t hyroid h ormone s ynthesis, s ecretion, a ction, r eceptors & a ntibodies emily brennan pgy-4,...
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THYROID HORMONE SYNTHESIS, SECRETION, ACTION,
RECEPTORS & ANTIBODIES
Emily BrennanPGY-4, EndocrinologyAugust 13, 2014
ObjectivesAt the end of this lecture, you will be able to understand:
• the synthesis of thyroid hormone• thyroid hormone transport and activation at local tissues• the regulation and feedback mechanisms of the thyroid axis• the downstream action of thyroid hormone on tissues• the role and value of thyroid antibodies
Recall: The Thyroid• The thyroid is the largest single
organ specialized for endocrine hormone production
• The thyroid’s function is to secrete an appropriate amount of the thyroid hormone primarily as T4
• The gland is composed of closely packed, spherical units termed follicles
Thyroid Hormone and Related Structures • Comprised iodinated
thyronines• two tyrosine molecules joined
by an ether linkage
• Produced only by the follicular cells of the thyroid
• Iodine is a key structural component of thyroid hormone
Thyrocyte
Thyroid Hormone Synthesis• Requirements for hormone production:• Sodium-Iodide Transporter (NIS)• Iodine• Thyroglobulin (Tg)• Thyroid peroxidase (TPO)
1. Trapping -- Active transport of iodide across the basement membrane into the thyroid cell
Sodium-iodide symporter (NIS)Location: basal membrane of the thyrocyte
Function: actively transports iodide from the blood; maintains concentration of iodide ~30x greater than plasma
Regulation: stimulated by TSH, suppressed by excess iodide
1. Trapping -- Active transport of iodide across the basement membrane into the thyroid cell
PendrinLocation: apical border of the thyrocyte
Function: transports iodide into the membrane-colloid interface to become substrate for thyroid hormonogenesis
Mutation: Pendred syndrome (goitre and congenital deafness)
Iodine• Essential micronutrient consumed in food and
water
• Recommended intake: • 150 mcg adults• 200 mcg pregnant/lactating women• 50-250 mcg children
• When iodine intake is less than 50 mcg/d, a normal-sized thyroid cannot sustain adequate hormone production
• The concentrating ability of the thyroid creates an intrathyroid pool (8-10 mg)
Thyroglobulin• A large glycoprotein molecule, composed of two subunits
• Structure: Includes 140 tyrosyl residues, but only four tyrosyl sites are sterically oriented for effective hormornogenesis in each molecule
• Role: serves in synthesis and storage of thyroid hormone
• Regulation: TSH regulates the expression of the thryoglobulin gene
Thyroglobulinthyroglobulin mRNA translated
into RER↓
glycosylated and transported to golgi apparatus
↓incorporated into exocytic
vesicles↓
fused with basement membrane↓
at apical-colloid border, Tg is iodinated
2. Organification -- oxidation of iodide and iodination of tyrosyl residues in thyroglobulin
Thyroid Perioxase (TPO)A membrane-bound glycoprotein
Function: catalyses both iodide oxidation and covalent linkage of iodine to the tyrosine residues of thyroglobulin
Location: Cell-Colloid Interface
Regulation: gene expression stimulated by TSH
Inhibition: methimazole, carbimazole, and propylthiouracil
3. Coupling -- linking pairs of iodotyrosine molecules within the thyroglobulin to form T3 and T4
The coupling of iodotyrosyl resides is also catalyzed by TPO
Thought to be an intramolecular process involving the oxidation of two iodotyrosyl residues brought into proximity by the tertiary and quatenary structures of thyroglobulin, and it involves a linkage and splitting to form an iodothyronine
Coupling of iodotyrosine molecules
Coupling of iodotyrosine molecules
4. Pinocytosis and then proteolysis of thyroglobulin with release of free iodothyronines and iodotyrosines into circulation
Proteolysis of Thyroglobulin and Thyroid Hormone Secretion
Colloid is engulfed in vesicles by pinocytosis and absorbed into the cell
↓Lysosomes then fuse with the vesicle
↓Releases T4 and T3 and inactive
peptides↓
The biologically active thyroid hormones T4 and T3 enter the
circulation ↓
DIT and MIT are deiodinated and their iodide conserved
5. Deiodination of iodothyroxines within the thyroid cell with conservation and reuse of liberated iodine
6. Intrathyroidal 5’-deiodination of T4 to T3
Summary
Thyroid Hormone Transport• Both T3 and T4 are poorly soluble in water, and therefore,
circulate bound to plasma proteins• 0.04% T4 unbound• 0.4% T3 unbound
• Three major transport proteins:• Thyroxine-binding globulin (TBG)• Transthyretin• Albumin
Thyroxine-Binding Globulin (TBG)• Liver-derived glycoprotein• Each TBG molecule has a single binding site for T4 or T3• It carries about 70% of circulating thyroid hormones (high
binding affinity for T4 and T3)
Conditions Affecting Binding of TBGCongenital• TBG deficiency, X-linked recessive; low total T4/T3, but free hormone levels are
normal; euthyroid• Congenital TBG excess: elevated total T4/T3, normal fT4/fT3, euthyroid
Physiologic• Pregnancy/estrogen – increase the sialic acid content of TBG, decrease metabolic
clearance and elevated TBG levels
Pathophysiologic• Estrogen secreting tumours, OCP, acute hepatitis (increased siailac acid)• Systemic illness – decrease TBG due to cleavage by leukocyte protease, and decrease
binding affinity; lowers hormone concentrations
Drugs• Salicylates, high dose phenytoin, furosemide – bind to TBG, displace T4/T3
Transthyretin• Formerly known as: thyroxine-binding prealbumin• Binds to 10% of circulating T4• Affinity for T4 is 10-fold greater than for T3
• Expressed in the choroid plexus• the major thyroid hormone-binding produced in the CSF
• The dissociation of T4 and T3 from transthyretin is rapid, so that transthyretin is a source of readily available T4
Conditions Affecting Binding• Congenital increased affinity for transthyretin binding for T4
• elevated total T4, but normal fT4• Ectopic production of transthyretin can occur w/ pancreatic and hepatic tumours
• causes euthyroid hyperthyroxinemia
Albumin• Binds to T4 and T3 with a lesser affinity than TBG or
transthyretin, but it has a high plasma concentration• 15% of circulating T4 and T3
• Rapid thyroid hormone dissociation rates from albumin make it a major source of free hormone to tissues
Conditions Affecting Binding• Hypoalbuminia (nephrotic syndrome, cirrhosis) is associated with
a low total T4 and T3, but normal free hormone levels • Familial dysalbuminemic hyperthroxiemia: AD where 25%
albumin has a higher than normal T4 binding affinity results in elevated total T4 level, but normal fT4 = euthyroid
Transport across Cell Membranes• Originally thought to be primarily passive; however, several
specific thyroid hormone transport proteins have been identified:• MCT8 • MCT10• Organic anion transporting polypeptide 1C1 (OATP1C1) – predominantly
in the brain, transports T4 preferentially
• In most cells, 90% of T3 is in the cytosol except the pituitary, where 50% of the T3 is in the nucleus
Metabolism of Thyroid Hormones• Most of the plasma pool of T3
(80%) is derived from:• 5’-deiodination of T4 in the liver,
kidney, and skeletal muscle
• Deiodination of the inner ring of T4 (5-deiodination) produces reverse T3 (metabolically inert)
*
*
Metabolisms of Thyroid Hormone • Three deiodinases enzymes catalyze these reactions:• D1• D2• D3
• permit local tissue and cellular modulation of thyroid hormone action
DeiodinationD1Location: most abundant form, found predominantly in liver, kidney, and lesser extent in thyroid gland, skeletal, and heart muscle, and other tissues
Function: major converter of T4 to T3
Inhibition: PTU (not methimazole), amiodarone and iodinated radiocontrast dye
DeiodinationD2Location: expressed in the brain and pituitary gland, where is maintains constant levels of intracellular T3 in the CNS
Function: maintenance of the level of intraacellular T3 and its neuronal cellular functions
D2 is very sensitive to circulating T4
DeiodinationD3Location: chorionic membranes of the placenta and glial cells in the CNS
Function: inactivates T4 by converting it to rT3 and it inactivates T3 by converting it to 3,3’-T2
Elevated in hyperthyroidism and decreased in hypothyroidism, may help to insulate the fetus and the brain from T4 excess of deficiency
Regulation of Thyroid Function
Regulation of Thyroid FunctionThyrotropin-Releasing Hormone (TRH)• Synthesized in hypothalamus (supraoptic and
supraventricular nuclei)
• Stored in the median eminence and then transported via the pituitary portal venous system to the anterior pituitary
• Role: controls synthesis/release of TSH in the anterior pituitary (binds to G-protein receptor on thyrotrophs to initiate pathway to stimulate for TSH release). Pulsatile secretion, peak midnight-4am
• Negative Control: • TRH gene expression negatively regulated
by T3 and T4• T3/T4 downregulate the TRH receptors in
the pituitary thyrotrophs
Regulation of Thyroid FunctionThyroid-Stimulating Hormone• Structure: Glycoprotein composed of
an alpha and beta subunit, that are noncovalently linked• The alpha subunit is common to FSH and
LH and hCG• The beta subunit is unique to each
hormone, conferring its specific binding properties and biologic activity
• Role: TSH controls thyroid cell growth and hormone production• binding to a specific TSH G-protein
receptor on the basolateral thyrocyte membrane
• activates a cascade responsible for promoting thyroid cell growth and hormone synthesis/release
Effects of TSH on the Thyrocyte• Changes in thyrocyte morphology• induces pseudopods at the follicular cell-colloid border, accelerating
thyroglobulin resorption, which increases thyroid hormone release
• Cell growth• individual cells increase in size, vascularity, and over time, thyroid enlargement
or goiter develops
• Iodine metabolism• TSH stimulates all phases of iodide metabolism (increased uptake, transport,
secretion); increased NIS expression
• Thyroglobulin and TPO mRNA expression• Results in increased incorporation of iodide into MIT, DIT, T3 and T4
TSH RegulationFeedback• T3 concentration in the hypothalamus within the thyrotrophs
cells regulates mRNA expression, TSH translation and T3/T4 release• TRH - controls postranslational glycosylation and release of TSH
Inhibitors of TSH• somatostatin, dopamine, dopamine agonists, high dose glucocorticoids
TSH Receptor Conditions
• Congenital• Familial hyperthyroidism – activating mutation of TSH
receptor• Familial gestational hyperthyroidism – due to structural
similar hCG hormone activated aberrant TSH receptor• Acquired• TSH receptor blocking antibodies (cause hypothyroid)• Graves’ (most common)• Autoantibodies bind and stimulate the TSH receptor
Iodine Autoregulation• The capacity of the thyroid gland to modify its function to the
availability of iodine, independent of pituitary TSH
• Allows maintenance of adequate/appropriate thyroid hormone secretion is varying intake of iodine
• Major adaptation to low iodide intake is preferential synthesis of T3 over T4
• Iodide excess inhibits many thyroidal functions including:• iodide trapping• thyroglobulin iodination (Wolff-Chaikoff)• thyroid hormone release from the gland
Mechanism of Thyroid Hormone ActionGenomic Actions• T3 interacts with its nuclear receptors to regulate gene activity• T3 binds to a specific nuclear thyroid hormone receptor (TR),
which in turns bind to DNA at specific sequences called thyroid hormone response elements (TREs)
• T3 has a 15-fold higher binding affinity for TRs than T4• There are tissue-specific preferences in expression of the various
TRs (difference expression in hypothalamus vs kidney, liver, brain and heart)
Nongenomic Actions• Nongenomic actions mediated by T3 and T4 occur with certain
enzymes
Thyroid Hormone Genomic Actions
Thyroid Hormone ActionsFetal Development• Iodide is found in thyroid tissue
and pituitary TSH appear in the fetus at about 11 weeks
• The high placental content of D3 inactivates most maternal T3 and T4, and very little free hormone reaches fetal circulation
• After 15-18 weeks, the fetus controls most of its own thyroidal secretion
Thyroid Hormone Action
Increases oxygen consumption and heat production by stimulation of Na-K-ATPase in all tissues
Oxygen, Heat, Free Radicals
Stimulates mitochondriogenesis, augmenting the cell’s oxidative capacity
Increases basal metabolic rate
Thyroid Hormone Action
Lowers peripheral vascular resistance, increase intravascular volume increased cardiac output
Increases the rate of myocardial diastolic relaxation
Increases the alpha receptors
Cardiovascular System
Increases the rate of depolarization and repolarization of the SA node, increasing heart rate
Positive inotropic and chronotropic effects on the heart with heightened adrenergic sensitivity
Thyroid Hormone Action
Maintains the ventilatory responses to hypoxia and hypocapnia in the brain stem respiratory center
Pulmonary System
Thyroid Hormone Action
Increased cellular demand for oxygen in hyperthyroidism leads to increased production of EPO and erythropoiesis
Hematopoeitic Effects
Increases the oxygen dissociation from hemoglobin and increases oxygen availability to the tissues
Thyroid Hormone ActionSkeletal System
Stimulates bone turnover and increases bone resorption and to a lesser degree, bone formation
Thyroid Hormone Action
Thyroid hormone promotes gut motility
Gastrointestinal System
Thyroid Hormone Action
In hyperthyroidism, there is increased protein turnover and loss in skeletal muscle
Changes in speed of muscle contraction and relaxation, noted as hyperreflexia or delayed DTR in hypothyroidisms
Neuromuscular System
Fine distal hand tremor is typical in hyperthyroidism
Hyperactivity and sluggishness can be striking
Thyroid Hormone Action
Thyroid hormone regulates the synthesis of pituitary hormones, stimulates growth hormone production, and inhibits TSH
Low thyroid hormones levels can cause delayed puberty by impairing GnRH secretion
Reproductive System
Thyroid Hormone Action
increases hepatic gluconeogenesis and glycogenlysis, and intestinal glucose absorption
?thyroid hormone mediated decreases in insulin sensitivity worse glycemic control
Lipids and Carbohydrate Metabolism
Increase cholesterol synthesis and degradation (largely due to an increase in hepatic LDL receptors, accelerating LDL clearance)
Thyroid Antibodies• Considered the hallmark of the autoimmune thyroid disorders
• Antibodies to thyroglobulin (Tg)• Antibodies to thyroid peroxidase (TPO)• Antibodies directed to TSH-receptor (TSHR)
TPO-Ab and Tg-Ab• Polyclonal antibodies, IgG class• Develop as a secondary response to thyroid injury• Not thought to cause the disease themselves (i.e. they cross the placenta
and do not cause disease in fetus)• Contribute to the disease mechanisms (complement fixing cytotoxicity
or T cell activation)• correlates well with thyroidal damage and lymphocytic infiltration
TPO-Ab and Tg-Ab• Almost all patients with autoimmune thyroiditis are associated with Tg-
Ab and TPO-Ab• May be present in Graves’ as well
• TPO-Ab has a higher affinity and occurs in higher concentrations
• Tg-Ab and TPO-Ab are more common in patients with sporadic goitre, multinodular goiter, or isolated thyroid nodules and cancer than the general population• Non-specific
• Not necessary for evaluation of thyroid function• may be helpful to predict progression of subclinical hypothyroidisms
TSHR-Ab• The presence of antibodies favours an autoimmune cause for
hyperthyroidism, but not sensitive or specific interpretable only as part of the clinical scenario
• Testing for TSHR Ab are the test of choice; they usually are thyroid receptor stimulating antibodies (compete with TSH for binding to its specific receptor site in the cell membrane)• can behave as stimulating, inhibitory or neutral
• Tg-Ab and TPO-Ab are also detectable in 50-90% of patients’ with Graves
Take Home:Requirements for Thyroid Hormone Synthesis:• Sodium-Iodide Transporter (NIS)• Iodine• Thyroglobulin (Tg)• Thyroid peroxidase (TPO)
Steps for Thyroid Hormone Synthesis:• Iodine trapping via the Sodium-Iodide Symporter• Oxidation and organification of thyroglobulin by TPO• Coupling pairs of MIT or DIT within the thyroglobulin molecule to form T3
and T4• De-iodination of MIT/DIT to conserve iodine• Release of T3/T4 into circulation
Take Home:• T3 and T4 are circulate bound to: thyroxine-binding globulin, transthyretinin,
and albumin
• T4 undergoes 5’-deiodination to form T3 (the more biologically potent hormone)
• T3 acts on the cell nucleus to regulate gene expression and protein synthesis.
• Thyroid hormone has targets of action in most tissues in the body
• TSHR-Ab is the most specific for Graves’ disease, and is the mechanism for the disease
• TPO-Ab and Tg-Ab are evidence of thyroidal injury, but are nonspecific for diagnostic purposes
References• Williams Textbook of Endocrinology 12th edition. • Greenspan’s Basic and Clinical Endocrinology.• Endocrine Physiology. Lange. • UpToDate• Boron WF (2003). Medical Physiology: A Cellular And Molecular
Approaoch. Elsevier/Saunders. p. 1300. ISBN 1-4160-2328-3.
Questions?