# Endocytosis in Adenoviral Ocular Pathogenesis

> **NIH NIH R01** · MASSACHUSETTS EYE AND EAR INFIRMARY · 2020 · $425,000

## Abstract

Project Summary
Corneal disease is one of the major causes of blindness. Corneal infections include epidemic
keratoconjunctivitis (EKC), caused by human adenoviruses (HAdV). EKC is characterized by a severe,
acute keratoconjunctivitis. HAdV species D, in particular, types HAdV-D8, 37, 64 (formerly type 19),
and the recently identified 53, 54 and 56, cause EKC, the only adenoviral infection with significant
corneal involvement. Detailed knowledge of the foremost events in infection, viral entry and intracellular
trafficking, are needed to design a specific therapy for the disorder. Our prior work focused on human
corneal fibroblasts, because of their role in corneal inflammation. We now extend our studies to corneal
epithelial cells, the first cells in the cornea to encounter virus. The Specific Aims of this proposal are to
test the hypotheses that 1) adenovirus entry and trafficking in corneal epithelial cells requires clathrin
mediated endocytosis, 2) adenovirus infection of corneal epithelial cells is controlled by dynamin 2, and
3) signaling molecules unique to clathrin vesicles and caveosomes determine adenovirus trafficking in
corneal epithelial cells and fibroblasts, respectively. Viral entry is a complex phenomenon, and while
governed by the tropism of the infecting virus, is also cell type dependent. Our preliminary data in
human corneal epithelial cells suggests a critical role for clathrin mediated endocytosis in HAdV-D37
infection. We will apply chemical inhibitors and siRNA against specific components of each viral entry
pathway to dissect and elucidate the mechanisms of HAdV-D37 entry and trafficking in primary human
corneal epithelial cells. Dynamin 2 shows a paradoxically negative effect on HAdV-D37 entry in corneal
fibroblasts, and preliminary studies suggest a similar effect in corneal epithelial cells. We will test viral
entry when overexpressing various dynamin 2 mutants in order to determine the mechanism by which
dynamin 2 controls infection. Finally, because endosomes are a platform for receptor mediated
signaling distinct from membrane signaling and play an essential role in antiviral responses, we will
purify endosomes from infected and non-infected corneal cells and perform mass spectrometry analysis
to elucidate the specific signalosomes activated by viral infection. The role of validated proteins in viral
trafficking and replication will be tested using inhibitors and siRNA techniques. The proposed studies
will take us closer to our goal of effective information-based therapies against EKC, a common affliction.
Therefore, the proposed study addresses a major public health concern.

## Key facts

- **NIH application ID:** 9933057
- **Project number:** 5R01EY021558-09
- **Recipient organization:** MASSACHUSETTS EYE AND EAR INFIRMARY
- **Principal Investigator:** James Chodosh
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $425,000
- **Award type:** 5
- **Project period:** 2011-09-30 → 2021-09-29

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9933057

## Citation

> US National Institutes of Health, RePORTER application 9933057, Endocytosis in Adenoviral Ocular Pathogenesis (5R01EY021558-09). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/9933057. Licensed CC0.

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