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Identification of the Adapter Molecule MTSS1 as a Potential Oncogene-Specific Tumor Suppressor in Acute Myeloid Leukemia.

Schemionek M, Kharabi Masouleh B, Klaile Y, Krug U, Hebestreit K, Schubert C, Dugas M, Büchner T, Wörmann B, Hiddemann W, Berdel WE, Brümmendorf TH, Müller-Tidow C, Koschmieder S - PLoS ONE (2015)

Bottom Line: High expression of MTSS1 predicted better clinical outcome of patients with normal-karyotype AML.Pharmacological treatment of AML cell lines carrying the FLT3-ITD mutation with the specific FLT3 inhibitor PKC-412 caused upregulation of MTSS1.Moreover, treatment of acute promyelocytic cells (APL) with all-trans retinoic acid (ATRA) increased MTSS1 mRNA levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Oncology, Hemostaseology, and Stem Cell Transplantation, Faculty of Medicine, RWTH Aachen University, Aachen, Germany.

ABSTRACT
The adapter protein metastasis suppressor 1 (MTSS1) is implicated as a tumor suppressor or tumor promoter, depending on the type of solid cancer. Here, we identified Mtss1 expression to be increased in AML subsets with favorable outcome, while suppressed in high risk AML patients. High expression of MTSS1 predicted better clinical outcome of patients with normal-karyotype AML. Mechanistically, MTSS1 expression was negatively regulated by FLT3-ITD signaling but enhanced by the AML1-ETO fusion protein. DNMT3B, a negative regulator of MTSS1, showed strong binding to the MTSS1 promoter in PML-RARA positive but not AML1-ETO positive cells, suggesting that AML1-ETO leads to derepression of MTSS1. Pharmacological treatment of AML cell lines carrying the FLT3-ITD mutation with the specific FLT3 inhibitor PKC-412 caused upregulation of MTSS1. Moreover, treatment of acute promyelocytic cells (APL) with all-trans retinoic acid (ATRA) increased MTSS1 mRNA levels. Taken together, our findings suggest that MTSS1 might have a context-dependent function and could act as a tumor suppressor, which is pharmacologically targetable in AML patients.

No MeSH data available.


Related in: MedlinePlus

MTSS1 is positively regulated by AML1-ETO in human AML cells.The gene expression of MTSS1 in human AML cells (Kasumi-1 without transduction (no Trans), scramble siRNA (ctr RNAi) or siRNA targeting the AML1-ETO translocation is shown (GEO accession numbers GSE15646; A). Similarly, the gene expression of MTSS1, DNMT3B and 3A are measured in a gene expression analysis of human cord-blood derived CD34+ cells and patient-derived AML samples with the AML-1 ETO translocation (GEO accession numbers GSE8023; B and C). Human AML cell lines carrying either a FLT3-ITD mutation (MV4-11), t(15;17) (NB4), t(8;21) (Kasumi-1) or not defined (U937) were transduced with a MTSS1 firefly / renilla construct and the MTSS1 promoter activity as a normalized relative ratio was measured (n = 3;D).
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pone.0125783.g004: MTSS1 is positively regulated by AML1-ETO in human AML cells.The gene expression of MTSS1 in human AML cells (Kasumi-1 without transduction (no Trans), scramble siRNA (ctr RNAi) or siRNA targeting the AML1-ETO translocation is shown (GEO accession numbers GSE15646; A). Similarly, the gene expression of MTSS1, DNMT3B and 3A are measured in a gene expression analysis of human cord-blood derived CD34+ cells and patient-derived AML samples with the AML-1 ETO translocation (GEO accession numbers GSE8023; B and C). Human AML cell lines carrying either a FLT3-ITD mutation (MV4-11), t(15;17) (NB4), t(8;21) (Kasumi-1) or not defined (U937) were transduced with a MTSS1 firefly / renilla construct and the MTSS1 promoter activity as a normalized relative ratio was measured (n = 3;D).

Mentions: Since MTSS1 expression was high in t(8;21) positive cell lines and primary AML samples, we next studied whether there was a direct link between MTSS1 and AML1-ETO. Using publicly available databases, we could show that there is a trend of MTSS1 reduction after siRNA-mediated knockdown of AML1-ETO in human t(8;21) positive AML cells (Kasumi-1) (Fig 4A). Similarly, in patient-derived AML1-ETO positive primary AML cells, MTSS1 mRNA levels were significantly increased compared to human cord-blood-derived CD34+ myeloid cells (Fig 4B). When assayed for expression, Mtss1 was low (blue squares) in CD34 positive normal cells but increased in t(8;21) cells (red squares) and this was also true for DNMT3A expression. Reversely, expression of DNMT3B was high (red squares) in CD34 positive normal cells but low in t(8;21) cells (blue and white squares (Fig 4C). Using our own experiments, we next tested the hypothesis if MTSS1 promoter activity is directly regulated by AML1-ETO, using the same Kasumi-1 cell line that had suggested a trend of AML1-ETO siRNA mediated downregulation of MTSS1 (Fig 4A). Luciferase promoter assay using a promoter region containing a fragment of -276 to -13 bp has previously been described to confer highest Mtss1 expression. The indicated nucleotide position -276 to -13bp is related to the 5´end of annotated Mtss1 mRNA sequence with the accession number (NM_014751.5) [21]. Our data demonstrate that MTSS1 promoter activity was significantly higher in human t(8;21) positive AML cells (Kasumi-1) when compared to various AML1-ETO negative cell lines (Fig 4D). This suggests that the AML1-ETO fusion molecule is a positive regulator of MTSS1.


Identification of the Adapter Molecule MTSS1 as a Potential Oncogene-Specific Tumor Suppressor in Acute Myeloid Leukemia.

Schemionek M, Kharabi Masouleh B, Klaile Y, Krug U, Hebestreit K, Schubert C, Dugas M, Büchner T, Wörmann B, Hiddemann W, Berdel WE, Brümmendorf TH, Müller-Tidow C, Koschmieder S - PLoS ONE (2015)

MTSS1 is positively regulated by AML1-ETO in human AML cells.The gene expression of MTSS1 in human AML cells (Kasumi-1 without transduction (no Trans), scramble siRNA (ctr RNAi) or siRNA targeting the AML1-ETO translocation is shown (GEO accession numbers GSE15646; A). Similarly, the gene expression of MTSS1, DNMT3B and 3A are measured in a gene expression analysis of human cord-blood derived CD34+ cells and patient-derived AML samples with the AML-1 ETO translocation (GEO accession numbers GSE8023; B and C). Human AML cell lines carrying either a FLT3-ITD mutation (MV4-11), t(15;17) (NB4), t(8;21) (Kasumi-1) or not defined (U937) were transduced with a MTSS1 firefly / renilla construct and the MTSS1 promoter activity as a normalized relative ratio was measured (n = 3;D).
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Related In: Results  -  Collection

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pone.0125783.g004: MTSS1 is positively regulated by AML1-ETO in human AML cells.The gene expression of MTSS1 in human AML cells (Kasumi-1 without transduction (no Trans), scramble siRNA (ctr RNAi) or siRNA targeting the AML1-ETO translocation is shown (GEO accession numbers GSE15646; A). Similarly, the gene expression of MTSS1, DNMT3B and 3A are measured in a gene expression analysis of human cord-blood derived CD34+ cells and patient-derived AML samples with the AML-1 ETO translocation (GEO accession numbers GSE8023; B and C). Human AML cell lines carrying either a FLT3-ITD mutation (MV4-11), t(15;17) (NB4), t(8;21) (Kasumi-1) or not defined (U937) were transduced with a MTSS1 firefly / renilla construct and the MTSS1 promoter activity as a normalized relative ratio was measured (n = 3;D).
Mentions: Since MTSS1 expression was high in t(8;21) positive cell lines and primary AML samples, we next studied whether there was a direct link between MTSS1 and AML1-ETO. Using publicly available databases, we could show that there is a trend of MTSS1 reduction after siRNA-mediated knockdown of AML1-ETO in human t(8;21) positive AML cells (Kasumi-1) (Fig 4A). Similarly, in patient-derived AML1-ETO positive primary AML cells, MTSS1 mRNA levels were significantly increased compared to human cord-blood-derived CD34+ myeloid cells (Fig 4B). When assayed for expression, Mtss1 was low (blue squares) in CD34 positive normal cells but increased in t(8;21) cells (red squares) and this was also true for DNMT3A expression. Reversely, expression of DNMT3B was high (red squares) in CD34 positive normal cells but low in t(8;21) cells (blue and white squares (Fig 4C). Using our own experiments, we next tested the hypothesis if MTSS1 promoter activity is directly regulated by AML1-ETO, using the same Kasumi-1 cell line that had suggested a trend of AML1-ETO siRNA mediated downregulation of MTSS1 (Fig 4A). Luciferase promoter assay using a promoter region containing a fragment of -276 to -13 bp has previously been described to confer highest Mtss1 expression. The indicated nucleotide position -276 to -13bp is related to the 5´end of annotated Mtss1 mRNA sequence with the accession number (NM_014751.5) [21]. Our data demonstrate that MTSS1 promoter activity was significantly higher in human t(8;21) positive AML cells (Kasumi-1) when compared to various AML1-ETO negative cell lines (Fig 4D). This suggests that the AML1-ETO fusion molecule is a positive regulator of MTSS1.

Bottom Line: High expression of MTSS1 predicted better clinical outcome of patients with normal-karyotype AML.Pharmacological treatment of AML cell lines carrying the FLT3-ITD mutation with the specific FLT3 inhibitor PKC-412 caused upregulation of MTSS1.Moreover, treatment of acute promyelocytic cells (APL) with all-trans retinoic acid (ATRA) increased MTSS1 mRNA levels.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Oncology, Hemostaseology, and Stem Cell Transplantation, Faculty of Medicine, RWTH Aachen University, Aachen, Germany.

ABSTRACT
The adapter protein metastasis suppressor 1 (MTSS1) is implicated as a tumor suppressor or tumor promoter, depending on the type of solid cancer. Here, we identified Mtss1 expression to be increased in AML subsets with favorable outcome, while suppressed in high risk AML patients. High expression of MTSS1 predicted better clinical outcome of patients with normal-karyotype AML. Mechanistically, MTSS1 expression was negatively regulated by FLT3-ITD signaling but enhanced by the AML1-ETO fusion protein. DNMT3B, a negative regulator of MTSS1, showed strong binding to the MTSS1 promoter in PML-RARA positive but not AML1-ETO positive cells, suggesting that AML1-ETO leads to derepression of MTSS1. Pharmacological treatment of AML cell lines carrying the FLT3-ITD mutation with the specific FLT3 inhibitor PKC-412 caused upregulation of MTSS1. Moreover, treatment of acute promyelocytic cells (APL) with all-trans retinoic acid (ATRA) increased MTSS1 mRNA levels. Taken together, our findings suggest that MTSS1 might have a context-dependent function and could act as a tumor suppressor, which is pharmacologically targetable in AML patients.

No MeSH data available.


Related in: MedlinePlus