Evaluation of AMG510 Therapy on KRAS-Mutant Non–Small Cell Lung Cancer and Colorectal Cancer Cell Using a 3D Invasive Tumor Spheroid System under Normoxia and Hypoxia

Abstract

A 3D tumor spheroid has been increasingly applied in pharmaceutical development for its simulation of the tumor structure and microenvironment. The embedded-culture of a tumor spheroid within a hydrogel microenvironment could help to improve the mimicking of in vivo cell growth and the development of 3D models for tumor invasiveness evaluation, which could enhance its drug efficiency prediction together with cell viability detection. NCI-H23 spheroids and CT-26 spheroids, from a non–small cell lung cancer and colorectal cancer cell line, respectively, together with extracellular matrix were generated for evaluating their sensitivity to AMG510 (a KRASG12C inhibitor) under normoxia and hypoxia conditions, which were created by an on-stage environmental chamber. Results demonstrated that NCI-H23, the KRASG12C moderate expression cell line, only mildly responded to AMG510 treatment in normal 2D and 3D cultures and could be clearly evaluated by our system in hypoxia conditions, while the negative control CT-26 (G12D-mutant) spheroid exhibited no significant response to AMG510 treatment. In summary, our system, together with a controlled microenvironment and imaging methodology, provided an easily assessable and effective methodology for 3D in vitro drug efficiency testing and screenings.

Replacement, Reduction, and Refinement of Animal Experiments in Anticancer Drug Development: The Contribution of 3D In Vitro Cancer Models in the Drug Efficacy Assessment

“…In addition, scaffold-based models, such as embedded-cultures of tumor spheroids within a hydrogel ECM, could provide useful in vitro tools for the evaluation of tumor invasiveness, whose assessment could enhance the spheroid prediction of drug efficiency. For example, Huang et al developed tumor spheroids embedded within a Matrigel-based ECM to monitor the drug responses of two invasive cell lines from non-small cell lung cancer and CRC to sotorasib (AMG510) under normoxia and hypoxia conditions [ 209 ].…”

Biomimetic Scaffolds—A Novel Approach to Three Dimensional Cell Culture Techniques for Potential Implementation in Tissue Engineering

“…One of the main advantages of using spheroids in cancer research is the ability to measure drug response. Multiple studies have been conducted involving spheroids derived from various types of cancer, including colorectal cancer [ 39 , 40 , 41 , 42 ], breast cancer [ 41 , 43 , 44 ], lung cancer [ 45 , 46 ], ovarian cancer [ 1 , 47 , 48 ] and even Ewing sarcoma cells [ 49 ]. It is important to note that 3D cultures, particularly spheroids, are a valuable and effective tool for studying cancer biology.…”

Assays on 3D tumor spheroids for exploring the light dosimetry of photodynamic effects under different gaseous conditions

“…To obtain uniform tumor spheroids with spherical morphology and large dimensions, we adjusted the traditional culture methods by integrating the hanging drop method with agar overlay technique and refined several critical technical parameters [16][17][18]. Figure 1A illustrates the schematic representation depicting the sequential steps of 3D tumor spheroid generation.…”