Classification of Patients Having Diffuse Large B-cell Lymphoma Based upon Gene Expression

Abstract

Methods and kits for classifying patients having diffuse large B-cell lymphoma (DLBCL) based upon expression of a plurality of genes are disclosed. Real-time quantitative RT-PCR can be used to measure expression values. Correlating expression values of the plurality of genes in a tumor sample from the patient to reference expression values obtained from DLBCL patients can stratify patients in the classification groups. The methods and kits can be used to predict overall patient survival.

1 - 13 . (canceled) 
     
     
         14 . A kit comprising:
 real time PCR assays for measuring expression of a plurality of genes in a sample from a patient having DLBCL;   wherein normalized expression of a plurality of genes in samples from DLBCL patients stratifies said DLBCL patients into classification groups, wherein said normalized expression of said plurality of genes classify said DLBCL patients in groups predictive of probability of survival;   wherein said plurality of genes consists essentially of LMO2, BCL-6, FN1, CCND2, SCYA3 and BCL-2.   
     
     
         15 . The kit according to  claim 14  further comprising a housekeeping gene normalizing said expression of said plurality of genes in said sample from said patient having DLBCL. 
     
     
         16 . The kit according to  claim 15  wherein said housekeeping gene is GADPH or PGK1. 
     
     
         17 . The kit according to  claim 14  further comprising a reference cell line to calibrate said real time PCR assays. 
     
     
         18 . The kit according to  claim 14  wherein at least one of said plurality of genes in said samples from DLBCL patients is selected from the group consisting essentially of LMO2, BCL-6, FN1, CCND2, SCYA3 and BCL-2. 
     
     
         19 . The kit according to  claim 14  further comprising a naturally occurring or recombinant reverse transcriptase enzyme. 
     
     
         20 . The kit according to  claim 14  further comprising Taq polymerase copying said plurality of genes in said sample. 
     
     
         21 . The kit according to  claim 14  wherein said plurality of genes in said samples from DLBCL patients is at least four genes. 
     
     
         22 . The kit according to  claim 14  wherein said plurality of genes in said sample from the DLBCL patient is at least four genes. 
     
     
         23 . The kit according to  claim 14  further comprising software stratifying said patient having DLBCL into one of said plurality of classification groups classified from low, medium and high overall probability of survival after anthracycline-based chemotherapy. 
     
     
         24 - 44 . (canceled) 
     
     
         45 . The kit according to  claim 23 , wherein said software stratifies said patient having DLBCL by:
 measuring expression of the plurality of genes in the tumor sample from the patient to produce measured expression values;   normalizing said measured expression values to produce normalized expression values;   correlating said normalized expression values to normalized reference expression values obtained for said plurality of genes from DLBCL patients grouped into survival groups; and   classifying the patient into one of said survival groups based upon weighed predictor Z in formula:
     Z =( A ×LMO2)+( B ×BCL6)+( C ×FN1)+( D ×CCND2)+( E ×SC-YA3)+( F ×BCL2) 
   wherein A is −0.03, B is −0.2, C is −0.2, D is 0.03, E is 0.2, and F is 0.6 and wherein LMO2, BCL6, FN1, CCND2, SCYA3 and BCL2 are log base 2 of normalized expression values for genes LMO2, BCL6, FN1, CCND2, SCYA3 and BCL2, respectively.   
     
     
         46 . The kit according to  claim 45 , wherein a Z value of less than −0.06 indicates high probability of survival, a Z value of from −0.06 to 0.09 indicates medium probability of survival and a Z value of greater than 0.09 indicates low probability of survival. 
     
     
         47 . The kit according to  claim 45 , further comprising: predicting survival of the patient wherein said plurality of genes are predictive of probability of survival and wherein said survival groups comprise groups of said DLBCL patients having known overall probability of survival.