Biomarkers of Exposure to Hazardous Substances (2017-2022)
The UC Davis Superfund Research Center conducts research to: a) Improve understanding of the mechanisms by which
hazardous chemicals produce adverse health affects, b) Develop, validate and integrate novel methods to evaluate
chemical exposures, levels of contamination, and health risks, and
c) Develops innovative remediation strategies to reduce hazardous substance exposure and toxicity.
These activities improve the ability of the National Superfund Program to address legacy and emerging contaminants and associated transformation products to more comprehensively protect the U.S. population from health risks posed by hazardous substances.
Biomarkers of Exposure to Hazardous Substances
Project PI 1. Optimizing Bioremediation Tom Young, Frank Loge
2. Nanosensing Platforms TingruiPan 3. Immunochemical BioMarkers Natalia Vasylieva
4. Cardiac Toxicity Nipavan Chiamvimonvat 5. Endoplasmic Reticulum Stress Fawaz Haj, Christophe Morisseau
Core C: Community Engagement - Dr. Beth Rose Middleton, PI
In response to intensive forestry management and illegal marijuana groves, collaborative research with the Yurok Tribe Environmental Program (YTEP) will:
• Conduct environmental sampling to identify contaminants and their concentrations
• Implement field deployable assays for use by YTEP partners • Collaboratively identify culturally and ecologically
appropriate remediation strategies
Community Engagement Core - Dr. Beth Rose Middleton, PI
The Community Engagement Core works to develop meaningful bi-directional communication strategies between university and tribal researchers and community partners to apply UCD Center research to address community concerns.
Broadly, the chemical detection technologies, remediation strategies and training opportunities aim to provide communities with autonomous methods for addressing environmental health problems within their community while training scientists on developing equitable, respectful, and responsible projects with community partners.
Core A: Analytical Chemistry Core - Dr. Jun Yang, PI
Develop analytical methods to detect hazardous chemicals for the variety of UCD-SRP projects.
Validate alternative analytical methods such as: • Immunoassays • Cell-based assays
PERK
sEH inhibition and EpFA block Endoplasmic Reticulum Stress (ER Stress)
ROS
ATF6
PP
Glucose
Nucleus
A2 Phospholipase
IRE1 Arachidonic Acid α P CYP450 P
OXBP1s O
H ERAD O 14,15 EET
sEH sEH inhibitor ATF6(N) P OelF2α
O
CHOP HATF4 H O 14,15 DHET Wagner etal.2017
O H
Project 5: Monitoring Endoplasmic Reticulum Stress Caused by Chronic Exposure to Chemicals, Dr. Fawaz Haj and Dr. Christophe Morisseau
Investigate new mechanistic insights into the effects of chronic exposure of Superfund (SF) chemicals on endoplasmic reticulum (ER) stress.
Effects of SF chemicals on ER stress by • Altering gene expression • Inhibition • Competition for catalysis • Increasing reactive oxygen species • BLOOD AND URINARY BIOMARKERS OF DISRUPTION OF
THE ER STRESS PATHWAY TO MONITOR XENOBIOTIC EXPOSURE AND POSSIBLY DRIVE THERAPEUTIC INTERVENTION.
Project 4: Critical Role of Mitochondrial Oxidative Stress (MOS) in Chemical Induced Cardiac Toxicity, Dr. Aldrin Gomes (mitochondria) and Dr. Nipavan Chiamvimonvat (heart)
Investigate molecular mechanisms of chronic exposure to Superfund chemicals on mitochondrial oxidative stress (MOS) and proteasome dysfunction
Target Analytes: • Pesticides • Antimicrobials • HaHs/PaHs • Commercial Chemicals • Pharmaceuticals • CELL, BLOOD AND URINARY BIOMARKERS OF
DISRUPTION OF MITOCHONDRIA TO MONITOR XENOBIOTIC EXPOSURE AND POSSIBLY DRIVE THERAPEUTIC INTERVENTION.
DEVELOP BIOMARKERS TO DETECT FUNDAMENTAL PROCESSES OF TOXICITY
DEPRESSION CANCER
TOOTHEpFA: EETS EEQS EDPS DECAY
THE MITOCHONDRIAL ROS ER STRESS AXIS
MPTP NEUROPATHIC TRICLOSAN PAIN
PARKINSON’SPARAQUAT INDOMETHACIN
CARBONTET
NITROPHENOLS DICLOFENAC
IBD FIBROSIS
DIABETES
HEART FAILURE INFLAMMATION
Project 4 - Monitoring Mitochondrial Oxidative Stress and Cardiac Toxicity Caused by Chronic Exposure to Chemicals
Dr. Nipavan Chiamvimonvat, Project Leader
Dr. Aldrin Gomes, Co-Leader
Overall aims Hypothesis: chronic exposure to xenobiotics and/or non-
steroidal anti-inflammatory drugs (NSAIDs) leads to mitochondrial oxidative stress (MOS) that results in proteasome dysfunction, apoptosis, tissue fibrosis and cardiac toxicity.
Focus: Heart health related diseases. Approach: used cell based assay and in vivo models to
test effect of exposure to SF chemicals and/or NSAIDs on mitochondrial stress, proteasome dysfunction, apoptosis, fibrosis and associated alterations of cell, plasma and urine profile as a biomarker.
Deliverable: Easier methods to monitor mitochondrial oxidative stress as a marker of xenobiotic exposure.
The MIT-ROS-ER stress axis
Effect of xenobiotics on cell viability, Reactive Oxygen Species (ROS) production, and mitochondrial
membrane permeability (MMP)
Cell viability in H9c2 cardiac cells incubated with 50µM CCl4, 100µM paraquat, 20µM naphthalene, 10 µM diclofenac (DIC) for 24 h. Pre-treatment with 20µM mito-Tempol (MT) prevented reduced cell vitality caused by CCl4. H202, 200 µM.
Con
trol
Cel
ecox
ib (3
0uM
) C
CL4
(50u
M)
Cel
ecox
ib (3
0uM
)
H2O
2 (2
00uM
)
+ C
CL4
(50u
M)
Effect of xenobiotics on Cardiac Cell Viability
Rel
ativ
e C
ell V
iabi
lity
(%)
0
20
40
60
80
100
120
**
**
**
**
β1 β2 β5
Xenobiotic exposure affects mitochondrial electron chain transport activity and proteasome activity
Mitochondrial complex l activity is decreased by naphthaline (20 µM) and paraquat (100 µM) but not CCl4 (20 µM) or DIC (20 µM). Lower figures show proteasome dysfunction occurs in hearts of ibuprofen treated mice
Com
plex
I A
ctiv
ity (A
bs 3
40nM
)
Reducing mitochondrial electron transport chain activity increases ROS and reduces cell viability
Ibuprofen treated mice 0.35
ROS 0.34
Com
plex
I A
ctiv
ity (A
bs 3
40nM
)
Female Heart 8D
** 0.33
0.32
0.31
0.30
0.29
0.28
0.27 0 20 40 60 80 100 120 140 160 180 200
Control Rotenone IB
0.36
0.35
0.34
0.33
0.32
0.31
0.30
0.29ROT – Rotenone (a Complex I activity inhibitor) 0.28
0 20 40 60 80 100 120 140 160 180
Male Heart 8D
Controkl Rotenone IB
creased
NSAIDs
Current Model
Cardiomyocytes dysfunction; Complex I and III inhibited
Δψ de
Oxidized proteins
UPS dysfunction
ROS
Mitochondrial
Cell Death
ER stress CARDIOTOXICITY
Proteasome Antioxidants Transfection
SF Chemicals
Conclusions • CCl4 naphthalene, paraquat induces cardiac toxicity, mitochondrial stress and proteasome dysfunction.
• Mitochondrial-stress is induced by other xenobiotics: diclofenac, ibuprofen, naproxen.
Future Directions • Expand target analysis (Pesticides, HaHs/PaHs, Commercial Chemicals and Pharmaceuticals).
• Determine cell, blood and urinary biomarkers of mitochondrial dysfunction to monitorXenobiotic exposureand possibly drivetherapeutic intervention.
Acknowledgements • Bruce Hammock
• Natalia Vasylieva (project 3) • Fawaz G. Haj (Project 5) • Christophe Morisseau (Project 5) • Jun Yang (core A) • Daniel Tancredi (core B)
Funding from: NIEHS/Superfund Research Program P42 ES004699
Project 4 - Monitoring Mitochondrial Oxidative Stress and Cardiac Toxicity
Caused by Chronic Exposure to Chemicals
Nipavan Chiamvimonvat, MD Division of Cardiovascular Medicine
Mortality rate of cardiovascular disease surpasses that of cancer
Cardiovascular Disease
Cancer
Circ 131(4): e29-322, 2015
Cardiac fibroblasts
• Cardiac fibroblasts account for ~75% of all cardiac cells, but contribute only ∼10-15% of total cardiac cell volume.
• The principal sources of extracellular matrix (ECM) proteins.
• A heterogeneous population. • Derived from various distinct tissue niches including
resident fibroblasts, endothelial cells, and bone marrow sources.
Roles of cardiac fibroblasts
Yue et al, Cardiovascular Res 2011
Molecular mechanisms leading to cardiac fibrosis
Wakili et al, 2011
Flow cytometric analysis of the isolated cells from mouse hearts
0
Recreating disease in a dish hiPSCs and hiPSC-CMs
SSEA4 DAPI Merged
Background Background Fluorescence Oct3/4 Specific Fluorescence SSEA-4 Specific
103 <1% 102102
99%
SS
EA
+ hi
PS
Cs
Oct
3/4+
hiP
SC
s
102
101
103
100 101 102 103 104 100 101 102 103 104 100 101 102 103 104 100 101 102 103 104
<1% 102
99%103
103
101101 101
0 0 0
Side Scatter Side Scatter
hiPSC-CMs Phalloidin
Troponin T Merged
DAPI
Activation of MAPK in hiPSC-CMs and hiPSC-fibroblasts by TNF-α
Novel Cell-in-Gel Platform
Novel 3D Cell-in-Gel
Spontaneous APs Single APs
20 m
V
1s
20 m
V
250 ms
Effects of mechanical stress on Ca2+ handling
Conclusions
• Generation of reliable platform for testing the effects of Superfund chemicals on cardiac myocytes and fibroblasts.
• Development of bioassays to test the effects of exposure.
Acknowledgements • Bruce Hammock • Aldrin Gomes (Project 4) • Fawaz G. Haj (Project 5) • Christophe Morisseau (Project 5) • Jun Yang (core A) • Ye Chen-Izu • Padmini Sirish
Funding from: NIEHS/Superfund Research Program P42 ES004699