# Delineating the functions of EKLF during mammalian terminal erythroid differentiation

> **NIH NIH K01** · CLEVELAND STATE UNIVERSITY · 2021 · $54,000

## Abstract

Project Summary
Erythroid terminal differentiation is comprised of 3-4 rapid terminal cell divisions also known as ‘differentiation
divisions’, which are coupled with morphological changes such as a dramatic decrease in cell and nuclear size.
The switch from self-renewal to terminal divisions in erythroid cells are peculiarly characterized by short G1
and S phases, and fast DNA replication. We do not yet understand the processes that regulate the timing,
integrity and the numbers of these rapid terminal divisions. When these divisions go awry, it leads to severe
anemias such as Congenital Dyserythropoietic Anemia (CDA), which arise due to failures in DNA replication
and/or cytokinesis, and are characterized by binucleate erythroblasts in the bone marrow and in some cases
chromatin bridges between erythroblasts.
EKLF/KLF1 is one the genes when mutated causes a type of CDA, CDA type IV. Although previous genome
wide studies have alluded to its roles in DNA replication/repair and cytokinesis, this has not been functionally
investigated. Interestingly, EKLF-/- erythroid cultures during terminal differentiation have increased proportions
of binucleate cells and chromatin bridges; similar to what has been observed in CDA disorders. Based on our
preliminary data, we hypothesize that EKLF transcriptionally upregulates the genes involved in the
maintenance of DNA replication fidelity (Aim1) and cytokinesis (Aim2) to accommodate the rapid pace of
terminal erythroid cell divisions; impairment of this regulation in EKLF-/- erythroblasts results in replication
stress, cytokinesis failure and the formation of binucleate erythroblasts.
I will study the role of EKLF in the maintenance of DNA replication fidelity by quantifying the levels of DNA
damage and replication stress, replication dynamics, and the extent to which unresolved DNA damage
perturbs cytokinesis. I will investigate the role of EKLF in cytokinesis by studying the formation, structure, and
the function of the midbody organelle, which forms between two daughter cells and is essential for abscission.
Finally, I will also examine the extent to which EKLF regulated candidate genes contribute to the observed
defects in EKLF-/- erythroblasts. These studies will reveal a specialized transcriptional regulation in erythroid
cells to ensure that the cell cycle machinery is able to accommodate the rapid pace of the terminal cell
divisions. They will also provide insights on the pathogenesis of severe anemias such as CDA, some of whose
etiology is unknown.
The studies proposed here along with career development plan described in my application, will enable me to
benefit from the mentorship of Dr. James Bieker, who discovered EKLF and has contributed immensely to the
field of erythropoiesis, forge collaborations to expand my expertise, and gain guidance on career and scientific
progression from my advisory committee. Overall, this will pave the way for my successful transition to
independence.

## Key facts

- **NIH application ID:** 10326466
- **Project number:** 3K01DK115686-04S1
- **Recipient organization:** CLEVELAND STATE UNIVERSITY
- **Principal Investigator:** Merlin Nithya Gnanapragasam
- **Activity code:** K01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $54,000
- **Award type:** 3
- **Project period:** 2018-03-01 → 2024-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10326466

## Citation

> US National Institutes of Health, RePORTER application 10326466, Delineating the functions of EKLF during mammalian terminal erythroid differentiation (3K01DK115686-04S1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10326466. Licensed CC0.

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