Free Radical Biology in Cancer Shared Resource Facility
The mission of the Free Radical Biology in Cancer Shared Resource Facility (FRBC SRF) is to provide expertise and services in free radicals, oxidative stress, mitochondrial function, and proteomics to basic and clinical investigators of the Markey Cancer Center (MCC) doing basic, pre-clinical, and clinical research.
Value-added Importance of FBRC SRF to MCC
Faculty and Staff
Hours of Operation
Cost and Chargebacks to Investigators
The role of free radicals and altered metabolism is increasingly apparent in various aspects of cancer biology and cancer therapy. Recognizing the importance of free radicals from programmatic meetings and retreats, MCC members advocated for the formation of the FRBC SRF. This demonstrates that the FRBC SRF emanated from member wishes that the MCC provide excellence in this important aspect of cancer biology.
Analysis of the roles of free radicals in cancer biology requires highly skilled ability to:
- Measure the damage induced by free radicals in various tissues, cells, and fluids associated with different cancers.
- Measure oxygen and nitrogen free radical species via biochemical and biophysical methods.
- Measure molecules responsible for free radical production, oxidant scavenging, and free radical damage.
- Use proteomics to identify proteins in tissues, cells, and fluids that have differential levels or have been oxidatively modified and to determine associated functional changes.
- Measure mitochondrial function, since mitochondria are a major source of free radicals in cells.
- Educate MCC investigators on proper sample handling and preparation so that reliable, precise, and artifact-free assay results are obtained.
Value-added Importance of FRBC SRF to MCC
As noted, the importance of free radical biology in multiple aspects of cancer biology and environmental carcinogenesis, including cancer etiology and cancer therapy, has become increasingly apparent. Consequently, the resources provided by the FRBC SRF are invaluable to investigators in each of the MCC Research Programs. For example, given the association of arsenic in Appalachia with the etiology of lung cancer and the known redox chemistry and biology that arsenic can undergo, the recent DOD grant on lung cancer (Dr. Susanne Arnold, PI) will employ the FRBC SRF in clinical research designed to increase the understanding of the role of arsenic in a cancer of high incidence in this population served by MCC.
Second, the NCI R01 grant to Dr. Jeff Moscow for a pilot study of MESNA as a possible preventative of doxorubicin-induced oxidative stress and TNF-α elevation in plasma is a direct result of oxidative stress studies provided by the FRBC.
Third, Clinical trials directed by Dr. John Hayslip designed to investigate nutritional intervention in hospital stay for colorectal cancer patients and led by Dr. Rachel Miller on chemotherapy prior to surgery in ovarian cancer on survival outcome, both of the Drug Discovery, Delivery, and Translational Therapy Program (DT), will provide many samples to the FRBC SRF for analysis of oxidative damage.
Also, Dr. Kyung-Bo Kim of the Cancer Cell Biology and Signaling Program (CS) uses FRBC SRF-resident proteomics analysis to investigate the immunoproteasome in cancer, and a new NCI R01 has been obtained. EPR analysis of DT-generated compounds as potential antioxidants can be accomplished easily by EPR.
Importantly, and unique to this region of the United States, the FRBC SRF is one of only a few cancer center-resident resources in the world with the knowledge and reagents necessary to provide the services needed by MCC members. Personnel in the FRBC are highly knowledgeable about free radicals, oxidative stress, sample handling and preparation, and the advantages and limitations of each assay employed, including bioenergetics and proteomics. These skill sets provide a distinct advantage to MCC researchers in sample use, analysis, and interpretation of results.
The FRBC SRF also works in conjunction with other MCC Shared Resource Facilities, such as Biostatistics, Cancer Research Informatics and Flow Cytometry and Cell Sorting.
Faculty and Staff
D. Allan Butterfield, PhD, Director: 859-323-0476, firstname.lastname@example.org »
Prof. Butterfield is the UK Alumni Association Endowed Professor of Biological Chemistry. His expertise is in detection of free radicals, measures of oxidative stress, and proteomics identification of oxidatively modified proteins. Prof. Butterfield serves as the director of the FRBC SRF.
Mihail Mitov, PhD, Research Associate: email@example.com »
Dr. Mitov determines indices of oxidative damage, separates and analyzes proteins for proteomics analyses, and assists with Seahorse technology-mediated determination of mitochondrial functions.
Haining Zhu, PhD, Director Proteomics Facility
Prof. Zhu is primarily responsible for overseeing proteomics analyses of proteins of interest to MCC investigators.
The FRBC SRF Technical Advisory Committee provides advanced expertise in oxidative/nitrosative stress, mitochondrial biology, EPR detection of free radicals, and mass spectrometry to guide MCC investigators in implementing available approaches to support their cancer studies.
- Allan Butterfield, PhD, UK Alumni Association Endowed Professor of Biological Chemistry; Director, FRBC SRF
- Daret St. Clair, PhD, James Graham Brown Professor of Toxicology; MCC Associate Director for Basic Research; Co-leader, RR Program
- Xianglin Shi, PhD, William A. Marquard Chair and Professor of Environmental Toxicology; member, RR Program
- Haining Zhu, PhD, Professor of Molecular and Cellular Biochemistry; Director of the UK Proteomics Core Facility; member, RR Program
The FRBC SRF Internal Advisory Committee monitors the effectiveness of the shared resource, including generation of grant proposals based on FRBC SRF assistance, recommends policy for fee structure for FRBC SRF services, and sets priorities for sample analysis if needed. The FRBC SRF Internal Advisory Committee, which meets semi-annually or on an ad hoc basis if needed, receives any concerns on quality or timeliness of service and resolves concerns.
- Susanne Arnold, MD, Professor of Medicine, MCC Associate Director; member, CP Program
- Allan Butterfield, PhD (ex officio), UK Alumni Association Endowed Professor of Biological Chemistry; Director FRBC SRF; member, RR Program
- Natasha Kyprianou, PhD, Professor of Surgery; member CS Program
- Guo-Min Li, PhD, Professor of Toxicology; member RR Program
- Rachel Miller, MD, Assistant Professor of Obstetrics and Gynecology; member DT Program
- Jeff Moscow, MD, Professor of Pediatric Oncology; Co-leader DT Program
- Mary Vore, PhD, Chair, Department of Toxicology; Co-leader, RR Program
- Zhou Zhang, MPH, PhD, Assistant Professor of Toxicology; member CP Program
- Haining Zhu, PhD (ex officio), Professor of Molecular and Cellular Biochemistry; Director of the UK Proteomics Core Facility; member, RR Program
If multiple PIs request simultaneous analyses, the Director, with input from the Executive Committee, decides priority of analyses based on the following criteria:
- Level 1: Samples from MCC members with NCI funding or cancer-related federally funded peer reviewed studies
- Level 2: Samples from MCC investigators preparing cancer-related, peer reviewed grant applications
- Level 3: Samples from MCC investigators with a cancer-related project funded by MCC pilot studies or non-peer reviewed source
- Level 4: Samples from MCC investigators without funding whose analysis would lead to preliminary data and the likelihood of a subsequent extramural proposal submission
Hours of Operation
The hours of operation of the FRBC are from Monday-Friday, 8:30 am-5:00 pm.
Costs and Chargeback to Investigators
A recharge center for the FRBC SRF was initiated in July 2012, with the following cost structure:
- Service 1: Analysis of markers of oxidative and nitrosative stress. Assays of total protein oxidation and lipid peroxidation ($10/per oxidative modification/per membrane used).
- Service 2: Molecular biological manipulation of biological systems with which to investigate redox signals, including measurements of mitochondrial function. Seahorse analysis ($25/sample). Molecular biological manipulation of redox signaling ($50/sample). Assay of redox enzymes ($10/sample).
- Service 3: Proteomics identification of proteins. Proteomics or redox proteomics analyses of protein expression or oxidatively or covalently modified proteins. The MCC subsidizes proteomics services for MCC members at a rate of 40%. For instance, LC-MS/MS analysis of protein modifications using Orbitrap is $90 for MCC members instead of $150. To comply with the University policy that all users should be charged with the same rate, MCC provides a $60 subsidy for this analysis. The commitment from MCC will enable the investigators to utilize the state-of-the-art technology in their cancer research programs.
- Service 4: EPR detection of free radicals. Via spin trapping ($25/sample).
These rates are competitive compared to commercial outsourcing. The investigator completes a Sample Submission Form on which an account number to be charged is indicated. Upon completion of the analyses, an invoice is generated and provided to MCC administration, which uses this account number for transfer of funds to MCC.