissive to MV, and availability of fresh human PM tissue is scarce. We therefore developed a humanized 3D "vascularized mesothelioma-on-chip" (VMOC) model using microfluidic chips. It comprises two perfusable endothelial-lined parental vessels flanking a central secondary microvascular network (MVN), generated using human umbilical vein endothelial cells (HUVECs) embedded in fibrin and co-cultured alongside PM cells and primary human lung fibroblasts (hLFs). We characterized the integrity and functionality of the endothelial compartment as well as the cellular heterogeneity in VMOC using single-cell RNA sequencing. After administration of MV via the endothelial network, we observed infection and death of PM cells in addition to a strong activation of the type I interferon pathway and production of multiple inflammatory mediators. The VMOC model enables in vitro study of both MV infection and TME reprogramming, paving the way for a better understanding of the role of the TME in the response to treatment and for supporting the development of more personalized, targeted therapies for PM.

cancer biology↗

bioRxiv2024

In vitro models to mimic tumor endothelial cell-mediated immune cell reprogramming in lung adenocarcinoma

Tumor endothelial (TECs) cells play a critical role in regulating immune responses within the tumor microenvironment (TME). However, the mechanisms by which TECs modulate immune cell population remain unclear, particularly in non-small cell lung cancer (NSCLC). Here, we investigated how NSCLC cells tweak normal endothelial cells (NECs) into TECs and the subsequent effects on immune regulation. NECs were cocultured with various NSCLC cell lines, using 2D and 3D coculture models to evaluate TEC-mediated effects on immune cells. We show that direct coculture led to significant transcriptomic, proteomic and kinomic alterations in TECs, especially in pro-inflammatory pathways. We identified a downregulation of the co-stimulatory molecule OX40L in TECs compared to NECs, suggesting impaired T-cell proliferation support. While TECs showed a limited effect on CD8+ T-cell activation, TECs supported CD4 T-cells polarization into Treg and Th22 subsets. Moreover, TECs also promoted M2-like macrophages polarization, thereby potentially contributing to the TME immunosuppression. State-of-the-art single-cell RNA sequencing of 3D multicellular tumor spheroids (MCTS) revealed distinct TEC subpopulations, including an inflammatory subset with UPR signature. The latter was absent in 2D-cultured NECs but present in freshly isolated and 2D-cultured TECs from NSCLC patients. Importantly, we also identified within MCTS a perivascular M2-like macrophage subset, predicted to interact with TECs with MIF and Midkine signaling. In conclusion, TECs in NSCLC tumors play a pivotal role in remodeling the TME immune landscape by promoting immune suppression. This study highlights the complex immunoregulatory functions of TECs within our different in vitro models that mimic aspects of the TME. Our data may provide new insights into potential therapeutic strategies targeting TECs or regulatory signaling to improve the efficacy of immunotherapy in NSCLC.

cancer biology↗