Nuclear receptors. Lecture 6. VDR and THR
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1 Nuclear receptors Lecture 6 VDR and THR
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4 Vitamin D and other diseases Vitamin D reduces colorectal epithelial cell proliferation and induces differentiation, which may be mediated through the VDR. Thus vitamin D may inversely associated with the risk of colorectal, prostate or breast cancers, but epidemiological data are contradictory Vitamin D supplementation may inhibit or stop the progression of multiple sclerosis (MS). Most MS patients suffer from vitamin D deficiency. MS prevalence increases with decreased exposure to the sun, and diets rich in fish, which is rich in vitamin D, may lower MS severity. Studies in Japanese population suggested an association between the less active form of VDR and frequency of MS. Experiments in animals suggest that vitamin D may be of use in treating the insulin-dependent diabetes and in suppressing transplant rejection.
5 Development of type I diabates - The greater prevalence of T1D at northerly latitudes than in sunnier climates. - Vitamin D supplementation in early childhood not only helps preventing rickets, but also leads to a reduction in the risk of type 1 diabetes
6 Development of type I diabates
7 Vitamin D and psoriasis
8 Treatment of psoriasis with vitamin D3
9 Clinical trials with vitamin D supplementation In a double-blind, placebo-controlled trial, volunteers aged >70 were given a daily tablet of 10 g (400 IU) vitamin D3 for up to 3.5 years to investigate whether this reduced the risk of fractures. Serum 25-(OH)-D levels were increased in the treated group. However, there was no difference in fracture incidence. No adverse effects were reported. In a trial of 800 IU per day vitamin D3 supplementation in 64 monozygotic twin pairs over two years no adverse effects were observed, nor was any significant difference in bone mass density. In a 3 year placebo-controlled study of 17.5 g/day vitamin D3 and calcium supplementation, bone mass density was significantly increased in the supplemented group. No adverse effects were reported. Short term supplementation of elderly women, with 800 IU/day vitamin D3 and calcium for 8 weeks, has been shown to decrease body sway and the incidence of fractures during a 1 year follow-up period.
10 Rickets Late closure of the fontanelle (craniotabes) is the earliest sign of rickets. It is detected in infants less than 12 months of age as round unossified areas in the skull. Beading of the ribs, termed rachitic rosary, is an almost consistent sign after 6 months of age. This is caused by the swollen cartilaginous ends of the ribs. The chest may be narrow and rather funnel shaped, described as pigeon chest, in severe cases this may interfere with breathing. When the child begins to toddle, putting weight on the legs results in the femur becoming bowed. Severe vitamin D deficiency reduces the growth rate and causes microcephaly, with reduction in brain growth and the eruption of teeth is delayed.
11 Rickets During the Second World War, it had become commonplace in Great Britain for dried milk and infant cereals to be fortified with ergocalciferol (vitamin D2), and to give supplementary concentrated vitamin D to infants. Additionally the cod-liver oil was supplied. Jedrzej Sniadecki Increases in vitamin D intake since the recognition of the aetiology of rickets, improved the nutrition, resulting in a dramatic reduction in the incidence of rickets. 1822: describes the method for treatment of rickets by exposure to sun
12 Rickets Insufficient vitamin D delivery e.g. too low exposure to sunlight, low-fat diet. Renal osteodystrophy kidney damage leading to loss of 1 -hydroxylase activity, low 1,25(OH) 2 D3, impaired intestinal calcium absorption (rickets/osteomalacia). Hereditary rickets type I inherited defect of kidney 1 -hydroxylase prevents formation of active hormone; treatable with exogenous calcitriol. Hereditary rickets type II end organ resistance due to mutation in the VDR receptor DNA or hormone binding domains; DNA binding domain mutations unresponsive to administered calcitriol; ligand binding domain mutations can sometimes respond to very high dose 1,25D3 treatment.
13 Vitamin D insensitivity Inherited vitamin-d3 insensitivity type II mutation in VDR * Inherited, recessive autosomal disease. * Insufficient mineralization of new-formed bones and ossified cartilages. * Symptoms of rickets start to be visible during the first few months of live: -painof bones - general weakness and weakness of muscles - sometimes convulsions resulting from too low level of calcium * Inhibited growth, underdevelopment of teeth. * Many children have alopecia both on the head and on the eyebrows or eyelash. * High doses of vitamin-d are not effective in children with alopecia. Better results can be obtained using high doses of calcium.
14 Vitamin D insensitivity Inherited vitamin-d3 insensitivity type II mutation in DBD of VDR * Biochemical features: - low level of calcium and phosphates in blood - increased level of alkaline phosphatase in blood -increasedlevelof PTH - high level of calcitriol (there is a difference comparing to rickets type-i, where level of calcitriol is diminished because of deficiency in 1α-hydroxylase).
15 Thyroid glands The thyroid gland is a strongly vascularized organ, located in the neck, in close approximation to the first part of the trachea. In humans, it has a "butterfly" shape, with two lateral lobes that are connected by isthmus. Most animals have two separate glands on either side of the trachea.
16 Thyroid glands Thyroid epithelial cells - the cells responsible for synthesis of thyroid hormones - are arranged in spheres called thyroid follicles. The follicle lumen is filled with a thick colloid, which predominantly contains thyroglobulin. Thyroglobulin is a highly glycosylated protein of two subunits, each of 330 kda. The subunit contains 115 tyrosine residues.
17 Thyroid hormones Thyroid hormones are derivatives of tyrosine bound covalently to iodine. The two principal thyroid hormones are: thyroxine (T4 or L-3,5,3',5'-tetraiodothyronine; major form released from the gland) triiodothyronine (T3 or L-3,5,3'-triiodothyronine). Although both T3 and T4 are important for normal growth and development and energy metabolism, T3 is ~10 times more active ligand of THR than T4. Thyroid hormones are basically two tyrosines linked together with the critical addition of iodine at three or four positions on the aromatic rings.
18 Physiologic effects of thyroid hormones Thyroid hormones regulate "big time processes (development, growth and metabolism). Metabolism: Thyroid hormones lead to an increase in basal metabolic rate (BMR). One consequence of this activity is increased heat production (nonshivering thermogenesis). Thyroid hormones induce expression of UCP-1. Mitochondrial ATP production in rats control hyperthyroid heart liver soleus plantaris
19 Processes modulated by THR action
20 Growth: Physiologic effects of thyroid hormones Thyroid hormones are necessary for normal growth of juveniles, as evidenced by the growth-retardation observed in thyroid deficiency. Development: A classical experiment in endocrinology was the demonstration that tadpoles deprived of thyroid hormone failed to undergo metamorphosis into frogs. Rana temporaria Xenopus laevis healthy lack of thyroid gland Rot-Nikcevic & Wassersug, J Exp Biol 2004.
21 Physiologic effects of thyroid hormones - Amphibian metamorphosis requires adaptation to oxygen rich environment. - TH is low during embryonic and early larva development and increases before metamorphosis. - Low TH can adapt salamander to low oxygen aquatic environment. -? Analogies in mammalian tisses after ischemia and reperfusion? Possibly yes. Pantos & Mourousis. Oxid Med Cell Longevity 2013
22 THR and cardiac regeneration - A decline in T3 levels occurs after myocardial infarction (AMI) or cardiac surgery. - At 6 months, recovery of cardiac function is correlated with T3 plasma levels. - Acute T3 (and not T4) administration after heart ischemia in rats or mice improved the postischemic recovery of function. - TR 1 is predominantly expressed in the myocardium and regulates genes related to cell differentiation, survival (through PI3K activation) and contractile function (e.g. TRE-mediated upregulation of connexin-43). Pantos & Mourousis. Oxid Med Cell Longevity 2013
23 Mammalian THRs are encoded by two genes, designated and, both can be alternatively spliced, generating different isoforms: * -1, -2, Thyroid hormone receptor (THR) * -1 (can bind cyclin D1 and p53) and -2. The -2 isoform has a unique C-terminus and does not bind T3 and may act as inhibitor of other THRs.
24 Thyroid hormone receptor (THR) THRs can bind to a TRE as monomers, as homodimers or as heterodimers with the retinoid X receptor (RXR). The heterodimer affords the highest affinity binding, and is thought to represent the major functional form of the receptor. The most stable binding occurs on the classical DR4 TRE. THRs bind to DNA regardless of whether they are occupied by T3. Binding of THR alone to DNA leads to repression of transcription, whereas binding of the thyroid hormone-receptor complex leads to activation of transcription.
25 Mode of THR action Ligand-free state: T3-free receptor, as a heterodimer with RXR, assumes a conformation that promotes interaction with a corepressor molecules. Binding of T3 induces a conformational change to bind the coactivator proteins. Increased by cyclin D1
26 Thyroid hormone receptor Reduced by p53
27 Hypothyroidism Causes: - Iodine deficiency - Thyroid disease (e.g. Hashimoto's disease, autoimmune disease leading to inflammation). healthy HS - Hypothyroidism is a common side effect of TKIs. - Sunitinib is an oral TKI with activity against VEGFRs, PDGFRs, and ckit. Onethird (14 85%) of patients on sunitinib treatment develop hypothyroidism.
28 Mutations in THR Mutations in the ligand-binding domain of THR can lead to generalized resistance to thyroid hormones. Clinically, such individuals show a type of hypothyroidism characterized by goiter, but also elevated serum concentrations of T3 and thyroxine and normal or elevated serum concentrations of TSH.
29 Causes: Too high secretion of thyroid hormones (less common than hypothyroidism). Graves disease, an autoimmune disease in which antibodies bind to and activate the thyroid-stimulating hormone receptor, leading to continuous stimulation of thyroid hormone synthesis. Thyroid cancer. Hamburger thyroxicosis (rare ). Hyperthyroidism Exophthalmos
30 Hamburger thyroxicosis Thyroid hormones are orally active, thus consumption of thyroid gland tissue can cause thyrotoxicosis. Several outbreaks of thyrotoxicosis have been attributed to a practice (banned), where meat in the neck region of slaughtered animals is ground into hamburger. Thyrotoxicosis in Minnesota (in 1987) and South Dakota was traced to thyroid-contaminated hamburger. A total of 121 cases were identified. The patients complained of sleeplessness, nervousness, headache, fatique, excessive sweating and weight loss.
31 Hamburger thyroxicosis Just one contaminated hamburger.
32 Physiologic effects of thyroid hormones Central nervous system: Both decreased and increased concentrations of THs alter the mental state. Too little thyroid hormone, and the individual tends to feel mentally sluggish, while too much induces anxiety and nervousness. Normal levels of THs are essential to the development of the fetal and neonatal brain. Congenital thyroid deficiency results in cretinism, which includes dwarfism and mental retardation (an impairment of abstract thought, with preserved vegetative, personal, social functions and memory).
33 Thyroid hormone and fetal brain development TH have most profound effects on the terminal stages of brain differentiation, including growth of dendrites and axons, and myelination (e.g. direct up regulation of myelin basic protein). Human fetuses acquire the ability to synthesize THs at 10 to 12 weeks of gestation. There is substantial transfer of maternal THs across the placenta. In the placenta the deiodinases converts T4 to T3. Purkinje cells kept for 14 days TH induced increase in Purkinje cell dendritic branching. A fetal rat brain produced by in situ hybridization with a probe for the rat thyroid hormone receptor.
34 Thyroid hormone and fetal development Thyroid deficiency is known to affect fetal development: * Isolated maternal hypothyroidism: Typically is not a cause of fetal disease because it usually is associated with infertility. When pregnancy does occur, there is increased risk of intrauterine fetal death and gestational hypertension. * Subclinical hypothyroidism: Mild maternal hypothyroidism, diagnosed only retrospectively from banked serum, may lead in children to such effects as slightly lower performance on IQ tests and difficulties with schoolwork. * Isolated fetal hypothyroidism: Failure of the fetal thyroid gland. Most children are normal at birth, because maternal thyroid hormones are transported across the placenta. What is absolutely critical is to identify and treat this condition very shortly after birth. If not, the child will become permanently mentally and growth retarded.
35 Isolated fetal hypothyroidism few months thyroid hormone replacement
36 THR: non-genomic activity THR after ligand binding can activate the PI3K pathway via phosphorylation of Akt/PKB (minutes aftet stimulation), leading e.g. to activation of enos and nnos. - T4 and T3 can bind to the integrin v 3, at sites denoted as S1 and S2: * S1 exclusively binds T3 at and leads to PI3K activation. * S2 binds T4, and to a lesser extent T3, and activates the ERK1/2 pathway. HIF-1 Increased production of TH is associated with progression of ovarian cancer, breast cancer and prostate cancer. Moeller & Fuhrer. Endocrin-Related Cancer 2013
37 THR and muscle differentiation deionidase Salvatore et al. Nat Rev Endocrinol 2014
38 THR and muscle differentiation Salvatore et al. Nat Rev Endocrinol 2014
39 Thank you and see you next week... What would be profitable to remember in June: - Types of rickens - Effect of vitamin D3 on immune system - Causes and symptoms of thyroid hormone deficiency - Splicing forms and mode of action of TR.
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