Cep55 regulation of PI3K/Akt signaling is required for neocortical development and ciliogenesis

Rashidieh, Behnam; Shohayeb, Belal; Bain, Amanda L.; Fortuna, Patrick R.J.; Sinha, Debottam; Burgess, Andrew; Mills, Richard J.; Adams, Ryan A.; Lopez, J. Alejandro; Blumbergs, Peter; Finnie, John; Kalimutho, Murugan; Piper, Michael; Hudson, James E.; Ng, Dominic C. H.; Khanna, Kum Kum

doi:10.1371/journal.pgen.1009334

Cited by 4 publications

(9 citation statements)

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“…Mouse embryonic fibroblasts (MEFs) were generated as described previously [ 8 ]. Briefly, primary MEFs were immortalized and transformed by E1A/Ras oncogene.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2023

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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.

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“…Mouse embryonic fibroblasts (MEFs) were generated as described previously [ 8 ]. Briefly, primary MEFs were immortalized and transformed by E1A/Ras oncogene.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2023

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“…IF was performed as described previously 28 Briefly, 50,000 cells were seeded on UV pre-treated coverslips within a 24-well plate. After heat shock (43 C), to induce Cryab expression, cells were fixed using 4% Paraformaldehyde (PFA) followed by permeabilization (0.5% Triton X-100 for 5 min) and blocking (2% BSA in PBS) before staining with primary and secondary antibodies(Table 2 below) and mounting on the glass cover slides using Prolong Gold antifade reagent (Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

“…Cells were prepared for lysis as described previously 28 with indicated antibodies (table below). The Super Signal chemiluminescent ECL-plus (Amersham) was used for signal detection.…”

Section: Immunoblottingmentioning

confidence: 99%

“…PCR was performed as described previously 28 using Genomic DNA from animal tissue or cell pellet was extracted using QuickExtract DNA solution (Gene target solutions, QE09050).…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…Mouse embryonic fibroblasts were generated as described previously 28 . Briefly, primary MEFs were immortalized by E1A/Ras transduction.…”

Section: Cell Culturementioning

confidence: 99%

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

Rashidieh

Bain

Tria

et al. 2022

Preprint

Self Cite

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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 reported tumor suppressor or tumor promoter (oncogene) roles for αB-Crystallin depending on cellular context and environmental conditions. To determine the causal relationship between CRYAB overexpression and cancer, we generated a Cryab overexpression knock-in mouse model. 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 mouse embryonic fibroblasts (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. 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.

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Proteome balance in ciliopathies: the OFD1 protein example

Morleo

Pezzella

Franco

2023

Trends in Molecular Medicine

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Cep55 regulation of PI3K/Akt signaling is required for neocortical development and ciliogenesis

Cited by 4 publications

References 58 publications

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

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

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

Proteome balance in ciliopathies: the OFD1 protein example

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