Methods for capturing, isolation, and targeting of circulating tumor cells and diagnostic and therapeutic applications thereof

Abstract

The invention relates to methods of detection, capture, isolation and targeting of cancer cells for example circulating tumor cells (CTCs) using carbohydrate recognition domain of a lectin. The invention relates to methods of diagnosis, prognosis and treatment of cancer.

What is claimed is: 
     
         1 . A method for capturing circulating tumor cells (CTCs) from a biological fluid of a subject who does not have a hematological tumor, the method comprising contacting the biological fluid with a lectin molecule attached to a surface. 
     
     
         2 . The method of  claim 1 , wherein the subject has a solid tumor. 
     
     
         3 . The method of  claim 1 , wherein the lectin molecule is a C-type lectin. 
     
     
         4 . The method of  claim 3 , wherein the C-type lectin molecule comprises a carbohydrate recognition domain (CRD). 
     
     
         5 . The method of  claim 4 , wherein the carbohydrate recognition domain (CRD) is a CRD of mannose binding lectin. 
     
     
         6 . The method of  claim 5 , wherein the C-type lectin molecule comprising a carbohydrate recognition domain (CRD) of mannose binding lectin (MBL) lacks a MBL collagen-like domain. 
     
     
         7 . The method of  claim 5 , wherein the carbohydrate recognition domain (CRD) of mannose binding lectin (MBL) comprises the sequence of SEQ ID NO: 3. 
     
     
         8 . The method of  claim 5 , wherein the carbohydrate recognition domain (CRD) of mannose binding lectin (MBL) does not activate complement or coagulation. 
     
     
         9 . The method of  claim 5 , wherein the lectin molecule comprises at least 80% amino acid sequence identity to human mannose binding lectin and retains at least 80% of the wild-type carbohydrate binding activity. 
     
     
         10 . The method of  claim 3 , wherein the C-type lectin molecule is a carbohydrate recognition domain (CRD) of mannose binding lectin (MBL). 
     
     
         11 . The method of  claim 1 , wherein the molecule further comprises the Fc region of an immunoglobulin. 
     
     
         12 . The method of  claim 11 , wherein the molecule is FcMBL. 
     
     
         13 . The method of  claim 1 , wherein the surface is a bead, hollow fiber, porous scaffold, particle, or well. 
     
     
         14 . The method of  claim 1 , wherein the surface is magnetic. 
     
     
         15 . The method of  claim 1 , wherein the CTCs express mannan carbohydrates on their cell surface. 
     
     
         16 . The method of  claim 1 , wherein the CTCs express carbohydrates containing D-mannose and L-fucose onto their cell surface. 
     
     
         17 . The method of  claim 1 , wherein the biological fluid is selected from a body fluid, such as whole blood, plasma, any cell-containing blood fraction, cerebrospinal fluid, bone marrow, cell sample, joint fluid, urine, tears or feces. 
     
     
         18 . The method of  claim 17 , wherein the bone marrow or cell sample is obtained before transplantation. 
     
     
         19 . The method of  claim 1 , further comprising isolation of the captured CTCs. 
     
     
         20 . The method of  claim 19 , wherein the isolation comprises passing the biological fluid containing captured CTCs through a microfluidic magnetic separation device. 
     
     
         21 . The method of  claim 1 , further comprising assaying captured cells for CTC markers. 
     
     
         22 . The method of  claim 21 , wherein the CTC markers are selected from GlcNAc, EpCAM, EphB4, HER2, EGFR, MUC-1, or a combination thereof. 
     
     
         23 . A method for generating cancer vaccine, comprising:
 (a) contacting a sample containing CTCs from a cancer patient not having a hematological tumor with lectin molecule attached to a surface, whereby the lectin molecule binds the CTCs in the sample, thereby providing captured CTCs;   (b) isolating the captured CTCs;   (c) combining the isolated CTCs or a component thereof with an adjuvant to generate a CTC-immunogen thereby producing a cancer vaccine.   
     
     
         24 . A method of treating cancer, the method comprising, administering to a subject not having a hematological tumor, a composition comprising a lectin molecule linked to an anticancer therapeutic molecule. 
     
     
         25 . A composition comprising lectin molecule linked to an anticancer therapeutic molecule.