Periodic Reporting for period 1 - MiTuMi (Bioengineered Matrices Mimicking the Lung Tumor Microenvironment)
Reporting period: 2021-05-01 to 2023-04-30
Lung cancer is the primary cause of cancer-related mortality despite advances in early diagnosis and next-generation therapeutics. Each year, more people die of lung cancer than of colon, breast, and prostate cancers combined. Non-small cell lung cancer (NSCLC) comprises over 80% of total lung cancer cases and affects both smoking and non-smoking populations. There is a vital need for a better understanding of the biological mechanisms that govern the onset and progress of this grave disease. Cancer research has been mainly led by animal models and conventional two-dimensional (2D) culturing of tumor cells in order to both investigate disease-mediating signaling events and testing of therapeutics. However, animal models do not always successfully predict the course of the disease or drug responses seen in humans. 2D culturing of tumor cells on the other hand, fail to represent the 3D structure and extracellular matrix (ECM) of native organs and tissues which have crucial roles in enabling cell-cell and cell-matrix interactions. The importance of the tumor microenvironment with tumor-specific ECM components and a crowded cast of different cell types which chaperon tumor cells has been recently appreciated in the field. This calls for engineering approaches which aim to develop biomimetic and physiologically relevant preclinical models for cancer research. This action aims to determine the key aberrant aspects of the lung tumor ECM and develop tissue engineered human in vitro lung tumor models which enable investigation of their biological roles on the formation and progression of disease.
Within the scope of this project, a 3D, biomimetic model of the lung tumor microenvironment has been established. In order to develop an organotypic biomaterial for use in lung tissue engineering, native lungs were decellularized via several methods and reconstituted into cyto-compatible hydrogels. These hydrogels were demonstrated to support viability and growth of lung tumor cells, healthy bronchial epithelial cells as well as lung organoids. Then organotypic double-network hydrogels were engineered which allow tunability of mechanical properties as well as cell instructive ligands. In these models, it was shown that lung tumor growth and invasive phenotype are regulated via a synergy of mechanical and biochemical extracellular cues. The project demonstrated that the effect of tissue stiffening, a hallmark of many tumor types, is dependent on the composition of the tissue matrix. Stiffness was shown to have opposing effects on cells in organotypic matrices when compared with tumor-mimetic matrices. Furthermore, drug responses of lung tumor cells within 3D biomimetic hydrogels were investigated. The study reveals that 3D drug responses of tumor cells are significantly different than cells cultured as monolayers. Moreover, presenting instructive cues in the extracellular microenvironment that mimic tumor tissues affect drug responses of lung tumor cells in 3D. The project examined whether tumor matrices affect how tumor cells interact with other cell types in the tumor microenvironment. It was found that tumor matrix characteristics play a vital role in regulation of cellular crosstalk in tumors. Lastly, the engineered hydrogels were established as a culturing platform for patient-derived lung tumor organoids which allowed long-term culturing of organoids. During the action, one peer-reviewed publication was produced, two oral presentations at international conferences were delivered, four invited talks at seminars/workshops were given.
This highly interdisciplinary MSCA project aimed to bring together bioengineering, material science, molecular biology and translational medicine to develop reliable, physiologically relevant in vitro human disease models. These models not only serve as a tool to investigate biological mechanisms within a complexity level approaching to that of native tumors but also offers a platform for testing therapeutics in a completely human setting. The findings demonstrate the importance of developing such biomimetic microenvironments for drug testing since drug responses of tumor cells differ drastically when compared to conventional culturing strategies. Patient-derived tumor organoids are one of the highlights of current translational cancer research where cellular and genetic heterogeneity of patients can be recapitulated. The models developed in this action bring together tissue engineering approaches with organoid research and allow tuning of the extracellular microenvironment and provide a robust culturing platform for long-term organoid culturing. Overall, the project findings will be a valuable contribution to the field of personalized medicine.