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| Status |
Public on Apr 01, 2018 |
| Title |
Expanded Genomic Profiling of Circulating Tumor Cells in Metastatic Breast Cancer Patients to Assess Biomarker Status and Biology Over Time (CALGB 40502 and CALGB 40503, Alliance). |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by RT-PCR
Genome variation profiling by genome tiling array
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| Summary |
We developed a novel approach to isolate tumor cells with high purity from blood which was subjected to immunomagnetic enrichment using EpCAM beads followed by fluorescence activated cell sorting (IE/FACS) to isolate EpCAM-positive cells away from leukocytes (CD45+). Duplicate samples of 20 cells were isolated from the same enriched blood from MBC patients and then subjected to DNA and RNA profiling in parallel.
For DNA profiling, sorted cells were subjected to BAC array comparative genomic hybridization analysis following whole genome amplification. For RNA profiling, QPCR analysis was performed on sixty four (64) cancer-related genes using Taqman® low density arrays.
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| Overall design |
Differential gene expression analysis between EpCAM-positive cells and matched leukocytes confirmed the up-regulation of EPCAM and other genes including MUC1 and KRT19 (adjusted p <0.05). In addition, EpCAM-positive cells showed a significant down-regulation of the leukocyte-specific marker PTPRC (encodes CD45) as well as CD44 and VIM, markers associated with stem cellness and epithelial to mesenchymal transition, respectively. Unsupervised hierarchical clustering analysis of RNA profiles showed that EpCAM-positive cells clustered away from the leukocytes. Genome-wide copy number analysis of EpCAM-expressing cells revealed gains (e.g. 1q and 8q), losses (e.g. 8p and 16q), and focal amplifications (e.g. on 8q and 11q including CCND1) frequently seen in primary breast cancers.
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| Contributor(s) |
Magbanua MJ, Scott JH, Hauranieh L, Olshen A, Sosa EV, Pinkel D, Roy R, Rugo HS, Eisenbud LE, Park JW |
| Citation(s) |
29311117, 30211312 |
| |
| Submission date |
Sep 05, 2012 |
| Last update date |
May 16, 2019 |
| Contact name |
Mark Magbanua |
| Organization name |
UCSF/Helen Diller Familiy Comprehensive Cancer Center
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| Department |
HemOnc
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| Lab |
Park
|
| Street address |
1450 3rd Street, PO Box 589001
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| City |
San Francisco |
| State/province |
CA |
| ZIP/Postal code |
94158-9001 |
| Country |
USA |
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| Platforms (2) |
| GPL6359 |
UCSF Cancer Center HumArray3.2 |
| GPL16020 |
Applied Biosystems TaqMan TLDA 64-Breast cancer associated-gene panel |
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| Samples (275)
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| Relations |
| BioProject |
PRJNA174414 |
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