Pilot Project Awards

Principal Investigators: Becky F. Antle, PhD, MSSW, LMFT
Co-Investigator: Luz Huntington-Moskos, PhD, RN, CPN
Title: Community Needs Assessment and Training to Address Health Disparities among Refugee/Immigrant Populations in Louisville
Description of Project: This project will assess the environmental health needs of two refugee/immigrant populations in the Louisville area through partnerships with See Forward Ministries (African populations), Backside Clinic/Learning Center and South Jefferson Neighborhood Place (Hispanic populations). Preliminary data from existing partnerships with these organizations shows community concerns regarding neighborhood safety and health, as well as overall challenges related to well-being indicators for these target groups. This project will conduct a needs assessment and develop training strategies for community members and providers to reduce health disparities and promote environmental justice.

Principal Investigators: Natalie DuPre, ScD; Sandy Kavalukas, MD
Collaborators: Lu Cai, MD, PhD
Title: Lifetime Environmental Exposure to Metal Carcinogens and Colorectal Cancer: A Clinical Case-Control Study
Description of Project: This pilot project will directly measure known environmental metal carcinogens in blood and urine in Louisville residents with colorectal cancer and without colorectal cancer from a colorectal cancer surgical clinic at UofL Health. It is unknown whether environmental metal carcinogens increase risk of colorectal cancer. However, in prior work, we observed colorectal cancer hotspots within Louisville and observed that higher exposure to ambient arsenic and ambient cadmium was associated with residing in these colorectal cancer hotspot neighborhoods. This pilot study will compare levels of environmental carcinogens in urine and blood samples of Louisvillians with colorectal cancer (i.e., “cases”) and without colorectal cancer (i.e., “controls”). It will provide necessary information to: 1) demonstrate our ability to develop a clinical case-control study in our community, and 2) compare levels of environmental carcinogens in patients with colorectal cancer and patients without colorectal cancer.

Principal Investigator: Jiapeng Huang, MD, PhD
MPIs/Co-Investigators: Lu Cai, MD, PhD (MPI); Lihui Bai, PhD (MPI); Xiaoyu Chen, PhD (MPI); Yash Kothari, MD (Co-I); Luz Huntington-Moskos, PhD, RN, CPN (Co-I)
Title: Metallomics and Machine Learning to Improve Risk Stratification and Optimize Treatment in Pulmonary Arterial Hypertension
Description of Project: In Louisville Regional Community, there are thousands of patients who suffer pulmonary hypertension (PH) due to bad genes, industrial pollutions, smoking, or lung and heart diseases. PH happens when the pressure in the blood vessels leading from the right heart to the lungs is too high and the right heart eventually fails over time. There is not enough unoxygenated blood going to the lungs causing low oxygen levels. PH is not reversible, and most patients die within 2 to 3 years after the diagnosis. Many PH patients have severe trouble breathing even at rest with poor quality of life. There is no cure for PH because we do not understand what causes PH in many cases. Our team found several metals are significantly different in PH patients when compared to controls in a pilot study for the first time. Metals could directly cause PH or change levels when patients develop PH. This project will evaluate 25 metal levels in the world’s largest collection of PH patients’ data and samples from many hospitals across USA. This large sample size enables us to draw solid conclusions confidently. We will use cutting edge mathematical prediction tools and decision support systems to help diagnose PH earlier, treat PH better, and find new medications targeting metals. The sooner appropriate PH treatments are started, the better the patient will do.

Principal Investigators: Timothy O’Toole, PhD
Co-I/Collaborators: Jingjing Zhao, PhD
Title: Exposure to Polystyrene Microplastics Exposure and Cardiovascular Outcomes
Description of Project: Modern society produces large amounts of plastics, much of which is discarded in landfills and water systems. In these ecosystems, there is a gradual breakdown of larger plastics into small particles (microplastics), by natural erosion and chemical forces. These microplastics can find their way into water supplies and the food chain where human consumption is inevitable, as evidenced by their detection in human blood, breast milk, and urine as well as in some organs like the lungs. The consumption of microplastics will be an ongoing concern in the future, given that globally increasing temperatures and UV exposures can accelerate the breakdown of large plastics. Furthermore, microplastics consumption is of a particular concern to residents of the greater Louisville area, given that our drinking water derives from the Ohio River, and that high levels of larger plastics and microplastics have been found up-stream in the Pittsburgh and Cincinnati areas.
The incidence of diabetes and obesity are considerable global health problems. Significantly, the Louisville area, and Kentucky in general, have some of the highest rates of diabetes and obesity in the US. These disorders are risk factors for larger cardiovascular disease and increase the probability of developing atherosclerosis and the incidence of heart attacks and strokes. Understanding why our population is more susceptible to these outcomes is an important area of research. Thus, in this proposal we will determine if there is any association between microplastics consumption and increased cardiovascular disease. This will be done in a mouse model, where polystyrene beads are supplied to these animals in drinking water. After this feeding, we will measure several markers of cardiovascular disease and also determine if these exposures alter the identity and abundance of bacterial species in the gut, which may also contribute to adverse outcomes. Results from this proposal will shed insight as to if and how consumption of microplastic particles impacts our health.

Principal Investigator: Kenneth Palmer, PhD
Collaborators: Daniel J. Conklin, PhD and Anna Gumpert, PhD
Title: Effects of Acrolein and Hypertension on COVID-19 (Sars-CoV-2) and Cardiopulmonary Injury in Mice 
Description of Project: From early in the COVID-19 pandemic, the National network of twelve regional biocontainment laboratories (RBL), including the University of Louisville Center for Predictive Medicine for Biodefense and Emerging Infectious Diseases (CPM) facilitated many biomedical innovations including novel COVID-19 diagnostics, vaccines and antiviral therapeutics. Our RBL rapidly developed preclinical models of SARS-CoV-2 infection and made major contributions to several community-engaged research projects. This has led to targeted investment from NIH/NIAID in enhancing our facilities and equipment. Since the University of Louisville is recognized as a distinguished Center of Excellence in Environmental Research, Dr. Palmer and Dr. Conklin are working together to build collaborations between infectious disease researchers in the CPM and environmental health researchers in the CIEHS and affiliated centers to better understand how environmental conditions impact emerging infectious disease problems. The common environmental pollutant acrolein is toxic to the lungs, heart, and the circulatory system and increases cardiovascular disease risk. Given the reality that COVID-19 outcomes are worse in patients with underlying cardiometabolic disease, we propose that studies focused on understanding how acrolein exposure impacts the severity of SARS-CoV-2 infection in mice will be a useful way to start integrating environmental exposure studies with emerging infectious disease research.

Principal Investigator: Anna Gumpert, PhD
Collaborator: Daniel Conklin, PhD
Title:  "Combined Effects of Air Pollution and Hypertension on Cardiovascular Remodeling"
Description of Project: Air pollution is ubiquitous, and exposure to air pollution is associated with 200,000 excessive deaths yearly in the U.S., of which the majority (60–80%) is due to cardiovascular disease. Although air pollution is typically higher in urban centers, unique sources of air pollution including particulate matter (PM) and gases (volatile organic compounds) can occur anywhere. One such phenomenon was observed in the past few years when widespread wildfires throughout North America contributed to increased air pollution both locally but also at distant locations. Additionally, rurally located industrial sources of air pollution can disproportionally contribute to local air pollution, as witnessed by the hemp processing plant located in western Kentucky. These exposure events demonstrate that air pollution is a health issue everywhere including in Kentuckiana and regionally. We will use Louisville’s particulate air pollution to perform controlled exposures to better understand the ways that air pollution influences cardiovascular health. Ultimately, we want to better protect the health of members of our regional community and worldwide with special emphasis on those who have pre-existing conditions such as high blood pressure – the most important modifiable risk factor of cardiovascular disease in the world.

Principal Investigator: Mikus Abolins-Abols, PhD and Ray Yeager, PhD
Title: "Songbirds as air pollution sentinels for public health: jointly testing the effect of pollution on avian and human stress and inflammation markers"
Description of Project: Exposure to air pollution has well documented negative effect on human health. However, assessing the health risk of air pollution at a local scale can be difficult due to many barriers to localized human studies. Using wild animals as bioindicators of pollution can solve these issues. Birds and mammals respond to pollution in very similar ways to humans, and large numbers of animals can be sampled more rapidly and affordably than humans. In this study, we will ask if American robins, common native urban songbirds, can serve as bioindicators for air pollution by comparing how air pollution affects health of both robins and humans in the same locations. This study has the potential to provide new insights on the burden of pollution exposure on cardiovascular disease across diverse communities in Louisville and other cities across the South and Midwest. This work could ultimately inform novel ways to identify, assess, and develop interventions to environmental health issues.

Principal Investigator: Haixun Guo, PhD
Co-Investigator: Lu Cai, MD, PhD
Title: "64Cu-PET metallomics for non-invasively tracing copper distribution in response to cadmium and/or HFD"
Description of Project: Cadmium has been known to cause a variety of ill effects from acute and chronic exposure, including nausea, vomiting, diarrhea, muscle cramps, liver injury, convulsions, and kidney failure. Long-term exposure to environmental contaminated metals, such as cadmium in drinking water and food has been linked to health problems such as liver, bone, and blood damage (US. Environmental Protection Agency, 2003). Although cadmium in Kentucky groundwater rarely exceeds the maximum contaminant level (MCL), Kentucky has been ranked number one as cadmium contamination by EPA, due to the coal mine and other industries. For instance, cadmium concentrations might expect to be higher in western Kentucky (specifically Caldwell, Crittenden, and Livingston Counties), where fluorspar was mined in the 1940’s, because of the association of barium and galena (lead ore) with fluorite. The Eastern Kentucky Coal Field also contains high amounts of sphalerite (zinc ore) associated with coal seams, which could cause higher cadmium levels. Therefore, to understand how cadmium affect our body is very important for us to develop the means to prevent cadmium-caused bad effect on our body. It is known that one of the reasons why cadmium exposure damage our body is due to its disturbance of other metals that are need to keep certain level for keeping us healthy. We already know cadmium exposure increases our body copper levels that also cause our tissue cell death. Therefore, we need to know which organ enriches the copper in the body. Therefore, there is an urgent need for tool and technology to non-invasively diagnosis, measure, and monitoring the cadmium exposures across the lifespan for Kentucky residents. The current project utilizes the non-invasive in vivo imaging technology, PET metallomics, to track the dynamic copper homeostasis response to the cadmium exposure in living bodies. The success of this project will provide a non-invasive imaging technology for monitoring the cadmium exposure-altered copper homeostasis in vivo.  Such imaging technology can be translated directly to cadmium-exposed patients by using the clinical PET scanner in Radiology for both diagnosis and therapeutic efficacy monitoring.

Principal Investigator: Banrida Wahlang, PhD
Collaborator: Shesh Rai, PhD and Michael Merchant, PhD
Title: "Sex-dependent Effects of Organochlorine Pesticides on Metabolic Diseases: Role of the Gut-Liver Axis"
Description of Project: Organochlorine pesticides (OCPs) were initially manufactured for agriculture and household purposes but were banned worldwide two decades ago when it was discovered that these chemicals elicited harmful effects on our ecosystem and wildlife. Despite this ban, OCPs continue to persist in our environment including our soils and rivers due to their resistance to chemical degradation, which is why OCPs are commonly referred to as “forever chemicals”. Exposures to OCPs still occur throughout the world, and not just in certain developing countries where these chemicals are still in use. The United States, too, has considerable exposure rates even though it was one of the first countries to ban these pesticides. In fact, significant levels of these chemicals were recently detected in residential areas in Louisville, Kentucky. The EPA reported that these detected OCPs likely migrated through the soil and sewer system, from the Black Leaf Chemical plant, which was already abandoned decades ago. The presence of these chemicals in the Ohio River and other Kentucky streams have also been reported. Importantly, marine organisms including fish can accumulate these “forever chemicals” in their bodies, and exposure in humans often occur through ingestion of pesticide-laden fish and water. Indeed, significant levels of these pesticides are still detected in blood samples in the American adult population. Scientific studies have shown that increased levels of these pesticides in human blood are directly correlated with numerous diseases, including obesity, high cholesterol levels and fatty liver. However, studies looking at how these chemicals can contribute to these diseases are still scarce and limited. Moreover, most published studies looking at health effects of pesticides in humans were done in agricultural workers, who were predominantly men. Therefore, this proposal seeks to understand “how” exposure to OCPs can contribute to such liver and metabolic diseases, and if the impact from these exposures is “different between males and females” (sex differences). Identifying how sex differences influence chemical exposure is of utmost importance because it may help explain why some diseases are more prevalent in women, while others are more prevalent in men. Furthermore, men and women may behave differently in their responses to chemical exposure due to their fundamental biological make-up and can have different levels of these chemicals in their bodies. Thus, the proposal, if funded, will provide ground-breaking insight into how pesticide exposure, which is relevant to the Louisville Regional Community, can contribute to metabolic diseases. The information, concepts and findings gained from this proposal will contribute to people in our community in the following ways: this research will 1)will help address sex and gender disparities in environmental health, 2) provide health authorities with further knowledge to identify environmentally-induced diseases in susceptible populations, 3)provide information for policy changes and decision-making, 4) help design sex-specific intervention strategies to better control future health care costs.

Principal Investigator: Barbara Clark, PhD and Carolyn Klinge, PhD
Consultant: Nichola Garbett, PhD
Title: "STARD5 and lipid dysregulation in toxicant-associated steatohepatitis (TASH)"
Description of Project: Non-alcoholic fatty liver disease (NAFLD) is a metabolic disorder that is characterized by an accumulation of fats in the liver, termed steatosis. NAFLD has increased in parallel with the rise in obesity over the past decade. Simple fatty liver (steatosis) can progress to fatty liver with inflammation (steatohepatitis or NASH). It is well-established that environmental chemicals, or toxicants, worsen obesity-induced NASH and this disease has been defined as toxicant associated steatohepatitis (TASH). Currently, there is no FDA-approved treatment for NAFLD-NASH/TASH, and agents that have shown beneficial results in phase III clinical trials appear to be effective on only a subset of patients. Therefore, there is a need to identify novel markers or druggable targets that may work on a subset of patients not responsive to the current therapies. We have identified a potential novel target, called STARD5, and the goals of this pilot project are to determine whether STARD5 contributes to TASH development. To address this question we generated a mouse that lacks STARD5 (Stard5-/-) and treated them with the endoplasmic reticulum (ER) stress inducer Tunicamycin. Chronic ER stress from over nutrition and environmental toxicants contributes to development of NAFLD/TASH and this pilot project will answer the question if the ER stress-induced liver NAFLD/NASH markers are different between the Stard5-/- and wild-type mice. We have also modeled polychlorinated biphenyls (PCBs), major environmental toxicants of concern, binding to STARD5. This pilot project will determine whether selected PCBs physically bind to STARD5. The potential for a STARD5-PCB interaction could alter the toxicity of PCB exposure in TASH development. Detection of PCBs in human populations is widespread and higher serum PCB levels have been associated with greater liver disease severity. PCB exposure is likely higher for residents surrounding the Arkema facility located in Marshall Co, KY (western KY region) that was a PCB manufacturing facility. The data generated from this pilot project will support future investigations on STARD5 as a risk factor for development of TASH.

Principal Investigator:   David Hein, PhD
Collaborator: Raul Salazar-Gonzalez, PhD           
Title: "Gene-environmental interactions of novel psychoactive chemicals substituting for illegal drugs of abuse"   
Description of Project: New psychoactive substances (NPS) include many different chemicals commonly sold as legal substitutes for classical drugs of abuse. NPS were mostly central stimulants in the phenylethylamine group but more recently the number of NPS is expanding. Public health institutions are closely investigating the epidemiology and toxicovigilance of NPS. The number of NPS has dramatically increased during the last few years. The information about the basic pharmacology of hallucinogenic phenylethylamines from the 2C series is scarce. Published binding studies show an affinity for G-protein–coupled 5-HT and adrenergic receptors, in which the 2C compounds may act either as agonists or antagonists. Toxicological effects including changes in mitochondrial membrane potential, decreased ATP and increased ROS production, increased levels of DNA damage and activation of apoptosis have been reported in preliminary studies. 
Arylamine N-acetyltransferase (NATs) are cytosolic enzymes, expressed as the two isoenzymes NAT1 and NAT2. NAT2 is a highly polymorphic gene, depending on the combination of NAT2 alleles, individuals can be categorized into rapid, intermediate or slow acetylators, and this NAT2 phenotype has been shown to have an influence on metabolism, and thus the efficacy and toxicity, of certain drugs. Interestingly, recent reports have demonstrated that NPS compounds, specifically 2C derivatives are N-acetylated by NAT2, but the influence of genetic polymorphism was not considered. In this pilot study we will: 1) evaluate the differences in metabolism of NPS between rapid, intermediate and slow acetylation phenotypes; and 2) investigate the effect of such differences using a comprehensive toxicology evaluation that includes DNA damage markers, apoptosis evaluation, ATP and mitochondrial function as well as oxidative stress markers, in order to assess the toxic effects by the N-acetylation of these compounds in the presence of different NAT2 genotypes. The aims of the pilot proposal are to evaluate 1), Is the N-acetylation of NPS NAT2-genotype-dependent? and 2) Are the toxic effects of the NPS modified by NAT2 genotypes? The evaluation of such gene-environment interaction is of high importance to public health and also have important implications to understand the toxic effects of these drugs of abuse.

Principal Investigator: Rachel Neal, PhD
Co-Investigator: Cynthia Corbitt, PhD      
Title: "Hepatic metabolic response to the fasting/refeeding transition in offspring exposed in utero to vaping"                   
Description of Project: This project will examine the impact of maternal vaping on key offspring liver nutrient utilization pathways in conjunction with weight gain, using two murine model systems. Outcomes of this study are critically important and necessary to characterize the potential impacts of maternal vaping on the fetal environment and infant health. 

Principal Investigator: Xiao-An Fu, PhD
Co-Investigator: Qunwei Zhang, PhD 
Title: "Analysis of harmful compounds in aerosols of electronic cigarettes to evaluate toxicity"                   
Description of Project: The dramatic increase of toxic chemicals, including formaldehyde, and acetaldehyde in aerosols of the newer generations of electronic cigarettes or e-cigarettes has raised significant public concern over the safety in using e-cigarettes. There were 4.47 million high-school and middle-school students used e-cigarettes in 2020 as reported in the National Youth Tobacco Survey.  Currently, the constituents in aerosols of e-cigarettes are not comprehensively measured and the toxic effect of the mixtures of toxicants in aerosols are not well studied.  Therefore, there is an urgent need to develop analytic approaches for comprehensive measurements of toxic chemicals in aerosols of e-cigarettes and to evaluate e-cigarette toxicity.   
This pilot project will develop a comprehensive approach for measurements of harmful chemicals in flavoring electronic liquids and e-cigarette aerosols and determine exacerbating toxicity of these chemicals through examining human lung cell cytotoxicity. Flavoring chemicals and thermal degradation of these flavoring chemicals and propylene glycol and vegetable glycerol form a mixture of toxicants in aerosols that will exacerbate toxic effect. This project will analyze all harmful organic compounds in aerosols of laboratory formulated e-liquids to understand the sources of toxic compounds. Constituents in five most popular flavoring electronic liquids and e-cigarettes used by teenagers and in aerosols of the e-cigarettes will then be analyzed to identify all toxicants. The toxic effects of e-cigarettes on human lung cells in vitro will be examined. The data obtained from this project will be informed to public by publications and to FDA for regulation of e-cigarettes. 

Principal Investigator: Jun Cai, MD, PhD
Co-Investigator: Lu Cai, MD, PhD, Gregory Barnes, MD, PhD 
Title: "Disruption of β-catenin destruction complex and ASD-like behaviors in whole-life cadmium exposure and postnatal obesity" 
Description of Project: Autism spectrum disorder (ASD) is a neurodevelopmental disease affecting young children and adults, which is characterized by communication, sensory and social deficits as well as repetitive behaviors. Both genetic vulnerability and environmental factors contribute to the etiology of ASD. However, it is rarely known how environmental factors trigger gene modifications for the development of ASD. The accumulating evidence indicates an association of either exposure to toxic metals including cadmium (Cd) or obesity with ASD or other neurodevelopmental disorders (NDD) in humans. Our preliminary study showed that high-fat diet (HFD) significantly increases the Cd accumulation in kidney, liver, heart and brain of the mice with a 24-week postweaning HFD feeding and whole-life exposure to low-dose Cd compared to those with Cd exposure only. The Wnt/β-catenin/GSK3β pathway, the vital hub in the onset/development of ASD/NDD, is disrupt in the cortexes of those offspring mouse brains. Cd exposure enhances the phosphorylation of serine 9 in GSK3β and HFD promotes transcriptional expression of GSK3β, probably altering the cellular accumulation of β-catenin. In this pilot project, we will test the hypothesis that whole-life low-dose Cd exposure disrupts the β-catenin destruction complex and develops the ASD-like behaviors in mice, which would be exacerbated by postweaning HFD. To test this hypothesis, we will collaborate with Drs. Lu Cai and Gregory N. Barnes (members of the CIEHS), who have an extensive expertise in the fields of cadmium toxicity, obesity, genetics, and autism. We propose 1) to examine the levels of Cd and essential metals (zinc, iron, copper, etc.) as well as neuropathological changes in different brain regions (cortex, hippocampus, and cerebellum) that are involved in ASD; 2) to identify the key factors involving in disrupting the β-catenin destruction complex and their cellular colocalization; 3) to assess the ASD-like behaviors in the offspring. We collaborated with Dr. Lu Cai and collected the offspring brain tissues at 10 and 24 weeks post-weaning for Aim 1 and Aim 2. A new batch of mice will be set up to test their ASD-like behaviors at 10 weeks post-weaning for Aim 3. Upon successful completion of these pilot studies, we will define the gene-environment interactions among cadmium toxicity, diet and ASD/NDD pathogenesis. These results will identify putative molecular target(s) for developing new interventions to treat ASD/NDD. The data achieved from this pilot project will provide strong supporting evidence for our hypothesis to apply for extramural funding.

Principal Investigators:    Rachel Neal, PhD and Luz Huntington Moskos, PhD, RN, CPN 
Title:  "Citizen Science Approach to Studying the Community Impact of Hemp Processing Facilities in Cadiz KY" 
Description of Project: The CIEHS community-engaged pilot project titled, “Citizen Science Approach to Studying the Community Impact of Hemp Processing Facilities in Cadiz KY” will involve a citizen scientist model of environmental monitoring. The aims of this work are to support community odor reporting, to disseminate information on the health impacts of PM 2.5 and PM 10, VOCs and, finally, to train community members in environmental sampling. The study of the environmental impact of hemp cultivation and processing is in its infancy. Nationwide, hemp processing plants represent a significant community environmental challenge due to the overwhelming odor profile that has led to citizen complaints to local and regional air quality boards. In Kentucky, there are multiple drying and cannabinoid extraction plants using a variety of industrial processes resulting in odors, reduced adjacent highway visibility, and lack of air pollution release permitting prior to operation. This community-engaged work will provide community members with their own data and strive to increase their environmental health literacy related to outdoor air quality.