mechanisms of thyroid toxicity kevin m. crofton neurotoxicology division national health and...

Mechanisms of Thyroid Toxicity

Kevin M. CroftonNeurotoxicology DivisionNational Health and Environmental Effects LaboratoryUS Environmental Protection Agency Research Triangle Park NC

Outline

1. Introduction to thyroid hormones2. Effects of perchlorate3. Effects of the disruption of thyroid hormones

Thyroid hyperplasia and neoplasiaDevelopmental abnormalities

4. Uncertainties

National Health and Environmental Effects Laboratory

LiverUDPGTs

T4-Gluc

Biliary Excretion

Hypothalamus-Pituitary-Thyroid-Liver Axis

Plasma/Blood

Free-TH

Bound-TH

PA/TBG

Hypothalamus

Pituitary

Thyroid

TRH

TSH+

+

T3 & T4

T3 & T4

Tissue deiodinase

T4 > T3

National Health and Environmental Effects Laboratory

Iodine and the Thyroid Gland

I

-IT I

-organification

T4 & T3

THYROID GLAND

National Health and Environmental Effects Laboratory

Thyroid Hormone Receptors and Thyroid Response Elements

Target Tissue Cell

T4-TTR

T4T3

deiodinase

Nucleus

T3 Response Element

RXR RAR TR

T3

AGGTCA.....

T3 Responsive Gene

RNA polymerase II

TATA....

National Health and Environmental Effects Laboratory

Targets for Environmental Chemicals

Thyroid Gland uptake processorganification release

Plasma Transport Proteins Tissue Deiodinases Hepatic Glucuronidation and Sulfation

National Health and Environmental Effects Laboratory

In Vitro Thyrotoxicity of Perchlorate125Inhibition of I-Uptake by Perchlorate

Perchlorate Concentration, uM

0 1 10 100 1000

Iod

ine

influ

x,

pm

ol/m

g p

rote

in/m

in (

me

an

±SD

)

0

10

20

30

40

50

60

70

80

(Gerard et al., 1994)

National Health and Environmental Effects Laboratory

14-Day In Vivo Perchlorate Study

Perchlorate dose (mg/kg/day)

0 0.125 0.5 1.25 2.5 5 12.5 25

TS

H C

on

cen

trat

ion

, pg

/dl

0

5

10

15

20

25

30

35

40

45

0 0.125 0.5 1.2 2.5 5 12.5 25

T3

Co

nce

ntr

atio

n, n

g/m

l

0

40

60

80

100

120

140

T4

Co

nce

ntr

atio

n, n

g/d

l

0

2

3

4

5

6

Male Female

0 0.125 0.5 1.2 2.5 5 12.5 25

T3 T4

TSH

(Caldwell et al., 1994)

National Health and Environmental Effects Laboratory

Effect of TH disruption Adult

–Main effect of prolonged depression of TH is upregulation of TSH which leads to thyroid gland hyperplasia

Developmental–Main effect is the disruption of developmental processes

National Health and Environmental Effects Laboratory

Main Symptoms/Effects of Hypothyrodism

Developmentaldelayed reflex ontogenyimpaired fine motor skillsdeaf-mutism, spasticitygait disturbancesmental retardationspeech impairments

transient disruption leads to permanent effects

Adultrun down, slow, depressed, sluggish, cold, tireddryness and brittleness of hairdry and itchy skin, constipationmuscle crampsincreased menstrual flow

transient disruption leads totransient effects

*thyroid tumors in rodents

National Health and Environmental Effects Laboratory

Data Deficiencies Long-term periodic exposures

Perchlorate kinetics Thyroid hormone homeostasis

Effects during development

National Health and Environmental Effects Laboratory

Uncertainties in Risk

1. Animal to Human Extrapolation

2. Age-Dependent Sensitivity

National Health and Environmental Effects Laboratory

Uncertainties Due to Potential Species Differences

AdministeredDose

TargetDose

Effect

AdministeredDose

TargetDose

Effect

Rat

Human

? ?

ADME studies

Mechanistic / SensitivityStudies1. Adult2. Developmental

National Health and Environmental Effects Laboratory

Mechanistic Studies

• Aid to quantitative interspecies extrapolation - basis to extend PBPK model

• Additional developmental studies to evaluate thyroid TH in fetal and post-natal periods

• Determine relative sensitivity of fetal/postnatal thyroid versus adult

• Determine relative sensitivity of rat versus human

National Health and Environmental Effects Laboratory

Do species differences exist? compare rodent and human sensitivitiesusing in vitro techniques

Resolving UncertaintiesMechanistic Studies

Perchlorate Concentration

0 1 10 100 1000

Iod

ine

Up

take

0

10

20

30

40

50

60

70

80

Human (?)

Rat

National Health and Environmental Effects Laboratory

2. Age-Dependent SensitivityDevelopmental effects can be permanent.Is the developing organism more sensitive?

Yes - need to carefully characterizeNo - protection of maternal compartment

Uncertainties in Risk

National Health and Environmental Effects Laboratory

ADME study

• Literature review of perchlorate discharge test

• Protocols proposed to evaluate perchlorate kinetics, iodine inhibition kinetics and thyroid hormone homeostasis

• Basis for development of physiologically-based pharmacokinetic (PBPK) model

National Health and Environmental Effects Laboratory

Resolving Uncertainty - Example2. Age-Dependent Sensitivity

Determine relative sensitivity of fetal/postnatal thyroid versus adult

Determine relative sensitivity of rat versus human

National Health and Environmental Effects Laboratory

National Health and Environmental Effects Laboratory

PBPK Model Schematic

Summary

Facts: Perchlorate cause hypothyroidism

Uncertainties Animal to Human extrapolation Age-dependent senstivity

National Health and Environmental Effects Laboratory