# DETERMINING THE ROLE OF THE REPLICATIVE HELICASE IN HUMAN NK CELL DEVELOPMENT

> **NIH NIH R01** · COLUMBIA UNIVERSITY HEALTH SCIENCES · 2021 · $364,500

## Abstract

PROJECT SUMMARY
This proposal builds upon exciting new paradigm defined by isolated human NK cell deficiency resulting from
hypomorphic mutations in multiple structural components of the eukaryotic DNA helicase. The discovery of
multiple patient cohorts with a strikingly similar NK cell phenotype, namely that of impaired terminal maturation
leading to susceptibility to severe viral infection and malignancy, underscores the impact of these mutations on
human NK cell development and function. Despite this compelling new insight driven by human disease, the
requirement for the DNA helicase complex specifically in NK cell function has not been defined. In the
proposed work, we will define this requirement in the context of NK cell differentiation and proliferation to make
important advances in both NK cell biology and human health.
 Despite their documented requirement in determining the outcome and success of transplantation, NK
cell development and acquisition of function is poorly understood. A powerful tool in defining requirements for
human NK cell differentiation is the study of patients with monogenic causes of impaired NK cell maturation
leading to isolated NK cell deficiency (NKD). Using this approach, we and others have identified novel NKD
resulting from mutations in the CDC45-MCM-GINS DNA helicase complex and associated proteins. In the
proposed work, we will define the requirement for the CMG complex and, more broadly, proliferation and cell
division in human NK cell development. Specifically, we will 1) define the effect of CMG helicase mutations
on peripheral blood subset heterogeneity using single cell RNA sequencing and unbiased quantitative flow
cytometry. Furthermore, we have 2) developed a model for patient mutations that will allow us to dissect
the effect of these mutations on NK cell differentiation from earliest precursor to mature cell. We will
use this, combined with our validated in vitro NK cell differentiation system, to dissect the mechanism by which
hypomorphic CMG mutations affect human NK cell development and acquisition of lytic function. Finally, we
will 3) define the effect of CMG helicase mutations on the human NK cell antiviral response. Using
careful analysis of gene expression, phenotype and function, we will determine the timing and the nature of
developmental deregulation in NK cell developmental intermediates with hypomorphic replisome mutations.
These aims we will advance a novel new paradigm in human NK cell differentiation, namely the specific
requirement for proliferation and cell cycle control for NK cell terminal maturation. These studies have
significant clinical importance for understanding the differentiation of NK cells in the unique milieu following
hematopoietic stem cell transplant. Defining the role of proliferation in the generation of mature NK cell
effectors will enable the better control of these cells to prevent graft vs. host disease and promote their natural
anti-tumor immunity.

## Key facts

- **NIH application ID:** 10232359
- **Project number:** 5R01AI137275-04
- **Recipient organization:** COLUMBIA UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** Emily Margaret Mace
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $364,500
- **Award type:** 5
- **Project period:** 2018-09-24 → 2023-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10232359

## Citation

> US National Institutes of Health, RePORTER application 10232359, DETERMINING THE ROLE OF THE REPLICATIVE HELICASE IN HUMAN NK CELL DEVELOPMENT (5R01AI137275-04). Retrieved via AI Analytics 2026-06-10 from https://api.ai-analytics.org/grant/nih/10232359. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*