Wednesday, January 27, 2010

7:30-8:15 am Breakfast Presentation

The RBM Approach to Biomarker Validation: Attention to Precision and Patterns
Ralph McDade, Ph.D., Strategic Development Officer, Rules-Based Medicine
The RBM approach to biomarker discovery utilizes DiscoveryMAP™ as a hypothesis-generating tool in the primary study.  Once identified, the biomarker pattern of interest is evaluated in subsequent studies to validate its utility.  At each step, assay precision and reproducibility are the keys to success.   RBM has developed a fully automated, multiplexed immunoassay platform analytically validated to clinical laboratory standards for this process; we provide this as a service to pharmaceutical, biotech and contract research organizations. In addition, we develop novel diagnostics in areas with an unmet medical need using the identical approach.  Case studies in oncology and CNS disease will be presented.

Detection of Circulating Tumor Cells

8:25-8:30 Chairperson’s Remarks

8:30-9:00 Can Circulating Tumor Cells Help Gain Insight into the Biology of Prostate Cancer?

Martin Fleisher, Ph.D., FACB, Chairman, Department of Clinical Laboratories; Chief, Clinical Chemistry, Memorial Sloan-Kettering Cancer Center

Circulating Tumor Cell (CTC) analysis is a clinically proven biomarker used in the management of patients treated with new targeted therapies and instrumental in evaluating new prospective clinical trials to address context of use. A major goal in the use of CTC is to accelerate the transition of laboratory discoveries to clinical practice. Our CLIA certified laboratory has adapted CTC enumeration, FISH analysis and gene expression analysis in the evaluation of new drugs designed to treat patients with castration resistant prostate cancer.

9:00-9:30 The Clinical Utility of Circulating Tumor Cell Analysis – What Have We Learned So Far?

Beverly C. Handy, M.D., M.S., Associate Professor, Clinical Chemistry, Department of Laboratory Medicine, The University of Texas M.D. Anderson Cancer Center

Metastatic cancers, even when of the same tissue type, show considerable variability in their clinical behavior. Prognostic indicators that are of proven value in predicting disease aggressiveness and response to therapy are therefore, extremely useful aides for optimizing individual treatment planning. Among these, a growing body of literature indicates that the enumeration of circulating tumor cells (CTC) from peripheral blood can serve as an independent prognostic marker for clinically managing patients with some types of metastatic cancers. In particular, the level of CTC has been shown to be predictive of survival in breast, colorectal, and prostate metastatic disease. It is likely that it will be useful in tumors of other organs as well. Additional potential applications include evaluation of treatment response and use in disease staging. Isolation of CTC also offer potential opportunities for further analysis and molecular characterization of these cells, which may allow further optimization of treatment.

9:30-10:00 Detection of Circulating Tumor Cells in the Blood of Cancer Patients using a Process which Only Depletes Normal Cells

Jeffrey J. Chalmers, Ph.D., Chemical and Biomolecular Engineering; Director, University Cell Analysis and Sorting Core, The Ohio State University

We have developed a process which utilizes a unbiased negative enrichment protocol that depletes the normal blood cells to give an enriched and relatively pure CTC cell suspension. In this presentation we demonstrate that our technique is capable of detecting a significant number of CTCs in the peripheral blood of Head and Neck Cancer and Breast Cancer patients with high levels of sensitivity. Since the outcome of our negative depletion, enrichment process is a cell suspension, the final product can be further analyzed. In almost all cases, cells that are positive for cytokeratins are also positive for vimentin, and CD44 or EGFR, and depending on the patient, we observe a significant number of patients that are cytokerain negative, but vimentin and either CD44 or EGFR positive.

10:00-11:00 Networking Coffee Break with Poster and Exhibit Viewing

11:00-11:30 Multiplexed Profiling of Individual Circulating Tumor Cells using a Hyperspectral Imaging System

Harold Garner, Ph.D., Executive Director, Virginia Bioinformatics Institute, Virginia Tech; Scientific Advisor, Xanapath LLC

A new hyperspectral microscope imaging system, named the Intelligent Single Cell Optical Profiling Engine (“I-SCOPE”), is capable of analyzing tumor marker expression levels in individual, intact cells. The system quantifies multiplexed cocktails of engineered fluorophores conjugated to up to 13 markers. Results from analyses of tumor cells in a variety of specimen types will be reviewed.

11:30-12:00 Multicenter Phase III Clinical Trial Validation of Circulating Tumor Cells in Blood as Prognostic Biomarkers in Melanoma Patients Receiving Immunotherapy

Dave S.B. Hoon, Ph.D., Molecular Oncology, John Wayne Cancer Institute, Saint John’s Health Center

Melanoma is often an aggressive disease with poor prognosis when metastasis occurs, although some patients have better outcome than others in treatment responses. Identification of these patients will help improve overall survival. Development of new therapeutics and monitoring patients during treatment requires critical assessment of patients with efficient blood biomarkers to identify those patients with good and poor prognosis. Circulating tumor cells (CTC) are a form of blood biomarkers that can potentially identify prognosis in patients. In a recently completed Phase III international multicenter clinical immunotherapy trial we validated mRNA CTC biomarkers correlation with disease outcome. CTC biomarkers before treatment as well as during treatment were significantly prognostic of disease-free and overall survival in AJCC stage IV melanoma patients. This presentation will discuss an international study that demonstrated the utility of CTC blood biomarkers.

12:00-12:30 Post Capture Enumeration and Molecular Analysis of CTCs using the CEE Platform

Farideh Bischoff, Ph.D., Senior Director, Translational Research and CLIA Development, Biocept, Inc.

The Biocept CEE (Cell Capture and Extraction) device consists of micro-channels designed to selectively capture and enrich target rare cells, including CTCs from blood.  Because the Biocept CEE device is attached to the surface of a glass slide, the system is particularly suited for direct and immediate single CTC morphologic assessment, in addition to immunochemical and genetic analysis. The processing of blood samples from advanced stage cancer patients, including breast, prostate and lung, have been optimized for CTC capture, enumeration and post capture analysis of protein levels (IHC) as well as chromosomal/DNA content within Biocept’s CLIA laboratory, which is accredited for diagnostic testing.

12:30-2:00 Lunch on Your Own

Novel Biomarker Assays for Cancer

2:00-2:30 The Right Biomarkers for the Right Targeted Therapies to Treat Cancer

Thomas Chan, Ph.D., Chief Scientific Officer, Research & Development, ArQule, Inc.

Many cellular processes exhibit significant cross talk. While emphasis has been placed on identifying downstream signals of inhibited pathways as biomarkers, or picking surrogate biomarkers from proteomic and/or genomic screens, relatively less effort has been applied to identifying cross talk biomarkers.

2:30-3:00 Developing Nanocytology as a Novel Biomarker for Personalized Cancer Management

Jianyu Rao, M.D., FCAP, Professor, Pathology & Epidemiology; Chief, Cytopathology; Director, Gynecological Pathology, Department of Pathology and Laboratory Medicine, Ronald Reagan Medical Center, University of California, Los Angeles

Cancer is deadly because cancer cells invade and metastasize. Studies from us and others have shown that cell nano-mechanical properties, e.g., cell elasticity and softness, is a new class of quantitative markers that is predictive of tumor cell behavior, whereas chemopreventive and therapeutic agents have profound modulating effect on cancer cell mechanics. Thus, combining cancer cell nano-mechanical analysis with cytomorphology and molecular expression profiling may produce functional diagnostic information that can be used for personalized management of cancer.

3:00-3:30 Lipidomics as a Diagnostic and Research Tool

Aaron Fernandis, Ph.D., Senior Research Fellow, Biochemistry, National University of Singapore Centre for Life Sciences

Lipids, an important regulator of cellular function if imbalanced, often leads to onset of pathology. Our long term goal is to profile various classes of lipids in body fluids (blood in particular) and compare these profiles against pathological disorders such as ovarian cancer which is the leading cause of death related to gynecologic cancer. We have measured the plasma concentrations of nearly 400 lipid species (including glycerophospholipids, sphingolipids) by electrospray ionization mass spectrometry and analyzed the data with multivariate statistics. Patients were differentiated from controls and benign from malignant cases with positive predictive values (PPV) of 92% and 87% and accuracy of prediction at 94% and 88%, respectively. Moreover, the altered lipids were then checked for their metastatic potentials by using various cell-based functional assays.

3:30 Close of Conference