The long-term objective of this grant is to identify how to modulate ?v?3 integrin signaling pathways in order to develop therapeutic targets to control intraocular pressure (IOP) in glaucoma. The glaucomas, which lead to irreversible loss of retinal ganglion cells, affect approximately 67 million people worldwide. They are commonly associated with elevated levels of intraocular pressure (IOP) due to a reduction in aqueous humor outflow from the trabecular meshwork (TM). One of the major risk factors that has emerged as an important regulatory mechanism for outflow facility is the actin cytoskeleton. It controls a number of key biological processes involved in maintaining normal outflow facility including contractility, phagocytosis, and deposition of the extracellular matrix. Integrins play a central role in regulating these cytoskeleton-mediated activities and our studies suggest that dysregulation of the ?v?3 integrin causes the major phenotypic changes associated with glaucoma including decreased phagocytosis, increased extracellular matrix deposition, CLAN formation and an elevation in IOP. We propose that this integrin is activated in glaucoma by elevated levels of TGF?2 or following treatments with glucocorticoids, like dexamethasone (DEX). In this grant, we plan to use RNAseq studies to identify the factors upregulated by DEX or TGF?2 that activate ?v?3 integrin and proximity ligation assays to determine if these factors are associated with the integrin adhesome. We also plan to demonstrate that an NRON/NFAT complex controls DEX-induced activation of ?v?3 integrin and the secondary glucocorticoid response involved in steroid-induced glaucoma. Finally, we plan to show that the responses to DEX (ECM formation, IOP elevation, and outflow facility) are affected by the activated state of ?v?3 integrin in vivo, not just its expression level. To test this last hypothesis we plan to use a tamoxifen inducible CreERcag-?3 integrinflox/flox mouse to knock down ?v?3 integrin expression in the mouse TM. Adenoviral (Ad5) vectors expressing 3 different activated states of ?v?3 integrin (wildtype, inactive and constitutively active) will be used to alter the activity levels of ?v?3 integrins in vivo. The proposed studies are the first to demonstrate that changes in a specific integrin signaling pathway can affect IOP, outflow facility, and ECM formation in vivo. They will enhance our understanding of how integrin signaling events are controlled in the TM and how this affects the cytoskeletal events (ECM deposition, cell contractility and phagocytosis) that regulate outflow facility. Understanding how integrins contribute to the regulation of these processes is important because it will enable us to provide new therapeutic targets to regulate the cytoskeleton in order to restore homeostasis and decrease IOP.

Public Health Relevance

Glaucoma is the second most common cause of blindness in the U.S. and the most common cause of blindness among African-Americans. An elevation in intraocular pressure caused by the restriction of aqueous humor outflow through the trabecular meshwork is a common risk factor. The goal of this project is to identify signaling pathways that contribute to the restriction in outflow facility and determine if inhibiting these mechanisms can be used as potential therapies to increase outflow facility.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY017006-12
Application #
10085164
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Liberman, Ellen S
Project Start
2006-09-15
Project End
2024-12-31
Budget Start
2020-01-01
Budget End
2021-12-31
Support Year
12
Fiscal Year
2021
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Pathology
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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Filla, Mark S; Dimeo, Kaylee D; Tong, Tiegang et al. (2017) Disruption of fibronectin matrix affects type IV collagen, fibrillin and laminin deposition into extracellular matrix of human trabecular meshwork (HTM) cells. Exp Eye Res 165:7-19
Filla, Mark S; Faralli, Jennifer A; Peotter, Jennifer L et al. (2017) The role of integrins in glaucoma. Exp Eye Res 158:124-136
Peotter, Jennifer L; Phillips, Jenny; Tong, Tiegang et al. (2016) Involvement of Tiam1, RhoG and ELMO2/ILK in Rac1-mediated phagocytosis in human trabecular meshwork cells. Exp Cell Res 347:301-11
Itakura, Tatsuo; Peters, Donna M; Fini, M Elizabeth (2015) Glaucomatous MYOC mutations activate the IL-1/NF-?B inflammatory stress response and the glaucoma marker SELE in trabecular meshwork cells. Mol Vis 21:1071-84
Faralli, Jennifer A; Clark, Ross W; Filla, Mark S et al. (2015) NFATc1 activity regulates the expression of myocilin induced by dexamethasone. Exp Eye Res 130:9-16
Lee, Eun Suk; Rasmussen, Carol A; Filla, Mark S et al. (2014) Prospects for lentiviral vector mediated prostaglandin F synthase gene delivery in monkey eyes in vivo. Curr Eye Res 39:859-70
Gagen, Debjani; Faralli, Jennifer A; Filla, Mark S et al. (2014) The role of integrins in the trabecular meshwork. J Ocul Pharmacol Ther 30:110-20
Filla, Mark S; Clark, Ross; Peters, Donna M (2014) A syndecan-4 binding peptide derived from laminin 5 uses a novel PKC? pathway to induce cross-linked actin network (CLAN) formation in human trabecular meshwork (HTM) cells. Exp Cell Res 327:171-82

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