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Rob Smart

Goodnight Distinguished Chair in Molecular Toxicology

Toxicology Building 2210


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Date: 08/01/20 - 8/01/22
Amount: $81,397.00
Funding Agencies: Lineberger Cancer Center

Despite advances in early detection and treatment strategies, the five-year survival rate for most squamous cell carcinoma (SCC) patients remains dismal (<10%). To identify new therapeutic targets, next-generation sequencing of SCCs revealed a significant number of activating mutations in the KEAP1-NRF2 pathway. NRF2’s activity, regulated by the ubiquitin ligase KEAP1, modulates a battery of cytoprotective genes against stress. Importantly, patients with cancers possessing NRF2 mutations display poorer survival and greater resistance to standard treatments. Thus, a need exists to elucidate the mechanisms by which mutant-NRF2 promotes cancer growth and survival. To discover mechanisms by which mutant-NRF2 drives SCC development, we generated a Nrf2E79Q/+ GEMM possessing one of the most common NRF2 activating mutations found in human SCC within the endogenous mouse Nrf2 locus. Using this model, we will determine the impact of mutant-NRF2 signaling on tumor development, progression and metastasis in the classic cutaneous SCC model. We hypothesize that mutant-NRF2 signaling will promote the progression of cutaneous SCC through novel downstream pathways. We will combine the expertise of 2 investigators, Dr. Robert Smart, a NC State Professor and Dr. Bernard Weissman, a UNC Professor, to perform the following aims: AIM#1: Determine the effects of mutant Nrf2E79Q expression on the development of squamous papillomas and their progression to SCCs and AIM#2: Identify transcription targets unique to mutant NRF2 in epidermis, papillomas and SCCs. Our research will help develop targeted drug therapies for human SCCs with frequent NRF2 mutations such as cutaneous, bladder, head and neck and esophagus.

Date: 07/01/77 - 6/30/22
Amount: $7,541,891.00
Funding Agencies: National Institutes of Health (NIH)

This is a continuation of Training in Biochemical and Environmental Toxicology. The main objective of this training grant is to provide didatic and research training to Ph.D. students in Toxicology

Date: 06/01/16 - 5/31/22
Amount: $1,692,571.00
Funding Agencies: National Institutes of Health (NIH)

Only ~3% of human genome encodes protein. The remaining 97% of the human genome is referred to as noncoding DNA. Initially, much of the intergenic noncoding sequence was referred to as “junk DNA” as it was considered to have no function. While some intergenic sequences contain DNA elements important in gene regulation, many intergenic sequences can be transcribed into RNA. In fact, ~85% of the human genome is transcribed into RNA. RNAs that lack protein coding function are referred to as noncoding RNAs (ncRNAs) and of these the long noncoding RNAs (lncRNAs >200 nt) represent the majority. LncRNAs are one of the largest and more diverse classes of cellular transcripts with over 10,000 lncRNA transcripts reported in the human genome; in most cases their biological function is unknown. Emerging evidence indicates they play an important role in regulating gene expression and are associated with human diseases such as cancer, Alzheimer’s and heart disease. There is a critical need to determine whether associations of lncRNAs with specific disease are functionally significant and to define and characterize the function of lncRNAs using in vivo disease model systems. Given that the etiology of most chronic human diseases involves interactions with environment, it is also important to determine how environmental factors impact the expression, activity and function of lncRNAs. Nonmelanoma skin cancer (NMSC) is the most common cancer in the United States. The majority of NMSCs is caused by solar UVB radiation. p53 plays a key role in the response of skin keratinocytes to UVB-induced DNA damage by inducing cell cycle arrest and apoptosis. In skin cancer, the incidence of p53 mutations ranges from 50-90%. UVB-induced mutation of p53 allows keratinocytes upon successive UVB exposures to evade apoptosis and cell cycle arrest and these defects have a critical role in skin cancer development. LincRNA-p21 is a lncRNA and was recently discovered to be a direct transcriptional target of p53 where it serves as a mediator of p53-dependent transcriptional repression. We observed that; i) lincRNA-p21 is highly inducible by UVB in the mouse skin in vivo and in human/mouse keratinocytes, ii) UVB-induction of lincRNA-p21 is p53-dependent and iii) lincRNA-p21 has a key role in UVB-induced apoptotic cell death in keratinocytes. Our plan is to characterize the regulation and function of lincRNA-p21 in keratinocytes and skin in vivo in response to UVB and to define the role of lincRNA-p21 in UVB-induced skin cancer. The central hypotheses are i) lincRNA-p21 is induced by UVB in keratinocytes through a p53-dependent pathway to produce apoptotic cell death and ii) lincRNA-p21 functions as a tumor suppressor in NMSC whereby the loss of lincRNA-p21 expression allows mutant p53 and non-mutant p53 keratinocytes to evade UVB-induced apoptotic death leading to skin cancer. The proposed studies are significant as they represent the first characterization of a lncRNA function in an in vivo disease model with a highly relevant environmental component; moreover if the hypothesis is correct this will be the first demonstration of a lncRNA functioning as a tumor suppressor in vivo.

Date: 05/11/20 - 3/31/22
Amount: $3,674,130.00
Funding Agencies: National Institutes of Health (NIH)

The vision of the Center for Human Health and the Environment (CHHE) is to become a global leader in environmental health sciences (EHS) along the continuum from genes to populations by building on NC State’s unique research strengths and resources in quantitative biology, -omics and analytical technologies, and diverse model organisms, as well as its emerging strength in human population science. Through the purposeful interfacing of different disciplines and a systems biology framework integrating all levels of biological organization - biomolecule, pathway, cell, tissue, organ, model organism, human, and human population - CHHE will elucidate fundamental mechanisms through which environmental stressors interface with pathways, the genome, and epigenome to influence human health outcomes. CHHE has made outstanding progress in the first funding cycle. CHHE has significantly: (1) advanced innovative multi-disciplinary EHS team research; (2) expanded its NIEHS grant base; (3) increased EHS capacity at its partner institutions, East Carolina University and North Carolina Central University; (4) cultivated the next generation of EHS leaders; and (5) developed multi-directional engagement with communities affected by exposure to toxic metals and per- and polyfluoroalkyl substances. During our first funding cycle, four Research Interest Groups (RIGs) evolved organically and the Emerging Contaminants, Environmental Epigenetics and Genetics, Pulmonary Health, and Behavior and Neuroscience RIGs now represent CHHE thematic areas. In the coming cycle, we have enhanced our three facility cores to increase the impact and the basic science and translational capacity of our membership. The Systems Technologies Core provides cutting-edge technologies involving genomics, metabolomics, metallomics, and proteomics. The Comparative Pathology Core provides pathologic phenotypic assessment of the many model organisms used by members and imaging support with links back to omics technologies. The Integrative Health Science Facility Core facilitates bidirectional translation between basic science and public health outcomes by providing data science analysis and visualization support for analysis of human population and multi-omic studies as well as population-based study expertise. As a land-grant university, NC State has a dedicated community engagement philosophy that augments CHHE’s Community Engagement Core and fosters relationships between CHHE and affected communities in NC which leads to collaborative interaction among researchers, educators, and citizens to enhance EHS knowledge, literacy, and health. A strong Career Development Program for early- and mid-career investigators is coordinated with a robust Pilot Project Program that supports collaborative and multidisciplinary EHS projects to enhance the research success of our members. Our CHHE mission is to continue to evolve as a premier NIEHS EHS Core Center and serve as the nexus of EHS research at NC State by providing focus, resources, and leadership for interdisciplinary research that will improve human health locally, nationally, and globally.

Date: 02/28/20 - 1/31/22
Amount: $3,938,845.00
Funding Agencies: National Institutes of Health (NIH)

Per- and polyfluoroalkyl substances (PFAS) are emerging as a major public health problem in North Carolina and across the United States. PFAS comprise a class of over 5,000 compounds. Their unique chemical properties have been harnessed to make consumer and industrial products more water, stain, and grease resistant; they are found in products as diverse as cosmetics and flame-retardants. PFAS are resistant to degradation, move easily through the environment, and accumulate in living organisms. Exposure to PFAS has been associated with health effects including cancer and toxicity to the liver, reproductive development, and thyroid and immune systems. Despite widespread detection in the environment and evidence of increasing human exposure, understanding about PFAS toxicity, its bioaccumulative potential in dietary sources such as aquatic organisms, and effective remediation remain notably understudied. The recent discovery by this proposed Center’s Deputy Director, Dr. Detlef Knappe, of widespread PFAS contamination in the Cape Fear River watershed in NC underscores that these compounds are in need of immediate investigation.. The goal of our Center is to advance understanding about the environmental and health impacts of PFAS. To meet this goal we are employing a highly trans-disciplinary approach that will integrate leaders in diverse fields (epidemiology, environmental science and engineering, biology, toxicology, immunology, data science, and advanced analytics); all levels of biological organization (biomolecule, pathway, cell, tissue, organ, model organism, human, and human population); state-of-the-art analytical technologies; cutting-edge data science approaches; a recognized track record in interdisciplinary, environmental health science (EHS) training; and well-established partnerships with government and community stakeholders.

Date: 04/20/15 - 3/31/21
Amount: $6,127,354.00
Funding Agencies: National Institutes of Health (NIH)

The mission of the Center for Human Health and the Environment (CHHE) is to advance understanding of environmental impacts on human health. Through a systems biology framework integrating all levels of biological organization, CHHE aims to elucidate the fundamental mechanisms through which environmental exposures/stressors interface with biomolecules, pathways, the genome, and epigenome to influence human disease. CHHE will develop three interdisciplinary research teams that represent NC State’s distinctive strengths. CHHE will implement specific mechanisms to promote intra- and inter-team interactions and build interdisciplinary bridges to advance basic science discovery and translational research in environmental health science along the continuum from genes to population. These teams are; - The Molecular/Cellular-Based Systems and Model Organisms Team will utilize cutting edge molecular/cellular-based systems and powerful vertebrate and invertebrate model organisms to define mechanisms, pathways, GxE interactions, and individual susceptibility to environmental agents. - The Human Population Science Team will integrate expertise on environmental exposures, epidemiology, genomics and epigenomics to identify key human pathways and link exposure and disease across populations. - Bioinformatics Team will develop novel analytics and computational tools to translate Big Data generated across high-throughput and multiscale experiments into systems-level discoveries To further increase the impact and translational capacity of these teams, CHHE will develop three new facility cores that will provide instrumentation, expertise, and training to facilitate basic mechanism- to population-based research. - The Integrative Health Sciences Facility Core will expand the ability of CHHE members to translate basic science discoveries across species and provide mechanistic insights into epidemiological studies by partnering with: a) NC State’s Comparative Toxicogenomics Database (CTD); b) East Carolina University Brody School of Medicine and c) NC Dept. of Health and Human Services. - The Comparative Pathobiology Core will be located at NC State’s top-ranked College of Veterinary Medicine and its nationally recognized veterinary pathology group to facilitate assessment of the effects of environmental stressors in the many model organisms utilized by CHHE members. - The Systems Technologies Core will introduce state-of-the-art proteomics capabilities and dedicated bioinformatics support to expand the ability of CHHE members to analyze the Next Generation Sequencing data involving the genome, transcriptome and epigenome. As a land-grant university, NC State has an extensive and active Cooperative Extension Service network throughout North Carolina. CHHE will utilize this unique network to develop a highly effective, multi-directional Community Outreach and Engagement Core to disseminate findings that will contribute to addressing disparity in exposures and health outcomes and to educate communities about environmental influences on health. A strong Career Development Core for early stage scientists that is coordinated with a robust Pilot Project Program will support cutting-edge, collaborative and multidisciplinary environmental health projects to enhance the research success and impact of our membership. Through these activities and the purposeful interfacing of different disciplines CHHE will build on NC State’s unique research and community outreach strengths to become a premier transformative and synergistic EHS Core Center.

Date: 11/01/17 - 10/31/20
Amount: $701,881.00
Funding Agencies: National Institutes of Health (NIH)

GenX is a perflourinated compound used and generated in the production of non-stick coatings. This chemical has been detected in the Cape Fear River in North Carolina, upstream of where the Cape Fear Public Utility Authority obtains drinking water for the City of Wilmington (population ~250,000), Brunswick County and Pender County, NC. In June 2017, community concern about this chemical in drinking water resulted in multiple public meetings of citizens trying to obtain information about their potential exposure and resulting health effects associated with consumption of drinking water. As a result of community concern, the chemical plant has reportedly stopped discharging GenX into the river. However community concern still exists. This project is designed to help address community concerns about GenX exposure and health effects. Little is known about how GenX is stored in the body, the toxicity of GenX, or how long the chemical will remain in the environment. To address these questions, we plan to conduct a community-based study of Wilmington area residents who are served by public utility water. We will work with community partners of the Cape Fear Riverkeeper and the New Hanover County Department of Health to help identify a representative sample of residents, to collect biological samples, and to keep the community informed about what is known about GenX and what the study finds. We plan to recruit ~400 Wilmington area residents (100 men, 100 women, 100 boys, 100 girls) to provide blood, urine, and drinking water samples and to complete a questionnaire on their water use history. We plan to analyze blood, urine, and drinking water for GenX and related perflourinated chemicals; blood and urine samples will also be used for clinical tests (lipid profile, thyroid function, liver function, and urinalysis). All results from the study will be shared with both the community as a whole and each individual participant. We will have a community advisory panel for the study to help advise about study protocols, methods of reporting back results to participants, and provide guidance on ongoing or new community concerns about GenX. This project leverages the expertise of NC State’s Center for Human Health and the Environment to respond to an emerging community concern.

Date: 01/01/17 - 3/31/19
Amount: $37,875.00
Funding Agencies: NCSU Center for Human Health and the Environment

The transcription factor p53 can be activated by several environmental stressors including UVB solar radiation, to regulate the transcription of genes involved in senescence, apoptosis and cell cycle arrest. These anti-tumor activities of p53 are essential to prevent cancer. p53 is a tumor suppressor and approximately half of all human cancers have p53 alterations that results in loss of p53 transcriptional activity. Recently, we discovered that deletion of the C/EBPβ transcription factor increased p53 protein levels and enhances p53 pro-apoptotic activity/apoptosis greater than 3-fold following UVB exposure. We have also found that UVB treatment induces a C/EBPβ-p53 complex and knockdown of C/EBPβ in UVB-treated keratinocytes increases p53 protein stability. These results suggest C/EBPβ is a negative regulator of p53 pro-apoptotic activity, through an unknown mechanism. C/EBPβ has a key role in an environmentally-induced disease as C/EBPβ-/- mice are resistant to UVB-induced skin cancer, however it is unknown if the enhancement of p53 pro-apoptotic activity prevents the onset of UVB-induced skin cancer in C/EBPβ-/- mice. Through utilization of ChIP-seq and RNA-seq we will map both C/EBPβ and p53 chromatin interactions and determine the effects of deletion of C/EBPβ on the p53 transcriptional network. These studies will provide mechanistic insight into an uncharacterized function of C/EBPβ in regulating p53 prop-apoptotic activity following UVB exposure and will enhance the foundation and fundability for our long-term goal to determine how C/EBPβ regulates p53 activity, stability and protein-protein interactions and the importance of this function of C/EBPβ in an environmentally-induced disease, UVB-induced skin cancer.

Date: 04/01/04 - 3/31/16
Amount: $2,440,806.00
Funding Agencies: National Institutes of Health (NIH)

Cytokines play important roles in human development and disease. Specificity and cross-talk of cytokine signaling pathways appear to be important for fine-tuning stress responses and cell fate decision during development. Transforming growth factor b (TGF-b) is involved in cell growth, differentiation, tissue remodeling, immune response and angiogenesis. Interleukin 1 (IL-1) pathway plays a central role in the generation of inflammatory responses. We have found that both TGF-b and IL-1 activate TGF-b activated kinase 1 (TAK1) MAPKKK. Active form of TAK1 can enhance both TGF-b- and IL-1-dependent transcription. In response to IL-1 stimulation, TAK1 activates transcription factors AP-1 and NF-kB. While TGF-b stimulation does activate TAK1, the role of TAK1 in TGF-b signaling pathway is not known. Recently, we found that TAK1 associates with a transcriptional repressor SnoN, a negative regulator of TGF-b signaling. TAK1 induces degradation of SnoN. We hypothesize that TGF-b activates TAK1 to induce phosphorylation of SnoN and targets SnoN for proteasomal degradation, thereby up-regulating TGF-b signal transduction. In addition, we hypothesize that TGF-b and IL-1 activate TAK1 in distinct manner via specific scaffold/regulatory proteins to induce their unique cellular responses. Thus, the overall objectives of this proposal are; to delineate the pathway and functional role of TAK1 in TGF-b signaling and to elucidate the mechanisms through which TAK1 regulates signal pathway specificity. To accomplish these objectives and to test our hypotheses we will; i) determine the mechanism and role of TAK1-induced SnoN degradation in TGF-b signaling pathway; ii) isolate and characterize molecules associated with TAK1 and iii) generate a skin specific knockout of TAK1 to characterize the in vivo role of TAK1 in a tissue in which TGF-b play important roles. These studies will address unsolved questions regarding the mechanisms of TGF-b and IL-1 family signaling and will provide an understanding of the physiological function of TAK 1 in vivo.

Date: 02/01/01 - 5/31/15
Amount: $2,496,522.00
Funding Agencies: National Institutes of Health (NIH)

This is a continuation to study the roles of C/EBP transcription factors in squamous differentiation and cancer.

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