Allen, Joseph G. (Assistant Professor, Environmental Health, Harvard T.H. Chan School of Public Health) “Cell Assays to Quantify Hormonal Potencies of Chemical Mixtures Collected on Silicone Wristband Personal Samplers of Global Cohort of Office Workers”
Birmann, Brenda M. (Assistant Professor, Channing Division of Network Medicine, Brigham and Women’s Hospital) “Association of geospatially modeled (non-occupational) exposure to dioxins and other hazardous air pollutants with lymphoid malignancies and lymphoma-related biomarkers: a pilot study”
Blomberg, Annelise (Post-Doctoral Fellow, Environmental Health, Harvard T.H. Chan School of Public Health) “A pilot study of associations between early-life exposures to per- and polyfluoroalkyl substances and lipoprotein subspecies”
Dominici, Francesca (Professor, Biostatistics, Harvard T.H. Chan School of Public Health) “Air pollution, race, and health outcomes for COVID-19 in the United States: data science methods and dissemination”
Golden, Christopher (Assistant Professor, Nutrition, Harvard T.H. Chan School of Public Health) “Does the Gut Microbiome Mediate Human Exposure to Methylmercury?”
Haber, Adam (Assistant Professor, Environmental Health, Harvard T.H. Chan School of Public Health) “Healthy homes: statistical surveillance of emergency departments for early identification of dangerous housing conditions”
James-Todd, Tamarra (Assistant Professor, Environmental Health, Harvard T.H. Chan School of Public Health) “The Role of Uncertainty in Environmental Health Literacy: Exploring the Relationships Among Phthalate Knowledge, Risk Perception, Risk Acceptability, and Protective Behaviors in Perinatal Mothers“
Moon, Michelle (Community Organizer, Civic Space Collaborative) “Heat Island Art: Research + Development”
Panganiban, Ronald (Research Scientisit. Environmental Health, Harvard T.H. Chan School of Public Health) “Extracellular Vesicle (EV) Protein Cargo, Metal Exposure, and Atherosclerosis”
Papatheodorou, Stefania (Lecturer, Epidemiology, Harvard T.H. Chan School of Public Health) “Effects of particle exposures on fetal growth”
Rawson, Jeff (Post-Doctoral Fellow, Chemistry and Chemical Biology, Harvard Faculty of Arts and Sciences) “Free radicals from electronic cigarettes for consuming extracts of cannabis”
Schwartz, Joel (Professor, Environmental Health, Harvard T.H. Chan School of Public Health) “Estimating Causal Effects of Multipollutant Mixtures on Hospital Admissions and differential effects on disadvantaged”
Shoaff, Jessica (Research Fellow, Channing Division of Network Medicine, Brigham and Women’s Hospital) “Assessing contextual environmental measures in a longitudinal birth cohort and associations with neurobehavior”
Zhang, Xuehong (Assistant Professor, Channing Division of Network Medicine, Brigham and Women’s Hospital) “A Pilot Study of Circulating Levels of Perfluoroalkyl Substances (PFASs) and Non-viral Liver Cancer Risk”
- Joanne Sordillo: Exposure to air pollutants and upper airway microbial communities in project via, a pre-birth cohort study
- Jin-Ah Park: Apoptosis priming in primary human airways epithelial cells in response to environmental exposures
- Johan Spetz, Chris Sarosiek: Dynamic regulation of apoptosis as a driver of cardiovascular disease caused by environmental lead exposure
- Carmen Messerlian: The role of environmental chemicals and ribosomal DNA age clock in female fertility
- Gary Adamkiewicz: Validating City Scanner: A low cost mobile air quality platform for cities
- Sneh Toprani, Zachary Nagel: Assessing DNA damage and repair capacity in airline flight crew
- Peggy Lai: Environmental chemicals, lung function, and the gut microbiome
- Shruthi Mahalingaiah: Menstrual Blood Metabolomics: Assay Development and Optimization
- Lindsay Jaacks: Cross validation of methods for measuring urinary metabolites of organophosphate pesticides in India
- David Itiro Kasahara, Ph.D., Research Scientist, Department of Environmental Health, Harvard Chan School of Public Health. “Transfer of Enhanced Responses to Air Pollution from Obese to Lean Mice via Gut Microbiota Transplant.”
- Emma Preston, M.P.H., Ph.D., Post-Doctoral Research Fellow, Department of Environmental Health, Harvard T.H. Chan School of Public Health. “Assessing Consumer Product Chemical Exposures in Daycare Facilities: Portable Fourier Transform Infrared (FTIR) as a Novel Forensic Tool.”
- Mary B. Rice, M.D., M.P.H., Assistant Professor, Division of Pulmonary & Critical Care, Department of Medicine, Beth Israel Deaconess Medical Center, “Sampling the Nasal Epithelial Lining Fluid as a Non-Invasive Indicator of Metal and Microbial Exposures and Immune Response sin People with COPD.”
- Joseph Allen, Assistant Professor, Department of Environmental Health, Harvard Chen School of Public Health “Impact of healthy material interventions in offices on reductions in fluorinated chemicals and endocrine-disrupting potency of indoor dust”.
- Jaime Hart, Assistant Professor, Channing Research Laboratory, Brigham and Women’s Hospital, “A pilot study of associations of multipollutant exposures on risk of anovulation”
- Ross Osgood, Post-Doctoral Fellow, Dr. Stephanie Shore’s Laboratory, Department of Environmental Health “The role of sex hormones in microbiome-dependent effects on pulmonary response to ozone”
- Sneh Toprani, Post Doctorial Research Fellow, Dr. Zachery Nagel’s Laboratory, Department of Environmental Health “Assessing DNA damage and repair capacity in airline flight crew”.
- Punyaganie de Silva, MBBS, MPH, Instructor, Dept of Gastroenterology, Hepatology and Endoscopy, Brigham’s & Women’s Hospital “The association between environmental endocrine disruptors and inflammatory bowel disease – A pilot study.”
- Francine Laden, ScD, Professor and Associate Chair, Department of Environmental Health, Harvard T.H. Chan School of Public Health. “Total organofluorine concentrations in drinking water samples from the Nurses Health Study cohort.”
- Maitreyi Mazumdar, MD, MPH, MS; Assistant Professor, Department of Neurology, Boston Children’s Hospital. “Building capacity for childhood lead poisoning prevention in Bangladesh.”
- Drew Michanowicz, DrPH, MPH, CPH; Postdoctoral Fellow, Center for Health and the Global Environment, Harvard T.H. Chan School of Public Health. “Piloting sensors without a pilot: A 3D gas-sensing system using drones and stationary monitoring to measure plume dispersion.”
- Kristopher Sarosiek, PhD, Department of Environmental Health, Harvard T.H. Chan School of Public Health. “Assessment of Apoptosis and DNA Repair: Bridging the Gap Between Cell Assays and Intact Animals/Humans.”
- Jaime Hart, ScD, Assistant Professor of Environmental Health, Channing Division of Network Medicine, Brigham and Women’s Hospital, Harvard Medical School. “High-Resolution Metabolomics in COPD: Elucidating Biological Responses to Multipollutant Indoor Air Exposures.”
- Joseph G. Allen, ScD MPH, Assistant Professor of Exposure Assessment Science, Department of Environmental Health, Harvard T.H. Chan School of Public Health. “Emission reductions and climate/health co-benefits from energy use reductions in Harvard‘s buildings.”
- Quan Lu, PhD, Associate Professor of Environmental Genetics and Pathophysiology, Department of Environmental Health, Harvard T.H. Chan School of Public Health. “Extracellular Vesicle (EV) MicroRNAs, Metal Exposure and Alzheimer’s Disease.”
- Tamarra James-Todd, PhD MPH, Mark and Catherine Winkler Assistant Professor of Environmental Reproductive and Perinatal Epidemiology, Department of Environmental Health, Harvard T.H. Chan School of Public Health. “Phthalates, PPAR-ϒ target gene expression, and maternal obesity measures in the perinatal period.”
2016 Round 1
- Diana Ceballos: Assessing Dermal Exposures to Reproductive Toxic Chemicals on Nail Salon Workers ($24,980)
- Wafaie Fawzi: Prevalence of aflatoxin contamination among pregnant women in Dar es Salaam, Tanzania, and subsequent effects on selected infant, child, and maternal health outcomes ($24,932)
- Qi Sun: Feasibility and reproducibility of glyphosate and 2,4-D (2,4-dichlorophenoxyacetic acid) assessments in 24-hr urine samples ($24,962)
- Akira Tsuda: ARDS mouse model for in vivo imaging of the lung’s gas exchange region ($25,000)
- Vishal Vaidya: Detection of Kidney Injury in Mexican Children Exposed to Environmental Toxicants ($25,000)
- Mahsa Yazdy: Temperature and humidity and the risk of preterm birth ($10,000)
2016 Round 2
- David Christiani: Exposures to polycyclic aromatic hydrocarbons and risk of Esophageal squamous cell carcinoma in Uganda ($20,000)
- Russ Hauser: Association between Dibutylphthalate and thyroid hormones in men in a crossover-crossback prospective study ($20,000)
- Peggy Lai: Effect of small-scale chicken farming on the indoor environmental and human microbiome: A Randomized controlled trial($20,000)
- Zachery Nagel: Exploratory DNA repair analysis using FM-HCR technology in crossover study of individual exposed to metal fumnes($20,000)
- Hadi Shafiee: A Smartphone-based sperm analyzer to facilitate studies on the effect of air pollution on semen quality ($20,000)
- Punyanganie de Silva: The association between environmental endocrine disruptors and inflammatory bowel disease – A Pilot Study ($20,000)
Massachusetts is home to approximately 14,000 nail salon workers, most of whom are female (94%) and part of a minority group (67%). These workers are exposed to a wide variety of chemicals. Epidemiological studies have suggested that nail salon workers experience an increased risk of pregnancy complications. To date, these epidemiological studies have used self-reported measures of exposure; however, we lack data on the types and levels of chemicals that may affect the reproductive health of nail salon workers. Further, little has been documented about the role skin absorption plays in the total exposure to these workers yet solvents, plasticizers, and metals are possibly absorbed through the skin. This study will collect data through a cross-sectional study on 10 nail salon workers to characterize the body burden of solvents (blood and exhaled breath) and metals (toenails). We will measure the amounts of metals in a representative sample of nail polishes to prioritize for metal biomarkers. We will measure inhalation exposure to solvents and estimate other potential routes of exposure. This study will inform future research for examining health effects and interventions to reduce exposures in this worker population.
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Wafaie Fawzi: Prevalence of aflatoxin contamination among pregnant women in Dar es Salaam, Tanzania, and subsequent effects on selected infant, child, and maternal health outcomes ($24,932)
Approximately 4.5 billion people, the majority of who live in low- and middle-income countries, are at risk of aflatoxin exposure. Aflatoxin’s effects on many health outcomes are still poorly understood due to a range of confounding factors, the expense of measuring biological aflatoxin concentrations, and the cross-sectional nature of most studies. Nonetheless, a growing body of epidemiologic evidence suggests that widespread, chronic exposure to aflatoxin may contribute significantly to the enormous burden of low birthweight and perinatal morbidity in sub-Saharan Africa and southeast Asia. This pilot will study this hypothesis using stored blood samples collected from pregnant women in Dar es Salaam, Tanzania. Aims of the study will be to determine the local prevalence of aflatoxin contamination among pregnant women giving birth to healthy weight and low birthweight infants using HPLC-fluorescence, and characterize associations between aflatoxin exposure in-utero and (i) maternal and perinatal outcomes, (ii) infant birthweight, and (iii) child growth and selected infectious disease among infants and their mothers between delivery and 2 years following, in which a causal interpretation of our findings is strengthened by a case-cohort approach. Results will inform a future aflatoxin study using this biorepository, involving a larger sample size and additional health outcomes.
Qi Sun: Feasibility and reproducibility of glyphosate and 2,4-D (2,4-dichlorophenoxyacetic acid) assessments in 24-hr urine samples ($24,962)
Abstract (Limit 13 lines) The use of herbicides containing glyphosate and 2,4-D has increased dramatically in the U.S. Glyphosate and 2,4-D are recently classified as potential carcinogens by International Agency for Research on Cancer (IARC), although U.S. Environmental Protection Agency (EPA) deems glyphosate and 2,4-D generally benign and has approved Enlist Duo, which is a new formulation comprised of glyphosate and 2,4-D. While there is a hot debate regarding the health and environmental consequences of the wide use of the herbicides, a consensus is that human evidence regarding the health effects of long-term exposures to the herbicides is incomplete. One major obstacle is the lack of sensitive assays for quantifying glyphosate levels in human urine samples. In this proposed pilot research, we will collaborate with Dr. Adrian Franke to develop a liquid chromatography mass spectrometry-based glyphosate assay at his well-established lab at the University of Hawai’i. We will further evaluate the reproducibility of excretion of glyphosate and 2,4-D in repeat 24-hr urine samples collected in the Women’s Lifestyle Validation Study. Data from this pilot study will serve as critical preliminary results for NIH grant applications aiming to collect multiple 24-hr urine samples in the dedicated Nurses’ Health Study II participants to facilitate investigations on long-term health effects of glyphosate and 2,4-D exposure.
Akira Tsuda: ARDS mouse model for in vivo imaging of the lung’s gas exchange region ($25,000)
The lungs are prestressed and their configuration and dynamics are maintained by a balance of tissue and surface forces. Taking advantage of rapid advancements in technology, we have recently visualized alveolar dynamics noninvasively in live mice using synchrotron-based x-ray imaging (Chang et al., 2015). We are currently using the unique opportunity of lung imaging in vivo to study Acute Respiratory Distress Syndrome (ARDS) – a life-threatening disease featured as alveolar tissue damage in an animal model. In this pilot project, we propose to perform experiments with mice with Lipopolysaccharide (LPS)-induced ARDS here at HSPH. We plan to ventilate these animals using a wide range of ventilatory conditions. Readouts of lung injury, such as albumin leakage into the alveolar space (BAL) and histolopathology will be recorded. With these systematic investigations we will select conditions that cause various degrees of organ damage. These studies at HSPH will form the basis of ARDS lung imaging in the Riken SPring-8 synchrotron Center in Japan. As the synchrotron beam time is limited we need to carefully select and design the experimental conditions we will use for synchrotron imaging in vivo of the ARDS mouse model.
Vishal Vaidya: Detection of Kidney Injury in Mexican Children Exposed to Environmental Toxicants ($25,000)
It is known that prolonged cumulative lifetime exposure to environmental kidney toxicants; in conjunction with other comorbid conditions accelerate the rate of deterioration in kidney function increasing the risk for chronic kidney disease (CKD). Previously, we found that children residents of Villa de Reyes, SLP, Mexico are exposed to environmental kidney toxicants such as arsenic, chromium and lead (n=83). Although, the kidney function evaluated by routine clinical tests was normal, we found that the levels of tubular injury biomarkers such as urine kidney injury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL) were high. Multivariable linear regression analysis adjusted for age, sex, BMI and urine specific gravity revealed significant association between children’s urine chromium and arsenic in tertiles and higher urine KIM-1 excretion. The associations were stronger in the highest tertile and moderate in the second tertile compared with the lowest tertile (reference). Therefore, the aims of this project are to expand these findings by 1) measuring serum KIM-1 and NGAL in order to assess kidney dysfunction apart from tubular injury and 2) to measure candidate micro-RNAs as mechanistic biomarkers for early detection of kidney injury in urine and serum samples. The goal of this project is to test our hypothesis that early detection of kidney injury using sensitive and specific mechanistic biomarkers will allow us to stratify children that are more susceptible to develop CKD later in life.
Mahsa Yazdy: Temperature and humidity and the risk of preterm birth ($10,000)
Climate change is expected to cause considerable changes in temperatures, which in turn can have substantial impact on human health. Due to physiological changes in pregnancy, one susceptible group to extreme temperatures is pregnant women. High temperatures and heat waves have been associated with triggering birth earlier than expected. The few studies that have assessed cold temperatures have not found an association with preterm birth, though the majority of these studies took place in cold climates where women may have adapted to cold weather. The aim of this study is to assess the association between temperature and humidity and preterm birth, as well as to identify susceptible subpopulations. Preterm birth is one of the leading causes of perinatal mortality and there are substantial disparities in rates of preterm birth, this study has potential to fundamentally advance our knowledge on contributors to preterm birth. The goal of this pilot study is to assess the feasibility of linking and estimating the association between publically available U.S. birth certificate data and climate data. If this proves successful there is an immense opportunity to use the birth certificate data, which contains data from all births in the U.S., for epidemiologic studies with detailed environmental exposures.
David Christiani: Exposures to polycyclic aromatic hydrocarbons and risk of esophageal squamous cell carcinoma in Uganda ($20,000)
Esophageal cancer is the eighth most common cancer and the sixth cause of cancer-related death worldwide. About 80 % of cases, predominantly the squamous cell variant, occur in developing countries. In East Africa, the esophageal squamous cell carcinoma (ESCC) hot spot of Africa, ESCC exhibits within country and ethnic variations suggesting a significant contribution of population-specific environmental, dietary, and genetic susceptibility.
In southwestern Uganda, our group showed risk factors of ESCC to be male gender, increasing age, smoking and alcohol use but no relationship was found with human papillomavirus. In this setting, ESCC presents at a younger age (> 40 years) and has poor treatment outcomes. We also found the population attributable fraction due to a combined effect of smoking and drinking alcohol to be lower than one-fourth compared to between two-thirds and three-quarters from other settings suggesting that smoking and alcohol contribute less than in other settings and other, unidentified risk factors are significant contributors to ESCC. We posit that exposure to polycyclic aromatic hydrocarbons (PAH) through biomass fuel and food processing such smoking of food may be significant contributors to esophageal cancer risk. These practices are rampant but understudied in Africa. If this interaction proves significant, the public health implications are broad given the billions of people affected by the intersection of the PAHs and esophageal cancer.
In the U.S. it is estimated that 20 million adults and children have thyroid disease (12% prevalence) with largely unknown causes. Untreated thyroid disease can lead to osteoporosis, cardiovascular diseases, and infertility. Recent experimental and human studies have suggested that environmental exposures, such as phthalates may impact the thyroid axis. Phthalates are commonly used plasticizers with ubiquitous general population exposures. Apart from low level background exposure to phthalates, there are specific sources of high exposure. This includes very high exposure to dibutyl phthalate (DBP) used in the coating of some medications, such as mesalamine used to treat inflammatory bowel disease (IBD). Some mesalamine formulations contain more than 1,000 times the median DBP exposure. Taking advantage of the difference in the mesalamine coatings, we conducted a prospective crossover-crossback study among men with IBD to investigate the impact of high DBP exposure on semen quality and reproductive hormones. Our study was not funded to measure thyroid hormones. Therefore the funds for the proposed pilot study will be used to leverage our recently completed study in which we archived serum samples. We are proposing to examine the association between high-DBP exposure with thyroid hormones.
Indoor air pollution leads to 3.9 million premature deaths a year globally and is one of the most important environmental risk factors contributing to the global burden of pulmonary disease. Women and children in low and middle income countries are disproportionately affected. Microbial exposures are an important component of indoor air pollution, and recent studies in asthma and chronic obstructive pulmonary diseases have demonstrated the importance of environmental and respiratory microbial communities on both disease exacerbation and development. What has not been clearly demonstrated is how acute changes in the environmental microbiome can alter the adult human microbiome and affect respiratory health.
This proposal is based on the “Clean Air Study” (PI: Lai), an ongoing randomized controlled clinical trial in Southwest Uganda where the intervention is the introduction of small-scale chicken farming to households. Chicken raising is associated with unique and high level microbial exposures, particularly for these participants who are subsistence farmers and cannot afford livestock. This proposal allows us to test our central hypothesis that a discrete change in the environmental microbiome will lead to changes in the human microbiome and
respiratory health. 81 women have already been enrolled and completed both baseline and 1 month follow-up testing. The approach is innovative, as a discrete change in the environmental microbiome has occurred as part of a randomized clinical trial, allowing for causal relationships to be identified. Results from this proposal are significant as they are expected to fundamentally advance the fields of pulmonary medicine and environmental health by defining a causal link between environmental microbial exposures, human microbial composition, and subsequent health. Although this study is taking place in Uganda, where a randomized controlled trial of this nature is feasible, results will have implications for over 5 million American workers who experience high level microbial exposures at work and for over 25 million Americans with asthma where there is an intense interest in understanding how microbial exposures affect asthma risk and severity.
Particulate air pollutants are mutagenic, carcinogenic and can cause DNA damage. Some particulates contain metals known to interfere with DNA repair. Results from our (Christiani lab) previous microarray analysis of genome-wide gene expression profiling in whole blood total RNA samples in response to occupational weldingfume exposure supported particulate-induced systemic inflammation, oxidative stress, and activation of cell death pathways. It is not known whether these responses include changes in DNA repair capacity. This proposal explores the functional dimension of local and systemic responses to welding-fume exposure using recently developed fluorescence based multiplex host cell reactivation (FM-HCR) technology. Comprehensive DNA repair profiling will be conducted on blood cells and nasal epithelial cells. We will use a previously developed self-controlled crossover study design with sequential plasma samples, which were collected at preworkshift (baseline), immediate post-workshift (~6 hours after baseline). All major DNA repair pathways will be analyzed. Hypothesis: exposure to respirable metal particulates induces changes in DNA repair capacity that can be detected in peripheral blood cells and nasal epithelial cells by FM-HCR. This work will have important implications for evaluating the host response to other forms of environmental exposures using FM-HCR technology in primary cells.
Sperm abnormalities account for approximately 50% of infertility and affect up to 12% of all men [1-5]. It has been reported that sperm quality has been decreased over the past decades [6, 7]. Several factors may negatively affect semen quality such as air pollution, obesity, overweight, smoking, drinking, stress, etc. Of particular interests are the effect of air pollution and other environmental exposures on semen quality [8, 9]. One of the major limitations on studies around the effect of air pollution on semen quality is the lack of access to semen samples from general population mainly due to difficulties in collecting semen samples. Studies based on samples from men who visit infertility clinics may introduce selection bias. In addition, semen sample collection through mail is limited as sperm total motility may significantly change during the transportation. A simple, rapid, inexpensive, home-based sperm analyzer can significantly facilitate studies on the effect of air pollution and other environmental exposures on sperm concentration, total sperm count, and total sperm motility. We propose to develop an inexpensive (<$5 in material costs), user-friendly, cellphone-based microchip assay, which we refer to as Fertilex, to measure sperm concentration, total sperm count, curvilinear and linear sperm velocities, and sperm motility using a semen sample (<50 μL) in less than a minute
Environmental endocrine disruptors (EDCs) are common risks to health, and hormonally active in low doses. Their risk to inflammatory bowel disease (IBD) is not well studied. IBD consists
of two main subtypes – ulcerative colitis (UC), a mucosal disease; and Crohn’s disease (CD) which involves trans-mural inflammation. The aim of this pilot study is to assess if short-term EDC exposure from parabens and phenols increases disease activity in established IBD.
2015 Round 1
- Andrea Baccarelli – DNA Conformational Changes and Arsenic Exposure
- David Christiani – Genetic Variants of Maternal DNA and Maternal Urinary Arsenic Methylation Species during Pregnancy in a Population Exposed to Inorganic Arsenic from Drinking Water
- Phil Demokritou – Systems and Methods for the Physico-Chemical & Toxicological Characterization of Emissions from Electronic Cigarettes
- Philippe Grandjean – Early life exposure to environmental persistent pollutants and blood concentrations of adipocytokines from birth through adolescence: Identification of relevant windows of exposure and susceptibility
- Elsie Sunderland and Francine Laden – Drinking water exposures to Per and Poly Fluorinated Alkylated Substances (PFASs) in the Nurses Health Study Cohort
2015 Round 2
- Jorge Chavarro – Pesticide residues in food: validation of an exposure assessment tool and relation to male fertility
- Alexey Fedulov – Effects of female hormone on talc phagocytosis
- Bernardo Lemos – rDNA copy number: a novel epigenetic modulator and marker of responses to environmental exposures?
- Ryan McCarthy: The role of iron in apolipoprotein E mediated clearance of amyloid-beta by microglia ($17,370)
- Chrystal North – Fine particulate matter and chronic HIV infection co-exposure: environmental determinants of pulmonary function in southwestern Uganda
- Joanne Sordillo: Environmental Triggers of Asthma Exacerbation and Gene Expression in an Airway In Vitro Model: Identification of common pathways and unique expression signals for individual exposure stimuli and exposure combinations ($25,000)
- Paula Tejera: Circulating extracellular vessicles-associaed microRNAs as novel biomarkers in acute lung injury ($25,000)
Project Abstracts: 2015 Round 1
DNA Conformational Changes and Arsenic Exposure
Andrea Baccarelli, et. al. ($24,300)
Epigenetic marks such as DNA methylation and histone modifications alter the 3-dimensional structure of DNA creating conformational states proximal to genes that are “open” or “closed.” Open conformations promote the transcription of genes while closed conformations inhibit transcription. Commonly measured epigenetic marks such as DNA methylation are proxy, rather than direct measures of conformation. A more direct measure of DNA conformation would be a major advance as investigators could assess effects of environmental exposures on gene “accessibility”. Using a recently developed assay of DNA accessibility – Assay for Transposase-Accessible Chromatin (ATAC) sequencing – we propose to pilot DNA accessibility measures in an ongoing longitudinal birth cohort with extensive measures of toxic exposures. The assay estimates the conformational state of genes across the genome. We will use arsenic exposure as our paradigm chemical exposure, as it is known to affect DNA methylation and histone modifications; therefore exposure would be predicted to alter DNA conformation. We will analyze data in a supervised (i.e., hypothesis driven) and unsupervised manner (i.e. non hypothesis driven) to identify genes that may be affected by arsenic exposure. This pilot will yield invaluable data to support larger studies on the cohort’s longitudinal phenotype measures.
Genetic Variants of Maternal DNA and Maternal Urinary Arsenic Methylation Species during Pregnancy in a Population Exposed to Inorganic Arsenic from Drinking Water
David Christiani et. al. ($24,555)
Prenatal inorganic Arsenic exposure was demonstrated associated with low birth weights by epidemiological evidence. The susceptibility to these negative health effects is possibly due to variations of inter-individual arsenic metabolism efficiency, and the related genetic variations. We aimed to examine whether genetic polymorphisms in inorganic As metabolism are associated with birth outcomes by altering the percentage of each urinary As metabolites in total As excretion. A reproductive prospective cohort study was conducted in two sub-districts of Bangladesh exposed to moderate and high levels of arsenic, respectively. We collected the urine sample in both the <18 weeks gestational age (<18GW) and second trimester (28GW) of 828 pregnant women and quantified the urinary As methylation metabolites (UAMs) are quantified, including inorganic trivalent As and pentavalent As (AsIII, AsV), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA). Maternal blood samples collected at <18GW would be genotyped for the candidate SNP panel (include SNPs in gene AS3MT, GWT family, etc.). Generalized linear model would be used to account for the relationship between genotypes and haplotypes of candidate SNPs, and different urinary arsenic profiles. We would further test the gene-environment interaction under total arsenic exposure and/or second hand smoke exposure. We thirdly propose to build a pathway model for the hypothesized causal connections utilizing structural equation model.
Systems and Methods for the Physico-Chemical & Toxicological Characterization of Emissions from Electronic Cigarettes
Phil Demokritou, et. al. ($22,500)
Understanding the potential hazards from inadequately characterized emissions from electronic cigarettes (e-cigs) is of great importance given the ocean of new tobacco products currently in the market utilizing this technology. Major knowledge gaps exist both in terms of the complex chemistries of both the various liquids used in e-cigarettes and the released aerosol and gaseous by products. Furthermore, evidence continues to grow linking emitted particulate matter and gaseous by products to adverse health effects. Towards this end, we propose to develop a realistic aerosol generation system for the physico-chemical characterization of aerosols emitted from e-cigarettes. State of the art instrumentation and analytical methods will be employed to measure the physico-chemical and morphological properties of by-products (S/TEM/EDX, EC/OC, NMR, etc). The developed exposure generation system will be coupled with in vivo and in vitro biological models in order to understand possible health effects. A number of widely used e-cigarettes will be assessed in order to test the versatility of the proposed methodologies and systems. The use of versatile and realistic generation systems along with the development of new in vitro screening methodologies and in vivo inhalation studies will improve our understanding on possible health risks associated with the use of e-cigarettes.
Early life exposure to environmental persistent pollutants and blood concentrations of adipocytokines from birth through adolescence: Identification of relevant windows of exposure and susceptibility
Philippe Grandjean et. al. ($22,400)
Early life exposure to environmental persistent pollutants e.g. dichlorodiphenyl-dichloroethylene, polychlorinated biphenyls and perfluoroalkyl substances has been linked with obesity and obesity-related conditions in epidemiological studies. Adipocytokines, such as leptin, adiponectin and resistin may play a key role in gene expression changes and hormone balance perturbations are thought to mediate the effects of persistent pollutants on adipogenesis, adipocyte differentiation and energy homeostasis. The integration of adipocytokine biomarkers in birth cohort studies may contribute to advance the mechanistic understanding and identify windows of susceptibility to the metabolic effects of exposure to persistent pollutants. Because blood concentrations of adipocytokines and their relationships to body size may vary through the different stages of childhood and adolescence the etiologically relevant time periods for sampling are unclear. We aim to explore the associations between prenatal and postnatal exposure to persistent pollutants and blood concentrations of adipocytokines measured at multiple time points from birth through adolescence and to identify windows of exposure and susceptibility. For this, we will rely on a well-characterized birth cohort study with already available data on persistent pollutant exposures, repeated body size measures and banked blood samples.
Drinking water exposures to Per and Poly Fluorinated Alkylated Substances (PFASs) in the Nurses Health Study Cohort
Elsie Sunderland and Francine Laden ($25,000)
Analytical advances have enabled detection of a suite of potent environmental chemicals in drinking water. The Nurses Health Study (NHS) collected 33,000 blood samples from women across the U.S. in 1989/1990. Blood samples were returned with tap water from each household and both were archived. Few data exist to compare drinking water exposures to human biomarkers and links to health outcomes are even more rare. We propose to measure a particularly potent class of environmental chemicals: per and polyfluorinated alkylated substances (PFASs) in these archived drinking water samples from the NHS. Recent research has revealed the most dramatic immunotoxic effects ever detected for an environmental contaminant associated with exposure to these compounds. Benchmark doses based on immunotoxicity suggest current guidelines for drinking water are 100 to 1000 fold too high. Serum concentrations of PFASs in NHS individuals are now being analyzed as part of a case-control study on associations with obesity and diabetes. We propose to measure PFASs in archived drinking water from a subset of these same individuals and quantify their contribution to measured serum concentrations. This will provide pilot data on the importance of drinking water exposures to PFASs in US individuals that could be expanded for a future R-01 application.
Project Abstracts – 2015, Round 2
Pesticide residues in food: validation of an exposure assessment tool and relation to male fertility
Jorge Chavarro ($25,000)
Occupational exposure to pesticides has been consistently related to lower semen quality. However, it is less clear whether exposure to pesticides through less intense but more common exposure routes could have similar effects. Diet appears to be the more common route of exposure in the general population. To facilitate research on thel health effects of exposure to pesticide residues through diet, we developed an exposure score based on surveillance data generated by the USDA and diet assessment tools used in epidemiologic studies. Using this method we found an inverse relation between exposure to pesticide residues in diet and semen quality, as a marker of male fertility. Here we propose to further develop and refine this method. Specifically, we propose to evaluate the validity of the dietary pesticide burden score by comparing it against urinary pesticide metabolites (Aim 1), and to evaluate the association of individual urinary pesticide levels with semen quality parameters and assisted reproduction treatment outcomes among men presenting to a fertility center (Aim 2). Addressing these aims will not only allow us to test the validity of our pesticide burden score but could also solidify the proposed approach allowing the broader scientific community to evaluate in an economical manner hypotheses regarding potential health effects of pesticide exposure.
Effects of female hormone on talc phagocytosis
Alexey Fedulov ($25,000)
Talc, a widely-used product considered inert, is a known cause of occupational asthma. More importantly, epidemiologic studies suggest that use of talc powders in genital hygiene increases risk for ovarian cancer. We believe our work to explain worsening of asthma during pregnancy may have relevance to this association. From both in-vitro and in-vivo studies, we showed that pulmonary macrophage phagocytosis of inert particles, like titanium dioxide, is impaired and airway inflammation enhanced by estradiol and progesterone, which are elevated during pregnancy. The relevance of our observations to the talc and ovarian cancer association is suggested by new epidemiologic data which found the association is stronger in premenopausal women and postmenopausal women who used menopausal hormonal therapy–compatible with the hypothesis that estrogen can affect macrophage uptake of talc particles, production of ROS, and subsequent signaling to produce inflammation and consequent carcinogenesis in the female genital tract. We therefore aim to test the effect of female hormones on macrophage uptake, production of ROS, and inflammatory responses of ‘inert’ talc particles including in-vitro and in-vivo approaches. Pilot data support the hypothesis and the feasibility of this proposal. Findings obtained from this pilot are intended to provide data to inform an R21 proposal to NIH.
rDNA copy number: a novel epigenetic modulator and marker of responses to environmental exposures?
Bernardo Lemos ($24,580)
Epigenetic inheritance requires the presence of proteins and RNAs that package the target DNA into a specialized epigenetic structure that can be activated, silenced, and transmitted between generations. The ribosomal DNA (rDNA) arrays are a key point of epigenetic control. Ribosomal DNA arrays give origin to the nucleolus, the site of rRNA synthesis and a major organizer of nuclear architecture. We have identified over 10-fold variation in ribosomal DNA copy number in humans with functional consequences to global epigenetic states. Our preliminary observations also suggest that rDNA copy number is potentially unstable upon environmental exposure. However, the hypothesis that rDNA CN is modulated by the environment is novel and the observations need to be carefully confirmed. Furthermore, the notion that rDNA CN can predict responses to a challenge is also novel. Hence this research is a required step towards testing two novel ideas: that the rDNA is a marker of response to exposure and that the rDNA might itself be modulated by the exposure. The project benefits from a new collaboration with an outstanding lung biology group at NIEHS (Dr. Steve Kleeberger) who is providing well-characterized samples to answer these questions using both in vitro and in vivo exposures. The research will provide required preliminary data towards larger analyses.
The role of iron in apolipoprotein E mediated clearance of amyloid-beta by microglia
Ryan McCarthy: ($17,370)
The metal theory of Alzheimer’s disease (AD) proposes that over time, exposure to metals like iron promotes late-onset AD. A major genetic risk factor for late onset AD is the apolipoprotein (ApoE) E4 allelic variant while the apoE2 variant allele appears to be protective. To explore gene-environment interactions between apoE and iron, our lab used isothermal calorimetry to demonstrate iron-binding to the apoE2 variant. This pilot study seeks to build on these preliminary data to determine binding affinities of different apoE isoforms, specifically testing whether metal interactions with apoE4 are altered. AD is characterized by amyloid-beta (Aβ) deposits in the brain. ApoE promotes Aβ clearance by microglia in an isoform-dependent manner (E2 > E3 > E4). Using immortalized microglia (IMG) cells, our recent in vitro studies have determined the presence of iron suppresses Aβ uptake but that iron-binding by apoE2 mitigates this effect. Based on these results, I postulate that the binding affinity of iron for ApoE is isoform-dependent and that iron binding by ApoE promotes clearance of Aβ by microglia. A corollary to this hypothesis is that impaired iron-apoE4 interactions fail to promote Aβ clearance, thereby increasing susceptibility to AD. This model will be tested by two specific aims: 1) To determine the binding affinity of iron for each of the three allelic variants of ApoE; and 2) To characterize the influence of iron binding by each of the ApoE isoforms on microglial Aβ clearance.
Fine particulate matter and chronic HIV infection co-exposure: environmental determinants of pulmonary function in southwestern Uganda
Chrystal North ($25,000)
Chronic obstructive pulmonary disease (COPD) is the third leading cause of death and disability globally, and in the top ten causes of death in sub-Saharan Africa. Primarily associated with cigarette smoking, a significant minority of people with COPD have never smoked, especially in the developing world. Globally, air pollution is the greatest risk factor for COPD. Likewise, chronic HIV infection has strong associations with COPD. Possible shared pathophysiologic mechanisms involve perturbations in pulmonary endothelial function and systemic inflammation. It therefore stands to reason that co-exposure to both fine particulate matter (PM2.5) and chronic HIV infection will impair lung function more than additively, yet the potentially synergistic exposures of air pollution and chronic HIV infection on respiratory morbidity are unstudied. If this interaction proves significant, the public health implications are broad given the billions of people affected by the intersection of the air pollution and HIV epidemics in sub-Saharan Africa.
Environmental Triggers of Asthma Exacerbation and Gene Expression in an Airway In Vitro Model: Identification of common pathways and unique expression signals for individual exposure stimuli and exposure combinations
Joanne Sordillo: ($25,000)
Asthma exacerbations significantly impact quality of life, and constitute a major public health burden. Although numerous environmental exposures have been individually associated with asthma severity, little is known about how exposure mixtures may trigger an exacerbation. Combinations of exposures may initiate portions of the airway inflammatory cascade that act synergistically, or may induce new signaling pathways not observed for individual exposures alone. In this pilot study, we aim to stimulate primary human bronchial epithelial cells cultured from asthmatics with exposure stimuli known to exacerbate asthma (endotoxin and dust mite allergen), to identify differential gene expression responses for these exposures both alone and in combination. Supernatants from stimulated bronchial epithelial cells will be incubated with airway smooth muscle cells, to determine the downstream impact of stimulated bronchial epithelial cells on the airway smooth muscle. Our ultimate goal is to incorporate information gained from multiple exposure-response expression networks into population studies, by using our airway in vitro model to identify gene sets most likely to interact with particular exposure combinations encountered by asthmatics. Identification of common gene “hubs” across exposures may significantly enhance power in gene by environment interaction models for asthma exacerbations, by providing the most relevant underlying gene network to test as a modifier for the complete environmental exposure profile (the “exposome” prior to the exacerbation). Successful completion of this pilot will yield data for a manuscript on cell stimulatory pathways up-regulated in response to this exposure combination (endotoxin/dust mite), and will also provide preliminary data for an R01 grant proposal on an expanded version of this in vitro model to identify synergistic effects across multiple different types of environmental exposures known to exacerbate asthma.
Circulating extracellular vessicles-associaed microRNAs as novel biomarkers in acute lung injury
Paula Tejera: ($25,000)
The inhalation of toxic gases (air pollution, cigarette smoke, noxious gases) as well as particles and microorganisms contribute to lung injury. A severe pulmonary inflammatory response to such insults results in epithelial and endothelial dysfunction and respiratory failure characteristics of Acute Respiratory Distress Syndrome (ARDS). ARDS is a condition still associated with high mortality and for which we lack effective medical therapies. Its complex pathophysiology, as well as the extensive heterogeneity of ARDS patients, dramatically limits the availability of diagnostic and prognostic biomarkers. In the current study we investigate extracellular vesicles (EVs) released by respiratory cells into the bloodstream and their cargo microRNA as novel biomarkers in ARDS. EVs play important roles in normal lung physiology and lung disease through intercellular communication in the human airway. miRNAs encapsulated in EVs are transferred to recipient cells where control the expression of target genes. In response to proinflammatory triggers, miRNA cargo of EVs is modified. We hypothesize that alterations in EVs-miRNA cargo may lead to ARDS development. In this current pilot project we will conduct an exploratory study aimed to generate EVs-miRNA profiles in plasma of ARDS patients that can serve as plasma biomarkers that facilitate early diagnosis, prognosis, and treatment of ARDS.
2014 Round 1
- David Christiani et al, Genome-wide microRNAs expression profiles and birth weight in umbilical cord serum among women and infants exposed prenatally to inorganic arsenic. ($22,500) Abstract
- John Godlleski and Alex Carll, Does Inhalation of Traffic-Related Particulates Impair Cardiac Performance? ($22,500) Abstract
- Mi-Sun Lee et al, Cardiac Autonomic Effects of Secondhand Exposure to Electronic Cigarettes (ECigarettes). ($20,000) Abstract
- Alex Lu et al, The interplay of sub-lethal neonicotinoids exposure, mitochondrial epigenetics, and Parkinson’s disease. ($22,500) Abstract
- Ronit Machtinger et al, BPA and Phthalate exposure, FF exosomes, and IVF outcome. ($15,000) Abstract
- Marc Weisskopf et al, Assessment of a novel portable x-ray fluorescence device to measure bone lead. ($17,500) Abstract
2014 Round 2
- Gary Adamkiewicz: Pilot Development of a Dorchester Air Quality Surveillance System (DAQSS). ($22,000) Abstract
- Joe Allen: Evaluation of inhalation exposure to diacetyl and other flavoring compounds in e- cigarette. ($22,000) Abstract
- Jennifer Bobb: A statistical approach for estimating the health effects of air pollution mixtures on multiple outcomes simultaneously. ($22,000) Abstract
- Rama Krishnan: Assessment of Environmental Toxins: Bridging the Gap Between Cell Assays and Intact Animals/Humans. ($22,000) Abstract
- Jim Shine: Assessment of heavy metals levels in soil and among a human population living near a copper smelter in Chile. ($22,000) Abstract
David Christiani et al, Genome-wide microRNAs expression profiles and birth weight in umbilical cord serum among women and infants exposed prenatally to inorganic arsenic. ($22,500)
About 130 million people world-wide are being poisoned by inorganic arsenic through drinking water with levels (>10 μg/L) higher than the guideline values defined by the World Health Organization . Arsenic at a modest level (<100 μg/L in urine) is known to induce toxic effect on the developing fetus and has been found to be associated with decreased birth weight, head and chest circumference and increased risk of infection in infants [2,3]. Low birth weight is one of the major causes of perinatal morbidity and mortality and also associated with an increased risk for certain diseases later in life . Both genetic and epigenetic mechanisms, including alteration of microRNA (miRNA) expression profiles are believed to play an important role in the pathogenesis of many arsenic associated disease . The goal of this pilot study is to explore miRNA expression profiles in cord blood serum to elucidate the molecular basis that underlie in utero arsenic exposure with birth weight and generate preliminary data for a full proposal. Since exposure data in our study population has already been collected, we propose to use this archived resource to explore how well maternal blood, urine, hair and nail arsenic correlate with the miRNA expression profiles in cord blood with a view to understand in-utero fetal exposure. The results of this pilot study will be used as preliminary data to apply for a five-year RO1 grant.
John Godlleski and Alex Carll, Does Inhalation of Traffic-Related Particulates Impair Cardiac Performance? ($22,500)
Traffic-derived particulate matter (TPM) exposure is linked to heart failure morbidity and mortality. Prior research suggests TPM may impair left ventricular (LV) performance (contractility and lusitropy) by enhancing the autonomic nervous system’s (ANS) sympathetic influence. Yet, the evidence supporting this mechanism is limited by unrepresentative exposures and a crude index of LV contractility. This project will: 1) develop chronic LV pressure (LVP) measurements; 2) test whether repeat exposure to real-world TPM at environmentally relevant concentrations impairs LV performance; and 3) determine if increased sympathetic tone mediates these effects. Rats will be analyzed during and after a 4-wk exposure to TPM from our novel highway tunnel exposure system (vs. clean air) using telemetry-derived chronic LVP, electrocardiogram (ECG), blood pressure (BP), heart rate, and heart rate variability (HRV, a measure of ANS balance). We will subsequently compare air- or TPM-exposed rats treated with a sympathetic inhibitor (guanethidine) or saline to test for sympathetic mediation of TPM-induced cardiotoxicity. Stress tests and baroreflex challenges will be used to unmask latent effects of TPM on LV performance and ANS regulation during increased cardiac demand. Echocardiography (Echo) and molecular assays will test for LV performance decrements and their potential mediators.
Mi-Sun Lee et al, Cardiac Autonomic Effects of Secondhand Exposure to Electronic Cigarettes (ECigarettes). ($20,000)
Electronic cigarettes, e-cigarettes or electronic nicotine delivery systems, are battery-operated devices that vaporize chemical mixtures composed of nicotine, propylene glycol and other chemicals. As the use of ecigarettes is becoming popular, secondhand smoke exposure from e-cigarettes is increasing among nonsmokers and children, but limited data is available about their safety, raising significant public health concerns. Recently, a few studies have shown that increased airway resistance and decreased pulmonary function are linked to e-cigarettes exposure, but, to our knowledge, it is not yet to be explored whether secondhand exposure to e-cigarette is associated with cardiovascular disease, which remains the leading cause of death worldwide. Our pilot study employs a repeated measures design to assess whether secondhand exposure to e-cigarette is associated with changes in cardiac autonomic function. Specifically, we will examine the association between selected airborne markers and biomarkers of nicotine and tobaccorelated toxicants from e-cigarettes, and heart rate variability (HRV) extended in lifelong nonsmokers. Findings from this study will be used as preliminary data to apply for a five-year RO1 grant to elucidate the chronic effects of e-cigarettes exposure.
Alex Lu et al, The interplay of sub-lethal neonicotinoids exposure, mitochondrial epigenetics, and Parkinson’s disease. ($22,500)
Parkinson’s disease (PD) is the most prevalent neurodegenerative movement disorder. Higher incidence of PD has been associated with farmers, and therefore agricultural pesticides have been implicated as the agents for PD progression. However the exact mechanisms on how pesticides affect PD are largely unknown. Data from our current study in honeybees have suggested a significant link of sub-lethal neonicotinoid exposure and the increase of mitochondrial DNA copy number (mtDNAcn) over generations. Here we aim to demonstrate how imidacloprid, the most persistent and commonly used neonicotinoid insecticide, affects PD using human neuroblastoma SH-SY5Y cell lines. The differentiated SH-SY5Y cell lines with Parkin- knockdown and overexpressing, provide a unique opportunity to examine the changes of mtDNAcn and the mtDNA methylation. This proposed study will expand our understanding on how neonicotinoids affect neurodegenerative disease such as PD, establish a cell line based protocol to examine the causal relationship, and pave the way to develop health-based biomarkers such as mtDNAcn and mtDNA methylation for future epidemiological research.
Ronit Machtinger et al, BPA and Phthalate exposure, FF exosomes, and IVF outcome. ($15,000)
Concerns are increasing that endocrine disrupting chemicals (EDCs) may have adverse effects on animal and human fertility. We have shown that in vitro exposure of immature human oocytes to bisphenol-A (BPA) impaired oocyte maturation and spindle formation. However, the mechanism by which bisphenol A and EDCs affect oocyte quality is unclear. The human oocyte is surrounded by and in bi-directional communication with cumulus granulosa cells which, in turn, are bathed in follicular fluid (FF) within the ovarian follicle. Innovative studies have proposed that intercellular transmission within the follicle might involve secretion and uptake of microRNAs (miRNAs) carried in membrane-bound extracellular vesicles such as exosomes. Our hypothesis is that EDCs such as BPA and phthalates in the FF might alter FF exosome profile and secretion of miRNAs and thus impair oocyte maturation, fertilization and day 3 embryo quality. We propose to isolate exosomes from the FF among women undergoing IVF. The goals of this pilot grant are to investigate whether there is a correlation between FF levels of BPA and phthalate metabolites with miRNA profiles of FF exosomes (Aim 1) and to determine the association of miRNA profiles of FF exosomes with oocyte maturation and day 3 embryo quality (Aim 2).
Marc Weisskopf et al, Assessment of a novel portable x-ray fluorescence device to measure bone lead. ($17,500)
The development of X-Ray Fluorescence (XRF) techniques to measure lead in bone led to a windfall of critical scientific research implicating cumulative exposure to lead in hypertension, heart disease, renal dysfunction, cognitive decline, Parkinson disease, cataracts, and psychiatric symptoms to name a few. However, the Kshell XRF (KXRF) using a 109Cd energy source which has been the principle method used for bone lead analysis has severe limitations such that all of this work has been essentially limited to only two research groups. Furthermore, the limitations make it impossible to use in many epidemiological studies, thereby limiting what questions can be asked. The KXRF approach requires a dedicated room; it requires at least one half hour of measurement time; and it uses a radioactive isotope in limited production as an energy source, which currently severely restricts the use of the technique because of regulatory demands. To address these limitations, we propose to conduct a pilot study to assess a portable XRF (pXRF) device to non-invasively measure lead in vivo in bone. We aim to compare results with this new approach against existing serial blood lead measurements and KXRF measurements in the Normative Aging Study, and an occupationally exposed population seen at the Cambridge Health Alliance. Demonstrating comparable results with the use of the pXRF could result in tremendous new research opportunities in settings among children or patients of diseases that affect their ability to come to a central research site(e.g. Alzheimer’s disease or ALS), as well as in field settings where exposures are often highest.
2014 Round 2
Gary Adamkiewicz: Pilot Development of a Dorchester Air Quality Surveillance System (DAQSS). ($22,000)
We propose to build a partnership between HSPH, MIT and Dorchester-based community groups to establish a prototype real-time community-based and community-scale air quality sensor network. This model is based on the web-based CLAIRITY monitoring network which is currently operational on the MIT campus. The proposed network will utilize low-cost and web-connected sensors, data storage and data visualization to allow for community and researcher engagement with quantitative air quality data collected across exposure-relevant spatial and temporal scales. Via the collaborative, we will work closely with community partners to design a real-time interface for dissemination of the data acquired with the network. Our aim is to provide the community with interactive tools that promote exploration and understanding of the factors controlling their air quality. Our expectation is the network will result in data-driven strategies to mitigate community-specific exposures, addressing topics that are central to the scientific and societal mission of the NIEHS center. We are confident that such a platform will have significant application to other studies and grant opportunities in the coming year, as evidenced by specific EPA and NIH RFPs.
Joe Allen: Evaluation of inhalation exposure to diacetyl and other flavoring compounds in e- cigarette. ($22,000)
The World Health Organization reports that $13 billion was spent on e-cigarettes in 2013, with sales expected to increase 17-fold in 15 years. A staggering number of children have begun to experiment with this product; CDC estimates that 1.75 million children tried e-cigarettes as of 2012, with 160,000 of them reporting that they had not used tobacco cigarettes. There has been very little research on the health implications from the constituents of e-cigarettes. Of that research, there is none that examines the flavoring chemicals used in e-cigarettes, despite over 7,000 flavors currently being marketed. Chemicals used in flavorings gained notoriety in the early 2000’s after workers at microwave popcorn were diagnosed with bronchiolitis obliterans, which was attributed to diacetyl exposure from inhalation of vapors off heated mixing vats. Diacetyl, a flavoring compound, and its replacements, 2,3-pentanedione and acetoin, are used in the manufacture of many foods for a wide range of flavors beyond butter (e.g., caramel, cream, pina colada, strawberry). Many of these flavors are common in e-cigarette flavor cartridges, and, similar to popcorn workers, users of e-cigarettes are directly inhaling heated flavoring compounds. We propose to quantify the concentrations of these flavoring compounds in e-cigarettes, estimate exposures for users of e-cigarettes, and compare these to health-based guidelines.
Jennifer Bobb: A statistical approach for estimating the health effects of air pollution mixtures on multiple outcomes simultaneously. ($22,000)
In environmental epidemiology, the most common approach is to select a few health outcomes hypothesized a priori to be affected by the exposure and then estimate the exposure-response function and associated risks. However, a key limitation of this approach is that other adverse outcomes that are clinically important but less expected or less common could be missed. In this study, we will develop a new scientific paradigm for analyzing a massive number of health outcomes simultaneously. We will apply this framework to identify the complete spectrum of health conditions that are most likely to occur during air pollution events, defined as days with high levels of several pollutants concurrently. This project will enable us to study, for the first time, the short-term effects of exposure to air pollution mixtures on more than 15,000 ICD-9 codes simultaneously, rather than on pre-specified individual conditions. Knowledge of the range of health responses that are affected by air pollution mixtures will provide insight into the physiological pathways by which pollution affects health, thereby informing public health approaches to prevention. Moreover, the statistical approach will enable future studies to estimate the impacts of exposures to mixtures of metals, organic chemicals, particulate matter, and other toxins on large constellations of human health outcomes.
Rama Krishnan: Assessment of Environmental Toxins: Bridging the Gap Between Cell Assays and Intact Animals/Humans. ($22,000)
Science and technology produce abundant new chemicals and particles. The NIEHS, NTP, and other government agencies are faced with identifying ones that have potential health and ecological threats. Currently, our NIEHS Center has a number of cell-based assays that emphasize high throughput screening. At the other end of the spectrum, we have lifetime animal studies in rodents, as exemplified by the classical NTP approach. There is even a smaller number of toxins with data from epidemiologic studies in humans. Between these oversimplified assays and the complexity and expense of studying intact animals and humans, there is an important gap. This pilot project intends to fill that gap with an assay that is capable of testing large numbers of environmental toxins, and yet better represents the complexity of intact animals and humans. We propose to improve and calibrate an assay, Precision Cut Lung Slices (PCLS), to investigate chemicals that are likely to be toxic. We take advantage of our experience with zinc chloride (ZnCl2) chemical exposures. Zinc (Zn) will serve as a model toxin to determine the predictive power of the proposed platform. We see this array of assays and especially the PCLS system as the foundation for multiple grants relating to novel uncharacterized materials, such as families of nanomaterials or organic chemicals.
Jim Shine: Assessment of heavy metals levels in soil and among a human population living near a copper smelter in Chile. ($22,000)
Chile is the largest copper producer in the world, and mining activities are conducted all over the country. In the area of Ventanas-Puchuncaví a copper smelter was established in 1962. Preliminary data show a mixture of different heavy metals (mainly lead, copper, arsenic, cadmium, mercury and manganese) in soils around the smelter and in surrounding residential areas. The goal of this project is to gather and analyze environmental and biological samples for generation of preliminary data assessing an association between the presence of a mixture of metals in soil with heavy metals in toenails and hair in 280 permanent residents for further epidemiologic studies. The next stages of this project will be an examination of metal related human toxicity, geoavailability and bioavailability, depending on variables related to different biological, chemical and geological processes (such as chemical state of metals, type and proportion of each metal, presence of other compounds, type of soil, proximity to slag piles, and biologic characteristics of exposed individuals). DNA samples are already available through a previous project funded in Chile, so we intend to study associations between metal levels and some biomarkers of epigenetic alterations and oxidative stress.
Pilot Grant Awardee: Andrea Baccarelli
Project Title: Flame retardants, plasma exosomal miRNAs in early pregnancy, and risk of gestational diabetes
Award Amount: $24,000
Description: Gestational Diabetes Mellitus (GDM) prevalence has increased up to 2.5 times in the past 20 years and is now diagnosed in 18% of all pregnancies. Polybrominated diphenyl ether (PBDE) flame retardants are endocrine disruptors that have been linked with insulin resistance and diabetes. This mechanistic project will build on a growing body of evidence suggesting that exosomes (extracellular vesicle) shed from placenta during gestation carry tissue-specific signals that reflect placental function and could be used to both predict pregnancy-related risks and characterize environmental exposures with placental toxicity. Exosomes are 30-100 nm vesicles encapsulated by a lipid bilayer and contain high concentrations of miRNAs. Our long term hypothesis is that PBDE exposures induce exosome-signaling from the placenta that mediates GMD development. We will isolate placenta-derived exosomes from maternal plasma in early pregnancy (8-21 weeks) in women at high GDM risk; measure their size and count; screen ~750 exosome-contained miRNAs; and prospectively determine their association with GDM [Aim 1]; evaluate the correlations of miRNAs in placental tissues with plasma exosomal miRNAs, as well as with GDM risk [Aim 2]; measure PBDEs in maternal plasma/placental samples and determine PBDE effects on exosomal/placental miRNAs and GDM risks [Aim 3].
Pilot Grant Awardee: Christopher Hug
Project Title: Does environmental arsenic exposure induce a novel form of cystic fibrosis?
Award Amount: $25,000
Description: The goal of this Harvard-NIEHS Center pilot program application is to conduct a cross-sectional study in Bangladesh to assess the relationship between environmental arsenic exposure, respiratory function and sweat chloride levels. Recent cell culture and biochemical studies have shown that arsenic promotes degradation of the cystic fibrosis transmembrane conductance regulator (CFTR), a chloride channel essential for normal function of the lungs, pancreas and intestines. Mutations in the CFTR gene cause cystic fibrosis, a rare autosomal recessive disease characterized by recurrent pneumonia and diagnosed by demonstrating an increased concentration of chloride in sweat. Our central hypothesis is that environmental arsenic exposure results in an induced cystic fibrosis phenotype in exposed individuals by causing degradation of CFTR. By demonstrating an association between arsenic exposure and abnormal sweat chloride levels, our project will establish a new molecular paradigm for the pathogenesis of arsenic-induced respiratory disease. The data from this pilot program will provide important preliminary data to support an R01 application in response to RFA HL-12-035, entitled “Early Cystic Fibrosis Lung Disease Studies in Humans.”
Pilot Grant Awardee: Peggy Lai
Project Title: Endotoxin, the airway transcriptome, and asthma
Award Amount: $25,000
Description: Asthma is a chronic inflammatory lung disease that affects an estimated 25 million Americans. The contribution of indoor endotoxin exposure to asthma morbidity has been extensively studied in the home environment, but little attention has been paid to indoor endotoxin exposure in schools. Studies show that gene expression changes due to environmental exposures in the lower airways (bronchial epithelium) are also mirrored in the upper airways (nasal epithelium), with the advantage that the latter is easily accessible. Endotoxin is a consistent predictor of acute changes in gene expression of the airway epithelium. Airway gene expression studies in tobacco-related lung disease have been used to identify mechanisms of disease and provide useful biomarkers for diagnosis, prognosis, and treatment. However, the transcriptional changes induced by chronic environmental endotoxin have not been defined, particularly in the field setting. The purpose of this pilot proposal is to identify longitudinal changes in the airway transcriptome in asthmatic children in response to school endotoxin exposure, and to identify a gene signature at baseline that predicts future asthma exacerbations in response to school endotoxin exposure.
Pilot Grant Awardee: Young-Ah Seo
Project Title: Manganese-induced neuronal cell apoptosis through ER stress
Award Amount: $18,000
Description: Chronic exposure to manganese (Mn) results in neurobehavioral deficits similar to Parkinson disease. Despite growing awareness of the problems associated with Mn neurotoxicity, particularly in children, little is known about the molecular mechanisms underlying Mn-induced neurotoxicity. We found that Mn induces cell injury and ultimately apoptosis both in vitro and in vivo. This apoptotic effect is synergized by iron deficiency. However, it is not clear how these two metals interact. This is particularly important because iron deficiency is the most prevalent nutritional deficiency in children, and Mn intoxication may produce greater effects during development. Accumulating evidence suggest that ER stress plays a significant role in neurodegenerative diseases because the unfolded protein response (UPR) leads to cell apoptosis. Thus, we postulate that Mn induces neuronal cell apoptosis through ER stress and this pathway is potentiated by iron deficiency. This hypothesis will be tested by two specific aims: 1) To determine whether Mn-induced ER stress initiates neuronal apoptosis in vitro. 2) To determine whether iron deficiency enhances Mn-induced neuronal apoptosis and/or ER stress in vitro and in vivo. These studies will provide insight into mechanisms of Mn-induced neurotoxicity and the environmental health risks from Mn exposures in iron-deficient children.
Pilot Grant Awardee: Vishal Vaidya
Project Title: Quantitative High-Throughput Screening Platform for Predictive Kidney Toxicology
Award Amount: $25,000
Description: Drugs and environmental chemicals play an important role in the high incidence and prevalence of kidney injury, which in many circumstances can be prevented or at least minimized by predictive toxicity screening. We propose to transform the traditional in vivo, dose-response based toxicity assessment by developing in vitro, high throughput, multi-dimensional signatures that map early biological perturbations of a damaged cell. This involves developing a ‘grid’ using quantitative dynamical systems analysis to integrate the chemical structure with dose and time dependent hemeoxygenase-1 (HO-1) response with high content imaging for perturbations in nucleus, endoplasmic reticulum and mitochondria following toxicant exposure. In Aim 1 we will develop and optimize homogenous time resolved fluorescence (HTRF) assay to allow high throughput measurement of HO-1 as a biomarker of toxicity. In Aim 2 the Library of pharmacologically active compounds will be screened and a multidimensional predictive toxico-response signature will be created by linking HO-1 response with structure, pharmacological activity and high content cell perturbation markers along dose-time axis by combining statistical, reverse engineering/inference and kinetic approaches. The ultimate goal is to develop a predictive kidney toxicity test to speed safety screening and risk assessment.
Pilot Grant Awardee: Konstantinos C. Makris
Project Title: Exposures to thyroid-disrupting chemicals and thyroid nodular disease: the case of Bisphenol A
Award Amount: $20,000
Description: There is an alarming increase in global incidence rates of thyroid nodules partially ascribed to the augmented use of ultrasonographs, while the effect of environmental chemicals, like bisphenol A (BPA) on the etiology of thyroid nodular disease remains poorly understood. Cyprus is topping the global list of countries with the highest thyroid cancer incidence rates. Our biomonitoring study showed very high BPA intake values for a Cypriot subpopulation during prolonged manifestation of adverse summer weather conditions (very high temperatures/UV index) that could enhance BPA leaching and thus, human exposures. Our aims are: (1) measuring urinary BPA and its chlorinated (mono-, and di-chloro BPA) and oxidative (4-methyl-2,4-bis(4- hydroxyphenyl)pent-1-ene, MBP) metabolite exposures in patients with thyroid nodular disease and perturbed or not thyroid hormonal status using a case-control design and an outcome-dependent sampling design in Cyprus, and (2) investigating relations between thyroid nodular disease cases, their dietary habits, and urinary BPA and its metabolite concentrations that show 10-1000x higher in-vitro estrogenic potency than that of BPA. The simultaneous urinary analyses for BPA and its derivatives could shed light on the importance of low-dose BPA health effects when parent compound partially transforms to more potent metabolites.
Pilot Grant Awardee: Stephanie Shore
Project Title: Impact of obesity and ozone on lung microbiota in mice
Award Amount: $24,000
Description: Obesity is a risk factor for asthma. Understanding the mechanistic basis for obesity-related asthma could lead to novel treatments. Gut microbiota are altered in obesity and these alterations appear to contribute to some obesity-related conditions. Microbiota also exist within the lungs and have been proposed to contribute to a variety of lung diseases. Hence, lung microbiota might also contribute to obesity-related asthma. Preliminary data indicate that both obesity and ozone (O3) exposure (a common asthma trigger) result in changes in lung concentrations of metabolites whose generation requires bacterial metabolism, suggesting that both obesity and O3 can impact lung microbiota. Such changes may result in lung metabolites that can affect airway function and trigger asthmatic symptoms. Hence, we propose to examine the hypothesis that both obesity and acute O3 exposure alter bacterial populations within the lungs. To do so, obese and lean mice will be exposed to room air or to O3. 24 h later, mice will be euthanized and a bronchoalveolar lavage (BAL) will be performed. Bacterial DNA will be extracted from BAL supernatant and 16-S pyrosequencing used to analyze taxa in the lungs. Both genetically obese db/db and genetically obese Cpefat mice will be examined to confirm that any observed changes are not specific to a particular genetic defect but a common result of obesity.
Pilot Grant Awardee: Akira Tsuda
Project Title: Rapid nanoparticle translocation across the neonatal alveolar air-blood barrier
Award Amount: $20,000
Description: The alveolar walls of infant lungs are likely more vulnerable than that of adults’ because the neonate lung undergoes significant morphogenetic and fluid balance changes. Thus, the ability of nano-size particles (NPs) to cross the alveolar air-blood barrier is also likely to be age-dependent during postnatal lung development. In the proposed project, we will perform systematic translocation experiments in neonate rats of different developmental stages to investigate, which physicochemical characteristics of the particles determine their ability to cross the alveolar air-blood barrier of infant lungs. Virtually nothing is known currently about NP translocation across the neonatal alveolar air-blood barrier. Therefore, the results of this project will provide us with fundamental knowledge on how NPs interact with the immature lungs of infants. This knowledge is essential for the assessment of nanotoxicology as well as for the optimal design of nanoparticles for systematic inhalation drug therapy for this vulnerable age group.
Pilot Grant Awardee: Yongyue Wei
Project Title: Maternal Metal Exposure, Cord Blood Metabolomics Biomarker, and Birth Outcome
Award Amount: $20,000
Description: Metals such as lead, manganese, and arsenic are of increasing concern since recent data demonstrate their biological toxicity to birth outcomes. However, the mechanisms of such toxicity are not well understood. Metabolomics are the end products of cellular regulatory processes, and their levels can be regarded as the ultimate response of biological systems to environmental changes, e.g. metal exposures. Systemic changes in the metabolome relate to particular outcomes, e.g. birth outcomes. However, few studies have investigated the toxic effects of maternal metal exposure on birth outcomes that may be mediated by metabolites. Human development reflects dynamic processes mediated by a dynamic interplay among genetic, epigenetic, metabolic and environmental determinants. Toxic chemicals, interactive genes and metabolites may compose a functional network and, ultimately produce adverse birth outcomes. Few studies have explored such toxic effects using a network analysis. Taking advantage of the Superfund Program Project (Project #2, PI: D. Christiani), we conduct this study to identify metabolic biomarkers that mediate the effect of maternal metal exposure to low birth weight, which would have impact on the understanding of how metals induce toxicity and provide biological insight for potential treatment and prevention of adverse reproductive outcomes.
Pilot Grant Awardee: Jonghan Kim
Project Title: Influence of FPN and HFE on clearance of manganese
Award Amount: $25,000
Description: Ferroportin (FPN) is an essential intestinal metal transporter and its expression is elevated in HFE-related hereditary hemochromatosis, one of the most common genetic disorders in humans. Hemochromatosis is caused by mutations in the HFE gene, resulting in hyperabsorption of intestinal iron. This promotes irreversible tissue damage including liver cirrhosis, cardiomyopathy, and premature death. In addition, we observed an increase in blood manganese clearance in HFE-deficient mice, suggesting a novel of role of FPN in manganese disposal. We postulate that FPN is responsible for Mn clearance by tissue deposition and/or by excretion from the liver into bile and these pathways are altered in HFE-associated hereditary hemochromatosis. This hypothesis will be tested by two specific aims: 1) To determine and compare the tissue distribution pharmacokinetics of 54Mn administered to FPN-mutant and wild-type “control” mice by intravenous and intraportal injection. 2) To characterize and compare the tissue distribution pharmacokinetics of 54Mn administered to HFE-deficient and wild-type mice by intravenous and intraportal injection. This work will enhance our knowledge of manganese homeostasis and the role of ferroportin in hemochromatosis and manganese neurotoxicity.
Pilot Grant Awardee: Chensheng (Alex) Lu
Project Title: Metabolomic Profiling for Dietary Pesticide Exposure in Children
Award Amount: $25,000
Description: “Environmental Metabolomics” is a newly emerging research focusing on the identification and quantification of the small molecule metabolites in biofluids. A key element in the environmental metabolomic research is to understanding what is “abnormal” in the targeted individuals under either well-defined or known exposure circumstances whose metabolic health can be interrogated using the metabolomic profiling approaches. The goal of this metabolomic analysis is to extract, identify, and quantify all of the metabolites, a global approach, in urine samples collected from the Children’s Pesticide Exposure Study (CPES). Since we will be looking into the non-targeted metabolomic profile changes in the same child who consumed organic and conventional foods during the study period, we should be able to minimize the potentially confounding influences, such as age, sex, race, and nutrition on interpretation of metabolomic results. Thus, this unbiased metabolomic analysis should provide robust insights into the influence of dietary pesticide intakes on the relative concentration of observed biochemicals. This proposed pilot study extends pesticide research to metabolomic analysis that links pesticide exposure to biochemistry-based health effects. Ultimately, we hope to identify biomarkers for pesticides with fingerprints on potential health effects through the metabolomic analysis.
Pilot Grant Awardee: Quan Lu
Project Title: Discovery of coding and non-coding RNA transcripts central to lead (Pb) neurotoxicity through Next-Gen deep sequencing
Award Amount: $25,000
Description: Lead (Pb) is a common environmental metal contaminant of great public health concern because it impairs neuronal function and adversely affects neurodevelopment in children. Despite overwhelming evidence from epidemiological as well as animal and cell-culture studies that show Pb is a neurotoxicant, the molecular mechanisms by which Pb impairs neuronal function remain poorly defined. Here we propose to use the powerful and now affordable Next-Gen deep sequencing technology to discover human gene transcripts, both coding and non-coding, that are critically involved in the neuronal response to Pb exposure. This proposal has two specific aims: 1) to identify messenger RNA (mRNA) and long non-coding RNA (lncRNA) expression changes through deep sequencing of polyadenylated RNAs from differentiated human neuronal cells exposed to Pb; and 2) to identify microRNAs (miRNAs) gene expression changes through deep sequencing of small RNAs from differentiated human neuronal cells exposed to Pb. Identification of these RNA transcripts will provide a comprehensive genetic framework critical for elucidating the mechanisms underlying Pb-induced neurotoxicity.
Pilot Grant Awardee: Joel Schwartz
Project Title: Particles, Metals, and changes in 5-OH-mC: a Pathway to Disease?
Award Amount: $18,000
Description: The methylation of cytosine is a key regulatory factor in the expression of genes, and is associated with environmental exposures. More recently attention has turned to processes that oxidize 5-methyl Cytosine into 5-hydroxymethyl Cytosine(5-OH-mC). While methylated Cytosine tends to repress gene expression, 5-OH-mC primes chromatin for expression. Moreover, oxidative stress is the key process driving the formation of 5-OHmC, and is also a response to particulate air pollutants and some heavy metals. Using a prospective, richly phenotyped cohort with excellent exposure markers, we propose to measure 5-OH-mC in 500 subjects and examine the association with short term and longer term exposure to particles, including particle components, whose relative toxicity has been identified by the National Academy of Sciences as a key research need. We will use a supersite located at HSPH, GIS and satellite based modeled exposure at subjects homes, and in home sampling to characterize exposure, giving us measures with flexible timescales, long term contrasts, and more personal exposure (but for a fixed time period). We will use toenail metal concentrations to assess exposure to As, Cd, Mn, Pb, and Hg. We will also examine associations of 5-OH-mC with intermediary biomarkers of cardiovascular disease (inflammatory markers, QT interval, and 8-OHdG).
Pilot Grant Awardee: Joanne Sordillo
Project Title: Development of a Molecular Method to Assess the Fungal Microbiome in Environmental Air and Dust Samples from Observational and Controlled Human Exposure Studies
Award Amount: $14,000
Description: Fungi in the environment influence human health in ways that are complex and incompletely understood. Current exposure assessment models may obscure relationships between fungi and health effects, either by grouping all fungi together (i.e. total colony forming units), or by focusing too narrowly on a few specific taxa. Alternatively, characterization of the fungal community in the environment (the “fungal microbiome”) has the potential to give a comprehensive look at all fungi present, while yielding important information on the unique mixture, or diversity, of an environmental exposure. Fungal, as well as bacterial diversity is posited to influence asthma risk, but despite its potential importance, the molecular and computational tools to assess fungal composition and diversity for epidemiologic purposes are far less well-developed than those for bacteria. Although culture methods have been used to assess fungal diversity to some extent, they require viable organisms, are time sensitive, labor intensive, and miss non-culturable fungi. This project aims to develop and validate a high-throughput molecular method to assess fungal diversity in various types of environmental samples. This method will utilize two regions of the fungal rRNA locus (ITS and LSU), with potential applications for environmental health studies of fungi, as well as gene by environment interaction studies.
Pilot Grant Awardee: Nancy Krieger
Project Title: Air pollution, racial discrimination, and blood pressure: an exploratory study
Award Amount: $22,000
Description: We will conduct a repeat cross-sectional exploratory and novel study that efficiently links 3 Boston-based data sets: 2 health studies (My Body My Story (N=1005) and United for Health (N = 1201)) and a unique spatiotemporal data set that enables precise estimation, to the latitude and longitude of a person’s residential address, of time-specific exposure to traffic-related air pollution, reflected by black carbon concentrations in PM2.5 (particulate matter with an aerodynamic diameter ≤ 2.5 μm). Our specific aims are:
- Aim 1: map and assess the social-spatial distribution of black carbon exposure, in relation to census tract sociodemographic characteristics and individual-level data on age, gender, race/ethnicity, exposure to racial discrimination, lifetime socioeconomic position, smoking, nativity, and body mass index; and
- Aim 2: test our hypotheses that: (1) higher exposure to black carbon is associated with increased risk of blood pressure (measured at the time of the exam, with analyses controlling for body mass index, based on measured weight and height, plus age, gender, nativity, smoking, and lifetime socioeconomic position), and (2) the effect of black carbon on blood pressure is modified by exposure to racial discrimination
Pilot Grant Awardee: Francine Laden
Project Title: Metabolomics to Identify Novel Biomarkers of Exposure to Traffic Exhaust
Award Amount: $24,500
Description: Vehicle exhaust is a mixture of toxicologically important chemicals, but there has not been a systematic effort to develop exposure biomarkers. “Exposome” technologies provide an opportunity for a data-driven approach to biomarker development. In particular, metabolomics techniques can identify chemical features of all small molecules in a blood sample, providing an opportunity to identify novel exposure and early effect markers by comparing samples from exposed and less exposed groups (1,2). The proposed study will demonstrate application of metabolomics technology to biomarker development for traffic exhaust using samples collected from workers in the trucking industry. We will identify chemical features that are associated with previously collected exposure measures, including job title, and area exposure levels of PM2.5, EC, OC, particle-bound PAH, and ultrafine particle surface area and urinary nitro-PAH metabolites. Additionally, we will determine if these features are associated with the following early effect markers: blood markers of inflammation (CRP, IL- 6), endothelial dysfunction and activation (ICAM-1, VCAM-1), and oxidative stress (8OHdG). If we successfully identify a novel biomarker of traffic exposure from the metabolome, the results from this pilot can be directly applied to future studies of chronic disease.
Pilot Grant Awardee: Jessica LaRocca
Project Title: Elucidating the effects of prenatal exposure to multiple endocrine disrupting chemicals on microRNA and mRNA expression in the human placenta
Award Amount: $25,000
Description: Recent evidence has suggested that early life exposure to environmental compounds, including endocrine disrupting chemicals (EDCs), may alter risk of disease development. A number of EDCs are found in many everyday products, such as plastic containers, canned goods and receipt paper. EDC exposure to developing children is of particular concern because early life chemical exposure to a hormonal sensitive organ can result in phenotypic organizational changes that are persistant throughout life. Several animal studies have demonstrated that prenatal exposure to EDCs can alter postnatal development, and may be linked to risk of developing adult disease. The health of the placenta, the hormonally active organ responsible for nutrient exchange between the fetus and the mother, is critical to the health of the developing child. Various environmental toxicants, including EDCs, have been shown to cross the placental barrier and alter gene expression. We hypothesize that prenatal exposure to EDCs will alter mRNA and miRNA expression in human placenta, which could potentially affect the health of the child. Using two Boston birth cohorts, we propose to examine if exposure to a number of EDCs during pregnancy alters mRNA and miRNA expression levels of the human placenta, and how this correlates with maternal complications and birth outcomes.
Pilot Grant Awardee: Maitreyi Mazumdar
Project Title: Neural Tube Defects and Environmental Arsenic Exposure
Award Amount: $24,000
Description: The scientific goal of this Harvard NIEHS Center pilot program application is to conduct a case-control study in Bangladesh to assess the relationship between environmental arsenic exposure and neural tube defects, which are common, serious birth defects that result in life-long disabilities. Secondary analyses will investigate genetic and nutritional factors that may account for individual differences in sensitivity to arsenic exposure. By combining investigations of toxic exposure with examination of genetic polymorphisms related to nutrition and metabolism, this proposal has the potential not only to demonstrate an association between environmental arsenic exposure and neurological defects in children but also to suggest means by which individual resistance to such exposure may be enhanced. The data from this pilot program project will provide important preliminary data to support an application in response to NIH RFA PAR-11-031 entitled “Brain Disorders in the Developing World (R21).” This RFA is soliciting applications for projects in developing countries that study disorders “that result from abnormal prenatal development or influences during the prenatal and perinatal period…including genetic and nutritional factors…and environmental toxins.” This future, prospective, active surveillance study will allow us to extend our evaluation of the gene-environment interactions explored here.
Pilot Grant Awardee: Stephanie Shore
Project Title: Metabolic profiling of obese and ozone exposed lungs
Award Amount: $18,000
Description: Obesity increases pulmonary responses to ozone, a common air pollutant and asthma trigger. We have established that ozone causes greater airway hyper-responsiveness (AHR), a characteristic feature of asthma in obese compared to lean mice. Our preliminary data indicate that IL-17A may be involved in obesity-related augmentation of ozone-induced AHR. Among its other activities, IL-17A has been shown to affect metabolism within adipocytes. Metabolomics is an emerging technology that simultaneously quantifies hundreds to thousands of small molecules generated from cellular metabolism. In mice, metabolomics has been successfully employed to identify strain related differences in lung cellular metabolism that may explain strain related differences in sensitivity to another environmental pollutant, acrolein. We propose to use a metabolomics platform developed by Metabolon, to 1) determine whether there are obesity-related differences in the effect of ozone on lung cell metabolism; and 2) determine whether IL-17A contributes to differences in metabolism. Understanding the metabolic “signatures” of ozone exposed mice and how they vary with a common risk factor (obesity) may ultimately lead to the development of biomarkers that can be used to assess human responses to ozone and identify other susceptibility factors.
Pilot Grant Awardee: Baccarelli, A
Project Title: Metal Exposure and DNA Retrotransposon Activation
Award Amount: $18,000
Description: Recent evidence suggests that noncoding DNA sequences (i.e. repetitive elements or retrotransposons) may actually be expressed, and if so can disrupt cellular functions. DNA methylation of these regions is an evolutionary response to inhibit their expression. Toxic metals, including lead, alter DNA methylation patterns and may induce hypomethylation of DNA. Methylation of retrotransposons can be lost in response to lead exposure. L1 sequences (also known as LINE-1) are the main group of viral-like retrotransposons that are still active in the human genome. L1 transcription during development might be reactivated by demethylation and thus elicit cell-wide inflammatory responses producing adverse health effects such as fetal growth restriction or preterm delivery. However, because <1% of the >500,000 L1 sequences are potentially active in the human genome, whether lead-induced L1 hypomethylation induces L1 retrotranscription, and whether such activation is responsible for lead toxic effects is undetermined. We propose to evaluate the role of L1 reactivation during pregnancy by measuring L1 mRNA in women selected from a large birth cohort in Mexico City. We will then explore the correlation between L1 methylation and L1 mRNA levels; the association of lead biomarkers with L1 encoded mRNAs; and possible mechanisms for demethylation.
Description: Prof. Racowsky and Dr. Machtinger conducted a study to evaluate whether exposure of immature human oocytes to BPA interferes with progression of the human oocyte through the normal stages of meiotic maturation. Immature oocytes (GV), otherwise discarded from patients undergoing IVF procedure, were used following patient signed informed consent for donation of their discarded oocytes for research. Previous documentation that BPA is present in human follicular fluid at a concentration of 1-2 ng/ml provided a guide for the work undertaken by Prof. Racowsky and Dr. Machtinger.
Two experiments were performed: The first experiment involved exposure of human oocytes to high concentrations of BPA (20µg/ml) in order to determine whether BPA could adversely effect meiotic progression. The maturation rates were compared to those of sibling oocytes incubated in a medium without BPA. This experiment included pairs of oocytes from 21 patients. We showed that oocytes exposed to 20µg/ml BPA underwent meiotic maturation (to metaphase II; MII) at a significantly lower rate compared with controls (77.8% vs. 16.7%, p=0.003). In the second experiment, lower concentrations of BPA (20ng/ml or 200 ng/ml) were tested. In cases of when there were only two GV oocytes available from a patient, one was incubated with 20ng/ml BPA and the other was incubated in control medium. In cases that three GV oocytes were available from the same patient, the third oocyte was incubated with higher concentration of BPA (200ng/ml).
This study is ongoing. Oocytes of 50 patients have been collected. In 20 of these cases, a third oocyte was available. Generalized estimating equations were used to combine the results from the two experiments to assess whether there is a dose response effect of BPA on oocyte progression of the human oocyte to MII. It was discovered that as BPA dose increased, there was a significant decrease in the likelihood of an oocyte reaching MII (p=0.003). We also showed that as BPA dose increased, there was a significant increase in the likelihood of an oocyte becoming activated (p=0.04).
Description: This project investigates the feasibility of predicting cholera outbreaks on a global scale using a variety of newly available earth observing datasets. Cholera affects between 3 and 5 million people each year, and has been re-emerging over the last decade as a significant public health problem. In the late 1970’s it was discovered that the causative agent of cholera, the bacterium Vibrio cholerae, naturally inhabits coastal and estuarine waters around the world in association with copepod zooplankton hosts (Colwell et al., 1977, Huq et al., 1983). Strong correlations between cholera outbreaks and variables such as rainfall anomalies and productivity (Chl a) have been demonstrated for particular regions over short periods of time (Lobitz et al., 2000, deMagny et al., 2008, Mendelsohn & Dawson, 2007). Although salinity is known to be important for Vibrio growth and survival (Lipp et al., 2002), no other research team has used global salinity data in their modeling efforts or attempted to conduct a global scale analysis. We are using 40-year assimilated meteorological data from the new-generation NASA GEOS-MERRA system to develop a physically based model of the distribution of Vibrio cholerae in the environment, validating the model using data on cholera outbreaks.
Pilot Grant Awardee: Weisskopf, M
Project Title: An Economic Approach to Linking Adoption and Use of Improved Cookstoves to Reduced Indoor Air Pollution and its Effect on Cognitive Function
Award Amount: $20,000
Description: Biofuels burned on traditional cookstoves produce high concentrations of indoor air pollutants that are toxic to health and the environment. Recent studies have shown that lung and cardiovascular dysfunction are linked to indoor air pollution (IAP), but it has yet to be explored whether IAP is associated with any cognitive outcomes, despite evidence that aspects of outdoor air pollution—at much lower concentrations—cause cognitive deficits. The health effects that have been documented thus far have been shown to be reduced when traditional cookstoves are replaced with improved cookstoves. Problematically, programs that distribute improved stoves have found low levels of adoption and use. This pilot study aims to address these issues by first exploring whether indoor carbon monoxide and PM2.5 are associated with cognitive performance in a rural population in China that uses traditional cookstoves. We will then use this group to conduct a feasibility study where we randomly distribute improved cookstoves to assess reductions in CO and PM2.5, and improvements in cognitive outcomes. We will also measure stove usage and qualitatively explore barriers to stove use and adoption through surveys and focus groups. All of the data gathered from this study will be used to apply for grants to fund a randomized controlled trial that looks at stove adoption and use, IAP exposure and cognitive effects.
Description: Mercury is a globally distributed toxin found in fish and shellfish, and there is ample evidence that exposure to methymercury via seafood consumption can have adverse health effects. The aim of this project is to test whether the stable isotopic content of mercury in human biomarkers (hair) can be used to infer the source of the seafood products leading to exposures. This will be tested in two populations, one from the Faroe Islands and a second from the Gulf of Mexico, where we have prior information on seafood consumption patterns and levels of mercury in hair biomarker samples. The isotopic composition of mercury in a given fish is the result of the sum total of fractionation processes occurring upstream. If we can further link the isotopic composition of hair to the consumed fish, we will open up the doors to future studies that can link mercury processing in the environment to human health outcomes. A primary goal of this pilot study is to bring together biomedical and non-biomedical researchers to look at this type of data and discuss how to use tools such as this to better link human health to the environment.
Description: Esophageal adenocarcinoma (EA) is a lethal cancer with a mortality rate greater than 85%, and for the past four decades its incidence has been increasing by approximately 500% in the United States1-6. Major risk factors for EA include gastroesophageal reflux disease (GERD), obesity, and smoking. Conversely, infection of Helicobacter pylori (HP) is associated with a 50% risk reduction of EA. Better understanding of the mechanisms by which HP protect against EA could lead to new prevention approaches for EA. Recent works suggest that HP may affect GERD and obesity through inducing atrophic gastritis (AG) and adipokine production. However, there is insufficient direct evidence for determinant roles of these mechanisms in HP-EA relationship. We propose to utilize an existing EA cohort to evaluate whether HP-induced AG and adipokine dysregulation mediate the HP-EA relationship. The results of this pilot study will be used as preliminary data to apply for a five-year RO1 grant to elucidate the protective mechanisms of HP infection on EA risk.
Description: As engineered nanoparticles (ENPs) swiftly proliferate and enter consumer products, humans increasingly become exposed. Our body’s first line of defense against ENPs is the epithelium. Cultured epithelial cells therefore provide a relevant and efficient model system for assessing health impacts. Most nanotoxicological assessments use epithelial cells that are healthy and intact, but when injuries arise cells might become more vulnerable to ENPs. Further, there is a particular lack of understanding on the effect of particulate matter on the epithelium of the eye, which is a primary site of environmental exposure and resulting irritation. To address this gap in knowledge, we hypothesize that ENPs impede the healing of a wounded epithelium. We are working on developing a novel functional wound healing assay to test this hypothesis on human corneal epithelial cells using a panel of industrially relevant ENPs, including metals and metal oxides generated in-house using the recently developed Harvard Versatile Engineered Nanomaterial Generation System (VENGES). Our work will help in developing a physiologically relevant framework for evaluating nanomaterial safety for epithelial wounds.
Pilot Grant Awardee: Gold, D
Project Title: Cardiopulmonary Response to Effective Particle Filtration with CPAP in Patients with Sleep- Disordered Breathing
Award Amount: $23,000
Description: Sleep-disordered breathing (SDB), the recurrent episodic disruption of normal breathing during sleep, affects as much as 17 percent of US adults. SDB and air pollution have each been linked to increased risk of autonomic dysfunction, pulmonary and systemic inflammation, elevated blood pressure, paroxysmal atrial fibrillation, ventricular arrhythmias, myocardial infarction, and cardiovascular mortality, but the influence of pollution on SDB is poorly understood. Many of the adverse cardiac effects of SDB are thought to be due in part to clinically significant apnea/hypopnea- induced hypoxemia and respiratory acidosis. However, there is only limited evidence that treatment of SDB with continuous positive airway pressure (CPAP) improves airway inflammation, blood pressure and other adverse cardiac outcomes. Although CPAP stabilizes airway patency and improves oxygenation, patients on CPAP are still exposed to pro-inflammatory allergens and air pollutants. We have demonstrated that current CPAP systems are not designed to efficiently filter either allergens or smaller respirable particles present in room air. We propose to pilot a randomized cross-over clinical trial of effective filtration of allergens and respirable particles during CPAP use, with the ultimate purpose to evaluate whether effective particle filtration improves airway inflammation, systemic inflammation, autonomic function, and blood pressure.
Pilot Grant Awardee: Korrick, S
Project Title: Conception with Assisted Reproductive Technology (ART), prenatal phthalate and bisphenol A (BPA) exposure and infant neurodevelopment
Award Amount: $15,500
Description: The long-term impact of Assisted Reproductive Technology (ART)-related exposure of gametes/embryos to physical and hormonal factors, including alterations in gonadal steroid exposures in early development, is largely unstudied. Early life exposure to phthalates and bisphenol A (BPA) has been associated with adverse development, especially that related to gonadal hormones. Prenatal exposure to gonadal steroids, in turn, is responsible for important aspects of brain development with impacts on cognition and behavior. The goal of this pilot is to assess the associations of ART, prenatal phthalate and BPA exposure, and (secondarily) their interaction, with infant neurocognitive development. Participants will be 30 infants (15 spontaneous, 15 ART conceptions) from a study of the relation of phthalates and BPA with infertility and pregnancy outcomes. Infants (age 12 months) will be assessed with standardized measures of cognition, behavior, and language. The relation of these tests with ART and phthalate/BPA exposure measures will be modeled. There is a notable paucity of information on the relation of phthalates, BPA and ART with childhood neurobehavioral development despite prevalent exposures and evidence suggestive of risk. The goal of this pilot is to begin to address this critical knowledge gap and to thereby develop the basis for a future more comprehensive study.
Pilot Grant Awardee: Tejera-Alvarez, P
Project Title: A Functional Study of Polymorphisms in the PI3 Gene, an inhibitor of Neutrophil Elastase in Inflammatory Lung Diseases
Award Amount: $22,650
Description: The inhalation of toxic gases (air pollution, cigarette smoke, noxious gases) as well as particles and microorganisms contribute to lung injury. Such insults exacerbate the pulmonary inflammatory response by increasing the concentration of pro-inflammatory mediators and target cells. Augmentation of neutrophils in the airways causes increased proteolytic enzymes, and results in the initiation and propagation of pulmonary damage. PI3 (pre-elafin) is a specific neutrophil elastase inhibitor. Our recent findings revealed that PI3 gene expression is down-regulated during the acute respiratory distress syndrome (ARDS) development. Recently, the polymorphism (SNP) rs2664581, located in exon 2 of pre-elafin, has been associated with increased ARDS risk and with PI3 circulating levels. However, because SNP rs2664581 is in high linkage disequilibrium (LD) with the remaining SNPs in the PI3 gene, it is difficult to distinguish between the casual SNP and those that are markers in LD with the functional SNP. This study is aimed at the functional characterization of the SNPs in PI3 gene. Our findings could contribute to a better understanding of the role PI3 as a neutrophil proteases inhibitor in the pathogenesis of inflammatory lung diseases. Morevover, the study brings together a multidisciplinary team, incorporating several experienced pulmonary researchers new to the field of environmental lung disease research.
Presentations of this work include: Tejera et al., 2010, Am. J. Respir. Crit. Care Med., 181: A6466. Tejera et al., 2011, Am. J. Respir. Crit. Care Med., 183: A5233
Pilot Grant Awardee: Wang, Z
Project Title: Application of Satellite Aerosol Remote Sensing Technology to Estimate the Health Impacts of Airborne Particles
Amount of Award: $25,000
Description: Numerous epidemiological studies have indicated that exposure to ambient-level fine particulate matter (PM2.5) is associated with increased morbidity and mortality from cardiopulmonary diseases and lung cancer. Exposure assessments are usually dependent on ground monitoring networks, which lack spatially complete coverage, especially in rural areas. With the rapid technological progress, satellite-based remote sensing provides a unique opportunity to monitor air quality from space at global, continental, national and regional scales. Studies have shown that satellite-derived aerosol optical depth (AOD) is correlated with ground-level PM2.5. Initial satellite-based environmental health studies revealed that the standardized county-level biennial mortality rates (2003-2004) of chronic heart diseases in the United States were associated with two-year average AOD raster data. Our preliminary study found that AOD is significantly associated with daily unscheduled hospital outpatient visits in Beijing, China for cardiovascular diseases, cerebrovascular diseases, and respiratory diseases, but not associated with cancer, injury, and skin diseases. We propose to expand our satellite-based study using newly acquired hospital admission data with much larger population coverage, which collected from all major hospitals in Beijing over a longer observational period (5 years, 2004 – 2008).
Pilot Grant Awardee: Deloid, G
Project Title: Novel screening assay for NLRP3 inflammasome activation by engineered nanoparticles
Award Amount: $10,000
Description: Engineered nanomaterials (ENMs) are widely and increasingly used in consumer products and industrial processes. It is therefore important to rigorously investigate potential effects of ENMs on health, including effects on the immune system. The multiprotein inflammasome complex is a central mediator of innate and adaptive immune responses. Inflammasomes have been implicated in the pathogenesis of a wide range of acute and chronic inflammatory disorders, including silicosis and asbestosis, and play a critical role in host defense against bacterial and viral pathogens, including Mycobacterium tuberculosis and Influenza.
Inflammasomes form in response to diverse pathogen and danger associated molecular patterns, and consist of NOD-like receptors (e.g. NLRP3), adaptor proteins (e.g. ASC), and inactive pro-caspase 1, which is cleaved within the inflammasome to active caspase 1, whose proteolytic activities mediate multiple inflammatory processes.
We will develop novel high-throughput fluorescence microscopy-based assays for detection of the NLRP3 inflammasome formation (association of NLRP3 and ASC, and ASC and caspase 1), and utilize these assays to assess the effects of a panel of ENMs on inflammasome activity in macrophages.
Pilot Grant Awardee: Godleski, J
Project Title: Exposure to Traffic Particles and DNA Methylation
Award Amount: $20,000
Description: The aim of this research is to define the role of epigenetic mechanisms specifically in vasoconstriction of blood vessels of the lung and heart after exposure to traffic-related ambient particles. Measuring the level of DNA methylation of three gene promoters involved in vascular processes (eNOS, NOTCH4 and endothelin-1) in rat heart and lung tissue will allow for comparison of PM exposed versus unexposed tissues. Differences in methylation could implicate changes in expression levels of these genes providing a mechanism by which particulate matter induces vasoconstriction. To date, sequencing has been conducted on a number of animal tissues in analyzing the eNOS promoter of rat heart and lung tissues and indications point to this being a very promising avenue of research. We hope to be able to expand this area of research on the NOTCH4 and endothelin-1 gene promoters.
Pilot Grant Awardee; McNeely, E
Project Title: Flight attendant exposures to flame retardants in airplanes
Award Amount: $20,000
Description: The evidence for flame retardants, such as polybrominated diphenyl ethers (PBDEs), to be chemicals toxicants has led to a ban on their manufacture in new products both in the U.S. and in the European Union (EU).Still, airplanes are a significant source of these compounds because of safety regulations that required their use. Recent studies have confirmed moderate levels of PBDE compounds in planes and one small study has noted higher passenger body burdens of these compounds after flight. Cabin crew continuously work in these environments, yet no study of their exposure has been done. This pilot proposes to evaluate exposure to these compounds after flight in a sample of flight attendants using two measures of exposure; serum levels of PBDEs and dust samples for PBDEs (via a newly developed hand-wipe method). This is the first study of these exposures in flight attendants, a workforce expected to grow to 106,000 workers by 2018. Aircraft exposures are particularly concerning for the majority of the U.S. flight crew who are female, many in their prime childbearing years, because new evidence shows PBDE exposures to be associated with decreases in thyroid hormones and that lower maternal circulating thyroid hormones have been associated with life-long adverse consequences on the nervous system, including cognitive function, in the offspring.
Pilot Grant Awardee: Mittleman, M
Project Title: Atrial fibrillation, ambient exposures and epigenetic mechanisms
Award Amount: $20,000
Project Description: Atrial fibrillation (AF) is a common arrhythmia responsible for a high mortality burden, but the etiology and risk factors are not well understood. Studies of familial aggregation have shown a complex pattern of inheritance, suggesting a potential role for polygenetic pathways, epigenetics and gene-environment interactions. However, these associations have not been explored. We propose to examine the relationship between DNA methylation in peripheral white cells and tissue from the right atrial appendage in a population of patients undergoing coronary artery bypass graft (CABG) surgery at Beth Israel Deaconess Medical Center. We will also look at the relationship between short and long term air pollution exposure and extent of methylation of the heart tissue and white cells. This pilot study will provide data that will allow for population-based case-control and cohort studies of the interaction between ambient air pollution exposure and DNA methylation on AF incidence outside of the post-CABG setting.
Atrial fibrillation (AF) is a common arrhythmia responsible for a high morbidity burden, but established risk factors account for only a portion of incident AF. Epigenetic mechanisms such as methylation of CpG islands in the DNA alter gene expression or phenotypic profile without modification to the DNA sequence and may influence atrial function, but this association has not been explored. Recent studies have demonstrated that environmental exposures to air pollution are associated with changes in the methylation of leukocytes. DNA-methylation can easily be assessed in peripheral white blood cells, but the relationship of leukocyte methylation to the methylation patterns in target tissue of the atrium is unknown.
We propose the following aims to be investigated in this pilot study of patients who undergo coronary artery bypass graft (CABG) surgery at Beth Israel Deaconess Medical Center. Subjects included in the study will be free of AF and other known atrial arrhythmias at the time of the procedure. We plan to collect preoperative clinical information and AF incidence in the postoperative period during the index hospitalization.
- To investigate the association between methylation in atrial target tissue and peripheral white cell methylation as measured by both global and genome-wide methylation analyses.
- To develop pilot data on the association between methylation at the site of the right atrial appendage and short and long-term exposures to ambient air pollutants.
If we find that peripheral leukocytes provide relevant information about methylation in atrial tissue, this will allow for population-based case-control and cohort studies of the role of air pollution on epigenetic changes leading to atrial fibrillation outside of the post-CABG setting.