Gregory Reed - US grants

Sulfur dioxide and the carcinogenic hydrocarbon benzo(a)pyrene (BP) are ubiquitous air pollutants and are components of tobacco smoke as well. Environmental exposure to these agents is unavoidable and social exposure remains far too prevalent. Epidemiologic data suggest an enhancing effect of sulfur dioxide on the formation of human respiratory tract cancers. Moreover, tumorigenicity studies in rats and hamsters have demonstrated an increased tumor yield and shortened latency period when BP and sulfur dioxide are administered together relative to those animals treated with BP alone. Sulfur dioxide itself is not carcinogenic. One way to enhance the carcinogenicity of BP would be to increase the formation of bay-region diolepoxides, the presumed ultimate carcinogenic derivatives, and to increase covalent modification of DNA by these activated forms of BP. This proposal describes a mechanistic investigation of the role of sulfur dioxide-derived oxidants on the conversion of 7,8-dihydroxy-7,8-dihydrobenzo(a)pyrene (BP-7,8-diol) to bay-region diolepoxides, i.e., the final activation step for BP as a carcinogen, and on the formation of specific deoxyribonucleoside adducts. The experimental system will be the intact hamster trachea in short-term organ culture, a model system for BP-induced pulmonary carcinogenesis. Sulfur dioxide exists in aqueous systems at neutral pH as its hydrated, ionized form, the sulfite anion. Spontaneous or enzymatic oxidation of sulfite produces peroxyl radical and peracid derivatives, and these highly reactive oxidants may be involved in the effects of sulfur dioxide on BP carcinogenesis. Sulfite-dependent effects on BP-7,8-diol metabolism will be determined, with particular attention paid to the role of sulfite oxidation products in the process. This project examines a likely target for the expression of sulfur dioxide-induced toxic effects in a mechanistic way. As exposure to sulfur dioxide is unavoidable, such an understanding of the mechanism of toxic action provides the only rational basis for intervention in the toxic process.

Sulfur dioxide, a ubiquitous air pollutant and a component of tobacco smoke, is a cocarcinogen for benzo[a]pyrene (BP) in the respiratory tract. We have investigated effects of sulfite, the physiological form of sulfur dioxide, on the metabolism and the genotoxicity of BP metabolites. This proposal focuses on interactions of sulfite with 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene (BP-7,8-diol) and 7r,8t-dihydroxy-9t,10t-epoxy7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE) to form isomeric bay-region BPT sulfonates. These derivatives, formed in high yield in incubations with either S. typhimurium tester strains or with hamster trachea in organ culture, represent a novel class of reactive intermediates. Although far more stable to hydrolysis than are bay-region diolepoxides, these sulfonates nevertheless bind covalently to DNA at levels comparable to those seen with the diolepoxides. Based on our findings, we have developed and will test the hypothesis that formation of genotoxic BPT sulfonates represents a mechanism by which sulfur dioxide enhances BP carcinogenicity. The overall goal of this application is to characterize the biochemical and biological factors affecting both the formation of BPT sulfonates and their biological activities. Chemical characterization will examine the reactivity of BPT sulfonates with DNA, RNA, and protein. Due to the importance of DNA modification in mutagenesis and carcinogenesis, the interactions of the sulfonates with DNA will be studied by both 32p-postlabeling and structural characterization of deoxynucleoside adducts. In all cases, behavior of the sulfonates will be compared with that of anti-BPDE. The ability of these BPT sulfonates to form in biological systems, and their effects on these systems will be examined using the V79 hamster lung cell line. V79 cells will be treated with anti-BPDE with and without sulfite, or with BPT sulfonates, and the resultant BP product profile and the induction of covalent binding to cellular macromolecules will be determined. Finally, the genotoxic activity of BPT sulfonates will be determined in the V79 system using selection for the hgprt phenotype. The primary aim of this project is to characterize the chemical and biological properties of BPT sulfonates, which represent a novel class of reactive intermediates. If these products are formed readily in intact mammalian cells, if they readily modify cellular nucleic acids, and if they exert genotoxic effects in this system, then a process consistent with the observed enhancing effect of sulfur dioxide on BP carcinogenesis will be established.

COBRE; CRISP; Center of Biomedical Research Excellence; Centers of Research Excellence; Chromatography, High Performance Liquid; Chromatography, High Pressure Liquid; Chromatography, High Speed Liquid; Computer Retrieval of Information on Scientific Projects Database; Core Facility; Disease; Disorder; FPLC; Fast Protein Liquid Chromatography, TM Pharmacia; Funding; Grant; HPLC; Health; High Pressure Liquid Chromatography; Institution; Instrumentation, Other; Investigators; Kansas; Medical; NIH; National Institutes of Health; National Institutes of Health (U.S.); Performance; Preparation; Programs (PT); Programs [Publication Type]; Research; Research Personnel; Research Resources; Researchers; Resources; Sampling; Source; Spectrophotometry; Training; United States National Institutes of Health; Work; analytical tool; disease/disorder; fast protein liquid chromatography; instrument; instrumentation; liver function; programs; quality assurance

[unreadable] DESCRIPTION (provided by applicant): St. John's Wort (SJW) is a widely used antidepressant. Both the antidepressant properties of SJW and its interactions with other drugs are related to its hyperforin content, not to hypericin. Hyperforin activates the Pregnane X Receptor (PXR), thereby inducing activities of drug metabolizing enzymes, cytochrome P450s (CYPs), and the transporter, P-glycoprotein (P-gp). Pharmacogenomics is a rapidly developing discipline whose objective is to customize drug therapy based on an individual's genotype. Thus far, the focus has been on adjusting doses of single drugs based on polymorphisms of drug metabolizing enzymes. As patients often take many drugs at once, our long term goal is to explore the genomics of pharmacokinetic (PK) drug interactions. Enhancement of drug metabolism by inducers varies by over 40 fold among individuals. We ultimately intend to explore genetic differences (Single Nucleotide Polymorphisms, SNPs) in CYPs and nuclear receptors (PXR, CAR [Constitutive Androstane Receptor], etc.) to develop genotype/phenotype relationships for individual responses to inducers. Probes are drugs used to study drug disposition. We have developed a probe drug cocktail that safely and conveniently probes for enzymes that metabolize and transport most clinically used drugs: caffeine (CYP1A2), losartan (CYP2C9), dextromethorphan (CYP2D6?also probes for CYP3A4), buspirone (CYP3A4), and fexofenadine (P-gp), in addition to cortisol, an endogenous CYPS A substrate. Initially, we propose to evaluate the feasibility of adding omeprazole to probe for CYP2C19 and for CYP3A4. (Multiple GYP3A4 substrates are desirable.) Then we will determine the ability of the probe drug cocktail to respond to enzyme inhibition and induction by SJW, using a product standardized to its hyperforin content. Initial genotyping of drug metabolizing enzymes and nuclear receptors will also be performed. Finally, we will determine if the drugs in the cocktail are susceptible to inhibition of metabolism by ketoconazole. The proposed studies will validate this well-tolerated probe cocktail for research into genomics of induction of enzymes by herbs like SJW. Development of such tools is a goal of the NIH Roadmap. We plan future use of these probes to look for human genomic markers to predict drug interactions. Relevance to public health: Saint John's wort, sold without prescription, affects the way that people respond to many prescription drugs. In addition, not all people are affected to the same extent. This project wjll test a procedure to better understand and predict the effects of Saint John's wort on prescription drugs in people. [unreadable] [unreadable]

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Analytical Core will support the investigators of the COBRE projects by providing instrumentation and expertise in its application to research problems. Specifically, the Analytical Core will provide and promote HPLC, LC-MS/MS, FPLC, and multi-mode spectrophotometry as analytical tools available to our investigators. This Core will provide COBRE investigators with modern instrumental approaches for the isolation, identification, and quantification of compounds of relevance to their studies of liver function in health and disease. We will validate instrument performance as a component of an overall quality assurance program for this core facility. This Core will assist and train COBRE investigators in appropriate sampling and sample preparation to take advantage of these instrumental approaches. Finally, the Analytical Core will develop working relationships with other Core Facilities on this campus and on other campuses in the state to further develop the capabilities and the efficiency of all Cores.

The objective of this proposal is to develop and verify a mixed unit life cycle analysis model of the US electric industry that includes environmental effects and the relationships to many major industries. The model will include both economic and energy/electricity flows. It is anticipated that this model will be a useful tool in energy and environmental policy decisions. Advances in the model will be due to including more interactions with other industries and a disaggregated (i.e. source-specific) power generation sector in the US. The model, if successful, will be an important tool for planners and policy makers.

Bruce K. Hamilton
2/24/10

This Partnerships for Innovation project proposes to develop multigraft copolymer-based thermoplastic elastomers by use of randomly spaced side chains, representing a more cost effective technique than the present technique which depends on difficult to produce evenly spaced side chains. Thermoplastic elastomers combine the desirable properties of crosslinked elastomers (like car tires) with low processing costs and recyclability of thermoplastics. This newly developed class of multigraft copolymer-based thermoplastic elastomers is called "superelastomers" and holds promise for improved processability and recyclability of rubbery materials used in many different products such as gloves, earbuds, condoms, nipples for baby bottles, etc. The superelastomers also have the potential to open up new applications for elastomer materials. These designer elastomers constitute a new concept where macromolecular architecture (the shape of the molecule) is used to create the desired property profile. Through partnerships and collaborations fostered by this project, the research team will progress from knowledge generated by fundamental research toward new products that will enhance U.S. competitiveness, and the partner companies will have access to new designer elastomer technology.

The broader impacts of this research are the creation of new jobs to stimulate the economy, and the reduction of the worldwide carbon footprint by developing a new class of highly tunable thermoplastic elastomers that have the potential to both be recyclable and reduce energy consumption during manufacturing.

Partners at the inception of this project are the University of Tennessee, Knoxville (Department of Chemistry, School of Business Administration, and Research Foundation), BBB Elastomers (small business, Knoxville, TN), Asius Technologies (small business, Longmont, CO), InaMei Skin Care (small business, Hadley, MA), MAPA GmbH (large business, Zeven, Germany), Fuji Film (large business, Dayton, TN), Technology 20/20 (non-profit, Oak Ridge, TN), and Venture Incite (regional venture capital firm, Oak Ridge, TN).

The investigators plan a three-year effort to improve understanding and prediction of surface melting on the Greenland ice sheet. Satellite remote-sensing, regional climate modeling and nonlinear analysis techniques will be used to: assess variability of observed surface melt occurrence; benchmark model-based melt proxies versus observed melt; diagnose synoptic-scale meteorological/sea-ice controls on melt; and assess future change in melt proxies based on regional models driven by CMIP5 (Coupled Model Intercomparison Project Phase 5) general circulation models (GCMs). Understanding surface melt on polar ice sheets is important because surface melt affects albedo, can produce useful paleoclimatic records, contributes to mass balance through runoff, and is a trigger for ice-shelf collapse leading to ice-flow acceleration and sea-level rise. Improved understanding of the synoptic-meteorological causes of melting, and of the ability of state-of-the-art models to simulate melting accurately, would help assess the effects of future warming on melting, ice-sheet flow and sea level rise. The investigators propose three main research themes to help address these issues: (1) regional-atmospheric-model skill assessments and diagnosis of present synoptic controls on surface melt; (2) application of results from the model skill assessments to GCM-based climate scenarios for estimates of future change; and (3) expanding satellite-based retrieval of surface melt state characterization through retrieval of melt magnitude using a novel fusion of passive microwave and optical/thermal satellite data. In addition to addressing key questions relating Greenland ice sheet balance to sea-level change, the project includes outreach to the public, primarily through a web site, and numerous educational impacts. The project would support undergraduate and graduate students, and involves investigators from a primarily undergraduate institution with a significant population of first-generation college students, and from a Hispanic-serving institution. All data and model results will be archived and made available through the ACADIS data center.

This is a project that creates an I-Corps Site at the University of Pittsburgh. NSF Innovation Corps (I-Corps) Sites are NSF-funded entities established at universities whose purpose is to nurture and support multiple, local teams to transition their technology concepts into the marketplace. Sites provide infrastructure, advice, resources, networking opportunities, training and modest funding to enable groups to transition their work into the marketplace or into becoming I-Corps Team applicants. I-Corps Sites also strengthen innovation locally and regionally and contribute to the National Innovation Network of mentors, researchers, entrepreneurs and investors.

This project aims to further Pitt's entrepreneurial competencies by creating an I-Corps site directed through the University of Pittsburgh Innovation Institute. This award builds on University efforts to instill innovation and entrepreneurship across the entire organization.

The main objectives of Pitt I-Corps include the following:
-Increase the number of entrepreneurial minded individuals by education, training and outreach to disciplines and individuals underrepresented in current commercialization processes;
-Enhance a recently deployed commercialization process focused on supplementing viable innovations from the laboratory with key steps required to move them to the market; and
-Improve connection to the regional entrepreneurial ecosystem for enhanced integration of Pitt opportunities into the local early-stage environment and similar networks beyond Pittsburgh.

Pitt Ventures is a new multi-step program that was created in the Innovation Institute to provide an experiential learning program enhancing the rate of commercial outcomes of Pitt discoveries. It is a lean launchpad engaging faculty, students, and local mentors in idea discovery and entrepreneurship as a means of turning concepts into products.

Through support provided in an I-Corps program, Pitt Ventures will be enhanced through a focus on disciplines and individuals that have been underrepresented in current innovation activities. With this outreach, the breadth of commercialized University innovations will be increased as its pipeline will contain a more diversified set of opportunities from a broader group of individuals.

Given Pitt's collection of resources and opportunities, Pitt I-Corps will play an important role in strengthening the entrepreneurial ecosystem in the Mid-West by providing more inventions for the region to work on. Through enhanced engagement with this network, the University will be better positioned to utilize the set of capabilities resident in the region that can help to convert Pitt ideas to marketed products. As such, the proposed I-Corps effort will aid the University in more effectively converting intellectual value conceived by its innovators into economic value realized through commercialization. The end result is more new job creation, increased local investment, enhanced economic growth and more products benefiting society reaching the market.

Clinical Pharmacology Shared Resource ? Project Summary Established as a University of Kansas Cancer Center (KUCC) developing shared resource in 2012 and selected as an established shared resource in 2015, the Clinical Pharmacology Shared Resource (CPSR) is led by Gregory Reed, PhD. This shared resource has three functional components. 1. Correlative Laboratories - directed by LaToya Berry. The Correlative Laboratories provide GXP-compliant acquisition, processing and storage or shipping of clinical research samples. Following either a sponsor's protocol or CPSR protocols, the staff efficiently and precisely prepares samples from blood (whole blood, serum, plasma or specific blood cell fractions), urine and saliva or processes tissue samples, and then stores or transfers those samples for analysis. The Correlative Laboratories are located near patient treatment areas at the KU Clinical Research Center (KU CRC), Westwood (The University of Kansas Health System's outpatient clinical facility) and at The University of Kansas Health System's main campus. The Correlative Laboratories also provide scientific and technical support to the community oncology sites and Midwest Cancer Alliance (MCA) sites to assist them in sample acquisition, processing and shipping. 2. Bioanalytical Laboratory - led by Reed. Located at the KU CRC, this GLP-compliant facility prepares biological fluids, cells or tissue samples, and analyzes them for concentrations of drugs, drug metabolites and other small molecule biomarkers. Analyses are performed on two UPLC-tandem quadrupole mass spectrometers, with a combined sample throughput of over 30,000 samples per year. 3. Pharmacokinetics/Pharmacodynamics (PK/PD) Unit ? also directed by Reed. Reed performs calculations and modeling using the Phoenix/WinNonlin® software to define and interpret the kinetics of drugs and their actions. In addition to these study-specific activities, the staff of the CPSR also play a major role in educating future physicians and researchers, as well as nurses and study coordinators currently involved in cancer clinical trials, on the theory and practice of clinical pharmacology and on how those applications of clinical pharmacology result in more powerful and informative results from clinical trials.

This University of Pittsburgh (Pitt) project builds on an existing I-Corps Site under the University's Innovation Institute.

NSF Innovation Corps (I-Corps) Sites are NSF-funded entities established at universities whose purpose is to nurture and support multiple, local teams to transition their technology concepts into the marketplace. Sites provide infrastructure, advice, resources, networking opportunities, training and modest funding to enable groups to transition their work into the marketplace or into becoming I-Corps Team applicants. I-Corps Sites also strengthen innovation locally and regionally and contribute to the National Innovation Network of mentors, researchers, entrepreneurs and investors. This is a Type II project, requesting a "renewal" of funding for an existing I-Corps Site. There are two types of I-Corps Site proposals. Type I proposals are submitted by institutions that have not had prior funding as an I-Corps Site. Type II proposals are submitted by institutions that have had prior funding as an I-Corps Site.

The main objectives of the Type II Pitt I-Corps Site include: -Increase the number of entrepreneurial minded individuals on campus through education/training with an emphasis on disciplines and individuals underrepresented in current commercialization offerings -Enhance the Pitt Ventures program through addition of a starter course focused on supplementing viable innovations from campus with key steps required to move them to market -Improve Pitt's connection to the regional entrepreneurial ecosystem for enhanced integration with the local early-stage environment and similar networks beyond Pittsburgh.