Quantitative centrosomal amplification score to predict local recurrence of ductal carcinoma in situ

A computer program product for determining the 10-year local recurrence risk profile of a patient diagnosed with a carcinoma in situ, the computer program product comprising a non-transitory computer readable storage having program instructions embodied therewith, the program instructions executable by a computer, to cause the computer to perform a method comprising:

• (a) acquiring three-dimensional optical sections of a sample of tumor tissue or cancer cells from the patient using confocal microscopy, wherein the sample is processed for visualization and demarcation of cell nuclei, individually distinguishable centrosomes (iCTRs) and megacentrosomes (mCTRs) in a region of interest (ROI) defined by a plurality of cell nuclei;
• (b) processing the optical sections using volume rendering software to generate three-dimensional volumetric data for each centrosome, wherein background subtraction and fluorescence intensity thresholds are applied to distinguish centrosomes from non-specific signals
• (c) determining a volume of each iCTR and mCTR in the ROI based on the three-dimensional volumetric data (d) determining numbers of iCTRs and mCTRs associated with each cell nucleus in the ROI, wherein a centrosome is categorized as an iCTR when the volume of the centrosome is within a range of volumes found in normal tissue, and a centrosome is categorized as an mCTR when the volume of the centrosome is greater than the range of volumes found in normal tissue:
• (e) determining severity and frequency of numerical and structural centrosome amplification present within the sample based on the determined volumes and numbers of iCTRs and mCTRs from the patient;
• and
• (f) determining at least one centrosome amplification score (CAS) value for the sample based on the determined severity and frequency of numerical and structural centrosome amplification in the sample, wherein the determined at least one CAS value provides a measure of a level of a 10 year risk of local recurrence associated with the carcinoma in situ.