We lack a complete understanding of the molecular mechanisms that drive the development of the lens fibrotic
disease, Posterior Capsule Opacification (PCO) in response to cataract surgery wounding. Our preliminary data
indicate a role for mitochondrial remodeling in driving lens fibrosis. Importantly, whether mitochondria play an
integral role in the development of PCO is not known. Mitochondria are recognized as central coordinators of
cell processes, as not only a major source of ATP for cells, but also through production of metabolites and
reactive oxygen species (ROS) that can impact cell signaling. The arrangement of mitochondria within a cell is
coordinated by fusion events and by fission to fragment mitochondria into transportable units that can travel
along microtubules, an efficient mechanism to provide energy and metabolic needs in a highly localized and
specific manner. Our unbiased transcriptome findings with a clinically relevant PCO model, provide strong
evidence that fibrosis is associated with gene reprogramming around remodeling mitochondrial function, and
metabolic rewiring that favors glycolysis and glutaminolysis. Our findings also support that acquisition to a lens
fibrotic phenotype is associated with an increase in mtROS, mitochondrial fission and mitochondrial trafficking
along microtubule cytoskeletal elements. Whether PCO development depends on this remodeling of
mitochondrial function, including elevated mtROS is not known nor do we know whether PCO development
depends on the repositioning of mitochondria within the cell to support the pro-fibrotic phenotype. The objective
of this proposal is to deliver new molecular insight into the role of mitochondrial remodeling in driving lens fibrotic
disease. The central hypothesis is that mitochondria become remodeled through changes in fission/fusion
dynamics and substrate metabolism to drive the acquisition of a pro-fibrotic lens phenotype. The central
hypothesis will be evaluated – in a clinically relevant model of PCO, as well as human lens cells and explants
derived from human cataract surgery patients - by the following two specific aims: 1) To determine how
mitochondrial substrate oxidation impacts mitochondrial ROS production and fibrosis development and 2) To
investigate whether modulating mitochondrial fission/fusion dynamics can prevent acquisition to a lens fibrotic
phenotype. These aims will be pursued utilizing cutting-edge techniques to investigate mitochondrial
bioenergetics and glycolytic flux, a combination of high-resolution confocal microscopy, time-lapse microscopy,
and a toolkit of small molecules that specifically target different mechanisms to effect changes in mitochondrial
form and function and thus enable mechanistic insights. The proposed research is significant as these studies
will provide a new comprehensive understanding about the role of mitochondrial form and function in driving lens
fibrosis. The long-term goal is to use this new understanding about mitochondrial remodeling as a foundational
resource to create new mitochondrial-targeted treatment strategies for preventing PCO.