Villenave, R. | Search bioRxiv

bioRxiv2026

Vascularized tumor organoids enable immunotherapy testing in tumor-remodeled stroma

Accurately predicting whether a new molecule will be safe and effective in humans remains a fundamental challenge in biomedical research, requiring models that can reliably inform clinical outcomes. In oncology, drug attrition rates remain disproportionately high compared to other therapeutic areas, largely due to the poor translatability of conventional preclinical models that fail to capture the complexity of human immune-tumor-stroma interactions (1, 2). Here, we describe human vascularized organoids (VascO), formed through self-assembly of an immune cell perfused capillary network and stromal compartment, interfacing with patient-derived colorectal cancer organoids that enable the combined safety and efficacy assessment of tumor-targeted immunotherapies. VascO generates functional and integrated vascular networks within tumor organoids that sustain organoid growth and recapitulate the morphology and functionality of tumor vasculature. Single-cell transcriptomics reveals that the tumor organoids-educated VascO niche reprograms otherwise healthy fibroblasts into canonical cancer-associated fibroblast states, adopting my-ofibroblastic or inflammatory signatures, and faithfully mirroring donor-specific tumor cues. In parallel, VascO tumor vessels display an expanded tip-cell program, aberrant morphology, and VEGF/ANG2-driven permeability, recapitulating defining hallmarks of tumor vasculature. Immune perfusion with a tumor-targeted T cell bispecific antibody induces dynamic immune trafficking, infiltration and engagement of effector cells with the tumor tissue, leading to its effective killing. A protease-activated variant restricts T cell activation to the tumor microenvironment, retaining potent tumoricidal activity while sparing donor-matched healthy colon organoids, and thereby improving the therapeutic index. VascO thus models a patient-specific, tumor-educated microenvironment that is amenable to assessing the combined safety and efficacy of new drug candidates and provides a scalable platform to improve the preclinical assessment of innovative cancer immunotherapies.

bioengineering↗

bioRxiv2021

Impact of charge patches on tumor disposition and biodistribution of therapeutic antibodies

This study explores the impact of antibody surface charge on tissue distribution into various tissues including tumor. Tumor-bearing mice were dosed intravenously with a mixture comprising three antibodies engineered to carry negative charge patches, a balanced charge distribution, or positive patches, respectively. Tissue levels were analyzed with a specific LC-MS/MS method. In addition, the antibody mix was administered to non-tumor bearing mice. Muscle and skin interstitial fluid were obtained by centrifugation and analyzed by LC-MS/MS. An in-vitro endothelium model was explored for its feasibility to mimic the observed distribution differences. A balanced charge distribution was optimal in terms of total tumor exposure, while in other tissues negatively charged and balanced charged antibodies gave similar results. In contrast, positive charge patches generally result in increased serum clearance but markedly enhance tumor and organ uptake, leading to higher tissue-to-serum ratios. The uptake and availability in the interstitial space were confirmed by specific assessment of antibody levels in the interstitial fluid of muscle and skin, with similar charge impact as in total tissue. The in vitro model was able to differentiate the transport propensity of this series of antibody variants. In summary, our results show the differential effects of charge patches on an antibody surface on biodistribution and tumor uptake. These insights may help in the design of molecules with biodistribution properties tailored to their purpose and an optimized safety profile.

cancer biology↗

bioRxiv2020

A Micro-Engineered Airway Lung-Chip Models Key Features of Viral-Induced Exacerbation of Asthma

RationaleViral-induced exacerbation of asthma remain a major cause of hospitalization and mortality. New human relevant models of the airways are urgently needed to understand how respiratory infections may trigger asthma attacks, and to advance treatment development. ObjectivesTo develop a new human relevant model of rhinovirus-induced asthma exacerbation that recapitulates viral infection of asthmatic airway epithelium, neutrophil transepithelial migration, and enables evaluation of immunomodulatory therapy. MethodsA micro-engineered model of fully differentiated human mucociliary airway epithelium was stimulated with IL-13 to induce a Th2-type asthmatic phenotype and infected with live human rhinovirus 16 (HRV16) to reproduce key features of viral-induced asthma exacerbation. Measurements and Main ResultsInfection with HRV16 replicated key hallmarks of the cytopathology and inflammatory responses observed in human airways. Generation of a Th2 microenvironment through exogenous IL-13 stimulation induced features of asthmatics airways, including goblet cell hyperplasia, reduction of cilia beating frequency, and endothelial activation, but did not alter rhinovirus infectivity or replication. High resolution kinetic analysis of secreted inflammatory markers revealed that IL-13 treatment altered the IL-6, IFN-{lambda}1, and CXCL10 secretion in response to HRV16. Neutrophil transepithelial migration was greatest when viral infection was combined with IL-13 treatment, while treatment with MK-7123, a CXCR2 antagonist, reduced neutrophil diapedesis in all conditions. ConclusionsThis micro-engineered Airway Lung-Chip provides a novel human-relevant platform for exploring the complex mechanisms underlying viral-induced asthma exacerbation. Our data suggest that IL-13 may impair the hosts ability to mount an appropriate and coordinated immune response to rhinovirus infection. We also show that the Airway Lung-Chip can be used to assess the efficacy of modulators of the immune response. NoteEmulate(R), Human Emulation System(R), S-1, ER-1, and ER-2 are trademarks of Emulate, Inc., and any other trademarks used herein remain with their respective holders. The technology disclosed in this document may be covered by one or more patents or patent applications, and no license to these is granted herein. You are solely responsible for determining whether you have all intellectual property rights that are necessary for your intended use of any of the disclosed materials, and whether you are required to obtain any additional intellectual property rights from a third party. Further information is available by contacting the authors. At a Glance CommentaryO_ST_ABSScientific Knowledge on the SubjectC_ST_ABSNew therapies for asthma exacerbations remain a significant unmet medical need. Development of human relevant preclinical models are needed to further elucidate the complex mechanisms underlying asthma exacerbation and investigate new therapeutic strategies. What This Study Adds to the FieldUsing a human Airway Lung-Chip model, we show here for the first time a live human rhinovirus (HRV) infection of the asthmatic epithelium that recapitulates complex features of viral-induced asthma exacerbation. The dynamic microenvironment of the chip enables the real-time study of virus infection, epithelial response, and immune cell recruitment under healthy and asthmatic conditions. The model reproduces key endpoints that have been observed in asthmatics and individuals infected with rhinovirus including the ciliated cell sloughing, altered cilia beating frequency, goblet cell hyperplasia, increased expression of adhesion molecules in microvascular endothelial cells, and inflammatory mediator release. High-resolution temporal analysis of secreted inflammatory markers enabled by dynamic sampling revealed alteration of IL-6, IFN-{lambda}1 and CXCL10 secretory phases after rhinovirus infection in an IL-13 high environment. Leveraging high-content imaging and analysis of circulating inflammatory cells, we demonstrated the efficacy of a CXCR2 antagonist to reduce adhesion, motility, and transmigration of perfused human neutrophils. Thus, this micro-engineered chip may offer a powerful addition to preclinical models for understanding mechanisms underlying asthma exacerbation pathology and developing new therapeutic strategies.

bioengineering↗