Mélanie Leroux Laboratory of Molecular and Cellular Responses to Xenobiotics Supervisor: Dr. Karine Andréau 2 nd European Doctoral College on Environment.

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Presentation transcript:

Mélanie Leroux Laboratory of Molecular and Cellular Responses to Xenobiotics Supervisor: Dr. Karine Andréau 2 nd European Doctoral College on Environment and Health

Composition (PM 2.5 ) : Organic compounds Biological compounds Metals Carbon Core Coarse particles (PM 10 ) ,5µm Coarse particles (PM 10 ) ,5µm Fine particles (PM 2.5 ) 2,5 - 0,1µm Fine particles (PM 2.5 ) 2,5 - 0,1µm Ultrafine particles (PM 0.1 ) ≤ 0,1µm Ultrafine particles (PM 0.1 ) ≤ 0,1µm Health effects of atmospheric particles

Composition (PM 2.5 ) : Organic compounds Biological compounds Metals Carbon Core Coarse particles (PM 10 ) ,5µm Coarse particles (PM 10 ) ,5µm Fine particles (PM 2.5 ) 2,5 - 0,1µm Fine particles (PM 2.5 ) 2,5 - 0,1µm Ultrafine particles (PM 0.1 ) ≤ 0,1µm Ultrafine particles (PM 0.1 ) ≤ 0,1µm o ↗ Cardiorespiratory morbidity and mortality o Oxidative stress o Genotoxicity o Inflammatory response o Modulation of apoptosis o Lung cancers o ↗ Chronic respiratory diseases : asthma, COPD (Chronic obstructive pulmonary disease)… o Accumulation of PM in the bronchial wall and airway remodeling Health effects of atmospheric particles

1  [PM] ≥ 100µg/cm² only found in high pollution periods 2  Cancer cells = resistance to cell death Atmospheric particles Lung cancer Epidemiologic studies Previous in vitro studies  PM 2.5 induce cell death (apoptosis)

Human Bronchial Epithelial cells (16HBE) 1- INDUCTION of APOPTOSIS ? Urban PM 2.5 (10 µg/cm²) 2- REDUCTION of APOPTOSIS ? Aim of our study 10µg/cm²= 5 days exposure to atmospheric pollution (79µg/m 3 ) 3- PM 2.5 COMPOUNDS INVOLVED? 24H

Particles Urban PM 2.5 Location of sampling Main sources identified Traffic Paris ring road (2003) Car traffic Background Parisian suburb (2003) Urban background pollution Wood burning Paris (Winter 2009) Wood-fired heating Mix Ouagadougou (Burkina-Faso 2009) Wood-fired cooking, Moped traffic

Methology: apoptosis measurement by flow cytometry Urban PM 2.5 particles are they toxic in human bronchial epithelial cells ? 1.  Membrane permeabilization  Membrane bleebing  MMP  Condensation of the chromatin  Fragmentation of DNA and nucleus  Modification of the cellular granularity  Reduction of the cell size Apoptosis  Oxydative stress

Methology: apoptosis measurement by flow cytometry Urban PM 2.5 particles are they toxic in human bronchial epithelial cells ? 1.  Membrane permeabilization  Membrane bleebing  MMP  Condensation of the chromatin  Fragmentation of DNA and nucleus  Modification of the cellular granularity  Reduction of the cell size Apoptosis  Oxydative stress Propidium Iodide (PI) Hydro Etydium (HE) DiOC6(3)

1.Urban PM 2.5 particles are they toxic in human bronchial epithelial cells ? Questions Human Bronchial Epithelial cells (16HBE) INDUCTION INDUCTION of APOPTOSIS ? Urban PM 2.5 (10 µg/cm²) 24 H

Urban PM 2.5 particles are they toxic in human bronchial epithelial cells ? 1. * p< vs Control n=3 Traffic Wood Burning Mix Control (Medium) Background 10 µg/cm²  Urban PM 2.5 do not induce apoptosis in 16HBE cells

1.Urban PM 2.5 particles are they toxic in human bronchial epithelial cells (16HBE) ? Questions Urban PM 2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells Urban PM 2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells

1.Urban PM 2.5 particles are they toxic in human bronchial epithelial cells (16HBE)? Questions Urban PM2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells Urban PM2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells 2.Urban PM 2.5 particles have they an antiapoptotic effect in 16HBE cells ? INITIATION PROMOTIONAPOPTOSISRESISTANCE DNA adducts Cancer cell Tumor formation : Initiated cell

Urban PM 2.5 particles have they an antiapoptotic effect in 16HBE cells ? 2. Apoptosis inducer: A23 (Calcium ionophore) ↗ CALCIUM Methology: PM 2.5 exposure before induction of apoptosis 20 H 16HBE Particles (10µg/cm²) 4 H v v v v v v v v v v v v v v v v v v v v v v v v Flow Cytometry Apoptosis induction by A23

Urban PM 2.5 particles have they an antiapoptotic effect in 16HBE cells ? 2.  Traffic PM2.5 have an antiapoptotic effect towards A23-induced apoptosis p< vs Control n=3 * Traffic 10µg/cm² Background 10µg/cm² Wood burning 10µg/cm² Mix 10µg/cm² + A23 Control (Medium) * A23-induced apoptosis (%)

1.Urban PM 2.5 particles are they toxic in human bronchial epithelial cells (16HBE) ? Questions Urban PM 2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells Urban PM 2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells 2.Urban PM 2.5 particles have they an antiapoptotic effect in 16HBE cells ? YES : Only Traffic PM 2.5

1.Urban PM 2.5 particles are they toxic in human bronchial epithelial cells (16HBE) ? Questions Urban PM 2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells Urban PM 2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells 2.Urban PM 2.5 particles have they an antiapoptotic effect in 16HBE cells ? YES : Only Traffic PM 2.5  Possible link with calcium 3.Which Traffic PM 2.5 components are involved in the antiapoptotic effect ?

↗ CALCIUM Organic compounds (PAH) Water-soluble Components (metals) Composition (PM 2.5 ) : Organic compounds Biological compounds Metals Carbon Core Which Traffic PM 2.5 components are involved in the antiapoptotic effect ? 3. Apoptosis inducer: A23 (Calcium ionophore) desorbtion

Control Tr affic Org. Ex. Aq. Ex. Washed * BC + A23 p<0.001 vs control n=3 Ferecatu et al. Particle and Fibre Toxicology 2010,7:18  Organic and Aqueous components contribute to the antiapoptotic effect of Traffic PM2.5 Which Traffic PM 2.5 components are involved in the antiapoptotic effect ? 3. * * * * A23-induced apoptosis (%) Composition (PM 2.5 ) : Organic compounds Biological compounds Metals Carbon Core

Ferecatu et al. Particle and Fibre Toxicology 2010,7:18 Which Traffic PM 2.5 components are involved in the antiapoptotic effect ? 3. a) Organic compounds : PAH ↗ CALCIUM Organic compounds PAH Apoptosis inducer: A23 (Calcium ionophore)

Ferecatu et al. Particle and Fibre Toxicology 2010,7:18  Heavy PAHs partially contribute to the antiapoptotic effect of Traffic PM 2.5 via the AhR Which Traffic PM 2.5 components are involved in the antiapoptotic effect ? 3. a) Organic compounds : PAH * * * * p< 0.05 vs Medium n=3 A23-induced apoptosis (%) Medium

Which Traffic PM 2.5 components are involved in the antiapoptotic effect ? 3. b) Aqueous compounds : Metals Metals (mg/g) : FerAluminiumCuivre Antiapoptotic effect Traffic 13,23,4 0,86YES Background 21,6 20,4 0,40 NO

Which Traffic PM 2.5 components are involved in the antiapoptotic effect ? 3. b) Aqueous compounds : Metals p< 0.05 vs Control n=6 * * * * * * Control (Medium) PM2.5 Fe2+ Fe3+ Al3+Cu2+ qs 10 µg/cm² % apoptosis induction (DiOC6(3) low HE high) + A23 Metals (mg/g) : FerAluminiumCuivre Antiapoptotic effect Traffic 13,23,4 0,86YES Background 21,6 20,4 0,40 NO

Which Traffic PM 2.5 components are involved in the antiapoptotic effect ? 3. b) Aqueous compounds : Metals p< 0.05 vs Control n=6 * * * * * * Control (Medium) PM2.5 Fe2+ Fe3+ Al3+Cu2+ qs 10 µg/cm² % apoptosis induction (DiOC6(3) low HE high) + A23  The content of transition metals in particles is involved in the antiapoptotic effect

1.Urban PM 2.5 particles are they toxic in human bronchial epithelial cells (16HBE) ? Urban PM2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells Urban PM2.5 (10 µg/cm²) do not induce apoptosis of human bronchial epithelial cells 2.Urban PM 2.5 particles have they an antiapoptotic effect in 16HBE cells ? YES : Only Traffic PM 2.5  Possible link with calcium 3.Which Traffic PM 2.5 components are involved in the antiapoptotic effect ?   Heavy PAHs via AhR   Metals (Fe, Al, Cu)   Heavy PAHs via AhR   Metals (Fe, Al, Cu)

1.Role of oxydative stress triggered by metals ? 2.Effect of metals on intracellular calcium ? 3.Role of mitochondria ? 4.Role of Caspases and Calpains ? Ferecatu et al. Particle and Fibre Toxicology 2010,7:18

Nicole Boggetto Ioana Ferecatu Aïda Bouharrour Emile Petit Armelle Baeza Karine Andreau Francelyne Marano Jean-Marie Dupret Ioana Ferecatu Aïda Bouharrour Emile Petit Armelle Baeza Karine Andreau Francelyne Marano Jean-Marie Dupret

Ioana Ferecatu Aïda Bouharrour Emile Petit Armelle Baeza Karine Andreau Francelyne Marano Jean-Marie Dupret Ioana Ferecatu Aïda Bouharrour Emile Petit Armelle Baeza Karine Andreau Francelyne Marano Jean-Marie Dupret Nicole Boggetto

BbF : Benzo[b]fluoranthrene iP : Indeno[1,2,3-cd]pyrene BghiP : Benzo[g,h,i]perylene DBahA : Dibenzo[a,h]anthracene BaP: Benzo[a]pyrene