Evidence that knock down of GSK-3β in Chronic Myelogenous Leukemia cells augments IFN-γ-induced apoptosis

Melissa R Kauffman; Saeed Nazemidashtarjandi; Davoud Ghazanfari; Abigail E Allen; Nathan M Reynolds; Ahmed Faik; Monica M Burdick; Kelly D McCall; Douglas J Goetz

doi:10.1016/j.leukres.2020.106464

Evidence that knock down of GSK-3β in Chronic Myelogenous Leukemia cells augments IFN-γ-induced apoptosis

Leuk Res. 2020 Dec:99:106464. doi: 10.1016/j.leukres.2020.106464. Epub 2020 Oct 21.

Authors

Melissa R Kauffman¹, Saeed Nazemidashtarjandi², Davoud Ghazanfari², Abigail E Allen¹, Nathan M Reynolds³, Ahmed Faik⁴, Monica M Burdick⁵, Kelly D McCall⁶, Douglas J Goetz⁷

Affiliations

¹ Department of Chemical and Biomolecular Engineering, Ohio University, Athens, OH 45701, USA; Biomedical Engineering Program, Ohio University, Athens, OH 45701, USA.
² Department of Chemical and Biomolecular Engineering, Ohio University, Athens, OH 45701, USA.
³ Department of Chemical and Biomolecular Engineering, Ohio University, Athens, OH 45701, USA; Translational Biomedical Sciences Program, Ohio University, Athens, OH 45701, USA.
⁴ Environmental and Plant Biology Department, Ohio University, Athens, OH 45701, USA; Molecular and Cellular Biology Program, Ohio University, Athens, OH 45701, USA.
⁵ Department of Chemical and Biomolecular Engineering, Ohio University, Athens, OH 45701, USA; Biomedical Engineering Program, Ohio University, Athens, OH 45701, USA; Molecular and Cellular Biology Program, Ohio University, Athens, OH 45701, USA; Translational Biomedical Sciences Program, Ohio University, Athens, OH 45701, USA.
⁶ Biomedical Engineering Program, Ohio University, Athens, OH 45701, USA; Molecular and Cellular Biology Program, Ohio University, Athens, OH 45701, USA; Translational Biomedical Sciences Program, Ohio University, Athens, OH 45701, USA; Department of Specialty Medicine, Ohio University, Athens, OH 45701, USA; The Diabetes Institute, Ohio University, Athens, OH 45701, USA.
⁷ Department of Chemical and Biomolecular Engineering, Ohio University, Athens, OH 45701, USA; Biomedical Engineering Program, Ohio University, Athens, OH 45701, USA; Molecular and Cellular Biology Program, Ohio University, Athens, OH 45701, USA; Translational Biomedical Sciences Program, Ohio University, Athens, OH 45701, USA. Electronic address: goetzd@ohio.edu.

Abstract

The role of interferon-gamma (IFN-γ) in Chronic Myelogenous/Myeloid Leukemia (CML) and in the treatment of CML remains unclear; specifically, the effect of IFN-γ on apoptosis. There is reported interplay between IFN-γ and glycogen synthase kinase-3 (GSK-3), a kinase which has been implicated in both cell death and, conversely, cell survival. Thus, we utilized the CML-derived HAP1 cell line and a mutant HAP1 GSK-3β knocked-down cell line (GSK-3β 31bp) to investigate whether GSK-3 modulates IFN-γ's action on CML cells. Significantly less GSK-3β 31bp cells, relative to HAP1 cells, were present after 48 h treatment with IFN-γ. IFN-γ treatment significantly decreased GSK-3β 31bp substrate adhesiveness (relative to HAP1 cells); an observation often correlated with cell death. Fluorescence microscopy revealed that IFN-γ induces a modest level of apoptosis in the HAP1 cells and that IFN-γ induced apoptosis is significantly enhanced in GSK-3β 31bp cells. Utilizing a complementary GSK-3β knocked-down cell line (8bp) we found, via flow cytometric analysis, that IFN-γ induced apoptosis is significantly enhanced in GSK-3β 8bp cells relative to HAP1 cells. Combined, our findings suggest that IFN-γ induces apoptosis of CML cells and that loss of GSK-3β significantly augments IFN-γ-induced apoptosis.

Keywords: Apoptosis; BCR-ABL; Glycogen synthase kinase-3; Interferon; Leukemia; Philadelphia chromosome.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Antineoplastic Agents / pharmacology
Apoptosis / drug effects*
CRISPR-Cas Systems
Cell Adhesion / drug effects
Cell Line, Tumor / drug effects
Codon, Nonsense
Drug Interactions
Flow Cytometry
Frameshift Mutation
Fusion Proteins, bcr-abl / antagonists & inhibitors
Gene Knockdown Techniques
Glycogen Synthase Kinase 3 beta / antagonists & inhibitors*
Glycogen Synthase Kinase 3 beta / genetics
Humans
Interferon-gamma / pharmacology*
Interferon-gamma / therapeutic use
Leukemia, Myelogenous, Chronic, BCR-ABL Positive / drug therapy*
Leukemia, Myelogenous, Chronic, BCR-ABL Positive / enzymology
Leukemia, Myelogenous, Chronic, BCR-ABL Positive / pathology
Male
Neoplasm Proteins / antagonists & inhibitors*
Protein Kinase Inhibitors / pharmacology*
RNA, Guide, CRISPR-Cas Systems
Spectrometry, Fluorescence

Substances

Antineoplastic Agents
Codon, Nonsense
IFNG protein, human
Neoplasm Proteins
Protein Kinase Inhibitors
Interferon-gamma
Fusion Proteins, bcr-abl
Glycogen Synthase Kinase 3 beta

Abstract

Publication types

MeSH terms

Substances

Grants and funding