WO2006130737A1 - Methode destinee a evaluer des carcinomes metastatiques a partir de cellules endotheliales circulantes et de cellules tumorales disseminees - Google Patents

Methode destinee a evaluer des carcinomes metastatiques a partir de cellules endotheliales circulantes et de cellules tumorales disseminees Download PDF

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WO2006130737A1
WO2006130737A1 PCT/US2006/021196 US2006021196W WO2006130737A1 WO 2006130737 A1 WO2006130737 A1 WO 2006130737A1 US 2006021196 W US2006021196 W US 2006021196W WO 2006130737 A1 WO2006130737 A1 WO 2006130737A1
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cells
blood
endothelial cells
circulating endothelial
cell
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PCT/US2006/021196
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Mark Carle Connelly
Gerald V. Doyle
Galla Chandra Rao
Leon W. M. M. Terstappen
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Immunivest Corporation
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Priority claimed from US11/202,875 external-priority patent/US20070037173A1/en
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Publication of WO2006130737A1 publication Critical patent/WO2006130737A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism

Definitions

  • the invention relates generally to cancer prognosis and survival in metastatic cancer patients, based on the presence of morphologically intact circulating cancer cells (CTC) in blood. More specifically, diagnostic methods, reagents and apparatus are described that correlate the presence of circulating cancer cells in 7.5 ml of blood of metastatic breast cancer patients with time to disease progression and survivability. Circulating tumor cells are determined by highly sensitive methodologies capable of isolating and imaging 1 or 2 cancer cells in approximately 5 to 50 ml of peripheral blood, the level of the tumor cell number and an increase in tumor cell number during treatment are correlated to the time to progression, time to death and response to therapy.
  • CTC morphologically intact circulating cancer cells
  • cancer patients are not killed by their primary tumor, but they succumb instead to metastases: multiple widespread tumor colonies established by malignant cells that detach themselves from the original tumor and travel through the body, often to distant sites.
  • metastases multiple widespread tumor colonies established by malignant cells that detach themselves from the original tumor and travel through the body, often to distant sites.
  • the most successful therapeutic strategy in cancer is early detection and surgical removal of the tumor while still organ confined. Early detection of cancer has proven feasible for some cancers, particularly where appropriate diagnostic tests exist such as PAP smears in cervical cancer, mammography in breast cancer, and serum prostate specific antigen (PSA) in prostate cancer.
  • PSA serum prostate specific antigen
  • many cancers detected at early stages have established micrometastases prior to surgical resection. Thus, early and accurate determination of the cancer's malignant potential is important for selection of proper therapy.
  • Optimal therapy will be based on a combination of diagnostic and prognostic information.
  • An accurate and reproducible diagnostic test is needed to provide specific information regarding the metastatic nature of a particular cancer, together with a prognostic assessment that will provide specific information regarding survival.
  • a properly designed prognostic test will give physicians information about risk and likelihood of survival, which in turn gives the patient the benefit of not having to endure unnecessary treatment. Patient morale would also be boosted from the knowledge that a selected therapy will be effective based on a prognostic test. The cost savings associated with such a test could be significant as the physician would be provided with a rationale for replacing ineffective therapies.
  • a properly developed diagnostic and prognostic data bank in the treatment and detection of metastatic cancer focusing on survival obviously would provide an enormous benefit to medicine (US 6,063,586). If a primary tumor is detected early enough, it can often be eliminated by surgery, radiation, or chemotherapy or some combination of those treatments. Unfortunately, the metastatic colonies are difficult to detect and eliminate and it is often impossible to treat all of them successfully. Therefore from a clinical point of view, metastasis can be considered the conclusive event in the natural progression of cancer. Moreover, the ability to metastasize is a property that uniquely characterizes a malignant tumor.
  • Soluble Tumor Antigen Based on the complexity of cancer and cancer metastasis and the frustration in treating cancer patients over the years, many attempts have been made to develop diagnostic tests to guide treatment and monitor the effects of such treatment on metastasis or relapse.
  • One of the first attempts to develop a useful test for diagnostic oncology was the formulation of an immunoassay for carcinoembryonic antigen (CEA). This antigen appears on fetal cells and reappears on tumor cells in certain cancers.
  • CEA carcinoembryonic antigen
  • PSA is a component of normal prostate tissue and benign prostatic hyperplasia (BHP) tissue.
  • BHP benign prostatic hyperplasia
  • An alternative approach incorporates immunomagnetic separation technology and provides greater sensitivity and specificity in the unequivocal detection of intact circulating cancer cells.
  • This simple and sensitive diagnostic tool as described (US6,365,362; US6,551 ,843; US6,623,982; US6,620,627; US6.645.731 ; WO 02/077604; WO03/065042; and WO 03/019141) is used in the present invention to correlate the statistical survivability of an individual patient.
  • a blood sample from a cancer patient (WO 03/018757) is incubated with magnetic beads, coated with antibodies directed against an epithelial cell surface antigen as for example EpCAM.
  • an epithelial cell surface antigen as for example EpCAM.
  • the magnetically labeled cells are then isolated using a magnetic separator.
  • the immunomagnetically enriched fraction is further processed for downstream immunocytochemical analysis or image cytometry, for example, in the CellTracks® System (Immunicon Corp., PA).
  • the magnetic fraction can also be used for downstream immunocytochemical analysis, RT-PCR, PCR, FISH, flowcytometry, or other types of image cytometry.
  • the CelITracks® System utilizes immunomagnetic selection and separation to highly enrich and concentrate any epithelial cells present in whole blood samples.
  • the captured cells are detectably labeled with a leukocyte specific marker and with one or more tumor cell specific fluorescent monoclonal antibodies to allow identification and enumeration of the captured CTCs as well as unequivocal instrumental or visual differentiation from contaminating non-target cells.
  • this assay allows tumor cell detection even in the early stages of low tumor mass.
  • the embodiment of the present invention is not limited to the Cell Spotter® System, but includes any isolation and imaging protocol of comparable sensitivity and specificity.
  • Endothelial cells line the luminal surface of blood vessels and are believed to be involved with the pathogenesis of multiple disease conditions including cancer, cardiovascular diseases, autoimmune diseases, infectious diseases, and various benign conditions. Endothelial cells can detach from their monolayer and end up in the circulation. The cause for the detachment, fate and role of these circulating endothelial cells (CECs) is not yet understood. The enumeration and characterization of CECs may however offer a unique opportunity to study the vasculature and improve our understanding of a variety of disease processes.
  • CECs Elevation of the number of circulating endothelial cells have been observed in a variety of pathological conditions such as cardiovascular diseases, inflammation, infection, autoimmune disease, and cancer.
  • CECs may increase in the circulation due to active angiogenesis or vascular damage due to tumor degeneration or as a sight effect of therapy.
  • the great variation in the reported ranges of CECs from 1 to 1000 per mL makes the interpretation of these reported results quite difficult if not impossible.
  • the large variation in the definition of CECs and the technologies used to measure the CECs are the main contributors. In addition little attention is paid to the characterization of the assays used to enumerate CECs.
  • CEC circulating endothelial cells
  • DTC disseminated tumor cells
  • MBC metastatic breast cancer
  • the present invention is a method and means for cancer prognosis, incorporating diagnostic tools in assessing time to disease progression, survival, and response to therapy based upon the absolute number, change, or combinations of both of circulating endothelial cells (CEC), circulating tumor cells (CTC) or disseminated tumor cells in bone (DTC) from patients with metastatic cancer.
  • CEC circulating endothelial cells
  • CTC circulating tumor cells
  • DTC disseminated tumor cells in bone
  • the system immunomagnetically concentrates the cells, fluorescently labels the cells, identifies and quantifies for positive enumeration.
  • the statistical analysis of the cell count predicts survival.
  • the present invention provides the apparatus, methods, and kits for assessing patient survival, the time to disease progression, and response to therapy in MBC.
  • the accurate cell enumeration provides a basisi for prediction of survival, based upon a threshold comparison of the number of cells in blood.
  • Figure 1 Classification of endothelial cell candidates. Seven rows of thumbnails of cell candidates from a blood sample. From right to left the columns show the DAPI, CD105 PE, DiOC16 and CD45 APC staining and a composite of DAPI and CD105 (green) staining. Row 1 and 2 show a DiOCI 6 prelabeled HUVEC cell staining with DAPI, CD105 and lacking CD45. Row 3, 4 and 5 show an endothelial cell staining with DAPI and CD105 but lacking CD45. Note that in row 4 the endothelial cell is surrounded by two leukocytes. Row 6 and 7 show leukocytes that express CD105. The checks in the boxes indicate the cell type and are tabulated by the software.
  • Figure 2 Gallery of CEC images. Nuclear staining of DAPI with CD105 staining.
  • Panel B shows the assay efficiency at low spike levels, 2-26 in 4 mL of blood.
  • Figure 4 Assay imprecision tested over a 20 day period at a low (48) and high (1014) cell spike level in 4 mL of blood
  • FIG. 5 Assay reproducibility.
  • Panel C shows the correlation between CECs enumerated from two 4 mL aliquots of blood from 72 samples.
  • the Bland-Altman plot of this data is shown in Panel D.
  • FIG. 6 Blood draw and CEC counts.
  • Figure 7 Prevalence of CECs in 4 mL of blood from167 healthy individuals and 206 patients with metastatic carcinomas. 50 breast cancer patients, 49 colorectal cancer patients, 35 lung cancer patients, 48 prostate cancer patients and a group of other carcinomas constisting of 8 ovarian/pancreatic, 3 renal, 2 bladder, 2 thyroid, 2 gastric, and 1 breast/colon, colon/prostate, esophageal, gastric, carcinoid tumor, squamous cell, tongue, and mandibular cancer patients.
  • An accepted method for collecting circulating tumor cells combines immunomagnetic enrichment technology, immunofluorescent labeling technology with an appropriate analytical platform after initial blood draw.
  • the associated test has the sensitivity and specificity to detect rare cells in a sample of whole blood and to investigate their role in the clinical course of the disease in malignant tumors of epithelial origin. From a sample of whole blood, rare cells are detected with a sensitivity and specificity to allow them to be collected and used in the diagnostic assays of the invention, namely predicting the clinical course of disease in malignant tumors.
  • CTC circulating tumor cells
  • the assay incorporates immunomagnetic sample enrichment and fluorescent monoclonal antibody staining followed by flowcytometry for a rapid and sensitive analysis of a sample.
  • the results show that the number of epithelial cells in peripheral blood of eight patients treated for metastatic carcinoma of the breast correlate with disease progression and response to therapy. In 13 of 14 patients with localized disease, 5 of 5 patients with lymph node involvement and 11 of 11 patients with distant metastasis, epithelial cells were found in peripheral blood. The number of epithelial cells was significantly larger in patients with extensive disease.
  • the assay was further configured to an image cytometric analysis. Using a fluorescence-based microscope image analysis system, visualization of events are easily obtain and the assessment of morphologic features to further identify objects is possible.
  • the CellTracks® System refers to an automated fluorescence microscopic system for automated enumeration of isolated cells from blood.
  • the system contains an integrated computer controlled fluorescence microscope and automated stage with a magnetic yoke assembly that will hold a disposable sample cartridge.
  • the magnetic yoke is designed to enable ferrofluid-labeled candidate tumor cells within the sample chamber to be magnetically localized to the upper viewing surface of the sample cartridge for microscopic viewing.
  • Software presents suspect cancer cells, labeled with antibodies to cytokeratin and having epithelial origin, to the operator for final selection..
  • While isolation of tumor cells for the CellTracks® System can be accomplished by any means known in the art, one embodiment uses the lmmunicon AutoPrep® System for isolating tumor cells using 7.5 ml of whole blood. Epithelial cell-specific magnetic particles are added and incubated for 20 minutes. After magnetic separation, the cells bound to the immunomagnetic-linked antibodies are magnetically held at the wall of the tube. Unbound sample is then aspirated and an isotonic solution is added to resuspend the sample. A nucleic acid dye, monoclonal antibodies to cytokeratin (a marker of epithelial cells) and CD 45 (a broad-spectrum leukocyte marker) are incubated with the sample.
  • cytokeratin a marker of epithelial cells
  • CD 45 a broad-spectrum leukocyte marker
  • the unbound fraction is again aspirated and the bound and labeled cells are resuspended in 0.2 ml of an isotonic solution.
  • the sample is suspended in a cell presentation chamber and placed in a magnetic device whose field orients the magnetically labeled cells for fluorescence microscopic examination in the CellTracks® System.
  • Cells are identified automatically in the CellTracks® System and candidate circulating tumor cells presented to the operator for checklist enumeration.
  • An enumeration checklist consists of predetermined morphologic criteria constituting a complete cell (see example 1 ).
  • the diagnostic potential of the CellTracks® System can provide a rapid and sensitive method for determining appropriate treatment. Accordingly in the present invention, the apparatus, method, and kits are provided for the rapid enumeration and characterization of tumor cells shed into the blood in metastatic and primary patients for prognostic assessment of survival potential.
  • the methods of the invention are useful in assessing a favorable or unfavorable survival, and even preventing unnecessary therapy that could result in harmful side-effects when the prognosis is favorable.
  • the present invention can be used for prognosis of any of a wide variety of cancers, including without limitation, solid tumors and leukemia's including highlighted cancers: apudoma, choristoma, branchioma, malignant carcinoid syndrome, carcinoid heart disease, carcinoma (i.e.
  • neoplasms i.e. bone, breast, digestive system, colorectal, liver, pancreatic, pituitary, testicular, orbital, head and neck, central nervous system, acoustic, pelvic, respiratory tract, and urogenital, neurofibromatosis, and cervical dysplasia.
  • Circulating endothelial cells (CEC) and circulating tumor cells (CTC) in patients with metastatic colorectal cancer Lack of validated surrogate endpoints is an impediment to developing new cancer therapy.
  • CTC and CEC could predict outcome in pts undergoing treatment for metastatic colorectal cancer.
  • Eligible patients for this multicenter study had metastatic colorectal cancer, and were initiating 1 st , 2 nd , or 3 rd -line systemic therapy.
  • Blood was obtained at baseline and 3-4 weeks after treatment initiation for enumeration of CTC/CEC.
  • CTC/7.5ml and CEC/4ml of blood were measured with the CellTracks® System.
  • CTC were immunomagnetically enriched targeting CD326 (EpCAM), stained with DAPI, cytokeratin 8,18,19, and CD45.
  • CEC expressing CD146 were immunomagnetically enriched and stained with DAPI, CD105, and counterstained with CD45. Cell morphology was confirmed in all cases.
  • Circulating Endothelial Cells in Peripheral Blood of Healthy Subjects and Patients with Metastatic Carcinomas In order to determine accuracy, precision, and linearity of endothelial cell enumeration in blood and compare CECs in healthy subjects and patients with metastatic carcinomas.
  • the CellTracks® System used for endothelial cell enumeration consists of a CellTracks Autoprep®, an Endothelial Cell Kit and a CellSpotterTM Analyzer.
  • the Endothelial Cell Kit consists of ferrofluids coated with CD146 antibodies to immunomagnetically enrich endothelial cells from 4 mL of blood.
  • CD146 is expressed on endothelial cells, smooth muscle cells and a subset of activated T- lymphocytes.
  • the enriched cells are labeled with the nuclear dye DAPI, CD105 conjugated to phycoerythrin and the pan-leukocyte antibody CD45 conjugated to allophycocyanin.
  • CD105 is expressed on endothelial cells, activated monocytes and pre-B-lymphocytes x . Buffers to wash, permeabilize and resuspend the cells are also included.
  • the CellTracks AutoPrep® is a fully-automated sample preparation system. Briefly, 4ml_ of blood is mixed with 1OmL of buffer, centrifuged at 80Og for 10 minutes, and placed on the CellTracks Autoprep®. The instrument aspirates the plasma / buffer layer and adds the ferrofluids. After incubation and subsequent magnetic separation the unbound cells and remaining plasma are aspirated. Next the staining reagents are added in conjunction with a permeablization buffer to fluorescently label the immunomagnetically bound cells.
  • the cells are resuspended in the MagNest®, a magnetic cell presentation device (Immunicon, Huntingdon Valley, PA). This device consists of a chamber and two magnets that orient the immunomagnetically labeled cells for analysis on the CelITracks Analyzer.
  • the MagNest is placed on the CelITracks Analyzer, a four color semi- automated fluorescent microscope. Image frames covering the entire surface of the cartridge for each of the four fluorescent filter cubes are captured. The captured images containing objects that meet predetermined criteria are automatically presented in a web-enabled browser from which final selection of cells is made by the operator.
  • CEC The criteria for an object to be defined as a CEC include variable morphology, a visible nucleus (DAPI positive), positive staining for CD105 and negative staining for CD45. Results of cell enumeration are expressed as the number of cells per 4 ml_ of blood.
  • CelITracks system Immunicon, Huntingdon Valley, PA
  • Cell Search Tumor Cell Kit Veridex, Warren, NJ
  • HUVECs human umbilical vein endothelial cell
  • the HUVEC cells were fluorescently labeled with DiOCI 6 before spiking to permit the discrimination of these cells from endogenous endothelial cells in the blood samples.
  • DiOCI 6 fluorescently labeled with DiOCI 6
  • 1280, 320, 80, and 20 HUVECs were spiked into 4ml_ aliquots of blood from 5 different donors and processed for CEC enumeration.
  • Assay precision was determined by spiking HUVECs labeled with DiOCI 6 at a concentration of ⁇ 48 and -1040 cells into 4 ml_ aliquots of blood from one normal donor sample for 20 days.
  • thumbnails represent the nuclear (DAPI), CD105 PE, DiOCI 6 labeled HUVEC cells (DiOCI 6) and CD45 (CD45-APC) staining.
  • the composite images shown at the left show a false color overlay of the nuclear (DAPI) and CD105-PE staining.
  • Check boxes beside the composite, CD45- APC and DiOCI 6 image allow the user to confirm that the images represented in the row are consistent with endothelial cells, stain with the leukocyte marker CD45 or are the DiOCI 6 labeled HUVEC cells.
  • the software tabulates the checked boxes for each sample and the information is stored in the database.
  • thumbnails in row 1 and 2 illustrate DiOCI 6 labeled HUVEC cells identified by their staining with DAPI, CD105 and DiOCI 6 and lack of staining with CD45.
  • Row 3, 4 and 5 show CECs identified by their staining with DAPI, CD105 and lack of staining with DiOCI 6 and CD45.
  • the endothelial cell presented in row 4 is surrounded by two leukocytes that stain with DAPI and CD45 but lack staining with DiOCI 6 and CD105.
  • Row 6 and 7 show two cells staining with DAPI, CD105 and CD45 but not DiOCI 6. The latter cells are most likely leukocytes either specifically or non-specifically staining with CD105. Circulating Endothelial Cells have a typical morphologic appearance.
  • Figure 2 shows a gallery of typical CEC images.
  • HUVEC cells were spiked into blood of five healthy donors at frequency of 5, 9, 78, 310 and 1241 cells and recovery was measured using the CellTracks system.
  • coefficient of variation (CV) increased as the number of cells spiked decreased, ranging from 12.5% at the 1241 cell spike to 37.3% at the 5 cell spike.
  • the average HUVEC cells recovered was 85.6%.
  • the analytical lower limit of detection was measured by spiking a low number of DiOCI 6 labeled HUVEC cells in the sample processing tube and verification of the actual number of spiked cells under an inverted fluorescence microscope before addition of 4 ml_ of blood to the sample processing tube.
  • the samples were processed and the spiked cells enumerated.
  • the assay efficiency or percentage of spiked cells recovered was determined in 60 experiments and is illustrated in Figure 3B.
  • the cell spike ranged from 2 to 26 (mean 12, SD 5) and the recovery ranged from 44 to 100% (mean 86%, SD14).
  • Endothelial cells were enumerated in different draw tubes to evaluate the effect of the vena puncture and the associated localized turbulence in blood flow on release of endothelial cells from the local vessel wall in the evacuated blood draw tube.
  • Blood was collected from 66 healthy individuals and the CEC counts from the first tube were compared to those from the second tube. Regression analysis showed a slope of 0.53 (95% confidence interval 0.39 to 0.68), an intercept of 4.64 (95% confidence interval -3.31 to 12.59), and a correlation coefficient (R 2 ) of 0.48.
  • Figure 6A shows the correlation between CECs in the first and second tube and Figure 6B shows the corresponding Bland-Altman plot.
  • the number of CECs in the first tube were significantly larger than those found in the second draw tube. To determine whether this increase in the number of CECs was due to the actual vena puncture blood was drawn from 100 donors and CECs were determined in the second and third blood draw tube. Regression analysis showed a slope of 0.61 (95% confidence interval 0.51 to 0.71), an intercept of 6.53 (95% confidence interval 2.82 to 10.24), and a correlation coefficient (R 2 ) of 0.60.
  • Figure 6C shows the correlation between CECs in the first and second tube and Figure 6D shows the corresponding Bland-Altman plot. The number of CECs in the second tube were not significantly larger than those found in the third draw tube.
  • CECs were enumerated in blood samples from 167 healthy individuals and 206 patients with various metastatic carcinomas.
  • Figure 6 shows a scatter plot comparing CEC counts from healthy individuals and carcinoma patients and Table 1 summarizes the CEC counts.
  • CECs and CTCs were enumerated in 124 metastatic carcinoma patients.
  • CECs ranged from 6 - 1546 (mean 140 SD 274, median 50) per 4 ml_ of blood and CTCs ranged from 0 - 13254 (mean 112 SD 1190, median 0) per 7.5 mL of blood.
  • CECs for patients with 0 CTCs ranged from 6-1546 (mean 161 +315, median 48).
  • CECs for patients with 1-4 CTCs ranged from 13 - 297 (mean 87 +82, median 79) and CECs for patients with 5 or more CTCs ranged from 14-246 (mean 80 +72, median 54).
  • Endothelial cells play a key role in the development and growth of tumors. During this process endothelial cells may be released into the circulation. Enumeration and characterization of these circulating endothelial cells (CECs) may provide insights into the nature of specific disease processes and/or tumor response to treatment. Unfortunately, the frequency of CECs is low and, as current assay methods are inadequate, results can be highly variable. Hence, automation is needed to provide more consistent results. Therefore, we developed the CellTracks AutoPrep System - an automated system for rare cell sample preparation and analysis to include the analysis of rare CECs. The system was used to determine the frequency of CECs in 4 ml_ of blood from healthy individuals and patients treated for metastatic carcinomas.
  • endothelial cells are defined as nucleated cells expressing S-endo1 / CD146, endoglin / CD105 and lacking the pan-leukocyte marker CD45.
  • CTCs Circulating Tumor Cells

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Abstract

L'invention concerne une méthode destinée à évaluer un cancer chez des sujets testés sur la base d'un comptage de cellules endothéliales circulantes et/ou de cellules tumorales disséminées chez le sujet testé. Cette méthode est utilisée pour quantifier des cellules tumorales disséminées. La réalisation de corrélations avec des cellules tumorales circulantes permet d'obtenir des informations de pronostic hautement précises pour l'évaluation du risque de récurrence chez des patientes présentant un cancer primitif du sein.
PCT/US2006/021196 2005-06-02 2006-06-01 Methode destinee a evaluer des carcinomes metastatiques a partir de cellules endotheliales circulantes et de cellules tumorales disseminees WO2006130737A1 (fr)

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US68670505P 2005-06-02 2005-06-02
US68670105P 2005-06-02 2005-06-02
US60/686,705 2005-06-02
US60/686,701 2005-06-02
US11/202,875 US20070037173A1 (en) 2005-08-12 2005-08-12 Circulating tumor cells (CTC's): early assessment of time to progression, survival and response to therapy in metastatic cancer patients
US11/202,875 2005-08-12

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101878429A (zh) * 2007-11-27 2010-11-03 维里德克斯有限责任公司 血液中循环黑素瘤细胞的自动化计数和表征
CN106932579A (zh) * 2017-03-21 2017-07-07 上海美吉医学检验有限公司 一种基于液体活检的肝癌检测的试剂盒
WO2018167312A1 (fr) * 2017-03-16 2018-09-20 Université Libre de Bruxelles Détection, quantification et/ou isolement de cellules tumorales circulantes sur la base de l'expression du marqueur cd321

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6365362B1 (en) * 1998-02-12 2002-04-02 Immunivest Corporation Methods and reagents for the rapid and efficient isolation of circulating cancer cells

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6365362B1 (en) * 1998-02-12 2002-04-02 Immunivest Corporation Methods and reagents for the rapid and efficient isolation of circulating cancer cells

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101878429A (zh) * 2007-11-27 2010-11-03 维里德克斯有限责任公司 血液中循环黑素瘤细胞的自动化计数和表征
CN101878429B (zh) * 2007-11-27 2015-05-20 维里德克斯有限责任公司 血液中循环黑素瘤细胞的自动化计数和表征
WO2018167312A1 (fr) * 2017-03-16 2018-09-20 Université Libre de Bruxelles Détection, quantification et/ou isolement de cellules tumorales circulantes sur la base de l'expression du marqueur cd321
KR20190142330A (ko) * 2017-03-16 2019-12-26 유니베르시테 리브레 드 브룩크젤즈 Cd321 마커의 발현을 기반으로 한 순환 종양 세포의 검출, 정량 및/또는 단리
KR102583603B1 (ko) 2017-03-16 2023-10-05 유니베르시테 리브레 드 브룩크젤즈 Cd321 마커의 발현을 기반으로 한 순환 종양 세포의 검출, 정량 및/또는 단리
EP4235179A3 (fr) * 2017-03-16 2023-11-15 Université Libre de Bruxelles Détection, quantification et/ou isolement de cellules tumorales circulantes sur la base de l'expression du marqueur cd321
CN106932579A (zh) * 2017-03-21 2017-07-07 上海美吉医学检验有限公司 一种基于液体活检的肝癌检测的试剂盒

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