# US 2017/0285035 A1 · Circulating tumor cell diagnostics for identification of resistance to androgen receptor targeted therapies > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2017/0285035 A1 - Document ID: US20170285035A1 - Status: Published application - Kind: A1 - Application number: 15/513,915 - Filed: 2015-09-24 - Published: 2017-10-05 - Earliest claimed: 2014-09-25 - Assignees: Epic Sciences, Inc. - Applicants: Epic Sciences, Inc. - Inventors: Ryan Dittamore - Web page: https://patentgrep.com/patent/US20170285035A1 ## CPC - G01N 33/57555: of the prostate - G16B 40/00: ICT specially adapted for biostatistics; ICT specially adapted for bioinformatics-related machine learning or data mining, e.g. knowledge discovery or pattern finding - G16B 40/10: Signal processing, e.g. from mass spectrometry [MS] or from PCR - G16B 40/20: Supervised data analysis - G01N 2333/4742: Keratin; Cytokeratin - G01N 2333/723: Steroid/thyroid hormone superfamily, e.g. GR, EcR, androgen receptor, oestrogen receptor - G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - G16B: BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY ## Abstract The disclosure provides a method of predicting de novo resistance to androgen receptor (AR) targeted therapy in a tumor of a prostate cancer patient comprising (a) performing a direct analysis comprising immunofluorescent staining and morphological characteristization of nucleated cells in a blood sample obtained from the patient to generate circulating tumor cell (CTC) data, wherein the analysis comprises determining a measurable feature of a panel of traditional and non-traditional CTC biomarkers for de novo resistance to androgen receptor (AR) targeted therapy, and (b) evaluating the CTC data to determine the probability of de novo resistance to the AR targeted therapy in the tumor of the prostate cancer patient. Further disclosed are the panel of traditional and non-traditional CTC biomarkers for the methods. ## Claims ### Claim 1 A method of predicting de novo resistance to androgen receptor (AR) targeted therapy in a tumor of a prostate cancer patient comprising (a) performing a direct analysis comprising immunofluorescent staining and morphological characteristization of nucleated cells in a blood sample obtained from the patient to generate circulating tumor cell (CTC) data, wherein the analysis comprises determining a measurable feature of a panel of traditional and non-traditional CTC biomarkers for de novo resistance to androgen receptor (AR) targeted therapy, and (c) evaluating the CTC data to determine the probability of de novo resistance to the AR targeted therapy in the tumor of the prostate cancer patient. ### Claim 2 (depends on 1) The method of claim 1, wherein the immunofluorescent staining comprises wherein the immunofluorescent staining of nucleated cells comprises pan cytokeratin, cluster of differentiation (CD) 45, diamidino-2-phenylindole (DAPI) and AR. ### Claim 3 (depends on 1) The method of claim 1, wherein the biomarkers comprise (1) CTC heterogeneity, (2) frequency of cytokeratin positive (CK+), AR N-terminal positive CTCs with prominent nucleoli morphology, and (3) frequency of AR C-terminal truncated CTCs. ### Claim 4 (depends on 3) The method of claim 3, wherein the CTC heterogeneity further comprises biomarkers selected from the group consisting of traditional CTCs, CTC clusters, CK− CTCs, small CTCs, nucleoli+ CTCs, CK speckled CTCs and the biomarkers listed in Table 1. ### Claim 5 (depends on 1) The method of claim 1 comprising an initial step of depositing the nucleated cells as a monolayer onto a slide. ### Claim 6 (depends on 1) The method of claim 1, wherein the prostate cancer is metastatic castration resistant prostate cancer (mCRPC). ### Claim 7 (depends on 1) The method of claim 1, wherein the CTC data is generated by fluorescent scanning microscopy. ### Claim 8 (depends on 7) The method of claim 7, wherein the microscopy provides a field of view comprising both CTCs and at least 200 surrounding white blood cells (WBCs). ### Claim 9 (depends on 1) The method of claim 1, wherein the CTC data is generated by assessing at least 4 million of the nucleated cells. ### Claim 10 (depends on 8) The method of claim 8, wherein the CTCs comprise distinct immunofluorescent staining from surrounding nucleated cells. ### Claim 11 (depends on 8) The method of claim 8, wherein the CTCs comprise distinct morphological characteristics compared to surrounding nucleated cells. ### Claim 12 (depends on 11) The method of claim 11, wherein the morphological characteristics comprise one or more of the group consisting of nucleus size, nucleus shape, presence of holes in nucleus, cell size, cell shape and nuclear to cytoplasmic ratio, nuclear detail, nuclear contour, presence or absence of nucleoli, quality of cytoplasm and quantity of cytoplasm. ### Claim 13 (depends on 1) The method of claim 1, wherein the identification of CTCs further comprises comparing intensity of pan cytokeratin fluorescent staining to surrounding nucleated cells. ### Claim 14 (depends on 1) The method of claim 1, further comprising an initial step of obtaining a white blood cell (WBC) count for the blood sample. ### Claim 15 (depends on 1) The method of claim 1, further comprising an initial step of lysing erythrocytes in the blood sample. ### Claim 16 (depends on 1) The method of claim 1, further comprising an initial step of depositing nucleated cells from the blood sample as a monolayer on a glass slide. ### Claim 17 (depends on 16) The method of claim 16, further comprising depositing between about 2 million and about 3 million cells onto the glass slide. ### Claim 18 (depends on 1) The method of claim 1, wherein the generation of the CTC data comprises enumeration of CTCs in the blood sample. ### Claim 19 (depends on 1) The method of claim 1, wherein the measurable features are analyzed using a predictive model. ### Claim 20 (depends on 19) The method of claim 19, wherein the model is a multivariate model.