Email this pageNovel Methodologies 2007 Awardees
Robert Block MD, MPH and Steve Georas MD
Project: Potent Lipid Mediator Measurement Methodology
Investigators: Robert C. Block, MD, MPH, Steve N. Georas, MD, Alan Friedman, PhD, (University of Rochester) and J. Thomas Brenna, PhD (Cornell University)
Brief Description of Project: This project will develop new assays for a variety of potent lipid mediators in a variety of human diseases, most notably (at first) heart disease and asthma. The accurate measurement and analysis of complex lipids is emerging as an important field in biomedical science. Although still in its infancy compared to other analytical disciplines, "lipidomics" is gaining momentum. For example, the NIH funded Lipid Metabolites And Pathways Strategy (LIPID MAPS) is providing an unprecedented picture of cellular lipid metabolism, cataloging several thousand structures (www.lipidmaps.org). Lipids are increasingly recognized for their role as potent mediators in the resolution of inflammation, cell growth, signal transduction, tissue protection and homeostasis.
Two examples at SMD: (1) Dr. Block is investigating the role of the "resolvin and protectin" family of lipids in patients with heart disease, testing the hypothesis that commonly used medications in patients with coronary artery disease, such as statins and aspirin, exert beneficial effects by generating the production of these lipid mediators; (2) Dr. Georas is exploring the role of the lipid mediator lysophosphatidic acid and its receptors in asthma. This compound is detectable in bronchoalveolar lavage fluids and may contribute to the resolution of allergic inflammation. Although other "-omic" technologies are well established at SMD, there is an absence of local expertise in measuring complex lipids. The goal of this proposal is to help establish this technology on-site.
Frances Eun-Hyung Lee MD and Marc Hellerstein MD, PhD
Project: Using Heavy Water in vivo Labeling to Measure Human Proliferation and Die-Away Kinetics-Development of the Core Facility at the University of Rochester
Investigators: Frances Eun-Hyung Lee, MD, Marc Hellerstein, PhD, Hulin Wu, PhD, Ignacio Sanz,MD, Tim Mosmann, PhD
Brief Description of Project: This project will establish a "human in vivo labeling" core facility at SMD that will be accessed by investigators in a diversity of disciplines such as Cancer biology, Microbiology, Immunology, Medicine, and Biodefense. Heavy water (2H20) or glucose (2H-glucose) labeling of DNA is the only safe and direct human measure of cell kinetics in vivo. Kinetics of rare immune populations has been difficult to study, however, due to extremely low frequencies. In this proposal, protocols will be developed for in vivo labeling, focusing particularly on techniques for studying extremely low frequency immune cell populations (approximately 50,000-100,000 cells). As an initial example, the immune kinetics of VH4-34 B cells, a marker of B cell tolerance, will be measured in vivo in patients with Systemic Lupus Erythematosis (SLE) and in healthy control subjects. This study will be the first to measure in vivo proliferation and die-away 
rates of these low frequency immune cells as a biomarker of autoimmune disease. This core facility will be instrumental in securing future NIH funding for many UR investigators.
Manish Kohli MD and Benjamin Miller PhD
Project: Arrayed Imaging Reflectometry-Based Detection of Circulatory Peptides in Prostate Cancer
Investigators: Manish Kohli, MD; Benjamin L. Miller, PhD.
Brief Description of Project: This proposal represents a collaborative effort to develop a novel diagnostic system for detecting and quantitating candidate pathway-specific biomarker proteins in prostate cancer. Drs. Kohli and Miller built on two unique resources developed in their laboratories: (1) a new chip-based diagnostic platform (Arrayed Imaging Reflectometry, or AIR) that can quantitatively detect concentrations of proteins with a sensitivity comparable to ELISA, but with significantly greater speed and lower cost; and (2) a bank of high quality blood and urine specimens collected from prostate cancer patients for the specific purpose of biomarker development. At present this on-going repository has collected over 350 samples of plasma and urine specimens on 200 unique subjects in four groups of prostate disease that are readily available for biomarker testing. The four research groups include (i) a control group of subjects with elevated PSA and without cancer; (ii) subjects diagnosed with local stage disease; (iii) subjects with rising PSA after primary prostatic therapy and (iv) patients with progressive disease during androgen deprivation therapy. Serial sampling is performed once every three to four months in all four groups, and serum/plasma from blood along with urine is stored at -80 °C.
AIR is a biosensor platform that enables the detection of any probe/target system by optically sensing molecular binding on a silicon chip. As a label-free technique, the assay procedure can be completed in less than 30 minutes, with nearly instantaneous readout. Comparable labeled techniques such as ELISA requires processing time of more than 2 hours. The AIR method is also amenable to multiplexing, potentially allowing 100s of different probes to be arrayed on the chip surface for simultaneous detection. In this first phase of biomarker development, Drs. Kohli and Miller will focus on the production of a chip capable of detecting several candidate angiogenesis and bone biochemistry biomarker proteins in patients with prostate cancer and controls, using specimens from the clinical groupings described above.