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Background: Tumor stroma percentage (TSP), as an independent, low-cost prognostic factor, could complement current pathology and act as a more feasible risk factor for prognosis. However, TSP hadn't been applied into TNM staging. Here, the objective of our study was to investigate the prognostic significance of TSP in a robust rapid multi-dynamic approach with the application of MATLAB and threshold Algorithm for Gray Image analysis. Methods: Using a retrospective collection of 1539 CRC patients comprising three independent cohorts; one SGH cohort (N=996) and two validation cohorts (N =106, N= 437) from 2 institutions. We investigated 996 CRC of no special type. According to our established thresholds, 357 cases (35.84%) were classified as TSP-high and 639 cases (64.16%) as TSP-low. We determined the gray image area as the stromal part of the WSI and calculated the stroma percentage with our proposed method on MATLAB software. Results: In both TSP-cad(50%) and TSP-cad(median), multivariate analysis showed the TSP-cad was an independent prognostic factor for the vessel invasion and tumor location. For OS, TSP-manual HR=1.512 (95% CI 1.045-2.187); TSP-cad HR=1.443 (95% CI 0.993-2.097) and TSP-cad(median) HR=1.632 (95% CI 1.105-2.410). Fortunately, TSP-manual and TSP-cad were also found independent prognostic factor in all the cohorts. It was found that TSP-cad had slightly higher HR and wider CI than TSP-manual. Conclusions: Our research showed that TSP was an independent prognostic factor in CRC. Moreover, threshold algorithm for the quantitation of TSP could be established. In conclusion, with this Rapid multi-dynamic threshold Algorithm for Gray Image counting of TSP, which showed a higher accuracy than manual evaluation by pathologists and could be a practical method for CRC to guide clinical decision making.

Rapid multi-dynamic algorithm for gray image analysis of the stroma percentage on colorectal cancer