# The Roles of Autophagy in Limbal/Corneal Epithelia

> **NIH NIH R01** · NORTHWESTERN UNIVERSITY · 2024 · $481,939

## Abstract

PROJECT SUMMARY/ABSTRACT
The long-term goal of this project is to understand the functions of selective autophagy in corneal inflammation
as well as corneal/limbal epithelial differentiation. Selective autophagy is a specialized form of autophagy and
can control signaling pathways in cells by specifically degrading key components of these pathways. Even
though our studies have advanced our understanding on the roles of non selective autophagy in corneal/limbal
epithelia, the roles of selective autophagy in corneal inflammation and corneal/limbal epithelial differentiation
remain unclear. Single-cell RNA-sequencing (scRNA-seq) methodologies enable the interrogation of relatively
rare cell populations (e.g., stem and early transit amplifying (TA) cells). This technology also allows the
comparison of different physiological and/or disease states. Thus, we took advantage of Beclin1+/- (Beclin1 het)
mice, a well-established mouse model with compromised autophagy and conducted a scRNA-seq analysis in
the ocular anterior segmental tissue from wild-type (WT) and beclin1 het mice. scRNA-seq data suggest that
autophagy plays a significant role in response to corneal inflammation, epithelial differentiation, and the
regulation of Notch signaling. Our preliminary data show that genetic attenuation of autophagy enhances
corneal inflammation. Interestingly, TRAF2, a positive regulator of inflammation, is regulated by NDP52-
mediated selective autophagy. Furthermore, we have demonstrated that attenuation of p62-mediated selective
autophagy markedly increased: (i) stem/TA cell enriched human limbal epithelial cell (HLEC) differentiation;
and (ii) Notch intracellular domain (Notch ICD, an active form of Notch) in HLECs in a proteosome independent
way. The scRNA-seq data also suggested that Wdfy1, which is preferentially expressed in the mouse limbal
epithelial basal layer, was positively regulated by autophagy. Knockdown of Wdfy1 enhanced HLEC
differentiation. Therefore, we will question if: (i) NDP52-mediated selective autophagy degrades TRAF2 and
consequently has an inhibitory role in the inflammatory response in both corneal epithelial cells and
macrophages (Aim1); and (ii) downregulation of p62-mediated selective autophagy allows for limbal epithelial
differentiation via increasing Notch ICD (active form) and thus decreasing Wdfy1 expression (Aim2). To
accomplish these goals, we will capitalize on our ability to conduct gain- and loss-of-function studies of
selective autophagy-related genes in submerged and 3D organotypic raft cultures of primary limbal/corneal
epithelial cells as well as mice. Knowledge gained from this study will provide molecular insights into how
selective autophagy regulates corneal inflammation and limbal epithelial differentiation. A better understanding
of the physiological importance of selective autophagy in corneal inflammation and differentiation will translate
into the development of novel therapies by targeting selective autopha...

## Key facts

- **NIH application ID:** 10802075
- **Project number:** 2R01EY028560-06A1
- **Recipient organization:** NORTHWESTERN UNIVERSITY
- **Principal Investigator:** Han Peng
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $481,939
- **Award type:** 2
- **Project period:** 2018-05-01 → 2029-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10802075

## Citation

> US National Institutes of Health, RePORTER application 10802075, The Roles of Autophagy in Limbal/Corneal Epithelia (2R01EY028560-06A1). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10802075. Licensed CC0.

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