This competitive renewal focuses on the regulation of ocular surface inflammation by lumican, an abundant extracellular matrix (ECM) protein at the ocular surface. Eight million people worldwide suffer from infections, allergies and autoimmune conditions leading to corneal scarring and blindness. The ECM has a significant role in modulating the local inflammatory milieu, while its breakdown can present immune cells with endogenous danger signals to drive excessive immune responses. However, the precise molecular mechanisms of ECM- immune interactions are not understood. In its most abundant form lumican is a structural protein that regulates collagen architecture to generate a transparent cornea. Recently, we identified its N-terminal domain to interact with leukocyte surfaces to promote toll-like receptor 4 (TLR4) mediated host response to gram-negative bacterial lipopolysaccharide (LPS) endotoxins. In a remodeling ECM during infections and inflammation, the immune-interactions of lumican can be a double-edged sword - needed for development of protective innate immunity, but unrestricted can drive chronic inflammation. Our current goals are to gain molecular insights into lumican - innate immune cell interactions, to develop strategies to strengthen protective immunity and diminish immune dysregulation using lumican-based peptides and antibodies. As our in vivo inflammatory setting we will use LPS and Pseudomonas aeruginosa (PA) keratitis models in wild type and lumican-deficient mice. Additional recombinant lumican variants, peptides, anti-lumican antibodies and cell cultures models have all been developed in the laboratory for this study. The following hypotheses will be tested in three aims.
Aim 1 : Hypothesis - Lumican promotes immune response by increasing pericellular LPS or modulates lipid rafts to enhance TLR4 recruitment to the cell surface. Lipid rafts are centrally important in innate immune signal transduction and clinically relevant as targets of therapy. We will use primary macrophages, cell lines, recombinant lumican, biochemical, confocal and FRET analyses to address lumican involvements at the cell surface.
Aim 2 : Hypothesis - The immunologically active N-terminal domain is unavailable for immune cell interactions in collagen-associated lumican. We will use recombinant lumican and three variants that either have the N-terminal domain or the central collagen binding domain to test this hypothesis. Collagen-pretreated lumican variants will be tested for their abilities to induced LPS response in culture. Secondly, the variants will be transiently expressed at the ocular surface of wild type and lumican deficient mice, and challenged with keratitis to address their role in modulating the cytokine milieu, innate and adaptive immune cells and ocular surface inflammation.
Aim 3 : Hypothesis - Ocular surface inflammation can be modulated by lumican-based peptides or anti-lumican antibodies. We will test if an antibody against the N- terminal domain or mimetic peptides can modulate lumican-immune cell interactions to ameliorate inflammation in the keratitis models to develop the premises for localized treatments of inflammation.
Most treatments of inflammation due to infections autoimmune dry eye diseases involve strong antimicrobial agents or immunosuppressants with harmful side effects. Our findings on immunomodulatory roles of lumican will lead to future alternative strategies for milder localized treatments of inflammation.
|Frikeche, Jihane; Maiti, George; Chakravarti, Shukti (2016) Small leucine-rich repeat proteoglycans in corneal inflammation and wound healing. Exp Eye Res 151:142-9|
|Stasiak, Marta; Boncela, Joanna; Perreau, Corinne et al. (2016) Lumican Inhibits SNAIL-Induced Melanoma Cell Migration Specifically by Blocking MMP-14 Activity. PLoS One 11:e0150226|
|Gowda, Ranjita N; Redfern, Rachel; Frikeche, Jihane et al. (2015) Functions of Peptidoglycan Recognition Proteins (Pglyrps) at the Ocular Surface: Bacterial Keratitis in Gene-Targeted Mice Deficient in Pglyrp-2, -3 and -4. PLoS One 10:e0137129|
|Hultgårdh-Nilsson, A; Borén, J; Chakravarti, S (2015) The small leucine-rich repeat proteoglycans in tissue repair and atherosclerosis. J Intern Med 278:447-61|
|Dupuis, Loren E; Berger, Matthew G; Feldman, Samuel et al. (2015) Lumican deficiency results in cardiomyocyte hypertrophy with altered collagen assembly. J Mol Cell Cardiol 84:70-80|
|Steinhart, Matthew R; Cone-Kimball, Elizabeth; Nguyen, Cathy et al. (2014) Susceptibility to glaucoma damage related to age and connective tissue mutations in mice. Exp Eye Res 119:54-60|
|Foster, James; Wu, Wai-Hong; Scott, Sherri-Gae et al. (2014) Transforming growth factor ? and insulin signal changes in stromal fibroblasts of individual keratoconus patients. PLoS One 9:e106556|
|Chen, Shoujun; Young, Marian F; Chakravarti, Shukti et al. (2014) Interclass small leucine-rich repeat proteoglycan interactions regulate collagen fibrillogenesis and corneal stromal assembly. Matrix Biol 35:103-11|
|Chaerkady, Raghothama; Shao, Hanjuan; Scott, Sherri-Gae et al. (2013) The keratoconus corneal proteome: loss of epithelial integrity and stromal degeneration. J Proteomics 87:122-31|
|Shao, Hanjuan; Scott, Sherri-Gae; Nakata, Chiaki et al. (2013) Extracellular matrix protein lumican promotes clearance and resolution of Pseudomonas aeruginosa keratitis in a mouse model. PLoS One 8:e54765|
Showing the most recent 10 out of 38 publications