Dataset Information

Assessment of global peptide abundance in TOE1 depleted myeloid leukaemia cells

ABSTRACT: Acute myeloid leukaemia (AML) is an aggressive haematological malignancy characterised by the clonal proliferation of myeloid progenitor cells in the bone marrow and peripheral blood. Dysregulation of the Wnt/β-catenin pathway has been implicated in the establishment and maintenance of leukaemic stem cells in AML, where higher expression of β-catenin promotes clonogenic capacity, drug resistance and inferior survival. The finding that low levels of Wnt signalling are necessary to maintain normal haematopoiesis makes β-catenin an attractive therapeutic target, however drug design has been hampered by a poor understanding of its molecular interactions in leukaemia cells. To address this, we previously characterised the β-catenin interactome in myeloid cells and identified a plethora of novel interacting proteins. One such validated interactor was Target of EGR1 (TOE1), a 510-amino acid member of the Asp-Glu-Asp-Asp (DEDD) family of deadenylases with previously uncharacterised function in haematopoietic cells. The β-catenin:TOE1 interaction was detected in the nuclear and cytosolic compartments of myeloid cell lines and primary AML samples, and β-catenin knockdown via short hairpin RNA (shRNA) was found to promote the cytosolic stabilisation of TOE1. Furthermore, TOE1 levels were found to be overexpressed in primary AML blasts versus normal cord-blood derived CD34+ haematopoietic stem and progenitor cells (HSPCs), suggesting that TOE1 levels may be dysregulated in leukaemia. TOE1 depletion using shRNA abrogated Wnt signalling capacity (TCF/LEF activity) through suppression of the Wnt transcription factor lymphoid enhancing factor 1 (LEF-1), likely via post-transcriptional mechanisms. TOE1 shRNA further suppressed the proliferation and survival of myeloid leukaemia cell lines (HEL and OCI-AML2) and primary human CD34+ HSPC, however this could not be fully explained through LEF-1 alone, since OCI-AML2 do not express LEF-1. Therefore, to explore a more unifying mechanism mediating the growth promoting phenotype of TOE1 we performed tandem mass tag (TMT)-labelling coupled to mass spectrometry analysis in TOE1 deficient HEL and OCI-AML2 cells. From this analysis we identified validated p21 (RAC1) activated kinase 2 (PAK2) as a consistently downregulated target across both cell lines, and demonstrated PAK2 knockdown alone was sufficient to reduce the proliferation (but not survival) of AML cell lines. Finally, expression of ectopic PAK2 was able to partially rescue the suppressed proliferation mediated by TOE1 depletion in myeloid cell lines, and primary human CD34+ HSPC. In summary, these data reveal TOE1 as novel interacting partner for β-catenin in haematological cells capable of modulating Wnt signalling output via LEF-1, and as a novel mediator of growth and survival in human HSPC and AML cells partly through PAK2 regulation.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Leukocyte, Blood

DISEASE(S): Acute Leukemia

SUBMITTER: Rhys Morgan

LAB HEAD: Rhys G Morgan

PROVIDER: PXD070891 | Pride | 2026-04-29

REPOSITORIES: Pride

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			Action	DRS
	140125_HEL_Fraction_1.raw	Raw
	140125_HEL_Fraction_10.raw	Raw
	140125_HEL_Fraction_11.raw	Raw
	140125_HEL_Fraction_12.raw	Raw
	140125_HEL_Fraction_13.raw	Raw

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TOE1 is a β-catenin interacting protein regulating the proliferation of hematopoietic cells through PAK2 modulation.

Park Hyun H Sevim Okan O Wagstaff Megan M Goff Aaron A Palmer David A DA Kim Bomee B Heesom Kate K Blair Allison A Newbury Sarah F SF Morgan Ethan L EL Towler Benjamin P BP Chevassut Timothy J TJ Morgan Rhys G RG

Stem cell reports 20260423

Acute myeloid leukemia (AML) is an aggressive hematological malignancy frequently exhibiting deregulated expression/activity/localization of the Wnt signaling mediator β-catenin. To derive more effective β-catenin targeting strategies, we previously interrogated its interaction network in myeloid cells and identified several putative novel interacting partners, including Target of EGR1 (TOE1); a deadenylase with unknown function in hematological tissue. This study aimed to define TOE1 function i ...[more]

PMID: 42030941

Similar Datasets

Project description:Background: Acute myeloid leukaemia is a disease characterised by the accumulation of large number of aberrant, immature myeloid cells in the bone marrow and peripheral blood as a result of a variety of genetic alterations in early haematopoietic stem cells. Normal stem cells, which have the ability to renew themselves, can undergo neoplastic transformation as a result of mutations in signalling pathway genes, changes in epigenetics or gene expression levels. The gene expression levels of beta-catenin, a key element of the Wnt signalling pathway, which is particularly active during embryogenesis and in various cancers, have been widely studied in the literature. In these studies, which were consistent with each other, serious differences in beta-catenin gene expression levels were found in AML patients. On this basis, our aim of this study was to determine beta-catenin gene expression levels in AML patients. In the next part of the study, it is planned to compare the genes between AML patient samples grouped according to beta-catenin gene expression levels by DNA microarray analysis. When the gene group identified in this way is combined with the genes in the healthy control group, the aim is to determine the genes and pathways that are associated with beta-catenin in AML patients. Material and Method: In this study, beta-catenin gene expression levels were determined in bone marrow/blood CD34+ cells of 19 AML patients and 3 healthy controls by qRT-PCR. Transcriptome analysis was performed on AML patient samples grouped according to beta-catenin expression levels. After transcriptome analysis, differentially expressed genes (DEGs) were identified using a 1.5-fold change threshold. DEGs were further investigated using the online analysis tools DAVID, GO, KEGG and STRING. Results: In this study, the mRNA analysis showed that beta-catenin levels varied between AML samples. Based on these results, the AML samples were divided into 2 groups: high and low beta catenin. The transcriptome profiles of the AML samples showed different molecular signature profiles according on their beta-catenin levels (high-low) when compared to the healthy control group. In our study, it was found that hub genes related to cell cycle, ribosome biogenesis and transcription factor pathways were beta-catenin-related genes and were prominent in the group of AML patients with high beta-catenin levels. Conclusion: It is suggested that the identification of beta-catenin-related gene profiles in AML may help to select new molecular therapeutic targets for the treatment of AML.