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CBER Research Projects
Project Title
Development and Application of Proteomic Tools for the Analysis of Human Cancer
Principal Investigator
Emanuel F. Petricoin III
Laboratory
Laboratory of Immunology; Division of Therapeutic Proteins; Office of Therapeutics Research and Review
Project Summary
Laser-capture microdissection (LCM), invented by Dr. Lance Liotta at the NCI, has enabled researchers for the first time to be able to analyze purified populations of cells directly from pateint tissue samples. These popluations include, but are not limited to, cancer epithelial cells- cells which comprise only 1-2% of the total population of cells found in any non-hematopoetic dervied tumor (the rest being stromal cells, fibroblasts, infiltrating lymphocytes, etc.). In a close collaborative effort, I have become the lead investigator for the proteomic analysis of cancer cells from a variety of tissue types (prostate, breast, lung, esophogeal, lung, brain, ovary, colon) derived from laser-capture microdissected cells. These cells have been analyzed via 2D-PAGE protein fingerprinting, signal pathway profiling, phophoprotein analysis, SELDI protein chip retentate mapping anlysis, multiplexed scanning immunoblot analysis. Additionally, serum and body fluid tumor marker identification is being performed via SELDI and 2D-PAGE analysis. We have found that we can successfully recover and analyze proteins from the LCM and have identified over one hundred proteins from ovarain, breast, lung, colon, prostate and esophogeal cancer that are differentially regulated (turned off or turned on). Over 150 proteins have been identified so far that track with the malignant phenotype. We are validating these now using a novel tool we have invented using high-throuput microarray-based lysates, antibdy arrays, and phosphoprotein pattern profiling. We have now taken these tools and are implementing them in actual human clinical trials for the first time ever.
We have successfully employed SELDI and phosphoprotein capture analysis on LCM derived cells, and invented a novel artificial intelligence-based algorithm for diagnostic analysis of cancer from SELDI-derived protein patterns using body fluids or LCM-procured cellular lysates. WE have found that we can discriminate with 96% sensitivity and 100% specificity prostate cancer from serum, irrespective of PSA values. We are currently using this approach to analyze serum from ovarian cancer patients and found we can detect early stage cancer nearly 100% of the time.
Publications
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Am J Pathol 2000 Feb;156(2):445-452
Sensitive Immunoassay of Tissue Cell Proteins Procured by Laser Capture Microdissection.
Simone NL, Remaley AT, Charboneau L, Petricoin EF 3rd, Glickman JW, Emmert-Buck MR, Fleisher TA, Liotta LA
Pub Med
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Clin Cancer Res 2000 Feb;6(2):353-6
Characterization of intracellular prostate-specific antigen from laser capture microdissected benign and malignant prostatic epithelium.
Ornstein DK, Englert C, Gillespie JW, Paweletz CP, Linehan WM, Emmert-Buck MR, Petricoin EF 3rd
Pub Med
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Mol Carcinog 2000 Mar;27(3):158-165
An approach to proteomic analysis of human tumors.
Emmert-Buck MR, Gillespie JW, Paweletz CP, Ornstein DK, Basrur V, Appella E, Wang QH, Huang J, Hu N, Taylor P, Petricoin EF 3rd
Pub Med
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Drug Dev Res 49: (1) 34-42 Jan 2000
Rapid protein display profiling of cancer progression directly from human tissue using a protein biochip.
Paweletz CP, Gillespie JW, Ornstein DK, Simone NL, Brown MR, Cole KA, Wang QH, Huang J, Hu N, Yip TT, Rich WE, Kohn EC, Linehan WM, Weber T, Taylor P, Emmert-Buck MR, Liotta LA, Petricoin EF
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Electrophoresis 2000 Jun;21(11):2235-42
Proteomic analysis of laser capture microdissected human prostate cancer and in vitro prostate cell lines.
Ornstein DK, Gillespie JW, Paweletz CP, Duray PH, Herring J, Vocke CD, Topalian SL, Bostwick DG, Linehan WM, Petricoin EF 3rd, Emmert-Buck MR
Pub Med
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Nat Rev Genet 2000 Oct;1(1):48-56
Molecular profiling of human cancer.
Liotta L, Petricoin E
Pub Med
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Cancer Res 2000 Nov 15;60(22):6293-7
Loss of annexin 1 correlates with early onset of tumorigenesis in esophageal and prostate carcinoma.
Paweletz CP, Ornstein DK, Roth MJ, Bichsel VE, Gillespie JW, Calvert VS, Vocke CD, Hewitt SM, Duray PH, Herring J, Wang QH, Hu N, Linehan WM, Taylor PR, Liotta LA, Emmert-Buck MR, Petricoin EF 3rd
Pub Med
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Proteomics 2001 Oct;1(10):1205-15
New approaches to proteomic analysis of breast cancer.
Wulfkuhle JD, McLean KC, Paweletz CP, Sgroi DC, Trock BJ, Steeg PS, Petricoin EF 3rd
Pub Med
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Proteomics 2001 Oct;1(10):1271-8
Proteomic profiling of the cancer microenvironment by antibody arrays.
Knezevic V, Leethanakul C, Bichsel VE, Worth JM, Prabhu VV, Gutkind JS, Liotta LA, Munson PJ, Petricoin EF 3rd, Krizman DB
Pub Med
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Urology 2001 Apr;57(4 Suppl 1):160-163
New technologies for biomarker analysis of prostate cancer progression: Laser capture microdissection and tissue proteomics.
Paweletz CP, Liotta LA, Petricoin EF
Pub Med
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Cancer J 2001 Jan-Feb;7(1):69-78
Cancer proteomics: From biomarker discovery to signal pathway profiling.
Bichsel VE, Liotta LA, Petricoin EF 3rd
Pub Med
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Dis Markers 2001;17(2):49-57
Cancer proteomics: The state of the art.
Herrmann PC, Liotta LA, Petricoin III EF
Pub Med
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Electrophoresis 2001 Jun;22(10):2120-4
Fingerprinting of signal transduction pathways using a combination of anti-phosphotyrosine immunoprecipitations and two-dimensional polyacrylamide gel electrophoresis.
Stancato LF, Petricoin EF 3rd
Pub Med
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FASEB J 15: (4) A38-A38, Part 1 MAR 7 2001
Proteomic analysis of lymph and plasma during cardiopulmonary bypass.
Leak LV, Petricoin EF, Liotta LA, Jones M
Last Updated: 4/1/2002
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Date created: September 25, 2003 |
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