# Elucidation of cellular reprogramming processes that drive lens regeneration in axolotl as a basis for future therapeutic approaches

> **NIH NIH K99** · HARVARD UNIVERSITY · 2020 · $128,574

## Abstract

The goal of the project is to harness the intrinsic ability of axolotl salamanders to regenerate ocular tissues and
unravel natural ways to improve human stem cell function for use in regenerative medicine. Axolotl’s ability to
regenerate the retina and the lens stems from the pigmented epithelium, which reprograms its cellular state
upon injury. To understand the mechanism of regeneration I will focus on iris pigmented epithelial (IPE) cells
and their capacity to reprogram to lens epithelial cells. This process is age-dependent in axolotls because
regeneration can only occur for a short time window of two weeks after hatching and is lost thereafter. This
allows me to both identify and functionally characterize the potency of candidate molecules to induce
regenerative responses in the same context. Studying axolotl IPE reprogramming will provide important insight
into how cellular plasticity has naturally evolved in organisms as a mean to regenerate lost organs. Modern
regenerative medicine can learn from this intrinsic regenerative mechanism and apply it to coax
reprogramming of any adult cell in a safer and more efficient way. In addition, its application could lead to
groundbreaking treatments for blind patients by inducing ocular regeneration. In Specific Aim 1, I will study the
role of Eya2 during IPE reprogramming. My preliminary data indicate that Eya2 is required for progenitor cell
survival upon genotoxic stress during regeneration. I will also study Eya2’s ability to facilitate IPE
reprogramming by regulating the activity of transcription factors required to maintain specific eye-tissue fates.
In Specific Aim 2, I will generate a transgenic axolotl model that will allow tracking of lens regeneration by
fluorescence in vivo. This transgenic animal will enable me to screen thousands of compounds in a high-
throughput manner for their ability to either inhibit or induce regeneration from the IPE. In Specific Aim 3, I will
study the cellular heterogeneity of the IPE with single cell RNA-seq. Pseudotime lineage trace analysis will be
used to identify whether an IPE-residing progenitor population is present before injury and if it is responsible for
regeneration. The in silico identification of a regeneration-poised IPE subpopulation will be further validated
and characterized using the gold-standard CreERT2/loxP system in which the recombinase will be targeted in
progenitor cell-specific genetic loci. My long-term career goal is to lead a research laboratory with central aim
to bridge the regenerative ability gap between salamanders and mammals. My experience working with newts,
axolotls and mice, in addition to the data I will acquire during the award training period in the Whited laboratory
at Brigham and Women’s Hospital and Harvard Medical School, will enable me to build a foundation for pilot
translational experiments in mammals in the near future. My access to and participation in a plethora of
meetings, journal clubs, seminars and wor...

## Key facts

- **NIH application ID:** 9918425
- **Project number:** 5K99EY029361-02
- **Recipient organization:** HARVARD UNIVERSITY
- **Principal Investigator:** Konstantinos Sousounis
- **Activity code:** K99 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $128,574
- **Award type:** 5
- **Project period:** 2019-05-01 → 2022-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9918425

## Citation

> US National Institutes of Health, RePORTER application 9918425, Elucidation of cellular reprogramming processes that drive lens regeneration in axolotl as a basis for future therapeutic approaches (5K99EY029361-02). Retrieved via AI Analytics 2026-06-01 from https://api.ai-analytics.org/grant/nih/9918425. Licensed CC0.

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