METHOD OF USING NON-RARE CELLS TO DETECT RARE CELLS
The invention provides seminal computational approaches utilizing data from non-rare cells to detect rare cells, such as circulating tumor cells (CTCs). The invention is applicable at two distinct stages of CTC detection; the first being to make decisions about data collection parameters and the second being to make decisions during data reduction and analysis. Additionally, the invention utilizes both one and multi-dimensional parameterized data in a decision making process.
1 . A method for detecting a cell in a sample from a subject comprising:
a) providing a sample suspected of having at least one rare cell and at least one cell that is present at a concentration that is at least 10 times that of the rare cell;
b) contacting the sample with at least one detectable agent;
c) performing cell imaging on the sample of (b) to generate a cell image; and
d) detecting the at least one rare cell as compared with other cells in the sample by analyzing the cell from the image of (c), thereby detecting the rare cell in the sample.
2 - 34 . (canceled)
35 . The method of claim 1 , wherein (d) further comprises measuring cellular sizes for the at least one cell, distribution of cellular sizes for the at least on cell, or combination thereof, and comparing the cellular sizes or distribution of cellular sizes for the at least one cell, to known cellular sizes and distributions.
36 . The method of claim 1 , wherein the analysis of (d) further comprises measuring nuclear sizes for the at least one cell, distribution of nuclear sizes for the at least one cell, contour patterns for the at least one cell, or combination thereof, and comparing the nuclear sizes, distribution of nuclear sizes or contour patterns to a putative rare cell to identify the suspected rare cell as a rare cell.
37 . The method of claim 1 , wherein the analysis of (d) further comprises determining the concentration of rare cells in a fluid of the subject from which the sample was taken.
38 . The method of claim 1 , wherein the detectable agent is a content marker and the analysis of (d) further comprises determining an expression level of the content marker.
39 . The method of claim 36 , wherein the expression level of the content marker is determined in the at least one rare cell and in the at least one cell and compared to the expression level of the content marker in a patient population.
40 . The method of claim 1 , wherein the rare cell is a circulating tumor cell (CTC).
41 . The method of claim 40 , wherein the CTC expresses a positive marker, has an intact nucleus, and is morphologically distinct from a normal WBCs, wherein the CTC is not positive for a negative marker.
42 . The method of claim 41 , wherein the positive marker is cytokeratin or EpCAM.
43 . The method of claim 41 , wherein the negative marker is CD45.
44 . The method of claim 40 , further comprising providing a diagnosis or prognosis to the subject.
45 . The method of claim 40 , wherein the subject is known to have cancer and is undergoing cancer therapy.
46 . The method of claim 45 , wherein the therapy is chemotherapy.
47 . The method of claim 46 , wherein a content marker is used to determine a chemotherapeutic agent.
48 . The method of claim 44 , wherein the subject is being administered a candidate agent.
49 . The method of claim 44 , further comprising determining the responsiveness of the subject to the cancer therapy.
50 . A method for diagnosing or prognosing cancer in a subject comprising:
a) performing the method of claim 40 ; and
b) analyzing the detected CTC to provide a diagnosis or prognosis, thereby diagnosing or prognosing cancer in a subject.
51 . A method for determining responsiveness of a subject to a therapeutic regime comprising:
a) performing the method of claim 40 ; and
b) analyzing the detected CTC, thereby determining the responsiveness of the subject to a therapeutic regime.
52 . The method of claim 51 , wherein the therapeutic regime is changed based on the results of b).
53 . A method for determining a candidate subject for a clinical trial comprising:
a) performing the method of claim 40 ; and
b) analyzing the detected CTC, thereby determining a candidate subject for a clinical trial.