Transient Expression of GATA3 in Hematopoietic Stem Cells Facilitates Helper Innate Lymphoid Cell Differentiation.

Dejene M Tufa,Renee Woods,Ashley M Yingst,Tyler Shank,Jessica Lake,Seonhui Shim,George Devon Trahan,Michael R Verneris,Kenneth L Jones

doi:10.3389/fimmu.2019.00510

Abstract

Helper Innate lymphoid cells (ILCs) are tissue resident lymphocytes that play a critical role in a number of biological processes. Several transcription factors are required for the differentiation of hematopoietic stem cells (HSCs) into ILCs. Recent studies demonstrate GATA3 as a transcriptional regulator that plays an essential role in ILC development. We aimed to modulate the differentiation of human cord blood-derived CD34+ cells into ILCs by transient and ectopic expression of mRNA encoding transcription factors known to be important for ILC lineage differentiation, including GATA3, TOX, NFIL3, ID2, and RORγt. Using this experimental protocol, only GATA3 significantly modulated HSCs to differentiate into helper ILCs. Transient overexpression of GATA3 drove the emergence of CD34+α4β7+ early ILC progenitors during the first few days of culture. These ILC progenitors further acquired IL-7Rα and CD117 to give rise to immediate ILC precursors. In support of these findings, analysis of the genes induced by GATA3 in HSCs showed an upregulation of those associated with ILC development. Moreover, we show GATA3 also acts on more committed progenitors and significantly shifts the differentiation of progenitors away from the ILC1/NK lineage to the ILC2 and ILC3 lineage. In summary, transient overexpression of GATA3 mRNA in CD34+ HSCs enhances the differentiation of HSCs into the helper ILC lineages, at the expense of NK cell development.

Highlights

Innate lymphoid cells (ILCs) are group of tissue-resident lymphocytes that have been grouped into three different cell types according to their transcription factor expression and cytokine production following stimulation [1, 2]
We cloned the genes for GATA3, ID2, NFIL3, RORC, TOX, and GFP and in vitro transcribed these into mRNA which was electroporated into CD34+ cells that had been cultured for 5 days
Murine ILCs are derived from a common lymphoid progenitor that give rise to a common ILC progenitor (CILP), which in turn has the capacity to differentiate into all helper ILC subtypes [1, 2]

Summary

Introduction

Innate lymphoid cells (ILCs) are group of tissue-resident lymphocytes that have been grouped into three different cell types according to their transcription factor expression and cytokine production following stimulation [1, 2]. Group 1 ILCs, which include natural killer (NK) cells, express T-bet and produce IFN-γ and some are cytotoxic upon activation [3, 4]. Group 2 ILCs express GATA3 and produce IL-5 and IL-13 upon stimulation, whereas group 3 ILCs express RORγt and produce IL-22 following stimulation [5,6,7,8,9]. All ILCs develop from common lymphoid progenitors (CLP), which further differentiates into the common ILC progenitor (CILP). The CILP in turn, differentiates into an NK progenitor (NKP), as well as the common “helper” innate lymphoid cell precursor (CHILP). The NK progenitors differentiate to NK cells, whereas the CHILP differentiates to all ILCs such as ILC1, ILC2, and ILC3, but not NK cells [1, 2, 11, 12]

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