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Cellular and Diagnostic Proteomics Laboratory

Head of laboratory

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Overview of research program

Introduction

The Cellular and Diagnostic Proteomics Laboratory analyses disease mechanisms by examining changes in the patterns of proteins in both cultured cells and patient tissues. The central technology platform is a protein chip mass spectrometer (MS), which uses a powerful new approach (surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry, or SELDI-TOF MS) to the analysis of complex protein mixtures such as serum and cell extracts by profiling hundreds of proteins simultaneously, thus creating characteristic patterns representing various cellular states or disease conditions. The other major analytical technique is two-dimensional difference gel electrophoresis (DIGE). The facility is managed by Dr Liping Chung, funded by a Cancer Institute NSW Infrastructure Grant. Many of our projects are collaborative, with various collaborating groups described below.

Recent outcomes

Current proteomic research in breast cancer has three strands. In the first, the identification of proteins in the tissue and serum of breast cancer patients that can be used as diagnostic or prognostic disease markers, a provisional panel of three proteins or peptides has been developed that allows cancer tissue to be distinguished from healthy tissue with very high reliability. When fully validated, measurement of these proteins might provide new information to help in the diagnosis of malignant breast tissue, and may be developed to assist with staging of the disease. The second area is a cell biology project to discover cellular proteins that indicate when breast cancer cells show an apoptotic (cell death) response to various chemotherapies. Dose-response curves for a range of therapies have been developed, and protein changes measured both by protein chip MS and DIGE. Proteins that reliably indicate cell death may be useful in determining when a patient has responded to chemotherapy. The third project is a biochemical investigation of the pattern of phosphorylation of IGF binding proteins in breast cancer cells. To date new IGFBP-5 phosphorylation sites have been discovered, providing information that may help to understand how this protein can cause cell death in breast cancer.

The other major cancer project is the continuation of a study on serum and tissue biomarkers in pancreatic cancer. This has resulted in further identification of proteins of possible diagnostic value. Among non-cancer projects underway in the proteomics lab, the analysis of protein biomarkers in white blood cell extracts, to help in the detection of growth hormone abuse in athletes, has yielded several related proteins that may have diagnostic utility. Further identifications are in progress. Finally, the laboratory, which aims to serve as a resource to researchers both outside and within the Kolling Institute, also hosts several external studies, under the guidance of Liping Chung.

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Major funding sources

  • Cancer Institute NSW: Infrastructure Grant
  • NHMRC: Project Grant
  • NSCCAHS: New Investigator and Project Grants
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Selected publications

Leong S, Christopherson RI, Baxter RC. (2007) Proteomic profiling of apoptotic changes in human breast cancer cells using SELDI-TOF mass spectrometry. Cell Physiol Biochem 20: 579-590.

Graham ME, Kilby D, Firth SM, Robinson PJ, Baxter RC (2007) The in vivo phosphorylation and glycosylation of human insulin-like growth factor binding protein-5. Mol Cell Proteomics 6: 1392-1405.

Scarlett CJ, Saxby AJ, Nielsen A, Bell C, Samra JS, Hugh T, Baxter RC, Smith RC (2006) Proteomic profiling of cholangiocarcinoma: Diagnostic potential of SELDI-TOF MS in malignant bile duct stricture. Hepatology 44: 658-666.

Scarlett CJ, Smith RC, Saxby A, Nielsen A, Samra JS, Wilson SR, Baxter RC (2006) Proteomic classification of pancreatic adenocarcinoma tissue using protein chip technology. Gastroenterology 130: 1670-1678.

Moscova M, Marsh DJ, Baxter RC (2006) Protein-chip discovery of secreted proteins regulated by the phosphatidylinositol 3-kinase pathway in ovarian cancer cell lines. Cancer Res 66:1376-1383.

Chung L, Clifford D, Buckley M, Baxter RC (2006) Novel biomarkers of human growth hormone action from serum proteomic profiling using protein chip mass spectrometry. J Clin Endocrinol Metab 91: 671-677.

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Major collaborations

  • Prof Richard Christopherson, School of Molecular and Microbial Bioscience, University of Sydney: Proteomic screening for apoptotic markers in breast cancer, supported by NHMRC 2007-2009.
  • Dr Katrina Moore and Dr Sabah Shibli, Department of Surgery, RNSH: Detection of cancer biomarkers in tissue and serum from breast cancer patients
  • Prof Ross Smith and Ms Aiqun Xue, Department of Surgery, University of Sydney: Detection of cancer biomarkers in tissue and serum from pancreatic cancer patients
  • Prof Phil Robinson and Dr Mark Graham, Children’s Medical Research Institute, Sydney: Phosphorylation analysis of IGF binding protein-5 in breast cancer cells
  • Prof Ken Ho and Dr Anne Nelson, Garvan Institute, Sydney: Detection of biomarkers of growth hormone abuse using white cell extracts
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Research project opportunities

Supervised by Professor Robert Baxter

* indicates the opportunity is full and unavailable.

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