Circulating tumor cell diagnostics for prostate cancer biomarkers
The present invention describes a method for detecting castration-resistant prostate cancer (CRPC) in a patient afflicted with prostate cancer comprising (a) performing a direct analysis comprising immunofluorescent staining and morphological characterization of nucleated cells in a blood sample obtained from the patient to detect circulating tumor cells (CTC), (b) determining prevalence of a CTC subpopulation associated with CRPC comprising detecting a measurable feature of each biomarker in a panel of morphological and protein biomarkers, and (c) comparing the prevalence of said CTC subpopulation to a predetermined threshold value, wherein the prevalence of the CTC subpopulation associated with CRPC above said predetermined threshold value is indicative of CRPC. In some embodiments, the CTC subpopulation associated with CRPC comprises CK− CTCs. In some embodiments, the CTC subpopulation associated with CRPC comprises small CTCs. In additional embodiments, the methods of the invention further comprise molecular analysis of the CTCs.
1. A method for detecting progression of prostate cancer to castration-resistant prostate cancer (CRPC) in a patient comprising (a) performing a direct analysis comprising immunofluorescent staining and morphological characterization of nucleated cells in a blood sample obtained from the patient to detect morphological and protein biomarkers of circulating tumor cells (CTC), wherein the CTCs are present among the nucleated cells subjected to the direct analysis at a frequency of less than 1 in 1,000 nucleated cells; (b) determining prevalence of a CTC subpopulation associated with CRPC comprising detecting a measurable feature of each of the biomarkers, wherein determining prevalence of the CTC subpopulation associated with CRPC comprises determining the prevalence of CTCs that have a nuclear size less than 67 μm 2 , and wherein the presence of the CTC subpopulation associated with CRPC is indicative of CRPC, (c) repeating steps (a) and (b), wherein an increase in the prevalence of the presence of the CTC population associated with CRPC indicates progression of prostate cancer to CRPC; and (d) treating the patient with either (i) androgen deprivation therapy (ADT) if the prevalence has not increased, or (ii) a cancer therapy other than ADT if the prevalence has increased.
2. The method of claim 1 , wherein the CRPC is metastatic castration-resistant prostate cancer (mCRPC).
3. The method of claim 2 , wherein said patient is undergoing androgen deprivation therapy (ADT).
4. The method of claim 1 , wherein the cancer therapy other than ADT comprises secondary hormonal therapy directed at AR inhibition.
5. The method of claim 4 , wherein the secondary hormonal therapy directed at AR inhibition comprises treatment with abiraterone, enzalutamide, ketoconazole, or an estrogen.
6. The method of claim 1 , wherein the ADT is a lueteinizing hormone-releasing hormone (LHRH) receptor agonist.
7. The method of claim 6 , wherein the LHRH receptor agonist is leuprolide or goserelin.
8. The method of claim 1 , wherein the cancer therapy other than ADT comprises treatment with chemotherapy, immunotherapy, or a radiopharmaceutical.
9. The method of claim 8 , wherein the cancer therapy other than ADT comprises treatment with chemotherapy selected from docetaxel, mitoxantrone, paclitaxel, and cabazitaxel.
10. The method of claim 1 , wherein said increase in the prevalence of the CTC population associated with CRPC predicts resistance to androgen deprivation therapy (ADT).
11. The method of claim 1 , wherein the nucleated cells subjected to direct analysis are not enriched for a subpopulation of the nucleated cells as a proportion of the nucleated cells relative to the blood sample originally obtained from the patient.
12. The method of claim 1 , wherein the nucleated cells subjected to direct analysis are not enriched for CTCs as a proportion of the nucleated cells relative to the blood sample originally obtained from the patient.
13. The method of claim 1 , wherein the immunofluorescent staining comprises staining for the presence of cytokeratin, CD45, and androgen receptor.
14. The method of claim 1 , wherein the CTCs are present among the nucleated cells subjected to the direct analysis at a frequency of less than 1 in 10,000 nucleated cells.