Five cores support the Center research projects and coordinate research activities:
The Administrative Core
The MU Center for Botanical Interaction Studies (CBIS) will foster interdisciplinary research and training to enable the systematic evaluation of the safety and efficacy of botanical dietary supplements. To coordinate, strengthen, and foster these and additional research and training efforts in the field of medicinal botanicals, the CBIS will:
- identify, characterize, and authenticate botanicals;
- assess the bioavailability and bioactivity of botanicals;
- identify active constituents in botanicals and explore their mechanisms of action; and
- conduct evaluations of the activities of botanicals in cell culture and in animal models of human disease.
Also, the Center will support three primary Research Projects and four research support Cores focused on interactions among five pathways with the primary focus being antioxidant signaling as it relates to the other four molecular mechanisms of action.
The Administrative Core will provide program leadership, daily administration, integration, and cohesion, while ensuring compliance with all applicable federal, NIH, and institutional regulations. Also, the Administrative Core will ensure that the Center's specific aims are met in a timely manner. This includes administering fiscal and human resources, convening meetings of the internal steering committee (ISC) and external advisory committees (EAC), interacting with other NIH Botanical Centers and with NIH Program Officers, coordinating the Pilot Research Project (PRP), administering the Training and Career Development Program, and coordinating visits of invited speakers.
The EAC will assist in meeting the aims of the three research projects, five research cores, training efforts, and pilot projects. The ISC will include all Research Project Leaders, all Research Core Leaders, and Lay Advisors and will meet monthly to facilitate internal communication and collaboration. Many activities to be supported and enhanced by the Center are already initiated, and the CBIS organization and infusion of new resources to these efforts, linked to pilot projects and training through the Center, will maximize their synergy and value.
Core A: Botanicals/Plant Genomics Core
The Botanicals/Plant Genomics Core will support the Research Projects by supplying some botanical test materials, ensuring that identity and quality of botanical materials are properly documented, and supplying samples for studies of infraspecific variation in species studied.
Specific aims include:
- identifying, documenting, and supplying botanical raw materials;
- collecting, cultivating, and supplying elderberry accessions for studies of infraspecific variation;
- collecting, cultivating, and supplying plant materials for apocynin screening;
- conducting plant genetic studies in soybean to elucidate the extent and genetic basis of variation in the level of bioactive compounds.
The Botanicals Core will identify and preserve voucher samples of all materials obtained as raw botanicals, including herbarium specimens to voucher samples collected by the Core. Elderberry and soy germplasm collections are already available, from which botanical test materials of desirable chemical composition will be supplied. Field work will be done to obtain more elderberry genotypes, and a horticultural study to determine the effect of nitrogen fertilization on chemical content will be performed. Native North American botanicals that might contain apocynin will be surveyed in an effort to locate a reliably obtainable, easily cultivated source of that compound.
A soybean breeding study will identify genotypes associated with high and low levels of compounds of interest. These studies will further the characterization of test materials used by the Research Projects and enhance the interpretation of their results relative to the broader universe of variation within those species. Additionally, they will facilitate the longer-term goals of selecting high-potency materials for future studies, including clinical trials, and superior genotypes for use in developing alternative crops.
Core B: Nutrition/Animal Core
This Core has two primary objectives:
- to formulate and produce all experimental diets required for animal feeding trials to be conducted within the Center;
- to produce, through breeding, mice of novel genotypes required for the studies designed to test hypotheses relating to underlying mechanism for the actions of botanicals.
Experimental diets will be formulated according to the most recent guidelines such that the quantity and quality of the nutrients provided is both appropriate and consistent throughout the life of the Center. Botanicals and bioactive components will be incorporated into experimental diets in amounts that follow principles of allometric scaling and the concept of hormesis. The goal being to test mice at levels of botanicals that will readily translate into human intake/exposure levels. This core will work closely with the Analytical Chemistry Core to conduct quality control analyses on all diets to assure consistency from batch-to-batch.
This Center will generate nearly 6,000 mice per year to investigate the bioactivity and possible modes of action for the botanicals under study. The impact of botanicals on prostate cancer development will be explored in TRAMP mice, a well-characterized mouse model for this disease. Initial studies in Projects #2 & #3 will use some common strains of mice (e.g., C57Bl/6) to investigate the impact of botanicals on ischemia-reperfusion injury and host response to bacterial and viral infection, respectively. The role of key signaling pathways for the actions of botanicals will be investigated by using tissue-specific gene deletion techniques (i.e., Cre-flox expression targeting) in all three projects.
The role of Nrf2-dependent gene expression in botanical-mediated changes in cell function and health outcomes will be explored using mice with astrocyte-, phagocyte-, and prostate-specific deletions of Keap1. Additional mice with neuronal- and astrocyte-specific deletions of NOX2, iNOS, and nNOS will be generated to support testing novel hypothesis for Project #2 as research findings dictate.
Core C: Analytical Chemistry Core
The Analytical Chemistry Core will provide analytical support for the Research Projects and the Botanical/Plant Genomics and Nutrition/Animal Cores. The facility will minimize equipment and personnel costs, provide expert technical support, training, and quality control.
The specific aims of the facility are to:
- develop and/or validate analytical procedures for quantifying active medicinal compounds and their metabolites;
- provide analytical quantitation for quality control of diets prepared by the Nutrition/Animal Core for the Research Projects and screening of botanical samples for the Botanical/Plant Genomics Core; and
- measure circulating levels of active medicinal compounds and their metabolites in body fluids of research subjects for the Research Projects.
The Analytical Chemistry Core will evaluate and validate analytical methodology for testing phytoestrogens in soy, S-sustituted cysteine derivatives in garlic, sutherlandiosides and sutherlandins in Lessertia frutescens (also known as Sutherlandia frutescens), apocynin in Apocynum and Picrorhiza species, and the phenolics in elderberry. All botanical test materials, commercial compounds, and diets will be routinely examined for purity and/or active or marker compounds. Elderberry, soy and apocynin-producing botanicals will be screened for active constituents to identify high and low-potency materials for research studies. Circulating levels of botanical active compounds or their metabolites will provide valuable information for the three Research Projects.
Core D: Interactions Core: Mega-sequencing/Proteomics/
The Interactions Core: Mega-sequencing, Proteomics, Informatics and Nitrosylation will provide proteomic analysis, mRNA mega-sequencing, bioinformatic analyses, and biostatistics to support the research projects. For proteomic analysis of antioxidant, anti-inflammatory, NADPH oxidase, hedgehog, and estrogen signaling pathway targets, affecting protein expression levels and redox-based protein post-translational modifications (PTM), particularly S-nitrosylation on specific protein cysteine thiols, we will collaborate with MU’s Charles W. Gehrke Proteomics Center to develop an integrative proteomics strategy including both gel- and LC/MS-based quantitative proteomics.
We will develop protein cysteine post-translational modification PTM site mapping using CID/ETD technology and provide our expertise to identify potential redox targets and aid in mechanistic studies. High-throughput mRNA mega-sequencing will also be completed by personnel in this core. We will work with the MU DNA Facility and its ultra-high throughput sequencing platform. Sequence data output will be routed directly to the bioinformatician for initial processing, library construction, and screening. In later years of the funding period, we will develop protocols to look at protein-level changes in plasma.
Both genomic and proteomic data will be integrated into a pathway interaction model that will provide hypotheses for experimental validation. Overall the Core will provide support to the research projects and work to develop a deeper understanding of how botanicals can influence signaling pathways.