Gas-Phase
Fractionation Data-Independent Acquisition
Analysis of 3D Cocultured Spheroid Tumor Model Reveals Altered Translational
Processes and Signaling Using Proteomics
posted on 2024-02-27, 19:27authored byAriana
E. Shannon, Claire E. Boos, Brian C. Searle, Amanda B. Hummon
Colorectal cancer (CRC) contains considerable heterogeneity;
therefore,
models of the disease must also reflect the multifarious components.
Compared to traditional 2D models, 3D cellular models, such as tumor
spheroids, have the utility to determine the drug efficacy of potential
therapeutics. Monoculture spheroids are well-known to recapitulate
gene expression, cell signaling, and pathophysiological gradients
of avascularized tumors. However, they fail to mimic the stromal cell
influence present in CRC, which is known to perturb drug efficacy
and is associated with metastatic, late-stage colorectal cancer. This
study seeks to develop a cocultured spheroid model using carcinoma
and noncancerous fibroblast cells. We characterized the proteomic
profile of cocultured spheroids in comparison to monocultured spheroids
using data-independent acquisition with gas-phase fractionation. Specifically,
we determined that proteomic differences related to translation and
mTOR signaling are significantly increased in cocultured spheroids
compared to monocultured spheroids. Proteins related to fibroblast
function, such as exocytosis of coated vesicles and secretion of growth
factors, were significantly differentially expressed in the cocultured
spheroids. Finally, we compared the proteomic profiles of both the
monocultured and cocultured spheroids against a publicly available
data set derived from solid CRC tumors. We found that the proteome
of the cocultured spheroids more closely resembles that of the patient
samples, indicating their potential as tumor mimics.