Alpha-B-Crystallin overexpression is sufficient to promote tumorigenesis and metastasis in mice

Behnam Rashidieh; Amanda Louise Bain; Simon Manuel Tria; Sowmya Sharma; Cameron Allan Stewart; Jacinta Ley Simmons; Pirjo M Apaja; Pascal H G Duijf; John Finnie; Kum Kum Khanna

doi:10.1186/s40164-022-00365-z

Alpha-B-Crystallin overexpression is sufficient to promote tumorigenesis and metastasis in mice

Exp Hematol Oncol. 2023 Jan 9;12(1):4. doi: 10.1186/s40164-022-00365-z.

Authors

Behnam Rashidieh^{1

2}, Amanda Louise Bain¹, Simon Manuel Tria^{1

3}, Sowmya Sharma¹, Cameron Allan Stewart^{1

3}, Jacinta Ley Simmons¹, Pirjo M Apaja^{4

5

6}, Pascal H G Duijf^{7

8

9

10

11

12}, John Finnie¹³, Kum Kum Khanna¹⁴

Affiliations

¹ QIMR Berghofer Medical Research Institute, 300 Herston Road, Herston, Brisbane, QLD, 4006, Australia.
² School of Biomedical Sciences, Faculty of Medicine, University of Queensland, Brisbane, QLD, 4006, Australia.
³ School of Environment and Science, Griffith University, 170 Kessels Road, Nathan, Brisbane, QLD, 4111, Australia.
⁴ South Australian Health and Medical Research Institute, Lifelong Health, Organelle Proteostasis Diseases, Adelaide, SA, 5000, Australia.
⁵ Department of Molecular and Biomedical Sciences, University of Adelaide, Adelaide, SA, 5000, Australia.
⁶ College of Public Health and Medicine, Flinders University, Bedford Park, SA, 5042, Australia.
⁷ School of Biomedical Sciences, Faculty of Health, Queensland University of Technology (QUT), Brisbane, QLD, 4000, Australia.
⁸ Centre for Data Science, Queensland University of Technology (QUT), Brisbane, QLD, 4000, Australia.
⁹ Centre for Genomics and Personalised Health, Queensland University of Technology, Brisbane, QLD, 4000, Australia.
¹⁰ Cancer and Aging Research Program, Queensland University of Technology, Brisbane, QLD, 4000, Australia.
¹¹ Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, 0372, Oslo, Norway.
¹² Department of Medical Genetics, Oslo University Hospital, 0450, Oslo, Norway.
¹³ Discipline of Anatomy and Pathology, Adelaide Medical School, University of Adelaide, Adelaide, SA, 5000, Australia.
¹⁴ QIMR Berghofer Medical Research Institute, 300 Herston Road, Herston, Brisbane, QLD, 4006, Australia. KumKum.Khanna@qimrberghofer.edu.au.

Abstract

Background: αB-Crystallin is a heat shock chaperone protein which binds to misfolded proteins to prevent their aggregation. It is overexpressed in a wide-variety of cancers. Previous studies using human cancer cell lines and human xenograft models have suggested potential tumor promoter (oncogene) roles for αB-Crystallin in a wide-spectrum of cancers.

Methods: To determine the causal relationship between CRYAB overexpression and cancer, we generated a Cryab overexpression knock-in mouse model and monitor them for development of spontaneous and carcinogen (DMBA)-induced tumorigenesis. In order to investigate the mechanism of malignancies observed in this model multiple techniques were used such as immunohistochemical characterizations of tumors, bioinformatics analysis of publically available human tumor datasets, and generation of mouse embryonic fibroblasts (MEFs) for in vitro assays (clonogenic survival and migration assays and proteome analysis by mass-spectrometry).

Results: This model revealed that constitutive overexpression of Cryab results in the formation of a variety of lethal spontaneous primary and metastatic tumors in mice. In vivo, the overexpression of Cryab correlated with the upregulation of epithelial-to-mesenchymal (EMT) markers, angiogenesis and some oncogenic proteins including Basigin. In vitro, using E1A/Ras transformed MEFs, we observed that the overexpression of Cryab led to the promotion of cell survival via upregulation of Akt signaling and downregulation of pro-apoptotic pathway mediator JNK, with subsequent attenuation of apoptosis as assessed by cleaved caspase-3 and Annexin V staining.

Conclusions: Overall, through the generation and characterization of Cryab overexpression model, we provide evidence supporting the role of αB-Crystallin as an oncogene, where its upregulation is sufficient to induce tumors, promote cell survival and inhibit apoptosis.

Keywords: Angiogenesis; Cryab mouse model; Heat-shock protein; Metastasis; Tumorigenesis.