# Defining the functional role of protein N-glycosylation on hematopoietic stem cell production from endothelium

> **NIH NIH R00** · DARTMOUTH COLLEGE · 2021 · $249,000

## Abstract

PROJECT SUMMARY
 Creation of hematopoietic stem/progenitor cells (HSPCs) early in development is a highly dynamic
process whereby flat hemogenic endothelial cells (ECs) change cell fate and emerge as rounded HSPCs with
self-renewing capacity and multi-lineage potential. Unfortunately, the molecular mechanisms controlling this
endothelial-to-hematopoietic transition (EHT) are still not well defined. This is underscored by the inability to
mass-produce long-term reconstituting HSPCs for blood-regenerative therapies. Identification of new
regulators of HSPC formation has been challenging in mammalian models likely due to the inability to visualize
EHT dynamics in utero. Recently, our lab has taken advantage of the transparency and external development
of zebrafish embryos to identify microRNA-223 as a novel inhibitor of HSPC formation. Zebrafish embryos
lacking miR-223 activity had an increased number of emerging HSPCs, resulting in mature HSPC expansion
from the onset and to later stages of hematopoiesis. How miR-223 controls HSPC production from the
endothelium at the cellular and molecular levels is currently unknown.
 miR-223 belongs to the microRNA (miRNA) class of small noncoding RNAs, which provide precision to
signaling pathways by post-transcriptionally repressing gene expression of complementary mRNA targets.
Comparative transcriptome profiling of zebrafish miR-223 mutant and wildtype endothelial cells revealed N-
glycosylation enzymes as candidate miR-223 molecular targets, and embryos treated with N-glycosylation
inhibitors phenocopy the HSPC expansion in miR-223 mutants. Thus, the goal of this proposal is to directly test
the hypothesis that miR-223 fine tunes N-glycosylation levels to control HSPC formation from the endothelium.
The proposed study will develop novel transgenic and mutant approaches in zebrafish to dynamically visualize
the cellular events controlled by miR-223 and to validate the N-glycosylation pathway as the direct target of
miR-223 during EHT. Altogether, findings from this study will contribute a novel mechanism of HSPC
formation, in which a miRNA dependent N-glycome regulates EC to HSPC cell fate transitions.

## Key facts

- **NIH application ID:** 10413509
- **Project number:** 4R00HL141687-03
- **Recipient organization:** DARTMOUTH COLLEGE
- **Principal Investigator:** Dionna Kasper
- **Activity code:** R00 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $249,000
- **Award type:** 4N
- **Project period:** 2021-09-01 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10413509

## Citation

> US National Institutes of Health, RePORTER application 10413509, Defining the functional role of protein N-glycosylation on hematopoietic stem cell production from endothelium (4R00HL141687-03). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10413509. Licensed CC0.

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