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Published on: June 16, 2022
A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase
Priya H Dedhia1,2,3, Hemamylammal Sivakumar4, Marco A Rodriguez4
1Division of Surgical Oncology, The Ohio State University and Arthur G. James Comprehensive Cancer Center, 816 Biomedical Research Tower, 460 W. 12th Ave, Columbus, OH, 43210, USA. priya.dedhia@osumc.edu.
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 and other small 3D in vitro models of tissues or tumors can model certain complexities of human in vivo biology; however, this technology has largely yet to be applied to ACC. In this study, we describe the generation of 3D tumor constructs from an established ACC cell line, NCI-H295R. NCI-H295R cells were encapsulated to generate 3D ACC constructs. Tumor constructs 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 NCI-H295R cells into a metastasis-on-a-chip microfluidic device platform. ACC tumor constructs showed expression of biomarkers associated with ACC, including SF-1, Melan A, and inhibin α. Treatment of ACC tumor constructs 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 tumor constructs by integrating fluorogenic MMP-sensitive peptide biosensors within the tumor constructs. Lastly, in our metastasis-on-a-chip device, NCI-H295R cells successfully engrafted in a downstream lung cell line-based construct, but invasion distance into the lung construct was decreased by MMP inhibition. These studies, which would not be possible using 2D cell cultures, demonstrated that NCI-H295R cells secreted active MMPs that are used for invasion in 3D. This work represents the first evidence of a 3D tumor constructs platform for ACC that can be deployed for future mechanistic studies as well as development of new targets for intervention and therapies.
Insights
This study introduces a novel 3D tumor construct for adrenocortical carcinoma (ACC), overcoming limitations of traditional models. The 3D ACC model accurately reflects disease characteristics and aids in understanding tumor invasion mechanisms.
Area of Science:
- Oncology
- Biotechnology
- Cancer Research
Background:
- Adrenocortical carcinoma (ACC) has a poor prognosis with limited therapeutic options due to a lack of effective preclinical models.
- Existing animal models and 2D cell cultures fail to replicate the complexity of human ACC in vivo.
- Three-dimensional (3D) in vitro models like organoids offer potential for more accurate disease modeling, but their application in ACC is nascent.
Purpose of the Study:
- To develop and characterize a 3D tumor construct using the NCI-H295R cell line for modeling adrenocortical carcinoma.
- To assess the utility of this 3D model in evaluating drug sensitivity and studying tumor cell invasion mechanisms, particularly matrix metalloproteinase (MMP) activity.
Main Methods:
- Generation of 3D ACC tumor constructs by encapsulating NCI-H295R cells.
- Assessment of construct biomarkers (SF-1, Melan A, inhibin α), viability, proliferation, and cortisol production.
- Evaluation of matrix metalloproteinase (MMP) functionality using biosensors and a metastasis-on-a-chip microfluidic device.
Main Results:
- The 3D ACC constructs successfully expressed key ACC biomarkers.
- Chemotherapeutic treatment showed reduced drug sensitivity in 3D constructs compared to 2D cultures.
- NCI-H295R cells in the 3D model secreted active MMPs, facilitating invasion, which was inhibited by MMP blockers in a metastasis-on-a-chip assay.
Conclusions:
- The developed 3D tumor construct platform is the first for ACC, offering a more physiologically relevant model than traditional 2D cultures.
- This 3D model enables the study of ACC invasion mechanisms involving MMPs and provides a valuable tool for future drug discovery and therapeutic target identification.
