As phagocytes, retinal pigment epithelial cells function as homeostatic regulators to maintain photoreceptor integrity and preserve visual function. Thus it is advantageous for the post-mitotic RPE cell to utilize catabolic pathways that result in degradation of phagocytosed cargo that under stress can handle ingestion of oxidized toxic substrates. We propose that phagosome maturation in the RPE includes LC3 lipidation in an LC3 associated phagocytic (LAP) process. We hypothesize that these processes are critical in RPE lipid homeostasis by providing a docasahexaenoic acid (DHA; 22:6) pool for Neuroprotectin D1 (NPD1) synthesis to promote RPE cell survival and in turn sustain photoreceptor integrity.
In specific aim 1 we will test the hypothesis that RPE phagosomes annex components of the autophagy pathway in LAP and assess the contribution of oxidized outer segments (OS) to this process.
In specific aim 2 we hypothesize that melanoregulin (MREG) mediated degradation involves LAP and will determine the role of MREG, an LC3 binding partner, in this process.
In Specific Aim 3, we focus our studies on understanding the protective role of LAP and MREG mediated degradation of OS lipids in maintaining RPE health. These studies will provide the ground-work for understanding how deficiencies in LAP dependent processes contribute to disease progression.

Public Health Relevance

As phagocytes, retinal pigment epithelial cells function as a homeostatic regulator to maintain photoreceptor integrity and preserve visual function. Impairment of photoreceptor clearance contributes to age-related retinopathies characterized by sub-retinal deposits as well as lipofuscinosis ultimately leading to blindness. In these studies we will investigate a novel degradative pathway utilized by the retinal pigment epithelia to digest toxic debris and provide precursors for neuroprotection. This pathway called, LC3 associated phagocytosis (LAP), utilizes components of two different albeit interconnected digestive processes, autophagy and phagocytosis. Critical for degradation by this pathway is the protein melanoregulin (MREG). Understanding how the retinal pigment epithelia recognizes debris to be degraded by LAP and the role of MREG in this process will allow us to develop new therapeutic approaches to enhancing degradation in disease progression.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY010420-18
Application #
8798656
Study Section
Special Emphasis Panel (ZRG1-CB-G (02))
Program Officer
Neuhold, Lisa
Project Start
1994-08-01
Project End
2017-01-31
Budget Start
2015-02-01
Budget End
2016-01-31
Support Year
18
Fiscal Year
2015
Total Cost
$457,739
Indirect Cost
$157,503
Name
University of Pennsylvania
Department
Biochemistry
Type
Schools of Dentistry
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Guziewicz, Karina E; McTish, Emily; Dufour, Valerie L et al. (2018) Underdeveloped RPE Apical Domain Underlies Lesion Formation in Canine Bestrophinopathies. Adv Exp Med Biol 1074:309-315
Dhingra, Anuradha; Bell, Brent A; Peachey, Neal S et al. (2018) Microtubule-Associated Protein 1 Light Chain 3B, (LC3B) Is Necessary to Maintain Lipid-Mediated Homeostasis in the Retinal Pigment Epithelium. Front Cell Neurosci 12:351
Dhingra, Anuradha; Alexander, Desiree; Reyes-Reveles, Juan et al. (2018) Microtubule-Associated Protein 1 Light Chain 3 (LC3) Isoforms in RPE and Retina. Adv Exp Med Biol 1074:609-616
Reyes-Reveles, Juan; Dhingra, Anuradha; Alexander, Desiree et al. (2017) Phagocytosis-dependent ketogenesis in retinal pigment epithelium. J Biol Chem 292:8038-8047
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Frost, Laura S; Lopes, Vanda S; Bragin, Alvina et al. (2015) The Contribution of Melanoregulin to Microtubule-Associated Protein 1 Light Chain 3 (LC3) Associated Phagocytosis in Retinal Pigment Epithelium. Mol Neurobiol 52:1135-1151
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Guha, Sonia; Coffey, Erin E; Lu, Wennan et al. (2014) Approaches for detecting lysosomal alkalinization and impaired degradation in fresh and cultured RPE cells: evidence for a role in retinal degenerations. Exp Eye Res 126:68-76
Brown, Angela C; Balashova, Nataliya V; Epand, Richard M et al. (2013) Aggregatibacter actinomycetemcomitans leukotoxin utilizes a cholesterol recognition/amino acid consensus site for membrane association. J Biol Chem 288:23607-21

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