Biomarkers of Exposure to Hazardous Substances (2017-2022) The UC - - PowerPoint PPT Presentation

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 Tingrui Pan

• 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

P E R K

sEH inhibition and EpFA block Endoplasmic Reticulum Stress (ER Stress)

ROS

ATF6 P P

Glucose

Nucleus A2 Phospholipase IRE1 Arachidonic Acid α P CYP450 P

O

XBP1s

O H

ERAD

O

14,15 EET sEH sEH inhibitor ATF6(N) P

O

elF2α

O

CHOP

H

ATF4

H O

14,15 DHET Wagner et al. 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 TOOTH

EpFA: EETS EEQS EDPS

DECAY

THE MITOCHONDRIAL ROS ER STRESS AXIS

MPTP NEUROPATHIC

TRICLOSAN PAIN

PARKINSON’S

PARAQUAT INDOMETHACIN CARBONTET NITROPHENOLS DICLOFENAC

IBD FIBROSIS DIABETES

HEART FAILURE

INFLAMMATION

Pr Project

• ject 4 - Monitoring

Mitochondrial Oxidative Stress and Cardiac Toxicity Caused by Chronic Exposure to Chemicals

• Dr. Nipavan Chiamvimonvat, Project

Leader

• Dr. Aldrin Gomes, Co-Leader

Ov Over erall all aims 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

• xidative 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.

Control Celecoxib (30uM) CCL4 (50uM) Celecoxib (30uM) H2O2 (200uM) + CCL4 (50uM)

Effect of xenobiotics on Cardiac Cell Viability

Relative Cell Viability (%)

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

Complex I Activity (Abs 340nM)

Reducing mitochondrial electron transport chain activity increases ROS and reduces cell viability

Ibuprofen treated mice

0.35

ROS

0.34

Complex I Activity (Abs 340nM)

Female Heart 8D

**

0.33 0.32 0.31 0.30 0.29 0.28 0.27

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 0.29

ROT – Rotenone (a Complex I activity inhibitor)

0.28

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

Con Concl clusion

• ns

s

• CCl4 naphthalene, paraquat

induces cardiac toxicity, mitochondrial stress and proteasome dysfunction.

• Mitochondrial-stress is induced by other

xenobiotics: diclofenac, ibuprofen, naproxen.

Fu Futu ture Dir Direction ections s

• Expand target

analysis (Pesticides, HaHs/PaHs, Commercial Chemicals and Pharmaceuticals).

• Determine cell, blood and urinary biomarkers of

mitochondrial dysfunction to monitor Xenobiotic exposure and possibly drive therapeutic 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

Recreating disease in a dish hiPSCs and hiPSC-CMs

SSEA4 DAPI Merged Background Background Fluorescence Oct3/4 Specific Fluorescence SSEA-4 Specific

103

<1%

102 102

99% SSEA+ hiPSCs

Oct3/4+ hiPSCs

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%

103 103 101 101 101

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 mV 1s 20 mV 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