The overall effort of the laboratory-based research is to provide mechanistic interpretation of the findings obtained from the clinical protocols developed and implements by our group. Specifically, the underlying research goal is the characterization of the role of thyroid hormone action and its local, tissue-specific modulation in the delivery of time- and tissue specificity of the signal. Our laboratory is currently characterizing in vitro and ex vivo the functional activity of common polymorphisms of type-2 deiodinase gene. In order to achieve this goal, we have established a cell culture-based type-2 deiodinase expression system. Standard biochemistry methods are utilized for the enzymatic activity assay;protein/protein and nucleic acids/protein interactions are tested by immunoprecipitation and mobility shift assay. A common polymorphism in the 5UTR region of the DIO2 gene (258 A/G DIO2) has been associated with a shift in the ratio of circulating T3/T4, suggesting an increased in the activity of the enzyme in vivo leading to a shift of the reaction equilibrium. Our in vitro and ex vivo data, consistent with others genotype/phenotype association studies, indicate that the 258 A/G DIO2 variant induces an increase in the transcription of the gene by displacing a putative repressor. Current efforts are aimed to characterize the putative repressor factor interacting with the polymorphism. Preliminary analysis of the data has indicated multiple candidate transcription factors. Our current efforts are aimed to the characterization of the repressor. Pre-adipocyte differentiation and culture. During the past two FYs we have focused our effort in developing a system of re-differentiation of human adipocytes from stromal cells obtained during adipose tissue biopsy. This system will become the platform to test in a controlled fashion the effects of specific genotypes on adipose tissue function independently of the metabolic status of the subject at the time of the sampling. We are currently using this experimental model to characterize the role of thyroid hormones in the differentiation of the adipocytes and in the modulation of their transcriptosome. Further effort is directed toward characterizing the transcriptional pattern of this system throughout the differentiation process with a particular focus on the expression of brown-fat specific genes. We are now characterizing the differential molecular signature of pre-adipocytes harvested from different anatomical locations, as well as their potential for differentiation in brown adypocytes. Type-2 deiodinase assay in primary culture of follicular thyroid cells. Collaborative work carried out with Dr. Gershengorns group (NIDDK-CEB) has led to the development of a reliable assay for the measurement of type-2 deiodinase activity in primary cultures of thyroid cells as a read-out of TSH-cAMP pathway. This system has been successfully utilized to test the activity of agonists of the TSH receptor. Development of a cell-based system to characterize the effects of substances modulating the type-2 deiodinase activity. We are currently developing cell line stably expressing type-2 deiodinase and a Thyroid Hormone Responsive Element (TRE)-driven reporter construct in order to evaluate the activity of small substances modulating the type-2 deiodinase activity. Effects of common polymorphisms in the modulation of the clinical presentation of the Resistance to Thyroid Hormone syndrome (RTH). In collaboration with Dr. Forreset we have characterized the role of common SNPs in the intron control region of the beta-2 thyroid hormone receptor gene. We have demonstrated that a common SNP causes a pituitary, tissue-specific, increase in transcription of the gene. This SNP is in cis with a pathogenic R338W mutation of the beta-receptor in the index case of pituitary selective RTH. Thus this combination causes a tissue selective over expression of the mutant gene ultimately generating a pituitary-selective dominant-negative state, recapitulating the peculiar phenotype of the index case. The results of this study have been recently published, and we are developing a translational protocol in collaboration with Dr. Douglas Forrest aimed to perform a haracterization of the role of polymorphisms in the regulatory region of the beta-2 thyroid hormone receptor gene in RTH syndrome patients with respect to their sensory system.

Project Start
Project End
Budget Start
Budget End
Support Year
9
Fiscal Year
2012
Total Cost
$442,082
Indirect Cost
City
State
Country
Zip Code
Vicinanza, Roberto; Coppotelli, Giuseppe; Malacrino, Carolina et al. (2013) Oxidized low-density lipoproteins impair endothelial function by inhibiting non-genomic action of thyroid hormone-mediated nitric oxide production in human endothelial cells. Thyroid 23:231-8
Lee, P; Werner, C D; Kebebew, E et al. (2013) Functional thermogenic beige adipogenesis is inducible in human neck fat. Int J Obes (Lond) :
Lee, Paul; Brychta, Robert J; Linderman, Joyce et al. (2013) Mild cold exposure modulates fibroblast growth factor 21 (FGF21) diurnal rhythm in humans: relationship between FGF21 levels, lipolysis, and cold-induced thermogenesis. J Clin Endocrinol Metab 98:E98-102
Wang, Ping-Yuan; Ma, Wenzhe; Park, Joon-Young et al. (2013) Increased oxidative metabolism in the Li-Fraumeni syndrome. N Engl J Med 368:1027-32
Sadowski, Samira M; He, Mei; Gesuwan, Krisana et al. (2013) Prospective screening in familial nonmedullary thyroid cancer. Surgery 154:1194-8
Lee, P; Brychta, R J; Collins, M T et al. (2013) Cold-activated brown adipose tissue is an independent predictor of higher bone mineral density in women. Osteoporos Int 24:1513-8
Lee, P; Linderman, J; Smith, S et al. (2013) Fibroblast growth factor 21 (FGF21) and bone: is there a relationship in humans? Osteoporos Int 24:3053-7
Peltsverger, Maya Y; Butler, Peter W; Alberobello, Anna Teresa et al. (2012) The -258A/G (SNP rs12885300) polymorphism of the human type 2 deiodinase gene is associated with a shift in the pattern of secretion of thyroid hormones following a TRH-induced acute rise in TSH. Eur J Endocrinol 166:839-45
Regard, Jean B; Cherman, Natasha; Palmer, Daniel et al. (2011) Wnt/β-catenin signaling is differentially regulated by Gα proteins and contributes to fibrous dysplasia. Proc Natl Acad Sci U S A 108:20101-6
Alberobello, Anna Teresa; Congedo, Valentina; Liu, Hong et al. (2011) An intronic SNP in the thyroid hormone receptor ýý gene is associated with pituitary cell-specific over-expression of a mutant thyroid hormone receptor ýý2 (R338W) in the index case of pituitary-selective resistance to thyroid hormone. J Transl Med 9:144

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