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Principal Investigator: Preethi Ganapathy
Organization: UPSTATE MEDICAL UNIVERSITY
Fiscal Year: 2024
Award: $251,295
Funding agency: National Eye Institute
PROJECT SUMMARY/ABSTRACT
Neuronal damage in glaucoma occurs following pathologic mechanical strains and fibrosis of
the optic nerve head (ONH). Despite an abundance of evidence implicating mechanical strain
and matrix fibrosis in glaucomatous damage, it is not known to what extent and how ONH cells
signal to each other to produce matrix fibrosis in response to mechanical strains. This is due, in
part, to a lack of adequate model systems to test mechanisms of matrix fibrosis following
mechanical strain. In this exploratory R21, we propose to overcome limitations of existing
models by establishing an in vitro 3D culture system that (a) accounts for native tissue stiffness,
(b) permits dynamic matrix remodeling, (c) includes human ONH astrocytes and lamina cribrosa
(LC) cells, and (d) allows for targeted manipulation of cell type-specific molecular pathways. We
will then use this system to examine fibrotic signaling pathways between ONH astrocytes and
LC cells in response to mechanical strain. Specifically, our preliminary data have identified the
extra domain A isoform of fibronectin (FN+EDA) – Toll-Like Receptor 4 (TLR4) signaling axis as
critical to matrix deposition by ONH cells. We will use our model system to test the role of the
FN+EDA-TLR4 signaling axis in mechanical strain-induced matrix fibrosis. The experiments
proposed in this R21 will advance our understanding of ONH mechanobiology by (i)
investigating whether crosstalk between ONH astrocytes and LC cells is critical to
mechanodysfunction, (ii) establishing quantitative models to test mechanisms of mechanical
strain-induced matrix fibrosis, and (iii) investigating the potential of ONH matrix proteins to act
as key signaling molecules to enhance ONH fibrosis. Our specific aims are:
Aim 1, Generate quantitative models of the fibrotic response of human ONH astrocytes and LC
cells after mechanical strain, in monocultures and co-cultures.
Aim 2, Test whether astrocyte-derived FN+EDA stimulation of LC cell TLR4 promotes
mechanical strain-induced matrix fibrosis.
Terms: <3-D><3-D modeling><3-Dimensional><3D><3D cell culture><3D culture><3D modeling><Address><Affect><Astrocytes><Astrocytus><Astroglia><Binding><Biologic Models><Biological Models><Bioreactors><Blindness><Body Tissues><Cell Body><Cell Communication and Signaling><Cell Signaling><Cells><Co-culture><Cocultivation><Coculture><Coculture Techniques><Cold-Insoluble Globulins><Collagen IV><Collagen Type IV><Cranial Nerve II><Custom><Data><Deposit><Deposition><Development><Dysfunction><Encapsulated><FN1><Fibronectin 1><Fibronectins><Fibrosis><Functional disorder><Future><Glaucoma><Goals><Homolog of Drosophila TOLL><Human><Hydrogels><In Vitro><Intracellular Communication and Signaling><Isoforms><Knowledge><LETS Proteins><Large External Transformation-Sensitive Protein><Measures><Mechanics><Model System><Modeling><Modern Man><Molecular><Molecular Interaction><Nerve Cells><Nerve Unit><Neural Cell><Neurocyte><Neurons><Opsonic Glycoprotein><Opsonic alpha(2)SB Glycoprotein><Optic Disk><Optic Nerve><Optic Nerve Head><Optic Papilla><Pathologic><Pathway interactions><Physiologic><Physiological><Physiopathology><Primates><Primates Mammals><Production><Protein Isoforms><Proteins><Rodent><Rodent Model><Rodentia><Rodents Mammals><Role><Second Cranial Nerve><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Signaling Molecule><System><TLR4><TLR4 gene><Testing><Time><Tissues><Toll Homologue><Transforming Growth Factors><Tumor Growth Factors><Type IV (Basement Membrane) Collagen><Work><alpha 2-Surface Binding Glycoprotein><astrocytic glia><biological signal transduction><cell type><customs><developmental><experiment><experimental research><experimental study><experiments><glaucomatous><in vivo><in vivo Model><knock-down><knockdown><mechanic><mechanical><microbioreactor><nerve damage><neural><neuronal><pathophysiology><pathway><pharmacologic><profibrotic cytokine><response><social role><three dimensional><three dimensional cell culture><three-dimensional modeling><toll-like receptor 4><transforming growth factors Animal growth regulators><vision loss><visual loss>