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Peripheral T-lymphocytes express WNT7A and its restoration in leukemia-derived lymphoblasts inhibits cell
Alejandra B Ochoa-Hernández1, Moisés Ramos-Solano, Ivan D Meza-Canales
1División de Genética, Centro de Investigación Biomédica de Occidente, Instituto Mexicano del Seguro Social, Guadalajara, Jalisco, Mexico.
Background:
WNT7a, a member of the Wnt ligand family implicated in several developmental processes, has also been reported to be dysregulated in some types of tumors; however, its function and implication in oncogenesis is poorly understood. Moreover, the expression of this gene and the role that it plays in the biology of blood cells remains unclear. In addition to determining the expression of the WNT7A gene in blood cells, in leukemia-derived cell lines, and in samples of patients with leukemia, the aim of this study was to seek the effect of this gene in proliferation.
Methods:
We analyzed peripheral blood mononuclear cells, sorted CD3 and CD19 cells, four leukemia-derived cell lines, and blood samples from 14 patients with Acute lymphoblastic leukemia (ALL), and 19 clinically healthy subjects. Reverse transcription followed by quantitative Real-time Polymerase chain reaction (qRT-PCR) analysis were performed to determine relative WNT7A expression. Restoration of WNT7a was done employing a lentiviral system and by using a recombinant human protein. Cell proliferation was measured by addition of WST-1 to cell cultures.
Results:
WNT7a is mainly produced by CD3 T-lymphocytes, its expression decreases upon activation, and it is severely reduced in leukemia-derived cell lines, as well as in the blood samples of patients with ALL when compared with healthy controls (p ≤0.001). By restoring WNT7A expression in leukemia-derived cells, we were able to demonstrate that WNT7a inhibits cell growth. A similar effect was observed when a recombinant human WNT7a protein was used. Interestingly, restoration of WNT7A expression in Jurkat cells did not activate the canonical Wnt/β-catenin pathway.
Conclusions:
To our knowledge, this is the first report evidencing quantitatively decreased WNT7A levels in leukemia-derived cells and that WNT7A restoration in T-lymphocytes inhibits cell proliferation. In addition, our results also support the possible function of WNT7A as a tumor suppressor gene as well as a therapeutic tool.
Insights
WNT7A levels are significantly decreased in leukemia. Restoring WNT7A in T-lymphocytes inhibits cell proliferation, suggesting its role as a tumor suppressor and potential therapeutic target.
Area of Science:
- Molecular Biology
- Oncology
- Hematology
Background:
- WNT7a, a Wnt ligand, is implicated in development and dysregulated in tumors, but its role in oncogenesis and blood cell biology is unclear.
- Investigating WNT7A expression and function in blood cells and leukemia is crucial for understanding its biological significance.
Purpose of the Study:
- To determine WNT7A gene expression in blood cells, leukemia cell lines, and patient samples.
- To investigate the effect of WNT7A on cell proliferation in leukemia.
Main Methods:
- Analysis of peripheral blood mononuclear cells, sorted lymphocytes, and leukemia cell lines.
- Quantitative Real-time Polymerase chain reaction (qRT-PCR) for WNT7A expression.
- WNT7A restoration via lentiviral system or recombinant protein; cell proliferation measured by WST-1 assay.
Main Results:
- WNT7a is primarily produced by CD3 T-lymphocytes and its expression decreases upon activation.
- WNT7A expression is significantly reduced in leukemia cell lines and Acute lymphoblastic leukemia (ALL) patient samples compared to healthy controls.
- Restoration of WNT7A inhibits proliferation in leukemia cells and Jurkat cells, without activating the canonical Wnt/β-catenin pathway.
Conclusions:
- This study provides the first quantitative evidence of decreased WNT7A levels in leukemia-derived cells.
- WNT7A restoration in T-lymphocytes inhibits cell proliferation, supporting its function as a tumor suppressor.
- WNT7A shows potential as a therapeutic tool for leukemia.
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