Competition and collaboration: GATA-3, PU.1, and Notch signaling in early T-cell fate determination

doi:10.1016/j.smim.2008.07.006

Seminars in Immunology

Volume 20, Issue 4, August 2008, Pages 236-246

https://doi.org/10.1016/j.smim.2008.07.006 Get rights and content

Abstract

T-cell precursors remain developmentally plastic for multiple cell generations after entering the thymus, preserving access to developmental alternatives of macrophage, dendritic-cell, and even mast-cell fates. The underlying regulatory basis of this plasticity is that early T-cell differentiation depends on transcription factors which can also promote alternative developmental programs. Interfactor competition, together with environmental signals, keep these diversions under control. Here the pathways leading to several lineage alternatives for early pro-T-cells are reviewed, with close focus on the mechanisms of action of three vital factors, GATA-3, PU.1, and Notch–Delta signals, whose counterbalance appears to be essential for T-cell specification.

Section snippets

Models for hematopoietic lineage choice

Hematopoietic stem cells can develop into any of at least 10 different kinds of effector cells, and these can be grouped broadly into lymphoid, myeloid, erythroid, megakaryocytic, and other types. Stem cells become particular cell types by passing through sequences of partially restricted, but still pluripotent intermediates. The outcome of competing regulatory forces at work on the cells during the partially restricted intermediate stages ultimately forms the basis for the commitment decision

Landmarks for early T-cell development

Early-stage T-cell precursors are partially restricted cells that migrate to the thymus either just after or immediately before losing the ability to give rise to B lymphocytes, but before they shut off molecularly distinct developmental alternatives including macrophage or dendritic-cell fates. Differentiation of these cells occurs in the thymic cortex over a period of many days and is accompanied by a flexible degree of proliferation. Fig. 2 summarizes the cell-surface and molecular

Control of early T-cell development by external signals

T-lineage specification is driven and maintained by signals from the environment. The transition from each DN stage to the next depends on the existing internal transcription factor complement in the cell together with changes in transcription factor function induced by external environmental signals. The most important of these signals, on current evidence, are cytokine signals, signals through the Wnt-β-catenin-TCF pathway, and repeated signals from the Notch pathway. As for other

Context-dependent modulation of Notch signals

Notch signaling is not a Boolean operator that determines T-lineage fate at a stroke, but a quantitatively modulated participant in protracted lineage decisions. A sustained succession of interactions with Delta-class target ligands, through all the cell cycles from ETP throughout DN2 stages, is needed to elicit full commitment to the T-lineage [35], [36], [44], [77]. Notch–Delta interaction can drive many different starting populations of hematopoietic precursors into T-cell development [21],

Essential roles for GATA-3 and PU.1 in early T-cell development

Both GATA-3 and PU.1 are essential for T-cell development. GATA-3, a double zinc finger (C4-type) transcription factor, is expressed in an almost T-lineage-specific pattern among hematopoietic cells. It is turned on among the earliest genes activated by Notch–Delta signaling in hematopoietic precursors, and is used throughout T-cell development from the ETP stage all the way to post-thymic responses to antigen. The ETS-family transcription factor PU.1 is almost the reverse, with virtually no

GATA-3-driven lineage infidelity: dosage and Notch sensitivity

With its T-lineage specific expression and recurrent T-lineage roles, GATA-3 has been an appealing candidate for a factor conferring T-lineage identity in early thymocytes (Fig. 1B). However, gain of function of GATA-3, in the presence or absence of Notch signals, fails to enhance either appearance or developmental progression of the earliest T-lineage cells [36], [106], [111], [112]. Only around the time of lineage commitment, from the DN3 stage or its human equivalent onward, does high-level

PU.1 and myeloid alternatives to the T-lineage program

Although needed to start T-cell development, PU.1 also helps to maintain a progenitor-like developmental plasticity in the ETP and DN2 cells. During commitment, expression of PU.1 and SCL (Tal1) precipitously shuts off [33], [124]. Forced gain of PU.1 function, beyond the stage when it is normally shut off [51], [52], [53], [54], [55], [106], provides evidence that endogenous PU.1 expression in earlier T-cell precursors is probably rate-limiting for their developmentally regulated access to

GATA-3 as antagonist of myeloid lineage options

The functions mobilized by Notch to preserve T-lineage identity before lineage commitment need to be identified. GATA-3 is an obvious candidate for one of the T-lineage gatekeeper functions that antagonizes myeloid fates under normal conditions. Myeloid lineage conversion of thymocytes by either C/EBPα or PU.1 is inhibited if GATA-3 is co-expressed [52], [127]. Furthermore, GATA-3 may be suspected of a role in lineage commitment itself, as overexpression of GATA-3 in early fetal thymocytes

Transdifferentiation, not dedifferentiation, in response to regulatory perturbation

Neither myeloid nor mast-cell programs are thought to be closely related to lymphocyte developmental programs, and indeed, conversion of thymocytes to dendritic cells by PU.1, inflammatory macrophages by C/EBPα, and mast cells by excess GATA-3 all involve a repression of T-lineage specific gene expression. Their diversionary activity is even more remarkable in view of the positive roles that two of these diversion-inducing factors normally play in T-cell development, when expressed at different

Conclusions

The results reviewed here show that T-cell precursor specification is not only a protracted process but a remarkably precarious one at the regulatory level. This makes sense of much biology that could otherwise seem counterintuitive. An increasingly rich literature confirms that most T-cell precursors naturally retain access to non-T and non-lymphoid programs well beyond the stage when they have lost B-lineage potential [16], [17], [18], [19], [77], [133], [134], [135]. The myeloid

Acknowledgments

We are most grateful to the primary authors of the work cited in this review, including members and former members of this laboratory whose results were discussed in detail. We especially thank Thomas Graf for patient encouragement and valuable insights, and Howard Petrie, Cornelis Murre, Hiroshi Kawamoto and Avinash Bhandoola for exciting discussions of work before publication. The authors’ work was supported by NIH grants CA90233, CA98925, and HL089123, the Albert Billings Ruddock

References (152)

J. Rhodes et al.
Interplay of pu.1 and gata1 determines myelo-erythroid progenitor cell fate in zebrafish
Dev Cell
(2005)
C. Nerlov et al.
GATA-1 interacts with the myeloid PU.1 transcription factor and represses PU. 1-dependent transcription
Blood
(2000)
R. Månsson et al.
Molecular evidence for hierarchical transcriptional lineage priming in fetal and adult stem cells and multipotent progenitors
Immunity
(2007)
M. Kondo et al.
Identification of clonogenic common lymphoid progenitors in mouse bone marrow
Cell
(1997)
J. Adolfsson et al.
Identification of Flt3⁺ lympho-myeloid stem cells lacking erythro-megakaryocytic potential a revised road map for adult blood lineage commitment
Cell
(2005)
G. Balciunaite et al.
The earliest subpopulation of mouse thymocytes contains potent T, significant macrophage, and natural killer cell but no B-lymphocyte potential
Blood
(2005)
L. Wu et al.
Development of dendritic-cell lineages
Immunity
(2007)
M. Zenke et al.
Towards an understanding of the transcription factor network of dendritic cell development
Trends Immunol
(2006)
H. Shigematsu et al.
Plasmacytoid dendritic cells activate lymphoid-specific genetic programs irrespective of their cellular origin
Immunity
(2004)
H.E. Porritt et al.
Heterogeneity among DN1 prothymocytes reveals multiple progenitors with different capacities to generate T cell and non-T cell lineages
Immunity
(2004)

S.I. Samson et al.

GATA-3 promotes maturation, IFN-γ production, and liver-specific homing of NK cells

Immunity

(2003)

C.V. Laiosa et al.

Determinants of lymphoid–myeloid lineage diversification

Annu Rev Immunol

(2006)

T. Graf

(2007)

M.K. Anderson

At the crossroads: diverse roles of early thymocyte transcriptional regulators

Immunol Rev

(2006)

Cited by (39)

GATA2-Mediated Transcriptional Activation of Notch3 Promotes Pancreatic Cancer Liver Metastasis
2022, Molecules and Cells
The liver is the predominant metastatic site for pancreatic cancer. However, the factors that determine the liver metastasis and the specific molecular mechanisms are still unclear. In this study, we used human pancreatic cancer cell line Hs766T to establish Hs766T-L3, a subline of Hs766T with stable liver metastatic ability. We performed RNA sequencing of Hs766T-L3 and its parental cell line Hs766T, and revealed huge differences in gene expression patterns and pathway activation between these two cell lines. We correlated the difference in pathway activation with the expression of the four core transcriptional factors including STAT1, NR2F2, GATA2, and SMAD4. Using the TCGA database, we examined the relative expression of these transcription factors (TFs) in pan-cancer and their relationship with the prognosis of the pancreatic cancer. Among these TFs, we considered GATA2 is closely involved in tumor metastasis and may serve as a potential metastatic driver. Further in vitro and in vivo experiments confirmed that GATA2-mediated transcriptional activation of Notch3 promotes the liver metastasis of Hs766T-L3, and knockdown of either GATA2 or Notch3 reduces the metastatic ability of Hs766T-L3. Therefore, we claim that GATA2 may serve as a metastatic driver of pancreatic cancer and a potential therapeutic target to treat liver metastasis of pancreatic cancer.
ETV6-NCOA2 fusion induces T/myeloid mixed-phenotype leukemia through transformation of nonthymic hematopoietic progenitor cells
2022, Blood
Citation Excerpt :
MEF2C is expressed in ETV6-NCOA2 PDX cells13 (supplemental Figure 9). The T-cell differentiation and the T/myeloid checkpoint are dependent on accurate regulation of NOTCH1 and PU.1/MEF2C pathways.32-35 For an early thymocyte to differentiate into a mature T cell, the NOTCH1 pathway should be activated whereas the PU.1/MEF2C pathway should be completely shut down (PU.1, MEF2C, and CSF1R).32-36
Mixed-phenotype acute leukemia is a rare subtype of leukemia in which both myeloid and lymphoid markers are co-expressed on the same malignant cells. The pathogenesis is largely unknown, and the treatment is challenging. We previously reported the specific association of the recurrent t(8;12)(q13;p13) chromosomal translocation that creates the ETV6-NCOA2 fusion with T/myeloid leukemias. Here we report that ETV6-NCOA2 initiates T/myeloid leukemia in preclinical models; ectopic expression of ETV6-NCOA2 in mouse bone marrow hematopoietic progenitors induced T/myeloid lymphoma accompanied by spontaneous Notch1-activating mutations. Similarly, cotransduction of human cord blood CD34⁺ progenitors with ETV6-NCOA2 and a nontransforming NOTCH1 mutant induced T/myeloid leukemia in immunodeficient mice; the immunophenotype and gene expression pattern were similar to those of patient-derived ETV6-NCOA2 leukemias. Mechanistically, we show that ETV6-NCOA2 forms a transcriptional complex with ETV6 and the histone acetyltransferase p300, leading to derepression of ETV6 target genes. The expression of ETV6-NCOA2 in human and mouse nonthymic hematopoietic progenitor cells induces transcriptional dysregulation, which activates a lymphoid program while failing to repress the expression of myeloid genes such as CSF1 and MEF2C. The ETV6-NCOA2 induced arrest at an early immature T-cell developmental stage. The additional acquisition of activating NOTCH1 mutations transforms the early immature ETV6-NCOA2 cells into T/myeloid leukemias. Here, we describe the first preclinical model to depict the initiation of T/myeloid leukemia by a specific somatic genetic aberration.
The Notch signaling pathway involvement in innate lymphoid cell biology
2021, Biomedical Journal
The role of Notch in the immune system was first described in the late 90s. Reports revealed that Notch is one of the most conserved developmental pathways involved in diverse biological processes such as the development, differentiation, survival and functions of many immune populations. Here, we provide an extended view of the pleiotropic effects of the Notch signaling on the innate lymphoid cell (ILC) biology. We review the current knowledge on Notch signaling in the regulation of ILC differentiation, plasticity and functions in diverse tissue types and at both the fetal and adult developmental stages. ILCs are early responder cells that secrete a large panel of cytokines after stimulation. By controlling the abundance of ILCs and the specificity of their release, the Notch pathway is also implicated in the regulation of their functions. The Notch pathway is therefore an important player in both ILC cell fate decision and ILC immune response.
Organoid-Induced Differentiation of Conventional T Cells from Human Pluripotent Stem Cells
2019, Cell Stem Cell
The ability to generate T cells from pluripotent stem cells (PSCs) has the potential to transform autologous T cell immunotherapy by facilitating universal, off-the-shelf cell products. However, differentiation of human PSCs into mature, conventional T cells has been challenging with existing methods. We report that a continuous 3D organoid system induced an orderly sequence of commitment and differentiation from PSC-derived embryonic mesoderm through hematopoietic specification and efficient terminal differentiation to naive CD3⁺CD8αβ⁺ and CD3⁺CD4⁺ conventional T cells with a diverse T cell receptor (TCR) repertoire. Introduction of an MHC class I-restricted TCR in PSCs produced naive, antigen-specific CD8αβ⁺ T cells that lacked endogenous TCR expression and showed anti-tumor efficacy in vitro and in vivo. Functional assays and RNA sequencing aligned PSC-derived T cells with primary naive CD8⁺ T cells. The PSC-artificial thymic organoid (ATO) system presented here is an efficient platform for generating functional, mature T cells from human PSCs.
Methylation of Gata3 protein at Arg-261 regulates transactivation of the Il5 gene in T helper 2 cells
2015, Journal of Biological Chemistry
Gata3 acts as a master regulator for T helper 2 (Th2) cell differentiation by inducing chromatin remodeling of the Th2 cytokine loci, accelerating Th2 cell proliferation, and repressing Th1 cell differentiation. Gata3 also directly transactivates the interleukin-5 (Il5) gene via additional mechanisms that have not been fully elucidated. We herein identified a mechanism whereby the methylation of Gata3 at Arg-261 regulates the transcriptional activation of the Il5 gene in Th2 cells. Although the methylation-mimicking Gata3 mutant retained the ability to induce IL-4 and repress IFNγ production, the IL-5 production was selectively impaired. We also demonstrated that heat shock protein (Hsp) 60 strongly associates with the methylation-mimicking Gata3 mutant and negatively regulates elongation of the Il5 transcript by RNA polymerase II. Thus, arginine methylation appears to play a pivotal role in the organization of Gata3 complexes and the target gene specificity of Gata3.
Background: Gata3 directly transactivates the Il5 gene.
Results: The methylation-mimicking Gata3 mutant at Arg-261 had a selective defect in the induction of IL-5 production via recruitment of Hsp60 to prevent transactivation.
Conclusion: Arginine methylation on Gata3 may play a critical role in the organization and function of the Gata3 transcriptional complex.
Significance: A new regulatory mechanism of Gata3-mediated Il5 transactivation is revealed.
E2Atranscription factors limit expression of Gata3 to facilitate T lymphocyte lineage commitment
2013, Blood
Citation Excerpt :
This hypothesis is consistent with previous studies that reported that GATA3 functions are concentration dependent and may be influenced by interactions with other transcription factors. For example, high concentrations of GATA3 antagonize PU.1 function in DN2s, whereas low concentrations of GATA3 synergize with PU.1 to produce a gene expression signature that allows for T-cell lineage commitment.6,45,46 Additional studies will be necessary to determine how the elevation of GATA3 influences the differentiation and commitment of T-lymphocyte progenitors.
The E2A transcription factors promote the development of thymus-seeding cells, but it remains unknown whether these proteins play a role in T lymphocyte lineage specification or commitment. Here, we showed that E2A proteins were required to promote T-lymphocyte commitment from DN2 thymocytes and to extinguish their potential for alternative fates. E2A proteins functioned in DN2 cells to limit expression of Gata3, which encodes an essential T-lymphocyte transcription factor whose ectopic expression can arrest T-cell differentiation. Genetic, or small interfering RNA-mediated, reduction of Gata3 rescued T-cell differentiation in the absence of E2A and restricted the development of alternative lineages by limiting the expanded self-renewal potential in E2A^−/− DN2 cells. Our data support a novel paradigm in lymphocyte lineage commitment in which the E2A proteins are necessary to limit the expression of an essential lineage specification and commitment factor to restrain self-renewal and to prevent an arrest in differentiation.

View all citing articles on Scopus

View full text

ReviewCompetition and collaboration: GATA-3, PU.1, and Notch signaling in early T-cell fate determination

Abstract

Section snippets

Models for hematopoietic lineage choice

Landmarks for early T-cell development

Control of early T-cell development by external signals

Context-dependent modulation of Notch signals

Essential roles for GATA-3 and PU.1 in early T-cell development

GATA-3-driven lineage infidelity: dosage and Notch sensitivity

PU.1 and myeloid alternatives to the T-lineage program

GATA-3 as antagonist of myeloid lineage options

Transdifferentiation, not dedifferentiation, in response to regulatory perturbation

Conclusions

Acknowledgments

Dev Cell

Blood

Immunity

Cell

Cell

Blood

Immunity

Trends Immunol

Immunity

Immunity

Curr Opin Genet Dev

Immunity

Immunity

Semin Immunol

Immunity

Immunity

Dev Biol

Curr Opin Immunol

Blood

Exp Hematol

Immunity

Immunity

Blood

Trends Immunol

Dev Comp Immunol

Blood

Blood

Blood

Immunity

Curr Biol

Immunity

Immunity

Determinants of lymphoid–myeloid lineage diversification

Annu Rev Immunol

Transcription factors that induce commitment of multipotent hematopoietic progenitors: lessons from the MEP system

Direct interaction of hematopoietic transcription factors PU.1 and GATA-1: functional antagonism in erythroid cells

Genes Dev

Negative cross-talk between hematopoietic regulators: GATA proteins repress PU.1

Proc Natl Acad Sci USA

Dynamic regulation of PU.1 expression in multipotent hematopoietic progenitors

J Exp Med

Regulation of lymphocyte development by Notch signaling

Nat Immunol

Regulation of lymphoid development, differentiation, and function by the Notch pathway

Annu Rev Immunol

Pax5: the guardian of B cell identity and function

Nat Immunol

Cell lineage regulators in B and T cell development

Nat Immunol

A clonogenic common myeloid progenitor that gives rise to all myeloid lineages

Nature

The earliest thymic progenitors in adults are restricted to T, NK, and dendritic cell lineage and have a potential to form more diverse TCRβ chains than fetal progenitors

J Immunol

T/NK bipotent progenitors in the thymus retain the potential to generate dendritic cells

J Immunol

Thymopoiesis independent of common lymphoid progenitors

Nat Immunol

Phenotypic plasticity of T cell progenitors upon exposure to Notch ligands

J Exp Med

Propensity of adult lymphoid progenitors to progress to DN2/3 stage thymocytes with Notch receptor ligation

J Immunol

Key factors in the organized chaos of early T cell development

Nat Immunol

Zoned out: functional mapping of stromal signaling microenvironments in the thymus

Annu Rev Immunol

At the crossroads: diverse roles of early thymocyte transcriptional regulators

Review
Competition and collaboration: GATA-3, PU.1, and Notch signaling in early T-cell fate determination