Institut für Experimentelle Endokrinologie IEÉ

Endokrinologisches Forschungs Centrum Charité

&

Hormon-Centrum Charité, Berlin

EnForCé

Josef Köhrle

Köhrle Charité

http://www.charite.de/enforce

http://www.charite.de/expendo

Institut Experimentelle EndokrinologieAscheim-Zondeck Haus

Die essentiellen Spurenelemente Selen & Jod: gezielte Prävention oder individuelle Selbstmedikation

mit Freiheit zur Mangel- oder Überversorgung ?

IEÉ ENFORCÉ

Topics

Trace Elements

Minerals

Iodine, Selenium and Thyroid

Mechanisms

Clinical Implications

Perspectives

Periodic Table

Twenty-five elements are essential to life

–Four of these make up about 96% of the weight of the human body

–Trace elements occur in smaller amounts

http://faculty.dbcc.cc.fl.us/viscoma/BSC1005Survey/notes/SurveyCH2.ppt

Standbild der Prinzessinnen Luise und Friederike. Luise war die Ehefrau Friedrich Wilhelms III., König von Preußen.

Ihre Schilddrüsenerkrankung verdeckte Sie mit Hilfe eines Schals

Schadow 1795

Goitre – Iodine Deficiency

V. Shashidar, Fiji School of Medicine

Iodine Deficiency

Enlarged Thyroids in GermanyIEÉ EnForCé

Including nodules

Direkte und Folgekosten für das deutsche Gesundheitswesen: 1.5 Milliarden € pro Jahr !

World-wide Iodine SupplyIEÉ EnForCé

Germany: at stage of developping countries

Adequate: Bahrain, South Africa (Cape town), Peru (coastal region), Switzerland)(> 100 to < 200 µg/d)

Moderate to mild deficiency: Germany, New Zealand, Australia, parts of USA(> 50 to < 100 µg/d)

Excess: coastal Hokkaido (> 500 µg/d)

Zimmermann et al. Am J Clin Nutr (2005) 81.840-4

Consequences of the world wide Iodine Deficiency

6 Million: Kretins

26 Million: Brain damage

655 Million: Goiters

1,6 Billion : Risk of impaired mental development

Quelle: WHO/UNICEF 1995

Iodine Deficient Populations

• Intelligence reduced

• by 13.5 IQ points

source: 18 studies (other countries)

Permanent!

Permanent!

The iodine nutrition status…. too little, enough, probably too much ...

Inpregnantwomen200- 250 µg/dis the

goal toachieve

In normal adults

< 50 µg/d overt ID50-100 µg/d relative restriction

100-150 µg/d probably OK150-200 µg/d certainly OK200-350 µg/d “grey zone”

> 350 µg/d iodine excess

Iodine supplementation programmes require prior assess-ment (and continuous monitoring) of the actual iodinenutrition situation to know « what needs to be implemented »

MIMVirchows Opinions on Cretinism

Goiter & Cretinism not relatedto iodine deficiencybut caused by 'Miasm':Adverse, bad, volatile influence of the environment and inadequateSocioeconomic status

Virchows Studies on Kretinism in Frankonian Spessart Region

Die Zwerge von Lhasa

Kashin-Beck Disease: Combined Iodine and Selenium Deficiency

Ein Leben lang nicht fähig zu arbeiten: Der 50-Jährige Bukchun (links) und eine Nachbarin. - Foto: maa Harald Maass 2007 TS

Kashin Beck Erkrankung

big joint disease

Osteoarthropathie, Störung der Knorpel- und Knochen-Differenzierung

Nahrungskontamination durch Pilzgifte (Getreide)

Nord-Ost-Asien

SELENIUM and the THYROID

•Thyroid: highest selenium content per mass unit

• expression of many selenoproteins

•Thyrocytes: life-long H202 production under control of TSH from fetal week 12 to death

• combined iodine and selenium deficiency leads to myxedematous cretinism in Central Africa

• association low serum Se and thyroid cancer

• Low serum Se associated with increased thyroid volume in females (SuViMax-study) Derumeaux et al EJE (2003)148:309

• selenite treatment reduces titers of thyroid auto-antibodies in Hashimoto‘s and postpartum thyroiditis

SELENIUM, THE THYROID GLAND & CLINICS

SELENIUM CONTENT OF HUMAN TISSUES[µg/g tissue]

Dickson and Tomlinson, 1967

infant1.5 y (pneumonia)

0.050.460.640.160.310.190.920.34

adult52 y (aneurisma)

0.130.471.240.270.220.170.630.39

pancreasgonadthyroidbrainintestinestomachkidneyliver

tissue

highest retentionof Se duringSe deficiency

in CNS and thyroid gland

EnForCé

Selenium in medicine and biology - or:a long way to overcome the first impression ...

1817 - Berzelius discovers Se 1957 - Schwarz & Foltz: Se - essential trace element1973 - Se: constituent of GSH-Peroxidase (GPx)1990 - an active residue in 5‘-deiodinase2003 – human selenogenome : 25 genes

Breite Varianz der täglichen Se-Aufnahme

Land Se [µg/day]Venezuela 200-350Kanada 113-220U.S.A. 60-160Japan 128Mexiko 61-73Germany 38-47UK, Scotland 31-43Finnland, -‘80 12-23““ , since –‘86 69-82

China, low 2-36““ , high 240-6990

Berzelius

RDA 2006:1 µg/kg BW

Low - lower - lowest

capillaryThyrocyteC-cell

Colloid-lumen

Capillary network Arteriole

NerveThyrocyte

C-cell

Organisation of Thyroid FolliclesEnForCéIEÉ

Köhrle, IEÉ, Charité

Thyroid gland – Iodide – TH Biosynthesis

TTF1/2, Pax8, AP1,NF-kB

Iodide

ThOx TPO

TSH

cAMP

NIS

Iodide

Dehal1Pendrin ?

Tg

Tg

2 Na+ Iodide

Thyroid Hormone BiosynthesisIEÉ EnForCé

3

5’

3’

5

DIT

DIT + DIT = THYROXINE (T4)3, 5, 3’, 5’-TETRAIODOTHYRONINE

MIT + DIT = TRIIODOTHYRONINE (T3)3, 5, 3’-TRIIODOTHYRONINE

Dohan et al. Endocr Rev 24 (2003) 48-77

Continuing, TSH-stimulated production of thyroid hormone during life depends on regulated H2O2generation and therefore requires antioxidativedefense mechanisms of thyrocytes.

Thyroid Hormone Biosynthesis

IEÉ ENFORCÉ

H2O2

GPx3

IEEEnForCé

Thyroid hormonesThyroid hormonesT4 (Thyroxine,

3,3′,5,5′-Tetraiodothyronine)

5´Deiodinases

T3 (3,3´,5-Triiodothyronine)

HO

I

O

I

I

OH

O

H2N

I

HO

I

O

I

I

OH

O

H2N

active hormone

Animation: Peter Hofmann, IEÉ

EnForCéIEÉ Selenium in proteins

seleno-cysteine

CH2 CNH3

+

HC

O

OSe

H

seleno-methionine

Se CH2 CH2 CNH3

+

HC

OOH3C

Functionally active & relevantRegulated biosynthesis21st codon

Not directly available,No specific function,Oxidized form Regenerated by MsrB(a selenoprotein) ?

2 H2Se

SeCys-tRNA

Seleno-Proteins

SeCys Se0

essential

3

(CH3)2Se

(CH3)3Se+

Seleno-sugars

Excretion

Steps in selenium (Se) metabolism I

seleno-methionine

Se CH2 CH2 CNH3

+

HC

OOH3C

SeMet

SeCys

Nutrient-Proteins

without special function

SeMet

SeMet-Proteine

1

SelenoproteineEnForCé

Selenoproteine stellen eine Erweiterung des klassischen genetischen Codes dar.Sie enthalten die 21te proteinogene Aminosäure Se-Cysteine (Sec).Der Einbau erfolgt co-translational:

UGA

eEFsecSECIS-BP

SeCys-tRNA

AUG 3´UGA5´

Sec eEFsec: Sec-spezifischer Elongations-Faktor SECIS: Sec-Insertions-Sequenz

Animation, Kostja Renko, IEÉ

The New World of Selenoproteins

Selenogenome:Human 25 genes, rodents 24C. elegans 1Drosophila 4

2003

selenoproteins withknown enzyme functions:GPx, TrxR, Dio

1973-2002

selenoproteome: 30-70:splicing, alternative start sites, posttranslationalmodification

2007-???

http://www.jb.man.ac.uk/~drl/

Selenoproteins25 genes encoding Se-proteins in the human genome

• 5 glutathione peroxidases• 3 deiodinases• 3 thioredoxin reductases• Selenoprotein P (SePP)• selenophosphate synthetase 2• methionine sulfoxide reductase• selenoproteins H, I, K, M, N, O, P, S, T, U, V, W

Se deficiency impairs the expression of Se proteins and is associated with increased vulnerability to "toxins " and changed thyroid hormone levels

Human Defect in Selenoprotein Biosynthesis –First Mutations in SBP2

IEÉ EnForCé

Two families with unknown cause of disturbed TH Feedback

Mildly elevated TSH and T4, elevated rT3; low T3

Inadequate T4- but normal T3-supression of TSH

Low Plasma GPx activity

Low Plasma Se

Low Plasma SePP

Impaired Type II 5'-Deiodinase activity in fibroblasts

Dumitrescu, …, Schomburg et al., Nature Genetics 16. Oct. 2005 aol

EnForCéMIMIEÉ SBP2 Mutation affects Selenoproteins

SBP2 mutation human: T4⇑ rT3⇑TSH ⇑;T3 ⇒

Dumitrescu, …., Schomburg et al, Nature Genetics 16. Oct. 2005 aol

Patients with hereditary SBP2-Defect

normal

defectnormal

defectl

A low SePP-concentration under normal nutrtion indicates a disturbed Selenium metabolism

Dumitrescu,A … Schomburg,L … Refetoff,S: Nature Genetics 2005.

EnForCéTissue-specific aspects of the hierarchical Se metabolism in mice

Relation between serum glutathione peroxidase activity (GPX) and serum selenium concentration after selenium supplementation in unsupplemented subjects ({blacktriangleup}), placebo-iodine-supplemented subjects ({circ}), and selenium-iodine-supplemented subjects (*)

Moreno-Reyes, R. et al. Am J Clin Nutr 2003;78:137-144

Copyright ©2003 The American Society for Nutrition

Selenium Excretion, Leipzig

Brauer VFH et al. European Journal of Endocrinology (2006) 155 807–812

TPOAb concentrations at the beginning of the study and 3 months after treatment with 200 mg L-selenomethionine/day

Turker O, Kumanlioglu K, Karapolat I, Dogan I. J Endocrinol. 2006 Jul;190(1):151-6

Percentage of patients who had PPTD (left) and hypothyroidism (right) develop in TPOAb(+) women who received Se (group S1) or placebo (group

S0), and in TPOAb(-) women (group C)

Negro, R. et al. J Clin Endocrinol Metab 2007;92:1263-1268

Copyright ©2007 The Endocrine Society

IEÉ EnForCéTreatment of AITD with Selenium

K.J. Schmidt, (1998) VitAminSpur 13:33-39100 µg selenium (yeast) for 6 weeks: M. Hashimoto, TSHdecrease, trend towards decreased thyroid volume, TPOAb, TRAK

R. Gärtner, JCEM 2002500 µg selenite for 3 to 6 months; M. Hashimotodecreased TPOAb, improved subjective feeling

L. Duntas, EJE 2003500 µg selenomethionine, 6 monthsdecreased TPOAb, improved subjective feeling

L. Duntas et al. ITC 2005, O 76200 µg selenomethionine, 6 monthsTRAB decreased, more rapid euthyroidism, mood effects

Turker et al. 2006: dose dependent effects in AITD, TPO-AbItalian Study 2007: benefit in postpartum thyroiditis

0

20

40

60

80

100

120

140

placebo Se

case

s

total cancerlung cancercolorectal cancerprostate cancer

Clark, Combs, et al. JAMA 1996

mean serum Se in USA: 125 µg/L(Kafai MR and Ganji V, J Trace Elem Med Biol 2003, 17, 13-18)

whereas mean serum Se in Germany: 67 µg/L(Oster O et al. Biol. Trace Elem. Res. 1998, 15, 47-81)… and mean serum in mice (C57BL/6): 320 µg/L (own data).

Reduction of Cancer Risk by Se

SELECT (Selenium and Vitamin E Clinical Trial)

• Double-blind, placebo-controlled, randomized, population-based trial in 32,400 men age ≥ 55.

• 2 x 2 factorial design: 200 µg Se (SeMet) x 400 mg α-tocopherol acetate daily for 7 years.

• 99% statistical power to detect 10% decrease in prostate cancer incidence.

• $180,000,000 for 12 years.

Cumulative incidence of type 2 diabetesCumulative Incidence Type II

Diabetes

Stranges, S. et. al. Ann Intern Med 2007;147:217-223

Sepsis

Selenbedarf bei Schwerkranken erhöht

Forceville et al, 1998 Crit. Care Med.

Plasmaselen bei schwerer Sepsis signifikant erniedrigt

Selenspiegel negativ mit Schwere der Erkrankung assoziiert

Selenium Deficiency Syndromes• Exudative diathesis in poultry (& Vit E).• Mulberry heart disease in swine (& Vit E).• Gizzard myopathy in turkeys (& Vit E).• White muscle disease (NMD) in sheep.• Pancreatic atrophy in poultry.• Liver necrosis in rats.• Keshan disease in humans (coxsackie B virus cofactor).• Kaschin-Beck Disease (osteoarthropathy) in humans (I?,

Mo?, B?, fungal toxins?).• Myxedematous cretinism in humans (iodine co-

deficiency).• Skeletal myopathy in TPN patients w/o added Se.

C. Hawkes et al.

IEÉ EnForCéAre There Reliable Indicators of Selenium Supply?

• blood selenium– erythrocytes– platelets

• plasma selenium• plasma selenoproteins

– pGPx– SePP

• tissue selenium content and selenoproteins

EnForCéNew Assay for SePP

SePP

Festphasen AB (CT)

Detektions AB (Tracer)

SePP

Schematische Darstellung des Sandwich-ELISAsim Coated-Tube Format zur SePP-Bestimmung.

hν

IEÉ EnForCé

Organ meats are another good source of the element.

From http://www.sciencenews.org/20030503/food.asp

The seafood in this bowl of sushi can be a rich source of selenium (and iodine, too! )

IEÉ EnForCé

Brazil Nuts are rich in Selenium

IEÉ EnForCéSummary & OutlookIEÉ

The trace element Selenium (Se)essential micronutrient for normal thyroid functionsignificant health benefits attributed to Sepromising cancer chemopreventive agentprevention of autoimmune thyroid disease

Active in Hashimoto‘s, Graves‘ and postpartum thyroiditisMechanism ???

Downregulation of expression of some selenoproteinsin (thyroid) cancer tissue

Mechanism ? DNA methylation (Dio1 promoter)

Biomarkers and indicators of adequate supply ? SePP

Interaction Se, Iodine, Iron?Köhrle Charité

IEÉ EnForCéSome Open IssuesIEÉ

Function of novel selenoproteinsThyroidal Se uptakeDifferent bioavailability & functions of various forms of SeSelenoprotein-specific drugs

Se & endocrine systemSe & immune system

Se in (chemo-)prevention & therapy of cancer

Mechanisms of Se toxicity

Köhrle Charité

Selenoproteins, ROS and DNA-damageIEÉ EnForCé

DNA-repair

defectiveDNA-repair enzymes or

cell cycle control-systems

Impaired antioxidativedefense systems e.g. Se-dependent GPx

Increased production of ROS: hydrogen peroxide, lipid- and phospholipid hydroperoxides

DNA-damage

IEÉ EnForCé

“The dose makes the poison”

Paracelsus

(1493-1541)

–“It is the task of chemistry to produce medicines for the treatment of disease since the vital functions are basically chemical in nature. All things are poisons, for there is nothing without poisonous qualities. It is only the dose, which makes things a poison.”

EnForCéIEÉ

Especially to the Deutsche Forschungsgemeinschaft DFG& Dt. Krebshilfe Dr. Mildred Scheel Stiftung & EnForCé

Institut für Experimentelle EndokrinologieMIMIEÉ EnForCé

Ulrich Schweizer

Lutz Schomburg

Florian Streckfuss

Marten Michaelis

Conny Riese

Kostya Renko

Jazmin Chiu

Vartiter Seher

Silke Kappler

Anita KinneDFG, Dt. Krebshilfe, Charité

IEÉ EnForCé

&

Institut Experimentelle Endokrinologie

Endokrinologisches Forschungs-Centrum der Charité EnForCé

IEÉ

IEÉ EnForCé

Fluorimetric Se-determination

HClSample HNO3/HClO4

Se IV

190° 150°

2 h 0,5 hSe xy Se VI

364 nm

Se IV-DAN

Se IV-DAN

2,3-DiAmino-Naphtalene

Cyclo-

hexan520 nm

Synthesis of thyroid hormones by follicular thyrocyte epithelial cells, storage of iodinated thyroglobulin in colloidal space and secretion of thyroid hormones

2I- + H2O2 2I0 + H2O + ½ O2O2 + 2e- +2H+ H2O2nI0 + Tg + nH2O2 Tg-DIT,Tg-MIT, Tg-T4,Tg-T3 + nH+ + H2O

DUOX

TPO

iodide

iodide

TgTg

T4 T3

NIS

PDS

1

2

3

5

6

4

Tg Tg

Tg Tg

7

MIT+NADPHI-+NADP+

8

TSHr

MCT8

pGPx

pGPx

9

1. basolateral iodide uptake by NIS

2. apical export by pendrin (PDS)

3. synthesis and apical secretion of thyroglobulin (Tg)

4. synthesis and apical insertion of thyroperoxidase (TPO) and dual oxidase (DUOX)

NADPH-dependent production of H2O2 by DUOX

5. iodide oxidation, iodination of Tg-tyrosyl residues and coupling of Tg iodtyrosine residues to iodothyronines iscatalyzed by TPO using H2O2 as cosubstrate

6. polymerisation and deposition of iodinated Tg in colloid

7. micropinocytosis, reduction and cathepsin catalyzedproteoloysis of Tg in secondary lysosomes

release of thyroid hormones T4 and T3 into the bloodby the transporter MCT8

8. dehalogenation of DIT and MIT and re-utilization of iodide for thyroid hormone biosynthesis

9. Secretion of pGPx (GPx-3) into the colloidal space fordegradation of excess H2O2

Enzymatic Deiodination of Thyroid Hormones by Deiodinase Selenoenzymes

T4

O

C O 2-H

N H3

I

I

I

I

-O

+

rT3

O

C O 2-H

N H 3

H

I

I

I

- O

+T3

O

C O 2-H

N H 3

I

I

H

I

HO

+

Dio1Dio2

5'-Deiodination 5-Deiodination

Dio3Dio1

Activation of prohormone T4 to active T3: Dio1 and Dio2Termination of T3 and T4 action: Dio3 and Dio1

Köhrle Charité

IEÉ EnForCéInstitut für Experimentelle Endokrinologie, Charité

Lutz Schomburg Ulrich SchweizerConny SchmutzlerBirgit MentrupBane RadovicFlorian StreckfussMarten MichaelisInka HamannConny RieseKostya RenkoPeter HofmannBirgit HollenbachSusi PiehlEva WirthStefan Roth

Vartiter SeherSilke KapplerAnita KinneAntje KunzeKatja SchreiberMetin YenilmezRosi GrossklausDFG, Charité, Dt. Krebshilfe, TSB & Zukunftsfonds Berlin

IEÉ EnForCé

Clark, Combs & Follow-upIEÉ

Se reduces incidence of lung cancer in persons with relatively low baseline plasma Se levels

Duffield-Lillico et al. 2002; Reid et al. 2002

Serum Se inversely related with risk of prostate cancerand a-tocopherol is higher in men with Serum Se in highest quartile

Vogt et al. Int J Cancer 2003 103:664

Se as chemopreventive agent for recurrent colon polyps(Se yeast alone and in combination with COX-2 inhibitor celeocoxib)

Organized by Alberts et al.

Köhrle Charité

Selenium & Iodine Status, Leipzig

Brauer VFH et al. European Journal of Endocrinology (2006) 155 807–812

Dietary Se Intake in Children New Sealand

n Infants (6–11.9 mo) n Toddlers (12–24 mo) P-value

Serum Se, µmol/L

Breast-fed 22

0.69 ± 0.27 7 0.61 ± 0.19 0.180

Nonbreast-fed 16

0.70 ± 0.34 91 0.57 ± 0.17 0.115

Se intake, µg/d 42

7.9 ± 6.2 188

13.7 ± 8.4 <0.001

Se density, µg/(MJ · d) 42

2.4 ± 1.7 188

3.2 ± 1.7 0.003

Energy intake, kJ/d 42

3227 ± 521 188

4162 ± 1099 <0.001

Protein intake, g/d 42

26 ± 7 188

36 ± 11 <0.001

1 Dietary selenium intakes exclude infants and toddlers currently breast-fed. 2 Values are means ± SD.

McLachlan SK et al. J. Nutr. 134:3290-3295, December 2004

Epidemiological criteria for assessing iodine nutrition based on median UI concentrations in school-age children

Median UI (µg/l) Iodine intake Iodine nutrition

< 20 Insufficient Severe iodine deficiency

20–49 Insufficient Moderate iodine deficiency

50–99 Insufficient Mild iodine deficiency

100–199 Adequate Optimal iodine nutrition

200–299 More than adequate Risk of iodine-induced hyperthyroidism within 5–10 years following introduction of iodized salt in susceptible groups

≥ 300 Excessive Risk of adverse health consequences(iodine induced hyperthyroidism,auto-immunethyroid diseases)

Source: WHO et al.

Population coveragea by UI surveys carried out between 1993 and 2003, by WHO region

Total number School-age Coverage of school-age children

children covered

WHO regionb (millions)c (millions) (%)

Africa 128.9 116.9 90.7

Americas 109.0 98.8 90.6

South-East Asia 242.4 239.4 98.8

Europe 81.2 70.5 86.8

Eastern Mediterranean 87.1 72.6 83.4

Western Pacific 199.4 183.0 91.8

Total 848.0 781.2 92.1a School-age children (6–12 years).

b 192 WHO Member States.

c Based on population estimates for the year 2002 (23).