Establishment of patient-derived three-dimensional organoid culture in renal cell carcinoma

Abstract

Renal cell carcinoma is a heterogeneous kidney cancer, and over 403,000 cases were reported worldwide in 2018. Current methods for studying renal cell carcinoma are limited to two-dimensional (2D) culture of primary cell lines and patient-derived xenograft models. Numerous studies have suggested that 2D culture poorly represents the diversity, heterogeneity, and drugresistance of primary tumors. The time and cost associated with patient-derived xenograft models poses a realistic barrier to their clinical utility. As a biomimetic model, patient-derived three-dimensional (3D) organoid culture can overcome these disadvantages and bridge the gap between in vitro cell culture and in vivo patient-derived xenograft models. Here, we establish a patient-derived 3D organoid culture system for clear cell renal cell carcinoma and demonstrate the biomimetic characteristics of our model with respect to both primary kidney cancer and conventional 2D culture. Materials and Methods: Normal renal tissues and tumor tissues were collected from patients with clear cell renal cell carcinoma. The dissociated cells were cultured as conventional 2D culture and 3D organoid culture. The biomimetic characteristic of the two cultures were compared. Results: Compared with 2D culture, the 3D organoid cultures retained the characteristic lipid-rich, clear cell morphology of clear cell renal cell carcinoma. Carbonic anhydrase 9 and vimentin were validated as biomarkers of renal cell carcinoma. Expression of the two validated biomarkers was more enhanced in 3D organoid culture. Conclusions: Patient-derived 3D organoid culture retains the characteristics of renal cell carcinoma with respect to morphology and biomarker expression.

Integration of Tumor Microenvironment in Patient-Derived Organoid Models Help Define Precision Medicine of Renal Cell Carcinoma

“…Na et al established a concrete protocol of PDO culture directly from surgical resected ccRCC sample. Such PDO preserves the morphology and biomarker expression of parental tumors ( 22 ). Grassi et al also reported an approach to establishing and passaging normal kidney organoids and RCC PDO from surgically resected tissues.…”

Patient-Derived Organoids as a Model for Cancer Drug Discovery

“…Crucially, the expression of characteristic biomarkers was more likely to be retained in PDOs than two-dimensional culture models [ 54 ]. For example, the response of colon cancer organoids to the inhibition of the histone methyltransferase EZH2 was correlated to expression of the pro-apoptotic BIK gene [ 55 ].…”

Organoid As a Novel Technology for Disease Modeling

“…The lacks of conventional cancer models are microenvironment, stromal compartments, organspecific function, immune system, genetic heterogeneity (2,13,14). The promising organoid technology bridges the gap between traditional in vitro and in vivo cancer models (15). Tumoroids have been generated from tissue biopsies, surgical resections, ascitic fluid, and circulating tumor cells (16)(17)(18)(19).…”