METHOD FOR CATEGORIZING CIRCULATING TUMOR CELLS
The present invention provides method for categorizing circulating tumor cells (CTCs) using various cellular markers and revealing or non-revealing assays which provide beneficial insights for clinical staging and therapy decision making in cancer patients.
1 . A method for prognosis of cancer in a subject, said method comprising:
a) contacting a sample suspected (CTCs) from a subject with a reagent specific for a first cell marker and a reagent specific for a second cell marker, wherein said first and second cell marker independently selectively bind to said CTCs;
b) analyzing said circulating tumor cells (CTCs) of said sample by detecting said reagent specific for said first cell marker and said reagent for said second cell marker; and
c) calculating a ratio of said first cell marker to said second cell marker in the sample, wherein a higher ratio indicates a negative prognostic indicator and predicts a poor response to therapy.
2 . The method of claim 1 , further comprising comparing said ratio with a prior calculated ratio of the subject or a known ratio.
3 - 5 . (canceled)
6 . The method of claim 1 , wherein the first and second cell marker are selected from the group consisting of: EGFR, HER2, ERCC1, CXCR4, EpCAM, E-Cadherin, Mucin-1, Cytokeratin, PSA, PSMA, RRMI, Androgen Receptor, Estrogen Receptor, Progesterone Receptor, IGF1, cMET, EML4, or Leukocyte Associated Receptor (LAR).
7 . The method of claim 1 , wherein the reagents are antibodies used to detect the cell markers.
8 . The method of claim 7 , wherein the antibodies are fluorescently labeled.
9 . The method of claim 8 , wherein the antibodies are directed to EpCAM, Cytokeratin, or a combination thereof.
10 - 19 . (canceled)
20 . The method of claim 1 , wherein the sample is about 200 microliters.
21 . The method of claim 1 , further comprising enriching the sample prior to analyzing.
22 . The method of claim 21 , wherein the sample is enriched immunomagnetically or by filtration.
23 . The method of claim 1 , wherein the analyzing comprises image analysis.
24 . The method of claim 23 , wherein the image analysis is performed by microscopy or flow cytometry.
25 . (canceled)
26 . The method of claim 1 , wherein the subject has cancer.
27 . The method of claim 26 , wherein the subject is undergoing cancer therapy.
28 . The method of claim 27 , wherein the therapy is chemotherapy.
29 - 31 . (canceled)
32 . A method for determining responsiveness of a subject to a therapeutic regime comprising:
a) contacting a sample suspected of comprising circulating tumor cells (CTCs) from a subject with a reagent specific for a first cell marker and a reagent specific for a second cell marker, wherein said first and second cell marker independently selectively bind to said CTCs;
b) analyzing said circulating tumor cells (CTCs) of said sample by detecting said reagent specific for said first cell marker and said reagent for said second cell marker;
c) calculating a ratio of said first cell marker to said second cell marker in the sample, wherein a higher ratio indicates poor responsiveness of said subject to said therapeutic regime.
33 . The method of claim 32 , further comprising comparing said ratio with a prior calculation ratio of the subject or a known ratio.
34 . (canceled)
35 . A method for determining effectiveness of a candidate agent in the treatment of cancer comprising:
a) contacting a sample suspected of comprising circulating tumor cells (CTCs) from a subject with a reagent specific for a first cell marker and a reagent specific for a second cell marker, wherein said first and second cell marker independently selectively bind to said CTCs;
b) detecting said first cell marker and said second cell marker in said sample, thereby identifying an amount of CTCs in said sample;
c) comparing said amount of CTCs in said sample to a control sample, wherein said control sample is a sample from the subject prior to said contacting or is a sample comprising a known amount of CTCs, wherein a decrease in said amount of CTCs in said sample compared to said control sample indicates said candidate agent is effective in the treatment of cancer.
36 . The method of claim 36 , further comprising comparing said ratio with a prior calculation ratio of the subject or a known ratio.
37 . (canceled)
38 . The method of claim 1 , wherein said first and second cell marker are selected from EpCAM and Cytokeratin.