This is a competing renewal application designed to continue studies on phosphate transport and homeostatic mechanisms during health and disease states. This renewal will focus on bone disease associated with inflammatory bowel disease (IBD) as it relates to phosphate homeostasis. To accomplish this goal, we are planning a series of in vitro and in vivo experiments to gain a deeper understanding of the mechanisms governing phosphate homeostasis, the gut-renal-skeletal axis, and immune-skeletal interactions in IBD at systemic, cellular and molecular levels. The field on phosphate homeostasis has recently seen major advances which challenge many of the established concepts. These advances include the discovery of phosphatonins and the Klotho gene. Therefore the central hypothesis to be tested in this proposal is that in acute and chronic colitis, TNFa is a central player in the disruption of the gut-skeletal- renal axis responsible for inorganic phosphate homeostasis and proper bone mineralization. To address this hypothesis, we are planning to investigate three specific aims: 1. To identify molecular mechanisms of TNFa-mediated down-regulation of Phex gene expression in osteoblasts. The hypothesis of this aim is that Phex, a gene indispensable for osteoblast mineralization and indirectly participating in inorganic phosphate reabsorption, is significantly down-regulated in acute and chronic colitis. TNFa mediates decreased Phex gene transcription through a novel molecular mechanism involving a poly-adenine region of the proximal Phex gene promoter. 2. To describe molecular mechanisms of downregulation of sodium-phosphate co-transporter (NaPi-IIa) expression at the transcriptional and post-translational levels by inflammatory mediators in acute and chronic colitis. The hypothesis which will be addressed in this specific aim is that in both acute (chemically induced) and chronic (in IL-10-/- mice) colitis, TNFa affects expression and cellular distribution of NaPi-IIa via distinct mechanisms involving protein trafficking and/or degradation and gene transcription. 3. To determine the effects of colitis and the role of TNFa in the regulation of Klotho gene expression. Here, we will investigate in vivo the effects of intestinal inflammation on expression of Klotho mRNA and protein, and characterize its endogenous expression and transcriptional regulation in vitro in immortalized mouse renal epithelial cells of distal convoluted tubules by TNFa. These studies are novel and will certainly lead to a better understanding of the effects of intestinal inflammation on the process of phosphate homeostasis as it relates to bone mineralization in disease states.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37DK033209-30
Application #
8215734
Study Section
Clinical and Integrative Gastrointestinal Pathobiology Study Section (CIGP)
Program Officer
Grey, Michael J
Project Start
1983-12-01
Project End
2013-03-14
Budget Start
2012-02-01
Budget End
2013-03-14
Support Year
30
Fiscal Year
2012
Total Cost
$314,489
Indirect Cost
$106,218
Name
University of Arizona
Department
Pediatrics
Type
Schools of Medicine
DUNS #
806345617
City
Tucson
State
AZ
Country
United States
Zip Code
85721
Larmonier, Claire B; Shehab, Kareem W; Laubitz, Daniel et al. (2016) Transcriptional Reprogramming and Resistance to Colonic Mucosal Injury in Poly(ADP-ribose) Polymerase 1 (PARP1)-deficient Mice. J Biol Chem 291:8918-30
Radhakrishnan, Vijayababu M; Kojs, Pawel; Ramalingam, Rajalakshmy et al. (2015) Experimental colitis is associated with transcriptional inhibition of Na+/Ca2+ exchanger isoform 1 (NCX1) expression by interferon γ in the renal distal convoluted tubules. J Biol Chem 290:8964-74
Ghishan, Fayez K; Kiela, Pawel R (2014) Epithelial transport in inflammatory bowel diseases. Inflamm Bowel Dis 20:1099-109
Radhakrishnan, Vijayababu M; Ramalingam, Rajalakshmy; Larmonier, Claire B et al. (2013) Post-translational loss of renal TRPV5 calcium channel expression, Ca(2+) wasting, and bone loss in experimental colitis. Gastroenterology 145:613-24
Larmonier, C B; McFadden, R-M T; Hill, F M et al. (2013) High vitamin D3 diet administered during active colitis negatively affects bone metabolism in an adoptive T cell transfer model. Am J Physiol Gastrointest Liver Physiol 305:G35-46
Ghishan, Fayez K; Kiela, Pawel R (2012) Small intestinal ion transport. Curr Opin Gastroenterol 28:130-4
Ghishan, Fayez K; Kiela, Pawel R (2011) Advances in the understanding of mineral and bone metabolism in inflammatory bowel diseases. Am J Physiol Gastrointest Liver Physiol 300:G191-201
Kiela, Pawel R (2010) Unraveling the pathophysiology of alcohol-induced thiamin deficiency. Am J Physiol Renal Physiol 299:F26-7
Majewski, Pawel M; Thurston, Robert D; Ramalingam, Rajalakshmy et al. (2010) Cooperative role of NF-{kappa}B and poly(ADP-ribose) polymerase 1 (PARP-1) in the TNF-induced inhibition of PHEX expression in osteoblasts. J Biol Chem 285:34828-38
Thurston, Robert D; Larmonier, Claire B; Majewski, Pawel M et al. (2010) Tumor necrosis factor and interferon-gamma down-regulate Klotho in mice with colitis. Gastroenterology 138:1384-94, 1394.e1-2

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