Loss of c‐Kit and bone marrow failure upon conditional removal of the GATA‐2 C‐terminal zinc finger domain in adult mice

Abstract

Heterozygous mutations in the transcriptional regulator GATA-2 associate with multilineage immunodeficiency, myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). The majority of these mutations localize in the zinc finger (ZnF) domains, which mediate GATA-2 DNA binding. Deregulated hematopoiesis with GATA-2 mutation frequently develops in adulthood, yet GATA-2 function in the bone marrow remains unresolved. To investigate this, we conditionally deleted the GATA-2 C-terminal ZnF (C-ZnF) coding sequences in adult mice. Upon Gata2 C-ZnF deletion, we observed rapid peripheral cytopenia, bone marrow failure and decreased c-Kit expression on hematopoietic progenitors. Transplant studies indicated GATA-2 has a cell autonomous role in bone marrow hematopoiesis. Moreover, myeloid lineage populations were particularly sensitive to Gata2 hemizygosity, while molecular assays indicated GATA-2 regulates c-Kit expression in multilineage progenitor cells. Enforced c-Kit expression in Gata2 C-ZnF-deficient hematopoietic progenitors enhanced myeloid colony activity, suggesting GATA-2 sustains myelopoiesis via a cell intrinsic role involving maintenance of c-Kit expression. Our results provide insight into mechanisms regulating hematopoiesis in bone marrow and may contribute to a better understanding of immunodeficiency and bone marrow failure associated with GATA-2 mutation.

Gata2 as a Crucial Regulator of Stem Cells in Adult Hematopoiesis and Acute Myeloid Leukemia

“…In this report, to explore the genetic requirement for Gata2 in adult hematopoiesis, we employed a conditional gene-targeting approach using the Mx1 -Cre system to specifically delete Gata2 in hematopoietic cells from adult mice. Consistent with data using ER -Cre-mediated deletion of the Gata2 C-terminal zinc-finger domain (Li et al., 2016), we demonstrate that acute genetic ablation of Gata2 leads to a rapid and complete cell-autonomous loss of adult HSCs and multi-lineage potential. Therefore, Gata2 joins a select list of TFs that are indispensable for adult HSC maintenance and survival (Hock et al., 2004, Kranc et al., 2009, Pajerowski et al., 2010).…”

Uncoupling key determinants of hematopoietic stem cell engraftment through cell-specific and temporally controlled recipient conditioning

“…We detected almost no contribution of Gata2 KO cells in the PB (Figure 1H) and BM (Figure 1I) of the recipients, while control cells showed a similar level of reconstitution as the WT competitor cells. Altogether, these results demonstrate a rapid and selective depletion of resident HSCs through poly(IC)-induced Gata2 deletion, consistent with previous reports where Gata2 has been depleted in adult mice (Li et al, 2016;Menendez-Gonzalez et al, 2019).…”

“…To this end, we established a model where HSCs can be selectively depleted by conditional deletion of Gata2 ( Haugas et al., 2010 ). GATA2 is a transcription factor that was identified as an essential regulator of HSC generation and survival during embryogenesis ( de Pater et al., 2013 ; Tsai et al., 1994 ), with an equally critical and dose-dependent role in regulation of adult hematopoiesis ( Li et al., 2016 ; Lim et al., 2012 ; Ling et al., 2004 ; Menendez-Gonzalez et al., 2019 ; Rodrigues, 2005 ).…”

Molecular regulation of dendritic cell development and function in homeostasis, inflammation, and cancer

“…Numerous transcriptional regulators mediate development of DCs from HSCs and specified hematopoietic progenitors (Table 1). Certain factors including PU.1, GATA-2, Ikaros, and Gfi1 are necessary for both pDC and cDC development, due to their essential activity in early hematopoietic progenitor subsets (Anderson et al, 2000;Carotta et al, 2010;Guerriero et al, 2000;Iwama et al, 2002;Li et al, 2016a;Onodera et al, 2016;Wu et al, 1997). Other transcriptional regulators play specific roles in one or more DC lineages.…”

“…Phenotypic markers and transcriptional regulators of murine and human DC subsets. (Bjorck, 2001;Nakano et al, 2001;Zhang et al, 2006) CD11c − CD123 + CD303 (BDCA-2) + CD304 (BDCA-4) + (Dzionek et al, 2000) TLR7, TLR9 (Dursun et al, 2016;Edwards et al, 2003;Ito et al, 2002;Jarrossay et al, 2001;Krug et al, 2001; Mashayekhi et al, 2011) GATA2 (Li et al, 2016a;Onodera et al, 2016) Spi-B (Sasaki et al, 2012) PU.1 (Anderson et al, 2000;Carotta et al, 2010;Guerriero et al, 2000;Iwama et al, 2002) Ikaros (Wu et al, 1997) Gfi1 (Rathinam et al, 2005) Ifd4 (Tamura et al, 2005) E2-2 (Cisse et al, 2008) Zeb2 (Wu et al, 2016) Bcl11a (Ippolito et al, 2014) Runx1…”