Quantitative methods and kits for providing reproducible ihc4 scores

Abstract

The present technology relates generally to determining a risk of recurrence of disease in a cancer patient. In particular, this approach to determining a risk of recurrence involves utilizing standardized quantitative assessments of the level of biomarker expression selected from estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and Ki-67 (Ki67) in a patient's tumor to determine the risk of recurrence, thereby allowing a caretaker to determine the best course of treatment for the patient.

1 - 24 . (canceled) 
     
     
         25 . A method of determining a risk of recurrence of disease in a cancer patient comprising:
 determining a level of expression of each of four biomarkers, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and Ki-67 (Ki67), in a tumor specimen taken from a cancer patient, the determination utilizing a technique that provides a continuous, quantitative score of the level of expression of each of said four biomarkers;   determining a cumulative score based on a score model comprising an algorithm (I), which is:
   IHC4 cs =94.7*[0.100ER 10 −0.079PR 10 +0.586HER2+0.240 ln(1+10*Ki67)]  (I)
 
   in which   ER 10  represents the normalized, continuous, quantitative score of the level of expression of ER;   PR 10  represents the normalized continuous, quantitative score of the level of expression of PR;   HER2 represents the dichotomized score from, continuous, quantitative scores for the level of expression of HER2; and   Ki67 represents the percent positive score of Ki67; and   determining a risk of recurrence of disease in the cancer patient based on the cumulative score.   
     
     
         26 . An assay kit for determining a risk of recurrence of disease in a cancer patient comprising reagents for determining a continuous, quantitative score of a level of expression, if desired, of each of four biomarkers, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and Ki-67 (Ki67), in a tumor specimen taken from a cancer patient. 
     
     
         27 . The assay kit of  claim 26 , which includes instructions for the use of said reagents. 
     
     
         28 . The assay kit of  claim 26 , in which said reagents include:
 a first stain specific for a nuclear compartment of a cell;   a second stain specific for estrogen receptor (ER);   a third stain specific for progesterone receptor (PR);   a fourth stain specific for human epidermal growth factor receptor 2 (HER2);   a fifth stain specific for Ki-67; and   a sixth stain specific for epithelial cytoplasmic subcellular compartment.   
     
     
         29 - 30 . (canceled) 
     
     
         31 . A non-transitory computer readable medium having program code recorded thereon that, when executed on a computing system, automatically processes immunofluorescence data, the program code comprising:
 code for processing a digital microscopy image of a stained tumor specimen taken from a cancer patient to extract data related to intensity values associated with one or more stains;   code for processing the extracted data to arrive at a value for intensity per pixel for each of the one or more stains;   code for processing pixel intensity of at least one stain for determining pixels associated with a preselected subcompartment and determining the area of the subcompartment for use as a denominator;   code for processing pixel intensity of a second stain for determining an expression level of a biomarker on a continuous scale and a value for total biomarker intensity in the same preselected subcompartment for use as a numerator;   code for calculating from the numerator and denominator a quantitative score of the biomarker expression per area;   optionally, code for processing the total area (pixels) stained by Ki67 as the numerator;   optionally, code for total tumor nuclear compartment area as the denominator;   code for collecting the quantitative score of each of at least three of the following four biomarkers, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and Ki-67 (Ki67); and   optionally, code for incorporating the continuous, quantitative scores of the at least three biomarkers into a score model for arriving at a cumulative score.   
     
     
         32 . The method of  claim 31  further comprising code for determining a risk of recurrence of disease in the cancer patient based on the cumulative score. 
     
     
         33 . The method of  claim 31  further comprising code for assigning the cumulative score to first, second, third, or fourth quartile. 
     
     
         34 . The method of  claim 31  further comprising code for determining a risk of recurrence of disease in the cancer patient based on the cumulative score by designating the position of the cumulative score of the patient on a curve of possible scores from a population of patients. 
     
     
         35 . A method of determining a risk of recurrence score in a cancer patient comprising:
 determining a level of expression of each of at least three of the following four biomarkers, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and Ki-67 (Ki67), in a tumor specimen taken from a cancer patient, the determination utilizing a technique that provides a continuous, quantitative score of the level of expression of each of the at least three of said four biomarkers; and   determining a cumulative score based on a score model in which the continuous, quantitative scores provided by the technique, one each for the at least three of said four biomarkers, are incorporated.   
     
     
         36 . The method of  claim 35  in which the continuous, quantitative score is intensity-based. 
     
     
         37 . The method of  claim 35  in which the continuous, quantitative score is both intensity-based and area-based. 
     
     
         38 . The method of  claim 35  in which the continuous, quantitative score is determined for each of said four biomarkers. 
     
     
         39 . The method of  claim 35  in which the technique includes an automated, quantitative image analysis procedure. 
     
     
         40 . The method of  claim 35  in which the technique includes an automated, quantitative, immunofluorescence image analysis procedure. 
     
     
         41 . The method of  claim 38  in which the automated, quantitative image analysis procedure is implemented by an automated, digital pathology system. 
     
     
         42 . The method of  claim 38  in which the automated, quantitative image analysis procedure is implemented by an automated, digital, immunofluorescence pathology system.