# Defining bacterial members of the ocular surface microbiome and assessing stability over time

> **NIH NIH U24** · UNIVERSITY OF PITTSBURGH AT PITTSBURGH · 2024 · $1,096,264

## Abstract

PROJECT SUMMARY
Recent studies have shown data suggesting that the ocular microbiome exists and influences ocular surface
health and disease. Despite this defining the “core” components of the ocular surface microbiome has been
difficult largely due to the low biomass nature of the eye, a lack of robust data sets, reliance only on next
generation sequencing (NGS), and the inability to show causal relationships between microbes and host
physiology. Herein, we have outlined a proposal that directly addresses these knowledge gaps and that will lay
the foundation for future studies aimed at investigating how the ocular surface microbiome influences health and
disease. We have recruited a multidisciplinary research team that consists of experts in clinical and basic
ophthalmology research, microbiome of low biomass sites, and immunology. With this team, we plan to first
characterize the healthy human ocular microbiome in two geographic locations (Pittsburgh and Miami)
longitudinally (0, 1 week, 3 months) using molecular techniques (DNA and RNA sequencing) and culturomics.
While molecular techniques will provide us with the broadest consortia of bacteria, culturomics will provide us
with viable bacteria that we plan to bank in the Campbell Laboratory, so that bacteria from healthy human eyes
can be a resource for the community in future studies. Next, because immunity is normally generated towards
components of the microbiome that remain associated with the host for extended periods of time, we plan to use
human immune responses directed against ocular bacteria to distinguish colonizing bacteria from transient
bacteria that are likely washed away. To further refine the understanding of ocular colonizing bacteria, we plan
to inoculate germ free (GF) and specific pathogen free (SPF) mice with human ocular bacteria. After an extended
period of time, we plan to re-isolate bacteria from the eyes of mice. Bacteria that are still present on the ocular
surface will be considered to have a higher likelihood of being an ocular surface colonizer in humans. The
development of a model such as this will allow others to assess the colonizing ability of other bacteria that may
be considered a core component of the ocular surface microbiome. In sum, data from our proposal will be a
resource for the community at large because we will have: 1) generated a robust set of ocular microbiome
sequencing data that will provide information on the stability and consistency of ocular microbiome signatures,
2) created a bank of bacteria that were acquired from healthy human eyes, 3) defined protocols to use human
immunity to measure the colonizing ability of bacteria, 4) developed an in vivo animal model to assess how
ocular bacteria can be inoculated and re-isolated from the eye. These resources will be free to use for the
community and will act as a base to further investigate how the ocular microbiome influences ocular surface
health and disease.

## Key facts

- **NIH application ID:** 10874789
- **Project number:** 5U24EY035102-02
- **Recipient organization:** UNIVERSITY OF PITTSBURGH AT PITTSBURGH
- **Principal Investigator:** Anat Galor
- **Activity code:** U24 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $1,096,264
- **Award type:** 5
- **Project period:** 2023-07-01 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10874789

## Citation

> US National Institutes of Health, RePORTER application 10874789, Defining bacterial members of the ocular surface microbiome and assessing stability over time (5U24EY035102-02). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/10874789. Licensed CC0.

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