Elsevier

Leukemia Research

Volume 32, Issue 6, June 2008, Pages 873-883
Leukemia Research

Overexpression of stem cell associated ALDH1A1, a target of the leukemogenic transcription factor TLX1/HOX11, inhibits lymphopoiesis and promotes myelopoiesis in murine hematopoietic progenitors

https://doi.org/10.1016/j.leukres.2007.11.001Get rights and content

Abstract

TLX1/HOX11 is an oncogenic transcription factor in human T-cell leukemia, however, the molecular basis for its transforming activity has remained elusive. The ALDH1A1 gene, whose product participates in retinoic acid synthesis, was previously identified as a TLX1-responsive gene. Here, we confirm regulation of ALDH1A1 transcription by TLX1 and show that ALDH1A1 can profoundly perturb murine hematopoiesis by promoting myeloid differentiation at the expense of lymphopoiesis. Together, these data demonstrate that ALDH1A1 plays a key role in normal hematopoiesis, and confirm ALDH1A1 as a TLX1 transcriptional target that may contribute to the ability of this homeoprotein to alter cell fate and induce tumor growth.

Introduction

Aldehyde dehydrogenase 1a1 (ALDH1A1) encodes a cytosolic enzyme whose main biological role has been reported to be the irreversible oxidation of retinal to the signalling molecule retinoic acid (RA) [1], [2]. Elevated levels of ALDH1A1 are a characteristic feature of hematopoietic stem cells (HSCs) [3], [4], [5], [6], while altered expression of this RA-synthesizing enzyme has been reported in a number of solid tumors and tumor cell lines [7], [8], [9]. Despite this, possible roles for ALDH1A1 in hematopoiesis and leukemic transformation have remained largely unexplored. This needs to be addressed since dysregulation of RA synthesis, which is tightly controlled by a select group of enzymes including ALDH1A1 and ALDH1A2, may interfere with processes regulating cellular differentiation and proliferation, thereby contributing to the tumorigenic phenotype. In a separate role, ALDH1A1 is of relevance for its ability to affect the sensitivity of stem and tumor cells to chemotherapeutic agents such as cyclophosphamide [10], [11], [12].

ALDH1A1 is additionally of interest because of its status as a transcriptional target of the homeoprotein TLX1/HOX11 [13], [14], [15]. Tlx1 has been shown to dictate the developmental fate of the spleen [16], [17] and of specific neurons within the spinal cord [18], while mice lacking Tlx1 additionally exhibit leukocytosis [16]. TLX1 is also a significant etiological factor in T-cell leukemogenesis since chromosomal translocations and other, as yet uncharacterised, molecular events involving the human 10q24 locus activate expression of the TLX1 oncogene in up to 50% of human T-cell acute lymphoblastic leukemia (T-ALL) cases [19], [20], [21], [22], [23]. The mechanism(s) by which TLX1 induces tumor growth and controls normal development remain to be fully determined, although TLX1 has been implicated in cell cycle progression [24], [25], predisposition to aneuploidy [26] and alteration of transcriptional regulator/co-regulator activity [27], [28].

TLX1 is a transcription factor and several studies have linked its expression to the perturbation of gene expression programs controlling cell proliferation and apoptosis [22], [25], [29], [30]. A plausible assumption, therefore, is that at least part of the oncogenic and/or normal activity of TLX1 is due to the direct or indirect transcriptional regulation of subordinate target genes. Aldh1a1, together with Fhl1/Slim1, which encodes a LIM-domain protein [31], [32], are two such genes previously identified as being TLX1-dependent in developing mouse spleen and/or murine cell lines [13], [14], [33]. Since RA is a key regulator of cellular differentiation, proliferation and apoptosis in diverse situations ranging from organogenesis to hematopoiesis, ALDH1A1 is of particular interest for its potential as a key mediator of normal and/or aberrant TLX1 function.

In this study, we demonstrate that ALDH1A1 is also a target for transcriptional regulation by TLX1 in human hematopoietic cells. Expression of ALDH1A1 was found in T-ALL cell lines that expressed TLX1, which was reminiscent of the correlation previously observed between SCL/LMO and ALDH1A2 expression in T-ALL [34]. A striking feature of TLX1 is its ability to impede hematopoietic cell differentiation [30], [35], [36], [37], [38]. We therefore employed retroviral-mediated transfer to enforce expression of ALDH1A1 in murine bone marrow progenitors. Using this approach we found that ALDH1A1 profoundly perturbed hematopoiesis by promoting myeloid differentiation at the expense of lymphopoiesis. These data implicate ALDH1A1 in the regulation of hematopoiesis, and suggest one mechanism by which TLX1 may accomplish its normal and/or oncogenic functions.

Section snippets

Cell culture and stable transfection

T-ALL cell lines were cultured in a humidified atmosphere at 37 °C with 5% CO2 in RPMI 1640 medium supplemented with 10% fetal calf serum (FCS; CSL, Melbourne, VIC, Australia), 0.03% l-glutamine, 0.03% pyruvate, 0.03% non-essential amino acids (Trace Scientific, Melbourne, VIC, Australia) and 1% penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA). The erythroleukemic cell line HEL, was likewise cultured without addition of pyruvate and non-essential amino acids. The cells were grown to a

ALDH1A1 gene expression is regulated by TLX1 in human hematopoietic cell lines

Aldh1a1 (formerly Aldh1 and also known as retinaldehyde dehydrogenase 1; Raldh1) gene expression is activated in murine cell lines and repressed in developing mouse spleen by TLX1 [13], [14], [15], [33], while a second putative target gene Fhl1/Slim1, was identified as a transcript induced by TLX1 expression in NIH 3T3 fibroblasts [13]. To assess whether ALDH1A1 and FHL1 expression could also be regulated by TLX1 in human hematopoietic cells, we used a T-cell line (PER-117) that does not

Discussion

In this study, ALDH1A1 was confirmed to be a subordinate target gene of TLX1 based on its up- and down-modulation in the human hematopoietic cell lines PER-117 and HEL, respectively. This adds to previous results showing that mouse Aldh1a1 is positively regulated by TLX1 in NIH 3T3 fibroblasts and J2E erythroleukemic cells [13], [33]. The ability of TLX1 to function as both an activator and repressor of endogenous ALDH1A1 transcription within multiple tissue types suggests that ALDH1A1 may be a

Acknowledgements

This work was supported by grants from the Australian National Health and Medical Research Council and Murdoch University. KLR was supported by an Australian Postgraduate Award. There are no conflicts of interest to declare.

Contributions. KLR, JF and AB performed most of the experiments and contributed to data analysis and interpretation. WKG, DJI and URK designed the study, interpreted the data, and critically revised the manuscript. In addition, WKG performed some of the experiments,

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