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Inflammation as a Co-Stressor
Jennifer Kay PhDEngelward Laboratory
MIT Superfund Research ProgramNortheast Superfund Meeting
4419
Inflammation and Cancer
Pancreatitis
Pancreatic Cancer
Liver Cancer
Colon Cancer
Gastric cancer
Lung cancer
Kidney Cancer
Inflammation Cancer
Crohnrsquos disease
Ulcerative Colitis
Hepatitis
Cirrhosis
Nephritis
H pylori infection
COPD
Inflammation and Superfund Chemicals
PFAS
Inflammation as a Co-Stressor
bull Inflammation is a strong promoter of cancer
bull Many chemical exposures cause inflammation
bull Inflammation is also modulated by
ndash Diet
ndash Behaviors
ndash Disease
bull As long as inflammation is happening there is
a greater risk for compounded consequences
of carcinogenic exposures
Multistep Model of Cancer
5
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Mutations
Inflammation
Inflammation Causes DNA Damage
Adapted from Dedon and Tannenbaum Arch Biochem Biophys 2004 6
NO
O2
ONOO-
ONOOCO2-
CO2
H2O2
N2O3O2
OH
NO2
CO3Macrophage
H2O2 + Cl-
NO2-
HOCl
NO2
Myeloperoxidase
H2O2 + Br- HOBr
Eosinophil
Eosinophil
peroxidase
Neutrophil
8oxoG
DNA Damage Causes Point Mutations
1280771279958oxoG C
8oxoG 8oxoG A
128078127995
G C
Mutation
Correct base pairing
DNA Damage Impedes DNA
Replication
8
Homologous Recombination(DNA Repair Pathway)
Double strand break
Replication Fork Breakdown
DNA replication
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Inflammation and Cancer
Pancreatitis
Pancreatic Cancer
Liver Cancer
Colon Cancer
Gastric cancer
Lung cancer
Kidney Cancer
Inflammation Cancer
Crohnrsquos disease
Ulcerative Colitis
Hepatitis
Cirrhosis
Nephritis
H pylori infection
COPD
Inflammation and Superfund Chemicals
PFAS
Inflammation as a Co-Stressor
bull Inflammation is a strong promoter of cancer
bull Many chemical exposures cause inflammation
bull Inflammation is also modulated by
ndash Diet
ndash Behaviors
ndash Disease
bull As long as inflammation is happening there is
a greater risk for compounded consequences
of carcinogenic exposures
Multistep Model of Cancer
5
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Mutations
Inflammation
Inflammation Causes DNA Damage
Adapted from Dedon and Tannenbaum Arch Biochem Biophys 2004 6
NO
O2
ONOO-
ONOOCO2-
CO2
H2O2
N2O3O2
OH
NO2
CO3Macrophage
H2O2 + Cl-
NO2-
HOCl
NO2
Myeloperoxidase
H2O2 + Br- HOBr
Eosinophil
Eosinophil
peroxidase
Neutrophil
8oxoG
DNA Damage Causes Point Mutations
1280771279958oxoG C
8oxoG 8oxoG A
128078127995
G C
Mutation
Correct base pairing
DNA Damage Impedes DNA
Replication
8
Homologous Recombination(DNA Repair Pathway)
Double strand break
Replication Fork Breakdown
DNA replication
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Inflammation and Superfund Chemicals
PFAS
Inflammation as a Co-Stressor
bull Inflammation is a strong promoter of cancer
bull Many chemical exposures cause inflammation
bull Inflammation is also modulated by
ndash Diet
ndash Behaviors
ndash Disease
bull As long as inflammation is happening there is
a greater risk for compounded consequences
of carcinogenic exposures
Multistep Model of Cancer
5
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Mutations
Inflammation
Inflammation Causes DNA Damage
Adapted from Dedon and Tannenbaum Arch Biochem Biophys 2004 6
NO
O2
ONOO-
ONOOCO2-
CO2
H2O2
N2O3O2
OH
NO2
CO3Macrophage
H2O2 + Cl-
NO2-
HOCl
NO2
Myeloperoxidase
H2O2 + Br- HOBr
Eosinophil
Eosinophil
peroxidase
Neutrophil
8oxoG
DNA Damage Causes Point Mutations
1280771279958oxoG C
8oxoG 8oxoG A
128078127995
G C
Mutation
Correct base pairing
DNA Damage Impedes DNA
Replication
8
Homologous Recombination(DNA Repair Pathway)
Double strand break
Replication Fork Breakdown
DNA replication
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Inflammation as a Co-Stressor
bull Inflammation is a strong promoter of cancer
bull Many chemical exposures cause inflammation
bull Inflammation is also modulated by
ndash Diet
ndash Behaviors
ndash Disease
bull As long as inflammation is happening there is
a greater risk for compounded consequences
of carcinogenic exposures
Multistep Model of Cancer
5
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Mutations
Inflammation
Inflammation Causes DNA Damage
Adapted from Dedon and Tannenbaum Arch Biochem Biophys 2004 6
NO
O2
ONOO-
ONOOCO2-
CO2
H2O2
N2O3O2
OH
NO2
CO3Macrophage
H2O2 + Cl-
NO2-
HOCl
NO2
Myeloperoxidase
H2O2 + Br- HOBr
Eosinophil
Eosinophil
peroxidase
Neutrophil
8oxoG
DNA Damage Causes Point Mutations
1280771279958oxoG C
8oxoG 8oxoG A
128078127995
G C
Mutation
Correct base pairing
DNA Damage Impedes DNA
Replication
8
Homologous Recombination(DNA Repair Pathway)
Double strand break
Replication Fork Breakdown
DNA replication
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Multistep Model of Cancer
5
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Mutations
Inflammation
Inflammation Causes DNA Damage
Adapted from Dedon and Tannenbaum Arch Biochem Biophys 2004 6
NO
O2
ONOO-
ONOOCO2-
CO2
H2O2
N2O3O2
OH
NO2
CO3Macrophage
H2O2 + Cl-
NO2-
HOCl
NO2
Myeloperoxidase
H2O2 + Br- HOBr
Eosinophil
Eosinophil
peroxidase
Neutrophil
8oxoG
DNA Damage Causes Point Mutations
1280771279958oxoG C
8oxoG 8oxoG A
128078127995
G C
Mutation
Correct base pairing
DNA Damage Impedes DNA
Replication
8
Homologous Recombination(DNA Repair Pathway)
Double strand break
Replication Fork Breakdown
DNA replication
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Inflammation Causes DNA Damage
Adapted from Dedon and Tannenbaum Arch Biochem Biophys 2004 6
NO
O2
ONOO-
ONOOCO2-
CO2
H2O2
N2O3O2
OH
NO2
CO3Macrophage
H2O2 + Cl-
NO2-
HOCl
NO2
Myeloperoxidase
H2O2 + Br- HOBr
Eosinophil
Eosinophil
peroxidase
Neutrophil
8oxoG
DNA Damage Causes Point Mutations
1280771279958oxoG C
8oxoG 8oxoG A
128078127995
G C
Mutation
Correct base pairing
DNA Damage Impedes DNA
Replication
8
Homologous Recombination(DNA Repair Pathway)
Double strand break
Replication Fork Breakdown
DNA replication
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
DNA Damage Causes Point Mutations
1280771279958oxoG C
8oxoG 8oxoG A
128078127995
G C
Mutation
Correct base pairing
DNA Damage Impedes DNA
Replication
8
Homologous Recombination(DNA Repair Pathway)
Double strand break
Replication Fork Breakdown
DNA replication
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
DNA Damage Impedes DNA
Replication
8
Homologous Recombination(DNA Repair Pathway)
Double strand break
Replication Fork Breakdown
DNA replication
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
DNA Repair of Inflammation-Induced DamageBase Excision Repair
Base lesion
Replication Fork Breakdown
Direct ReversalBase lesion
εA3rsquoOH
εA
9
A
Homologous RecombinationDouble strand break
Glycosylase
AP lyase APE1
Polymerase
Ligase
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
10
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Inflammation and Cancer
11
Inflammation Cancer
Proliferation
Cytokines chemokines growth
factors
Survival
Migration
Cytokines chemokines growth
factors
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Inflammation Causes Proliferation
Orsolya Kiraly et al PLoS Genetics 2015
Every time a cell divides it has the opportunity to acquire new mutations
Ki67 A protein expressed during cell division
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Inflammation
Mutations
Proliferation
Multistep Model of Cancer
13
DNA Damage
3 n
Cell division and additional mutationsNormal cells
1 2
Initiating mutation
Cancer
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
RaDR Mice A Tool to Measure Mutations In Situ
Fluorescent spot=
Mutated cell(s)
RaDR Mouse Liver
Replication fork restoration
Sukup-Jackson et al PLoS Genetics 2014
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
DNA Damage During Proliferation Synergistically Increases Mutations
MNU = DNA damaging agentInf = InflammationProlif = Cellular proliferation following inflammation
Bright spot =
Mutant cell(s)
Orsolya Kiraly PLoS Genetics 2015
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
Chemical Exposures Damage DNA
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Chemical Exposures Damage DNA
bull Many hazardous chemicals are metabolized to form DNA-reactive speciesndash N-nitrosamines (eg NDMA)
ndash Polycyclic aromatic hydrocarbons (PAHs eg Benzo[a]pyrene)
ndash Polychlorinated bisphenyls (PCBs)
ndash Many more PCB Adducts
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
NDMA Contaminates Water Near the Olin Chemical Superfund Site in Wilmington MA
National Polychemicals Inc 1967
Alarming rates of child brain cancer Hodgkins disease non-Hodgkins
lymphoma and leukemia
Ground Water
Soil
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
NDMA-Induced DNA Lesions Promote Mutations
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
DNA Repair of NDMA-Induced DamageBase Excision Repair
Base lesionDouble strand break
Homologous Recombination
Direct ReversalBase lesion
O6meG
3rsquoOH
3meA
G
Aag
Replication Fork Breakdown
20
AagTg animals overexpress Aag
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
Pan-nuclear γH2AX = Hyperactive signaling
suggests overwhelming DNA damage
γH2AX Foci = Localized at DNA DSBs
Cell is ldquoγH2AX+rdquo if foci ge 5
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
NDMA Induces DNA DSBs
Wild Type
AagTg
Saline NDMA
With Joshua Corrigan
γH2AX staining at DNA double strand breaks24 hours after NDMA
exposure
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
NDMA Causes Proliferation
With Joshua Corrigan
AagTg
Saline NDMA
Ki67 A protein expressed during cell division
Wild Type
Ki67 shows increased cell division 24 hours after
NDMA exposure
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
NDMA Causes Inflammatory Tissue Damage
Saline NDMA
AagTg
WT
Liver 20xWith Sebastian Carrasco
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Cancer
25
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Acknowledgments
P42 ES027707
Prof Bevin Engelward
Joshua Corrigan
Christy Chao
Dr Norah Owiti
Dr Simran Kaushal
Yuge Ji
Aimee Moise
Dr Orsolya Kiraly
Dr Michelle Sukup-Jackson
Sebastian Carrasco
T32-ES007020
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more
Cancer
27
Reactive Oxygen and
Nitrogen Species
DNA Damage Response
Inflammation DNA Damage
DNA repair
Inflammation DNA Damage and Cancer
BER (Aag--)
BER (AagTg)
DR(Mgmt--)
NDMA ampSuperfund Chemicals
NDMA PCBs PAHs more