Abstract
Organotypic, three-dimensional (3D) cancer models have enabled investigations of complex microtissues in increasingly realistic conditions. However, a drawback of these advanced models remains the poor biological relevance of cancer cell lines, while higher clinical significance would be obtainable with patient-derived cell cultures. Here, we describe the generation and data analysis of 3D microtissue models from patient-derived xenografts (PDX) of non-small cell lung carcinoma (NSCLC). Standard of care anti-cancer drugs were applied and the altered multicellular morphologies were captured by confocal microscopy, followed by automated image analyses to quantitatively measure phenotypic features for high-content chemosensitivity tests. The obtained image data were thresholded using a local entropy filter after which the image foreground was split into local regions, for a supervised classification into tumor or fibroblast cell types. Robust statistical methods were applied to evaluate treatment effects on growth and morphology. Both novel and existing computational approaches were compared at each step, while prioritizing high experimental throughput. Docetaxel was found to be the most effective drug that blocked both tumor growth and invasion. These effects were also validated in PDX tumors in vivo. Our research opens new avenues for high-content drug screening based on patient-derived cell cultures, and for personalized chemosensitivity testing.
Highlights
Despite some recent therapeutic advances, lung cancers remain a leading cause of cancer-related deaths
It becomes increasingly apparent that both established tumor cell lines, and standard mouse models based on such cell lines, only poorly recapitulate the morphology and heterogeneity seen in clinical lung cancer biopsies
non-small cell lung carcinoma (NSCLC) tumors from patients were cut into pieces, grown as patient-derived xenografts (PDX) tumors in mice, and cell suspensions for these tumors were directly transferred to organotypic 3D cultures
Summary
Despite some recent therapeutic advances, lung cancers remain a leading cause of cancer-related deaths. The tumor mass can be increased as PDX cultures from which subsequently a larger number of tumor cells can be isolated for culture in vitro Such PDX cultures represent natural co-cultures of human cancer and mouse fibroblast cells, which form complex epithelial tissue-like structures (organoids), surrounded by mesenchymal murine fibroblasts (the tumor stroma), when embedded in extracellular matrix (ECM). We have demonstrated that the complex, heterogeneous and dynamic interaction of tumor and stromal compartments can be simultaneously investigated by tracking their growth, morphogenesis and movement over time and space[16] Another 3D co-culture setup of human prostatic tissues was presented, where their behavior was assessed through systematic measurements of morphology and mobility[26]. These findings and recent contributions by others[27, 28], argue for the biological relevance of 3D tumor-fibroblast co-cultures in cancer research and personalized medicine, and highlight the need for further development both in the laboratory and in the analysis methodology
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