Joel Kaufman finds a connection between air pollution and heart disease

Joel Kaufman, Professor of Medicine, Environmental and Occupational Health Sciences, and Epidemiology and Deputy Director of the University of Washington Center for Ecogenetics and Environmental Health, was recently featured on a National Institutes of Enviromental Health Sciences (NIEHS) podcast, Air Pollution and Your Heart. NIEHS podcasts are part of the Partnerships in Public Health (PEPH) program of the NIEHS Division of Extramural Research and Training.

Most people think of air pollution as a cause of respiratory problems like coughing and asthma. It turns out that air pollution is also a significant risk factor for developing heart disease, the #1 cause of death in the United States (see statistics from CDC). It appears that high air pollution can trigger heart attacks and strokes in those who are at risk. As evidence, there is often a spike in the number of heart attacks on and after days with bad air pollution.

And now there is growing evidence that living in areas with higher air pollution puts people at risk for heart disease in the long term. Lab studies show that exposure to air pollution can lead to atherosclerosis (hardening of the arteries), high blood pressure, changes in the size and structure of the heart, and increased likelihood of the blood clots that trigger heart attacks and strokes.

There are ways to reduce your risk. One example is to avoid exercising near roads with heavy traffic or at high traffic times. The benefits of being outdoors, exercising and being active outweigh the risks from air pollution, so if you have to choose, choose to be outdoors and exercise. You can reduce inside air pollution by keeping windows closed or by installing a HEPA air filter. Those who have a heart condition can check daily air quality and stay indoors on bad air days.

Yet finally, air pollution is a societal issue which has to be addressed not by individuals but by society. Joel observes that over the past 40 years, the United States has been extremely successful at reducing air pollution. The story is a public health triumph. There is, however, research still to be done. We know that further reducing air pollution levels would reduce negative health effects. We don't know if there a level where air pollution is no longer a health concern. Also, we measure air pollution by measuring fine particulate matter while traffic exhaust is a mix of pollutants. One or a few agents in the mix may cause the worst health effects; if these can be identified, we could target our public health dollars more specifically and perhaps more effectively.

For more on Joel Kaufman's and NIEHS air pollution research, see this video of the Virtual Forum, Near Roadway Pollution and Health, with NIEHS Director Linda Birnbaum and other NIEHS researchers.

--Marilyn Hair

Kidney toxicity screening tool developed by National Toxicology Program Collaborative Team

Headshot of Vishal Vaidya
Vaidya received the 2015 Achievement Award from the Society of Toxicology, for his significant early career contributions to the field of toxicology. In 2011, he received an NIEHS Outstanding New Environmental Scientist award. (Photo courtesy of Vishal Vaidya)
Headshot of Auerbach
Auerbach is with the NTP Molecular Toxicology and Informatics Group, where he focuses on the analysis and interpretation of multivariate datasets and oversees the DrugMatrix database. (Photo courtesy of Steve McCaw)
A team of researchers including CEEH members Ed Kelly and Jonathan Himmelfarb, along with other NIEHS grantees and National Toxicology Program (NTP) scientists, developed the first method to test for kidney toxicity using high throughput screening (HTS). Because drugs and environmental chemicals can cause injury to kidneys, the new test could help reduce a significant health burden for patients and decrease the financial risk for pharmaceutical and chemical companies.
The team, led by Vishal Vaidya, Ph.D., of Harvard Medical School, and including molecular toxicologist Scott Auerbach, Ph.D., from NTP, published their approach Aug. 10 in the Journal of the American Society of Nephrology. “This high-throughput assay allows, for the first time, rapid and robust screening of kidney toxic compounds, to support chemical risk assessment and facilitate elimination of drug candidates early in the process before they reach humans,” Vaidya said.
High throughput screens were lacking
HTS, which has been advanced by the Tox21 collaboration, is rapidly becoming the standard tool for predictive toxicology, which aims for early detection of adverse health effects from chemical compounds, including medicines.
“The lack of adequate models to accurately predict human toxicity contributes to an underestimation of the kidney toxic potential of new therapeutic candidates,” the scientists wrote, “which also explains why nephrotoxic effects in patients are often only detected during late phase clinical trials, or in some cases, after regulatory approval.”
Using the right cells
The authors reported three important advances. First, they confirmed that the cells they used were suitable for the job. A particular type of kidney cell, called human proximal tubular epithelial cells (HPTECs), is the predominant target of most substances that are toxic to the kidney.
The researchers characterized the structure and function of HPTECs. The cells were shown to possess characteristics of differentiated epithelial cells, which made them desirable for use in in vitro systems.
Finding a biomarker
Next, the team identified a biomarker more sensitive than the currently used assays — cell viability and cell death — to indicate toxicity before changes occurred that might damage the cells.
“A few years ago at the Society of Toxicology annual meeting, Vishal stopped by the NIEHS booth, where we were demonstrating DrugMatrix,” Auerbach said. DrugMatrix is a molecular toxicology database that contains toxicogenomic profiles for hundreds of compounds. “DrugMatrix enabled us to provide a list of prototype kidney toxicants for him to test,” he said.
After exposing the HPTEC cells to nine kidney toxins and analyzing the expression of 1,000 genes, the researchers found that expression of one gene, HO-1, was significantly increased in the presence of the toxins.
The team validated the HO-1 biomarker in two ways. University of Washington researchers Edward Kelly, Ph.D., and Jonathan Himmelfarb, M.D., found that, following a kidney toxicant challenge, HO-1 was induced in a kidney-on-a-chip system that uses living tissue to accurately model organ function. Auerbach, collaborating with Dan Svoboda, Ph.D., of Sciome, LLC, examined two rat toxicogenomic databases and identified a significant association between HO-1 expression and kidney injury.
Additional analyses helped the team refine the assay. “Sensitivity and specificity can be improved even further by combining the readout for HO-1 concentration and the total cell number, measured in the same well,” reported the authors.
Developing and validating the test
Finally, the team developed a new assay to measure HO-1 in a rapid and cost-efficient manner. They used a test called a homogeneous time resolved fluorescence (HTRF) assay (see image below). “The HO-1 levels obtained in response to most of the [39 tested] compounds … correlated well with immunofluorescence,” the authors wrote.
The authors noted some limitations to the method. For example, a chemical might undergo modification in the liver and produce a metabolite toxic to the kidney. This could be missed by the new tool.
However, the authors remarked that it is an important step forward, providing a test that is robust — it detects kidney toxicants from multiple classes of chemicals; it is sensitive, because it picks up toxicants at a high rate; and it is specific, which means it correctly identifies compounds that are not kidney toxicants.

CitationAdler M, Ramm S, Hafner M, Muhlich JL, Gottwald EM, Weber E, Jaklic A, Ajay AK, Svoboda D, Auerbach S, Kelly EJ, Himmelfarb J, Vaidya VS. 2015. A Quantitative Approach to Screen for Nephrotoxic Compounds In Vitro. J Am Soc Nephrol; doi:10.1681/ASN.2015010060 [Online 10 Aug 2015].
Scheme of the HTRF assay performed in a 384-well plate
Scheme of the HTRF assay performed in a 384-well plate. When the acceptor labeled antibody and the donor labeled antibody      bind to HO-1, the two dyes are brought into close proximity with each other. Excitation of the donor with a light source triggers a fluorescence resonance energy transfer toward the acceptor. The emission fluorescence can be detected after incubation for four hours. This signal is proportional to the amount of human HO-1 present in the cell lysate. (Photo courtesy of Vishal Vaidya)

--Marilyn Hair
Based on an article in Environmental Factor by Kelly Lennox

Protect Yourself from Wildfire Smoke

The 2015 wildfire season in the Pacific Northwest is the worst yet. Hundreds of fires are burning in Washington, Idaho, Montana, northern California, Oregon and British Columbia. In Washington, it's the largest wildfire complex in state history.

Smoke is much worse this year because of the huge size of the wildfires and an inversion layer that is preventing the smoke from dissipating. Choking smoke has blanketed much of Central and Eastern Washington and some days has even traveled west across the Cascade Mountains. The smoke stings the eyes and throat and causes coughing, runny nose, and headache. These symptoms may happen sooner for children and older adults, those with heart or lung disease, and outdoor workers. People are concerned.

Last year we created a downloadable fact sheet, How to Protect Yourself From Wildfire Smoke with information about exposure to smoke and what to do.

These two websites give current information about smoke and air quality in Washington:

Washington Smoke Information
State air quality map, Washington Department of Ecology

As I write in late August, rain is forecast. Let's hope rain will slow the wildfires, help clear the smoke, and give firefighters and community members a chance to catch their breath.
-- Marilyn Hair

Center participates in National Conference of State Legislators Summit

Center for Ecogenetics & Environmental Health (CEEH) members and staff participated in the Environmental Health Sciences (EHS) Pre-Conference Session of the National Conference of State Legislators (NCSL) Summit on August 3rd at the Washington State Convention Center in Seattle. NCSL is a bipartisan public-policy organization. About 5,000 state lawmakers and staffers attended, including more than 70 Washington legislators.

Those who attended included from CEEH included Center members Terry Kavanagh, Gretchen Onstad, Kelly Edwards, and Rose James, as well as UW faculty Steve Gilbert, and Clarita Lefthand-Begay, and CEEH staff members Liz Guzy and Marilyn Hair. State legislators from California, Montana, Connecticut, Wisconsin, Arkansas, New York, and Maryland, among others, and a legislator and staff from Western Australia attended the EHS Pre-Conference Session. The American Chemical Council (ACC) and Scott's MiracleGro were also represented.

Dr. Onstad participated on a panel that addressed Water and Health: Quality and Quantity. Other panelists were Professor David Osterberg of the University of Iowa Environmental Health Sciences Research Center, Rep. Gary Scherer of Ohio, Assemblyman Adam Gray from California, and Ann Aquillo, Vice-President of Scotts Miracle-Gro. Dr. Lefthand-Begay served on a panel focused on Environmental Justice, along with Robin Fuchs-Young from the NIEHS Environmental Health Sciences Center at Texas A&M University, Johnnye Lewis from the University of New Mexico, and Elena Craft of the Environmental Defense Fund. A third panel presentation addressed Hydraulic Fracturing and Environmental Health.

Much of the session was devoted to audience interaction. Legislators were concerned about environmental health issues in their districts, and the concerns of their constituents, among them air quality; health effects of living near major roadways; silica sand mining in Wisconsin, industry-sponsored research on the health effects of fracking in North Dakota, toxic algal blooms from fertilizer runoff in the midwest, and increasing water capacity in central California's agricultural region, in the face of a continuing drought. The legislators were well informed and almost universally sympathetic to protecting public health.

Roundtable discussions were held over box lunches on five topics: Environmental Public Health Labs, PCBs in Schools, US-Mexico Border Environmental Health Concerns, Mining and Health, and Toxics and Chemicals.

Drs. Terry Kavanagh and Steve Gilbert
I joined the Toxics and Chemical discussion, facilitated by Terry Kavanagh. Three staff from the American Chemical Council, an industry group, joined the table. A dialog ensued between ACC representatives and EHS researchers regarding disclosing the components of products such as flame retardants and hydrofracking fluid. The ACC spoke up for propriety rights of industry, while the toxicologists defended the public's right to know what they're being exposed to. The ACC's defense of propriety rights was based on economics; they said information needs to be protected to keep others from using trade secrets and suggested we should wait and see what happens to people who live and work near fracking sites. Steve Gilbert shot back, "Who's going to be in that study?" and "We need to protect children from these chemicals so they can grow up to their full potential." The discussion was civil and respectful and it was good to have different perspectives talking to each other. But from a public health perspective, let's try it, wait and see, and trade secrets aren't good enough for protecting people's health.

The goal of the EHS workshop was to open a dialog between leading environmental health sciences researchers and outreach directors, and staff legislators and staff, with the intent to increase the level of scientific input into policy development and decisions. The workshop was supported by a grant from NIEHS, Dr. Robin Fuchs-Young, Principal Investigator.
--Marilyn Hair

Introducing Dr. Libin Xu

Dr. Libin Xu joined the Center for Ecogenetics and Environmental Health this year as an Affiliate Member. Dr. Xu is a researcher and Assistant Professor in the Department of Medicinal Chemistry in the UW School of Pharmacy. 

Libin studies lipids, or fat molecules (cholesterol, vitamin A and vitamin E are examples), and their oxidation, a chemical reaction that generates free radicals that can damage DNA and body tissues. Having too many free radicals leads to oxidative stress, but this process can be prevented by antioxidants made in our own body or absorbed from diet. The Xu Lab studies how certain lipids are oxidized and metabolized, and how oxidation during lipid metabolism might be involved in human diseases.

The Xu lab is studying a rare disease called Smith-Lemli-Opitz syndrome or SLOS. SLOS is a cholesterol disorder that causes problems in the development of the central nervous system starting at embryonic stages. People who have SLOS have birth defects, intellectual disability and behavior problems such as autism.

SLOS is caused by a mutation in a gene (DHCR7) that makes an enzyme (3β-hydroxysterole-Δ7-reductase) that turns one lipid molecule (7-DHC) into another (cholesterol). The mutation means that SLOS patients have a lot of 7-DHC and not enough cholesterol. And it turns out that 7-DHC is extremely reactive toward oxidation, leading to the formation of free radicals and toxic oxidation products called oxysterols. In addition, some oxidative enzymes (cytochrome P450) can also turn 7-DHC into toxic oxysterols. This may be the problem underlying SLOS.

The goal of the work in Dr. Xu’s lab is to figure out the exact biochemical processes that lead to the broad phenotype of SLOS, and to find therapies to improve SLOS and other diseases caused by disrupted lipids. This research also will help scientists understand other intellectual and developmental disabilities that are associated with lipid metabolism problems and affect the function of the brain.

The research tools being developed in Dr. Xu’s lab to study lipids, based on advanced mass spectrometry, can also be used to study the side effects that common drugs might have on the metabolism of lipid molecules.

Working with the CEEH Community Outreach and Engagement Core (COEC), Libin Xu and Andrew Dinh in the Xu Lab have created a handout of interest to the general public: Fast Facts about Fat-Soluble Vitamins. Libin has also agreed to talk about lipid oxidation and his research at a Public Health Café next year.
Dr. Xu graduated with a B.Sc. in Chemistry from Nankai University, Tianjin, China. He earned his PhD in Organic Chemistry from the University of Illinois at Chicago, and did a post-doc at Vanderbilt where he became interested in lipid peroxidation and its role in human diseases.  He received the Young Investigator Award from the Society for Free Radical Biology and Medicine in 2011 and the NIH Pathway to Independence Award from the National Institute of Child Health and Human Development in 2012. We welcome Libin Xu to the CEEH.                                                         
                                                                                                                                    --Marilyn Hair

Celebrating the new Port Gamble S'Klallam Environmental Health Children's Book, taʔt̕ə́wəsnaʔ

Author and Port Gamble S'Klallam tribal member Tleena Ives organized a Book Release for her children's book, taʔt̕ə́wəsnaʔ Star. 

Tleena's book was the result of her collaboration with the Center for Ecogenetics and Environmental Health (CEEH) Native TEACH Project.

New center to use novel method to screen chemicals’ toxicity

In-vitro, 3-D chip approach will enable faster evaluation and reduce need for animal models.
Scientists prepare cell cultures to 
test chemicals for their potential 
risk to humans. 

Tens of thousands of chemicals are currently in use, with more introduced every year. Scientists, however, have discerned the toxicity of only a fraction of these because the traditional method of testing is time- and cost-prohibitive – thus the need for better mechanisms to screen chemicals for potential health impacts.
The Environmental Protection Agency today said that it would provide $6 million in seed funds for a Predictive Toxicology Center at the University of Washington – one of three such facilities identified. It is intended to enable researchers to develop more accurate, higher capacity in vitro models – organ-mimicking cell cultures – to test chemicals' potential risk to humans.
Elaine Faustman and Terrance Kavanagh will co-direct the new center.
 “This research is important for establishing novel methods that reflect the complexity of biological systems, yet allow us to evaluate the large number of new chemicals for their potential health impacts before they are put on the market,” said Elaine Faustman. She and Terrance Kavanagh, both UW professors of environmental and occupational health sciences, will co-direct the new center. 
“These systems can help reduce the need for animal models while better replicating and more faithfully mimicking what happens in the human body, including differences in genetics and susceptibility” Kavanagh said. 
Researchers from the Schools of Public Health, Pharmacy, and Medicine will collaborate to develop these three-dimensional cell cultures for the kidney, liver, lung and testis to better model how a person would respond to a chemical exposure. 

A hallmark of the center’s research will be microfluidic chip technology developed by Nortis, a biotech founded by former UW faculty. Less than half the size of an index card, the microfluidic chip offers a platform for cells to be seeded onto the device, which then serves as a unique growing environment and can be connected with the other devices for each organ system, mimicking human circulation.

After validating the three-dimensional cell cultures using known chemical toxicants, researchers will investigate biomarkers of cell injury or altered function that indicate an adverse outcome from an exposure to chemicals for which insufficient data exists. 
Elijah Weber, Department of Pharmaceutics
Graduate students in the School of Pharmacy demonstrate how constant flowing media will go through microphysiological systems connected to pumps outside an incubator.
They will then use modeling techniques to assess and predict human health risks, linking laboratory data with human-exposure pathways.
Testing metals and metal-based nanomaterials will be a focus of the center. 

“There’s been a lot of interest in the biomedical field to use nanomaterials as therapeutics and bioimaging agents,” Kavanagh said, but their unintentional impacts in the general population are less well studied.
For example, inhalation is a primary route of exposure to nanomaterials, so developing a lung system is especially relevant, said William Altemeier, UW associate professor of pulmonary and critical care medicine. He will lead the center's lung cell culture development.  
Faustman will lead the testing of adverse reactions in testicular development and reproduction. Kavanagh will co-manage the liver-testing model with David Eaton, professor of environmental and occupational health sciences. Edward Kelly, UW associate professor in of pharmaceutics (Pharmacy), will manage the kidney cell culture project.
Read the full news release about the Predictive Toxicology Center.

--Elizabeth Sharpe

Meet new CEEH member Dr. Anne Manicone

The CEEH Career Development and Mentoring Core welcomed Dr. Anne Manicone as a junior investigator early this year. Anne is an Assistant Professor in the Department of Medicine, Division of Pulmonary and Critical Care. She will join the CEEH Cardiopulmonary & Metabolic Disease Collaborative Research Team (CRT) headed by Drs. Mike Rosenfeld and William AltemeierAnne presented her research at the monthly CEEH Breakfast Club on January 28 in an informal talk titled Regulation of Macrophage Polarization: Implications in Lung Injury and Repair. She is currently focused on the role of MMP28 in regulating macrophage influx and activation, and the role of macrophage subpopulations in regulating lung injury and its resolution.

More specifically, Anne is characterizing the role of matrix metalloproteinase 28 (MMP28) in regulating the inflammatory response to tobacco smoke-induced emphysema in a mouse model. MMPs comprise a family of extracellular proteinases that function in various processes of innate immunity. MMP28 is one of the newest members of this family, and it is expressed by both macrophages and epithelial cells. Her work suggests that MMP28 inhibits inflammation and promotes reparative macrophage function. In her preliminary studies, MMP28 was shown to be protective in chronic emphysema from cigarette smoke; and her hypothesis is that this is mediated by MMP28-dependent effects on macrophage polarization.  Ongoing work is aimed at uncovering the mechanism and substrates by which MMP28 functions. 

Dr. Manicone's work is pertinent to Chronic Pulmonary Obstructive Disease (COPD) and pulmonary fibrosis, conditions brought on by environmental exposures such as smoking, asbestos, and air pollution, as well as asthma and genetics. Anne indicates she is always interested in collaborations.

The Career Development and Mentoring Core (CDMC), directed by Mike Rosenfeld, works to ensure that the University of Washington recruits, supports, and retains for DEOHS and other Center-affiliated departments (Epidemiology, Genome Sciences, Pharmaceutics), a new generation of high-caliber EHS-focused investigators with a strong interest in gene-environment interactions. The CDMC provides mentorship to new and current CEEH junior faculty to ensure they achieve their full potential as scientists, teachers, and communicators, as well as provides unique career development opportunities and resources to advance their environmental science careers by incorporating a gene-environment approach into their research projects.                                                                                              

                                                                                - Marilyn Hair