A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity

Abstract

Adrenocortical carcinoma (ACC) has a poor prognosis, and no new drugs have been identified in decades. The absence of drug development can partly be attributed to a lack of preclinical models. Both animal models and 2D cell cultures of ACC fail to accurately mimic the disease, as animal physiology is inherently different than humans, and 2D cultures fail to represent the crucial 3D architecture. Organoids are small 3D in vitro models of tissues or tumors, which can model certain complexities of human in vivo biology; however, this technology has yet to be applied to ACC. In this study, we describe the generation of organoids from an established ACC cell line, H295R. H295R cells were encapsulated to generate ACC organoid constructs. Organoids were assessed for biomarker expression, viability, proliferation, and cortisol production. In addition, matrix metalloproteinase (MMP) functionality was assessed directly using fluorogenic MMP-sensitive biosensors and through infusion of H295R cells into a metastasis-on-a-chip microfluidic device platform. ACC organoids showed expression of biomarkers associated with ACC, including SF-1, Melan A, and inhibin alpha. Treatment of ACC organoids with chemotherapeutics demonstrated decreased drug sensitivity compared to 2D cell culture. Since most tumor cells migrate through tissue using MMPs to break down extracellular matrix, we validated the utility of ACC organoids by integrating fluorogenic MMP-sensitive peptide biosensors within the organoids. Lastly, in our metastasis-on-a-chip device, H295R cells successfully engrafted in a downstream lung cell line-based organoid construct, but invasion into the lung organoid construct was decreased by MMP inhibition. These studies, which would not be possible using 2D cell cultures, demonstrated that H295R cells secreted active MMPs that are used for migration in 3D. This work represents the first evidence of a 3D organoid platform for ACC that can be deployed for future mechanistic studies as well as development of new targets for intervention and therapies.

Preclinical Models of Adrenocortical Cancer

“…Although not yet peer reviewed, Dedhia et al released promising organoid models of ACC, studying metastasis through matrix metalloproteinase experiments in organoids and microfluidic models [99].…”

In vitrodifferentiation of mouse pluripotent stem cells into glucocorticoid-producing adrenocortical cells

“…In other systems, 3D cultures have been shown to be more conducive to differentiation by providing molecular cues from neighbouring cells as well as mechanical stimulation 31,32 . We therefore tested the aggregation of aIM/LPM cells in ultra-low attachment plates, or in microwell plates, the latter allowing the formation of hundreds of small spheroids in a single well.…”