System for immunotherapy targeting tumor propagation and progression

Abstract

Finding biologically relevant cancer markers is key to developing an effective treatment. Once specific antigens have been identified that are present on the cell population responsible for propagating tumors, T cells can be genetically altered to target these antigens and then used for personalized T cell therapy. A method to identify and select peptide antigens that effectively associate with and are presented by host HLA surface molecules originating from tumor cells responsible for the persistence and propagation of a cancer has been developed. The system of using these data to produce T cells engineered to express T cell receptors recognizing the peptide antigens is used in the production of a personalized adoptive T cell therapy for cancer that eliminates the cells capable of tumor propagation and cancer progression. The system is especially useful in the production of cancer treatments to achieve complete durable remission of cancers of epithelial origin.

We claim: 
     
         1 . A method of identifying C-RC cell-specific T cells, comprising
 (a) obtaining a tumor sample from an individual;   (b) cultivating the tumor sample under conditions that induce a stress response in differentiating and differentiated cells but permits C-RC cells to propagate and which activate a regenerative response;   (c) isolating the dominant actively expanding, most rapidly dividing population of cells from step (b); and   (d) culturing the cells to 60 to 95% confluence to obtain a population of 51% to 100% C-RC   mixing C-RC cells from step (d) with isolated CD8 + T cells from normal or cancer donors,   isolating T cells that react with the treated C-RC cells, wherein the reactive T cells comprise T cell receptors that target a C-RC-specific antigen.   
     
     
         2 . The method of  claim 1 , comprising treating the C-RC cells with an interferon gamma agonist to enhance expression of HLA-peptide complexes. 
     
     
         3 . The method of  claim 1  further comprising adding between initial lymphocyte isolation and the selection method an in vitro activation protocol selecting for CD8 + T cells that are reactive to appropriately presented peptide antigen in the context of patient-relevant HLA molecules. 
     
     
         4 . The method of  claim 1 , further comprising isolating the C-RC-specific antigen. 
     
     
         5 . The method of  claim 4 , further comprising making antibodies or molecular probes to the C-RC-specific antigen. 
     
     
         6 . The method of  claim 4 , further comprising sequencing the root-specific antigen. 
     
     
         7 . The method of  claim 4 , comprising comparing the C-RC-specific antigen, or antibodies or probes thereto, to antigen from non-tumor cells or cells of different types of tumors. 
     
     
         8 . The method of  claim 1 , further comprising cloning the T cell receptors from the reactive T cells to produce a cloned TCR construct. 
     
     
         9 . The method of  claim 8 , further comprising transducing a population of T cells with the cloned TCR construct and testing the transduced T cells for their ability to respond to expressed HLA-restricted peptide complex. 
     
     
         10 . The method of  claim 8  comprising forming a banked TCR panel. 
     
     
         11 . The method of  claim 4  further comprising digesting the C-RC-specific antigen to produce a plurality of peptides. 
     
     
         12 . The method of  claim 12  comprising presenting one or more of the peptides to CD8 + T cells and selecting T cells that are reactive to the one or more peptide.