Research Infrastructure Core

Health Disparities in the US are widespread, severe and unacceptable. Health disparities have been attributed to both socioeconomic and biological factors. Approaches to examine the biological factors that contribute to health disparities have been rapidly evolving due to the technological advances in biomedical sciences. Through the utilization of both in vitro and in vivo experimental models, these advances have led to the understanding of the molecular mechanism of some health disparities giving rise to the translation of research to meaningful improvements in health outcomes. Over the years, NCCU has developed multiple resources for health-related research within two research institutes, the Julius L. Chambers Biomedical/Biotechnology Research Institute (BBRI) and the Biomanufacturing Research Institute and Technology Enterprise (BRITE) along with other departments in the College of Arts and Sciences (CAS), and College of Behavioral and Social Sciences (CBSS) to conduct basic and behavioral biomedical research. We have detailed our commitment to biomedical sciences and health disparities research in the overall section. To maintain and expand our health disparity research activities at NCCU and support RCHDR investigators, we propose to develop a Research Infrastructure Core (RIC) within this RCMI grant that will facilitate those goals (Figure).

This proposed Core will develop infrastructure that will function in a synergistic way to maximize the quality and productivity of the three research projects proposed in this current proposal, future pilot projects and other relevant research projects at NCCU. The RIC will also integrate strategic collaborations with researchers at area institutions and other RCMI centers to enhance our research capacity in health-related science, in particular, minority health and health disparities. As indicated in our letters of support from partners from Duke, NCSU, UNC and Wake forest (details in Investigator Development Core), we will also leverage the resources of neighboring institutions to enhance our capacity. To achieve these goals, we propose three specific aims as follows:

Specific Aim 1: To establish new resources and integrate them with existing resources for health science research.

Specific Aim 2: To support three research projects in the current proposal and future health science projects.

Specific Aim 3: To interact with area institutions and RCMI Translational Research Network (RTRN).

In summary, RIC will provide synergistic environment to support to research projects in this proposal and future health disparity research projects at NCCU and promote intra- and inter-institutional collaborations.

• The primary goal of RCHDR is to develop/strengthen the research infrastructure at NCCU and conduct cutting edge health disparities research and foster the next generation of minority biomedical researchers. The RCHDR is pleased to announce a voucher program where Principle Investigators will be provided support for his or her research program in the form of services. The vouchers are aimed to provide reagents and resources to generate preliminary data utilizing RCHDR resources and foster interdisciplinary collaborations. Junior investigators/postdoctoral scientists are encouraged to work with a faculty mentor and will be given preference for the voucher program.

• To review the request for proposal (RFP) click here. Once you are ready to apply please visit InfoReady Review.

• Contact: Mr. Marc Xavier at [email protected]; Ms. Kendra Cardwell at [email protected]

Leaders:

• TinChung Leung, PhD. Email: [email protected]; Phone: 740-250-5728

Services:

1. Experimental design of animal experiments: Researchers will be provided consultation about experimental design including statistical analysis. Senior faculty with extensive experience of animal research will be available to support junior faculty, staff and students. The in vivo unit with zebrafish and rodent expertise will help NCCU researchers to explore the feasibility of genetically engineered zebrafish and rodent models to study human diseases and to help other scientists in the NCCU research community access models or to work with Unit leaders to develop models for specific needs.
2. Breeding and genotyping of genetically modified mice and rats: The AV plus experienced faculty and animal technicians will help the project PIs develop breeding schemes, genotyping methods, and generate animals for the proposed research work.
3. Treatment of mice and rats though diet feeding, oral gavage, subcutaneous injection, intraperitoneal injection, tail vein injection, etc.: The AV will develop treatment plans in consultation with the project PI that will be executed by the ARC staff.
4. Development of proper animal models, such as xenograft model, orthotopic, carcinogenesis model, etc.: Based on the literature, research expertise, and specific research project, animal models will be developed and maintained.
5. Survival surgery, such as partial hepatectomy, ovariectomy, gastroesophageal reflux surgery, etc.: Surgical expertise on campus will be shared to help new investigators conduct survival surgery on animals. Customization is necessary to fit specific needs.
6. Sampling of animal tissues and inventory storage: When animals are sacrificed, tissues will be sampled, and stored in PBS-buffered formalin or snap frozen, according to specific research needs.
7. Coordination with other facilities including the existing Diagnostic & Molecular Histopathology Facility for analyses of animal tissues: Tissue samples from animals may be shared or transferred if they need to be analyzed by other units in this Core.

Rodent Behavioral Phenotype Core

1. Co-leaders

   • Dr. Somnath Mukhopadhyay (BBRI)

   • Dr. Bin Xu (BRITE)

2. Tests:

   • Motor function test

   • Learning and memory test

   • Sensorimotor/pain test

   • Anxiety & depression-like behavior test

   • Social behavior/interaction test

   • Abnormal repetitive behavior (autism-related) tests

3. Current users

   • Two BBRI labs (Mukhopadhyay, Cheng)

   • One MTSB lab (Marshall)

   • Five BRITE labs (Li, Lopez, Xie, Ibaneu, Xu)

Leaders:

• Somnath Mukhopadhyay, PhD. Email: [email protected]; Phone: 919-530-7762
• Emmanuel Awumey, PhD. Email: [email protected]; Phone: 919-530-7644

Services

1. Cell culture specialists will be dedicated to support the NCCU researcher in their cell & tissue culture needs. Cells can be provided in single dishes or grown to large-scale in roller bottles, shaker flasks, or spinner flasks according to research needs. Services will include but not be limited to (a) mycoplasma testing; (b) cell banking; (c) transfection services (stable cell line production); (d) large-scale cell expansion; (e) specialized multiple-layer units for anchorage cells; (f) hollow-fiber bioreactors for cellular metabolomics study; and g) complete cell cryopreservation. A centralized cell repository will be developed on site. Initially approximately 100 most commonly used neuronal, immunological, endothelial and cancer cell lines will be available, along with computerized catalogues (American Type Culture Collection, NIGMS Human Genetic Mutant Cell Repository). Individualized consultation, training sessions, and training materials in cell and tissue culture techniques will be provided, including media and systems for in vitro propagation, testing, cryopreservation and troubleshooting.
2. Proteomic service will be provided for the analysis of proteins from tissues, cells or other biological samples. Services will include: (a) protein identification; (b) protein characterization; (c) phosphoproteomics; (d) large-scale proteomic profiling; and (e) protein-protein interaction analyses using quantitative labeled or label-free approaches. Comprehensive data analysis will also be provided through a variety of database search engines (Mascot, Sequest) and software packages including Protein Pilot, Proteome Discoverer, and MaxQuant in collaboration with the Biostatistics and Bioinformatics unit.
3. Genomic services will be provided to generate genetically modified cell and animal models of human disorders/polymorphic traits using CRISPR-CAS9 and TALEN and the related gene-targeting technologies. Genomic services will also include random-insertion-based transgenesis to rapidly create transgenic cell lines/animals. In particular, the genomic services will generate not only indel mutation-based functional knockouts, but also deliver functional genetic elements to a given genomic locus using the genome editing tools: targeted genomic modification with specific SNPs, knock-in with dominant mutation sequences or functional elements such as epigenetic modifying elements or reporter genes, etc. Successfully generated lines will be screened using the Surveyor nuclease assay and verified by Sanger sequencing.

Leaders:

• Kevin Williams, PhD. Email: [email protected]; Phone: 919-530-7726
• Ju-ahng Lee, PhD. Email: [email protected]; Phone: 919-530-7003

Services

The Phenotypic Screening unit has broad experience in adapting a variety of biochemical and cell-based assay formats for HTS. Technologically, this unit has available a variety of state-of-the–art detection platforms to facilitate the implementation of assays for HTS. It has the capability to adapt assays to 96-well, 384-well, and 1536-well plate formats. The impact of these technology platforms has been to ensure a high technical probability of success in executing robust high-quality screens for assays that were originally developed in formats not amenable to HTS. The screening center routinely implements assays using fluorescence platforms including fluorescence intensity (FI), fluorescence polarization (FP), time-resolved fluorescence (TRF), fluorescence resonance energy transfer (FRET), homogeneous time-resolved fluorescence (HTRF) and the fluorometric microvolume assay technology (FMAT) using Applied Biosystems 8200 cellular detection system (CDS). The unit also employs luminescence platforms in the form of lamp-based luminescence and laser-based AlphaScreen technologies. Whole cell assays to measure activities of intracellular and transmembrane proteins can be developed using the new generation FLIPRTETRA, and the 8200 CDS, whereas, quantitative analyses of protein trafficking are measured by high content assays in the Pathway imaging system through BRITE’s imaging core. A good deal of the assay development and HTS knowledge gathered from industry has found its way into the academic screening centers such as the one at BRITE and much of assay development has become standardized.

Capabilities to utilize a wide array of 3D physiologically relevant models: (a) Primary human tissue – isolate cells using AutoMacs Bead Sorter; (b) Whole organism – zebrafish; (c) Spheroid; (d) Colony formation; (e) Pseudo-islets; (f) Patient-derived cancer cell lines; (g) Skeletal muscle; (h) Scratch assay; (i) Cell paint assay. We propose to expand our current HTS and bioimaging capabilities to include high throughput live imaging of Zb models. This will be a collaborative effort with the Animal Core to enable high-throughput zebrafish phenotyping. Hardware to support this project consists of our current VAST Bioimager. The VAST BioImager platform automatically, precisely and reproducibly orientates Zb larvae in the field of view for organ-level bright field imaging, enabling us to image large numbers and avoid manual manipulation of each larva during large screens. Linking phenotype to behavior is a critical attribute of using the Zb model. Hence, to complement the VAST system and also undertake behavioral monitoring of Zb in high throughput we propose to add the DanioVision system. The DanioVision includes an observation chamber (IR sensitive camera, white light, tapping stimulus), that allows for the tracking of up to 96 individual fish simultaneously and the capability to assess drug effects, safety pharmacology, behavioral genetics and other research areas. Using the DanioVision system the entire 96 larvae's behavior can be captured within less than a minute, providing high throughput data from a large number of larvae treated with different chemicals. With the included tactile and light stimulus units, we can for example, monitor the effects of small molecules on motor system development.

Leaders:

• Irene Doherty, PhD. Email: [email protected]; Phone: 919-530-7014
• Qing Cheng, PhD. Email: [email protected]; Phone: 919-530-5416
• Weifan Zheng, PhD. Email: [email protected]; Phone: 919-530-6752

Services:

The analysis services provided by the core will include high throughtput ‘omic technologies (i.e. RNA-Seq and ChIP-Seq), development of translational data resources, integration with analytical core to streamline the processing and analysis of ‘omic samples from collection and preprocessing to analysis and interpretation. This unit will utilize and provide a suite of software including standard and customized R tools, Qiagen Ingenuity, SAS tools and software products, Stata software, SPSS, and Qualtrics Survey software for biostatistical analysis. The core will also develop statistical analysis plans and semantic modeling data resources. The integration with chemical biology database which is based on the Chem2Bio2RDF includes, but is not limited to, drugbank, pubchem data, side effect data, pharmacology data (pharmgkb), uniprot and PDB data as well as kegg pathway data. We will create links between these databases so that cross-domain queries can be easily performed, and hidden relationships can be mined and discovered. This integrated framework will afford a translational capability that connects basic genotype information to targets/diseases and drugs, and ultimately help discover population-specific medicines. The unit will provide access for external investigators to unique data management and analysis tools and tool development services as an integral and vital component of the RTRN Network.

Leaders:

• Undi Hoffler, PhD. Email: [email protected]; Phone: 919-530-5140
• Deepak Kumar, PhD. Email: [email protected]; Phone: 919-530-7017

Services:

1. Animal Subjects Research: NCCU operates its animal care and use program under an approved Assurance from the Office of Laboratory Animal Welfare. Additionally, the University has maintained full accreditation with AAALAC International since November 2010.
2. Human Subjects Research: NCCU has been granted a Federal-wide Assurance and the University’s IRB is registered with the Office for Human Research Protections. NCCU is committed to advancing its human research protections program and will be seeking Association for the Accreditation of Human Research Protection Programs (AAHRPP) accreditation during the life of this proposed study.
3. Financial Conflicts of Interest: NCCU is in compliance with federal regulations 42 CFR 50 Subpart F and 45 CFR 94. These regulations apply to all persons responsible for the design, conduct, or reporting of PHS-funded research conducted under the auspices of the University.
4. Research Integrity: NCCU maintains an active Assurance with the Office of Research Integrity detailing the University’s policies and procedures for addressing allegations of research misconduct.
5. Responsible Conduct of Research: In response to NOT-OD-10-019, NCCU has developed formal instruction in responsible conduct of research. This instruction delivered on-line and face-to-face is designed to equip researchers at all stages of their careers, including students and postdoctoral fellows, with a fundamental understanding and working knowledge of research ethics.
6. Intellectual Property (IP) and Technology Transfer: NCCU recognizes that IP sometimes arises in the course of innovative research conducted by its employees and students using University facilities. The ORCTT will seek protection (if needed) of IP developed from this grant and facilitate opportunities for the commercialization of new technologies. These opportunities include options, licenses and startup venture development