Integrating Tumor and Healthy Epithelium in a Microphysiology Multi-compartment Approach to Study Renal Cell Carcinoma Pathophysiology
The advent of micro-physiological systems (MPS) in biomedical research has enabled the introduction of more complex and relevant physiological into in vitro models. The recreation of complex morphological features in three-dimensional environments can recapitulate otherwise absent dynamic interactions in conventional models. In this study we developed an advanced in vitro Renal Cell Carcinoma (RCC) that mimics the interplay between healthy and malignant renal tissue. Based on the TissUse Humimic platform our model combines healthy renal proximal tubule epithelial cells (RPTEC) and RCC. Co-culturing reconstructed RPTEC tubules with RCC spheroids in a closed micro-perfused circuit resulted in significant phenotypical changes to the tubules. Expression of immune factors revealed that interleukin-8 (IL-8) and tumor necrosis factor-alfa (TNF-a) were upregulated in the non-malignant cells while neutrophil gelatinase-associated lipocalin (NGAL) was downregulated in both RCC and RPTEC. Metabolic analysis revealed that RCC prompted a shift in the energy production of RPTEC tubules, inducing glycolysis, in a metabolic adaptation that likely supports RCC growth and immunogenicity. In contrast, RCC maintained stable metabolic activity, emphasizing their resilience to external factors. RNA-seq and biological process analysis of primary RTPTEC tubules demonstrated that the 3D tubular architecture and MPS conditions reverted cells to a predominant oxidative phosphorylate state, a departure from the glycolytic metabolism observed in 2D culture. This dynamic RCC co-culture model, approximates the physiology of healthy renal tubules to that of RCC, providing new insights into tumor-host interactions. Our approach can show that an RCC-MPS can expand the complexity and scope of pathophysiology and biomarker studies in kidney cancer research. Overall design: This study aimed to reconstruct renal tubules using RPTEC_TERT1 cells embedded in a collagen type I matrix and to generate RCC spheroids employing CAKI-1 cells in an agar/collagen matrix within the Humimic chip2 Microphysiology System (MPS). Additionally, the impact of dynamic culture on RPTEC physiology was assessed by culturing primary RPTEC cells in both conventional 2D formats and the MPS without RCC spheroids. The experiment included three conditions: (1) Renal tubules only, (2) RCC spheroids only, and (3) Renal tubules and RCC spheroids in co-culture, all cultured in the MPS for 5 days. Cell and supernatant samples were collected from each condition for analysis. The anticipated outcomes include successful renal tubule reconstruction, RCC spheroid formation, and insights into the influence of dynamic culture on RPTEC physiology.