The objecfive is to study the funcfion of sulfated glycosaminoglycans (GAGs) in infiammation, with particular emphasis on the sulfafion pattern ofthe heparan, chondroitin and dermatan sulfate. We have successfully created conditional mouse mutants altered in heparan sulfate biosynthesis and showed that altering overall sulfation diminishes infiammatory responses in the context of neutrophil rolling and infiltrafion. In contrast, altering heparan sulfate in leukocytes does not affect these processes and has only mild effects on adaptive immune responses. In this proposal, we plan to complete our studies of endothelial heparan sulfate by examining the impact of altered uronic acid 2-0-sulfafion and glucosamine 6-O-sulfation using mice with conditional alleles in these enzymes, and then extend this analysis to chondroitin-4-sulfate and dermatan sulfate. Because we anticipate that systemic mutations in chondroitin/dermatan sulfate biosynthesis will result in embryonic or perinatal lethality, we are preparing conditional alleles ofthe target genes and will alter their expression selectively in endothelial cells and myeloid cells. Specifically, we propose to: (1) Create mouse lines with conditional alleles of chondroifin polymerase-1, chondroifin 4-0-sulfotransferase-1, chondroitin 4-0-sulfotransferase-2 , and dermatan 4-0-sulfotransferase and in collaboration with Project 1 a conditional targeting construct to express Hyal4, an endo-N-acetylgalactosaminidase that can degrade chondroifin;(2) Examine the role of GAGs in leukocyte rolling, firm adhesion, and diapedesis in vitro using isolated endothelial cells derived from the mutants;(3) In collaboration with Projects 2 and 4, examine the in vivo consequences of altering endothelial GAGs on inflammafion induced chemically, sterilely and by bacterial infecfion;and (4) Examine the consequences of altering endothelial glycosaminoglycans on vascular permeability. By focusing on inflammatory reacfions that take place in the vasculature, the proposed studies build on a strong base of preliminary data, which together validate GAGs as potential targets for treafing chronic inflammation and ischemic disease. Addifionally, the project will provide model organisms for other invesfigators interested in the physiological function of glycosaminoglycans in other organ systems.

Public Health Relevance

Inflammation is the body's response to injury and infection. The central hypothesis of Project 3 is that glycosaminoglycans affects one or more steps in the inflammatory response and that their function can be ascertained by geneticaly manipulating their biosynthesis in endothelial cells, neutrophils, and macrophages.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL107150-04
Application #
8669098
Study Section
Special Emphasis Panel (ZHL1-CSR-H)
Project Start
Project End
Budget Start
2014-06-01
Budget End
2015-05-31
Support Year
4
Fiscal Year
2014
Total Cost
$243,446
Indirect Cost
$86,011
Name
University of California San Diego
Department
Type
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Gordts, Philip L S M; Nock, Ryan; Son, Ni-Huiping et al. (2016) ApoC-III inhibits clearance of triglyceride-rich lipoproteins through LDL family receptors. J Clin Invest 126:2855-66
Varki, Ajit (2016) Biological Roles of Glycans. Glycobiology :
Yamaguchi, Masaya; Hirose, Yujiro; Nakata, Masanobu et al. (2016) Evolutionary inactivation of a sialidase in group B Streptococcus. Sci Rep 6:28852
Secundino, Ismael; Lizcano, Anel; Roupé, K Markus et al. (2016) Host and pathogen hyaluronan signal through human siglec-9 to suppress neutrophil activation. J Mol Med (Berl) 94:219-33
Thacker, Bryan E; Seamen, Emylie; Lawrence, Roger et al. (2016) Expanding the 3-O-Sulfate Proteome--Enhanced Binding of Neuropilin-1 to 3-O-Sulfated Heparan Sulfate Modulates Its Activity. ACS Chem Biol 11:971-80
Zhang, Ling-Juan; Sen, George L; Ward, Nicole L et al. (2016) Antimicrobial Peptide LL37 and MAVS Signaling Drive Interferon-β Production by Epidermal Keratinocytes during Skin Injury. Immunity 45:119-30
Schwarz, Flavio; Springer, Stevan A; Altheide, Tasha K et al. (2016) Human-specific derived alleles of CD33 and other genes protect against postreproductive cognitive decline. Proc Natl Acad Sci U S A 113:74-9
Sato, Emi; Muto, Jun; Zhang, Ling-Juan et al. (2016) The Parathyroid Hormone Second Receptor PTH2R and its Ligand Tuberoinfundibular Peptide of 39 Residues TIP39 Regulate Intracellular Calcium and Influence Keratinocyte Differentiation. J Invest Dermatol 136:1449-59
Miles, L A; Baik, N; Lighvani, S et al. (2016) Deficiency of Plasminogen Receptor, Plg-RKT, Causes Defects in Plasminogen Binding and Inflammatory Macrophage Recruitment in vivo. J Thromb Haemost :
Zaiss, Anne K; Foley, Erin M; Lawrence, Roger et al. (2016) Hepatocyte Heparan Sulfate Is Required for Adeno-Associated Virus 2 but Dispensable for Adenovirus 5 Liver Transduction In Vivo. J Virol 90:412-20

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