Capture, identification and use of a new biomarker of solid tumors in body fluids

Abstract

A new sensitive cell biomarker of solid tumors is identified in blood. This biomarker can be used to determine presence of solid tumors, rapid determination of treatment response, early detection of cancer, early detection of cancer recurrence, and may be used to determine therapy.

We claim: 
     
         1 . A method of screening a subject for cancer, comprising detecting circulating Cancer Associated Macrophage-Like cells (CAMLs) in blood of a subject, wherein said detecting comprises:
 (a) isolating intact cells of between 20 and 300 micron in size from a biological sample obtained from a subject using a means selected from the group consisting of size exclusion methodology, immunocapture, red blood cell lysis, white blood cell depletion, FICOLL separation, electrophoresis, dielectrophoresis, flow cytometry, magnetic levitation, microfluidic chip, and a combination thereof, and   (b) selecting cells isolated in (a) having
 (i) a large atypical polyploid nucleus of about 14-64 μm in size or multiple nuclei, and 
 (ii) a morphological shape selected from the group consisting of spindle, tadpole, round, oblong, two legs, more than two legs, thin legs, and amorphous, 
   thereby detecting CAMLs in a biological sample from a subject,   wherein when CAMLs are detected in the biological sample, the subject is determined to have cancer.   
     
     
         2 . The method of  claim 1 , wherein the cancer is carcinoma or solid tumor. 
     
     
         3 . The method of  claim 1 , wherein the cancer is breast, prostate, lung, pancreatic, or colorectal. 
     
     
         4 . The method of  claim 1 , further comprising detecting circulating tumor cells (CTCs) in the biological sample. 
     
     
         5 . The method of  claim 1 , wherein the cells are isolated in (a) using size exclusion methodology. 
     
     
         6 . The method of  claim 5 , wherein the size exclusion methodology is use of a microfilter having pores ranging in size from 5 to 20 microns. 
     
     
         7 . The method of  claim 1 , wherein the blood is peripheral blood. 
     
     
         8 . A method for confirming a diagnosis of cancer in a subject, comprising detecting CAMLs in a biological sample from a subject previously diagnosed with cancer, wherein said detecting comprises:
 (a) isolating intact cells of between 20 and 300 micron in size from blood obtained from a subject using a means selected from the group consisting of size exclusion methodology, immunocapture, red blood cell lysis, white blood cell depletion, FICOLL separation, electrophoresis, dielectrophoresis, flow cytometry, magnetic levitation, microfluidic chip, and a combination thereof, and   (b) selecting cells isolated in (a) having
 (i) a large atypical polyploid nucleus of about 14-64 μm in size or multiple nuclei, and 
 (ii) a morphological shape selected from the group consisting of spindle, tadpole, round, oblong, two legs, more than two legs, thin legs, and amorphous, 
   thereby detecting CAMLs in a biological sample from a subject,   wherein when CAMLs are detected in the biological sample, the diagnosis of cancer in the subject is confirmed.   
     
     
         9 . The method of  claim 8 , further comprising detecting CTCs in the biological sample, wherein when CAMLs and CTCs are detected in the biological sample, the subject is diagnosed with cancer. 
     
     
         10 . The method of  claim 8 , wherein the cells are isolated in (a) using size exclusion methodology. 
     
     
         11 . The method of  claim 10 , wherein the size exclusion methodology is use of a microfilter having pores ranging in size from 5 to 20 microns. 
     
     
         12 . The method of  claim 8 , wherein the cancer is breast, prostate, lung, pancreatic, or colorectal. 
     
     
         13 . The method of  claim 8 , wherein the blood is peripheral blood. 
     
     
         14 . A method for monitoring efficacy of a cancer treatment, comprising (a) determining the number of CAMLs in a biological sample from a subject having cancer before treatment of the subject for cancer, and (b) comparing the number of CAMLs determined in (a) to a number of CAMLs determined from a similar biological sample from the same subject at one or more time points after cancer treatment, wherein the number of CAMLs in a biological sample is determining by:
 (a) isolating intact cells of between 20 and 300 micron in size from blood obtained from a subject using a means selected from the group consisting of size exclusion methodology, immunocapture, red blood cell lysis, white blood cell depletion, FICOLL separation, electrophoresis, dielectrophoresis, flow cytometry, magnetic levitation, microfluidic chip, and a combination thereof, and   (b) selecting cells isolated in (a) having
 (i) a large atypical polyploid nucleus of about 14-64 μm in size or multiple nuclei, and 
 (ii) a morphological shape selected from the group consisting of spindle, tadpole, round, oblong, two legs, more than two legs, thin legs, and amorphous, 
   to obtain CAMLs from the biological sample.   
     
     
         15 . The method of  claim 14 , further comprising (c) determining the number of CTCs in the biological sample of (a), and (d) comparing the number of CTCs determined in (c) to a number of CTCs determined from the biological sample of (b). 
     
     
         16 . The method of  claim 14 , wherein the cells are isolated in (a) using size exclusion methodology. 
     
     
         17 . The method of  claim 16 , wherein the size exclusion methodology is use of a microfilter having pores ranging in size from 5 to 20 microns. 
     
     
         18 . The method of  claim 14 , wherein the cancer is breast, prostate, lung, pancreatic, or colorectal. 
     
     
         19 . The method of  claim 14 , wherein the blood is peripheral blood. 
     
     
         20 . A method for detecting CAMLs in a biological sample from a subject, comprising:
 (a) isolating intact cells of between 20 and 300 micron in size from blood obtained from a subject, and   (b) detecting cells isolated in (a) having
 (i) a large atypical polyploid nucleus of about 14-64 μm in size or multiple nuclei, and 
 (ii) a morphological shape selected from the group consisting of spindle, tadpole, round, oblong, two legs, more than two legs, thin legs, and amorphous, 
   thereby detecting CAMLs in a biological sample from a subject.   
     
     
         21 . The method of  claim 20 , wherein the subject has cancer. 
     
     
         22 . The method of  claim 21 , wherein the cancer is carcinoma or solid tumor. 
     
     
         23 . The method of  claim 20 , further comprising detecting CTCs in the biological sample. 
     
     
         24 . The method of  claim 20 , wherein the cells are isolated in (a) using a means selected from the group consisting of size exclusion methodology, immunocapture, red blood cell lysis, white blood cell depletion, FICOLL separation, electrophoresis, dielectrophoresis, flow cytometry, magnetic levitation, microfluidic chip, and a combination thereof. 
     
     
         25 . The method of  claim 24 , wherein the size exclusion methodology is use of a microfilter having pores ranging in size from 5 to 20 microns. 
     
     
         26 . The method of  claim 20 , wherein the blood is peripheral blood.