# Developing Remyelination Strategies for Demyelinating Optic Neuropathies Using Human Pluripotent Stem Cells

> **NIH NIH K99** · JOHNS HOPKINS UNIVERSITY · 2021 · $88,947

## Abstract

PROJECT SUMMARY/ABSTRACT
Optic nerve demyelinating diseases such as Optic Neuritis (ON) cause acute reduction of visual acuity and often
chronic visual loss. The estimated incidence of ON in the USA is 6.4/100,000, and 50% of the 2.5 million multiple
sclerosis (MS) patients worldwide are estimated to develop one or more episodes of ON during the course of
their disease. Myelination of the optic nerve is carried out by a specialized cell, the oligodendrocyte (OL), that
coats and protects axons and promotes neural conduction. Demyelination can cause apoptosis of the OLs,
axonal damage, and neuronal cell death. Remyelination-based treatments have the potential to help protect
retinal ganglion cell (RGC) neurons and reduce chronic vision loss, and perhaps help restore lost vision. The
long-term goal of this project is to identify molecular pathways involved in differentiation and maturation of OLs
that can be exploited for promoting remyelination of the demyelinated optic nerve. To this end, the PI has
developed human pluripotent stem cell (hPSC) reporters for RGCs and OLs, and performed single cell RNA-
sequencing and a preliminary high-throughput screen (HTS) to identify regulators of OL differentiation and
maturation. Based upon this preliminary data, the PI proposes to: 1) expand the screening effort to identify
additional compounds and pathways involved in OL maturation, 2) examine the transcription factors and
pathways identified from the preliminary work for their role in OL differentiation, and 3) further identify and
systemically characterize microRNAs (miRNAs) involved in OL specification and maturation. To perform the
proposed studies, the PI will develop innovative tools such as hPSC-reporters for OL differentiation with inducible
CRISPRi (interference) and CRISPRa (activation) system, and a functional in-vitro hOL/hRGC co-culture system
for assessing hRGC myelination by hOLs. In the mentored phase, in Dr. Don Zack’s lab, the PI will carry out
genome-editing to generate the CRISPRi and CRISPRa lines and acquire training at Wilmer’s HCS facility where
small molecule screening to probe for signaling pathways involved in OL maturation will be performed. In the
second year, under the guidance of Dr. Jay Baraban (an expert in microRNA biology), the miRNA studies on
OLs will be initiated. The mentored phase will also be supplemented by training with Ingo Ruczinski (biostatistics
collaborator), who will provide assistance with the pathway analysis and hit validation of HTS. Furthermore, the
PI’s co-mentor, Dr. Peter Calabresi, who is a leading scientist in demyelinating diseases, will regularly meet,
advise and help him prioritize the genes and pathways for further examination. Additionally, during the mentored
phase, the PI will regularly meet with his advisory committee, attend scientific conferences, and continue his
career development. The PI is in an ideal environment for the proposed research and for his career development
as Dr. Zack has ...

## Key facts

- **NIH application ID:** 10395288
- **Project number:** 3K99EY029011-02S1
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Xitiz Chamling
- **Activity code:** K99 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $88,947
- **Award type:** 3
- **Project period:** 2019-05-01 → 2021-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10395288

## Citation

> US National Institutes of Health, RePORTER application 10395288, Developing Remyelination Strategies for Demyelinating Optic Neuropathies Using Human Pluripotent Stem Cells (3K99EY029011-02S1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10395288. Licensed CC0.

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