Memorial Sloan-Kettering Cancer Center
Assessment of serum peptide profiling to detect cancer specific patterns
Team Leader: Paul Tempest, Ph.D.
Overall Project Goal:
The goal of this project is to evaluate and document whether serum peptide patterns, or custom-designed protease assays, have diagnostic value for cancer detection, mark a given clinical outcome, or distinguish clinically insignificant from significant cancer. Such a test could, for example, identify patients with newly diagnosed cancer who might safely avoid surgery or radiation. Investigation of analytical platform robustness and reproducibility is critical to assess feasibility of future clinical studies.
Laboratory Studies:
In addition to performing inter-laboratory studies within the CPTC, this laboratory also utilizes unique approaches to study cancer specific proteases. Highlights of these studies:
- Creation of mirror site at New York University to allow comparative performance.
- Use of robotic sample handler to eliminate variability.
- Establish systematic methods to standardize proteomic protocols and data analysis for use among multiple laboratories in CPTC and the scientific community
- Use of magnetic beads to enrich peptides prior to MS
- Use of MS platforms to evaluate sample repository for direct measure and in assays
Specifically, the mirror site at New York University uses an identical robotic sample handler and MALDI-TOF MS, which allows for direct comparative performance and reproducibility assessment. The use of robotics has the potential to eliminate handler variability and induced errors associated with protein measurements from clinical samples. This laboratory also utilizes a unique sample fractionation with magnetic beads that allows for the capture of peptides prior to MS analysis. This enrichment technique could allow for the capture of peptides relevant to cancer by increasing the depth of proteome coverage. A secondary platform, LC-MS, as well as an alternative, MS based functional proteomics platform, has been developed for evaluation of cancer specific protease panels in plasma. Both platforms have been evaluated using a repository of non-degradable, isotopically-labeled reference peptides for direct measurements and in assays. A repository of 1400 reference samples and 120 reference and assay substrate peptides has been established.
Team Expertise:
Team Leader:
Paul Tempst, Ph.D.
Member & Professor
Director, Protein Center
Memorial Sloan-Kettering Cancer Center
Professor of Molecular Biology
Weill Graduate School, Cornell University
Dr. Tempst leads a proteomics research team specializing in R&D of protein / peptide microanalysis, with applications in chromatin dynamics research and cancer biomarker discovery. He is a Member of the Sloan-Kettering Institute and Professor at the Gerstner Sloan-Kettering Graduate School of Biomedical Sciences, as well as Professor of Molecular Biology at the Weill Graduate School of Medical Sciences, Cornell University. He provides executive oversight of the microchemistry, proteomics, genomics and engineering resource laboratories at his host institution. He has over 25 years of experience in protein chemistry and biochemistry, and mass spectrometry, and he has collaborated with investigators worldwide to identify and characterize novel and unknown interacting proteins and protein complexes.
His research is documented in over 230 peer-reviewed papers on proteomics methods and applications, mostly to study the mechanisms of transcriptional regulation. He is an inventor on several patents, including microfluidics-based automated chemistries, a continuous nano-electrospray ion source for mass spectrometry, Gallium-affinity chromatography of phosphopeptides, a protein-nitrosylation assay, and antibody micro-arrays with multi-layered affinity detection. Recently, his lab developed a novel platform for serum peptidome analysis and advanced the model that cancer type-specific peptide patterns are surrogate markers for changing exoprotease panels.
Dr. Tempst obtained a B.S. (1976) and Ph.D. (1981) at Ghent University in his native Belgium. He received a NATO Research Fellowship to conduct postdoctoral studies (1982-85) in the laboratory of Dr. Leroy Hood at the California Institute of Technology, where he was involved in the development of the earliest microchemistry systems. He later received a Leukemia Society of America Special Fellow Award and the Irma Hirschl Trust Career Scientist Award for the application of these novel technologies in molecular biology, genetics and medicine. He was a faculty member at the Harvard Medical School (1986-90) before taking up his current positions in New York in 1991.
Proteomic Technology Specialists
Paul Tempst, Brian Chait*, David Fenyo, Phuong-Van Luc, Sofia Waldemarson, Thomas Neubert* and Josep Villaneueva
Clinical Cancer Researchers
James Eastham, Clifford Hudis, Hans Lija*, Martin Fleisher*, Larry Norton*, Peter Scardino*, Howard Scher* and Mark Robson
Bioinformaticians
Alex Lash* and Nicolas Socci
Biostatisticians
Adam Olshen, Irina Ostrovnaya, and Andrew Vickers*
Biologists
Brett Carver, Pier Paolo Pandolfi*, Charles Sawyers*
* Denotes a member of the Internal Advisory Committee
Participating Institutions:
Memorial Sloan-Kettering Cancer Center
New York University Langone Medical Center
Other Essential Personnel:
Amine Benchick, Richard Macchia, Ricardo Toledo-Crow, Yongbiao Li, Lin DeNoyer, John Philip, Kevin Lawlor, Arpi Nazarian, Steven Blais, Paul Fearn, John Major
About Memorial Sloan-Kettering Cancer Center:
Memorial Sloan-Kettering Cancer Center is one of the world’s premier cancer centers. Memorial hospital is one of the nation’s oldest cancer hospitals, and is one of 39 National Cancer Institute designated Comprehensive Cancer Centers. It is committed to exceptional patient care, leading-edge research, and superb educational programs. The physician and scientists work in collaboration to not only provide the best care, but also to discover methods to prevent, treat and cure cancer.
