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Endoplasmic reticulum chaperone GRP78/BiP is critical for mutant Kras-driven lung tumorigenesis

Oncogene volume 40, pages 3624–3632 (2021)Cite this article

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Abstract

Lung cancer is the leading cause of cancer mortality worldwide and KRAS is the most commonly mutated gene in lung adenocarcinoma (LUAD). The 78-kDa glucose-regulated protein GRP78/BiP is a key endoplasmic reticulum chaperone protein and a major pro-survival effector of the unfolded protein response (UPR). Analysis of the Cancer Genome Atlas database and immunostain of patient tissues revealed that compared to normal lung, GRP78 expression is generally elevated in human lung cancers, including tumors bearing the KRASG12D mutation. To test the requirement of GRP78 in human lung oncogenesis, we generated mouse models containing floxed Grp78 and Kras Lox-Stop-Lox G12D (KrasLSL-G12D) alleles. Simultaneous activation of the KrasG12D allele and knockout of the Grp78 alleles were achieved in the whole lung or selectively in lung alveolar epithelial type 2 cells known to be precursors for adenomas that progress to LUAD. Here we report that GRP78 haploinsufficiency is sufficient to suppress KrasG12D-mediated lung tumor progression and prolong survival. Furthermore, GRP78 knockdown in human lung cancer cell line A427 (KrasG12D/+) leads to activation of UPR and apoptotic markers and loss of cell viability. Our studies provide evidence that targeting GRP78 represents a novel therapeutic approach to suppress mutant KRAS-mediated lung tumorigenesis.

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Fig. 1: Characterization of the adenovirus-Cre mouse model.
Fig. 2: Comparative analysis of the lungs of K78+/+, K78f/+, and K78f/f mice following adenovirus-Cre treatment.
Fig. 3: Imaging of K78+/+, K78f/+, K78f/f mouse lungs.
Fig. 4: Analysis and characterization of CK78+/+, CK78f/+, and CK78f/f mice following SPC-Cre activation and effect of GRP78 knockdown in a human lung cancer cell line.

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References

  1. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87–108.

    Article  Google Scholar 

  2. Jackson EL, Willis N, Mercer K, Bronson RT, Crowley D, Montoya R, et al. Analysis of lung tumor initiation and progression using conditional expression of oncogenic K-ras. Genes Dev. 2001;15:3243–8.

    Article  CAS  Google Scholar 

  3. Cancer Genome Atlas Research Network. Comprehensive molecular profiling of lung adenocarcinoma. Nature. 2014;511:543–50.

    Article  Google Scholar 

  4. Hancock JF, Magee AI, Childs JE, Marshall CJ. All ras proteins are polyisoprenylated but only some are palmitoylated. Cell. 1989;57:1167–77.

    Article  CAS  Google Scholar 

  5. Hancock JF, Paterson H, Marshall CJ. A polybasic domain or palmitoylation is required in addition to the CAAX motif to localize p21ras to the plasma membrane. Cell. 1990;63:133–9.

    Article  CAS  Google Scholar 

  6. Dai Q, Choy E, Chiu V, Romano J, Slivka SR, Steitz SA, et al. Mammalian prenylcysteine carboxyl methyltransferase is in the endoplasmic reticulum. J Biol Chem. 1998;273:15030–4.

    Article  CAS  Google Scholar 

  7. Luo B, Lee AS. The critical roles of endoplasmic reticulum chaperones and unfolded protein response in tumorigenesis and anticancer therapies. Oncogene. 2013;32:805–18.

    Article  CAS  Google Scholar 

  8. Ni M, Lee AS. ER chaperones in mammalian development and human diseases. FEBS Lett. 2007;581:3641–51.

    Article  CAS  Google Scholar 

  9. Pobre KFR, Poet GJ, Hendershot LM. The endoplasmic reticulum (ER) chaperone BiP is a master regulator of ER functions: Getting by with a little help from ERdj friends. J Biol Chem. 2019;294:2098–108.

    Article  CAS  Google Scholar 

  10. Lee AS. Glucose-regulated proteins in cancer: molecular mechanisms and therapeutic potential. Nat Rev Cancer. 2014;14:263–76.

    Article  CAS  Google Scholar 

  11. Wang M, Kaufman RJ. The impact of the endoplasmic reticulum protein-folding environment on cancer development. Nat Rev Cancer. 2014;14:581–97.

    Article  CAS  Google Scholar 

  12. Zhang Y, Liu R, Ni M, Gill P, Lee AS. Cell surface relocalization of the endoplasmic reticulum chaperone and unfolded protein response regulator GRP78/BiP. J Biol Chem. 2010;285:15065–75.

    Article  CAS  Google Scholar 

  13. Ni M, Zhang Y, Lee AS. Beyond the endoplasmic reticulum: atypical GRP78 in cell viability, signalling and therapeutic targeting. Biochem J. 2011;434:181–8.

    Article  CAS  Google Scholar 

  14. Reddy RK, Mao C, Baumeister P, Austin RC, Kaufman RJ, Lee AS. Endoplasmic reticulum chaperone protein GRP78 protects cells from apoptosis induced by topoisomerase inhibitors: role of ATP binding site in suppression of caspase-7 activation. J Biol Chem. 2003;278:20915–24.

    Article  CAS  Google Scholar 

  15. Fu Y, Li J, Lee AS. GRP78/BiP inhibits endoplasmic reticulum BIK and protects human breast cancer cells against estrogen starvation-induced apoptosis. Cancer Res. 2007;67:3734–40.

    Article  CAS  Google Scholar 

  16. Rutkowski DT, Arnold SM, Miller CN, Wu J, Li J, Gunnison KM, et al. Adaptation to ER stress is mediated by differential stabilities of pro-survival and pro-apoptotic mRNAs and proteins. PLoS Biol. 2006;4. 10.137/journal.pbio.0040374.

  17. Chae YC, Caino MC, Lisanti S, Ghosh JC, Dohi T, Danial NN, et al. Control of tumor bioenergetics and survival stress signaling by mitochondrial HSP90s. Cancer Cell. 2012;22:331–44.

    Article  CAS  Google Scholar 

  18. Uramoto H, Uchiumi T, Izumi H, Kohno K, Oyama T, Sugio K, et al. A new mechanism for primary resistance to gefitinib in lung adenocarcinoma: the role of a novel G796A mutation in exon 20 of EGFR. Anticancer Res. 2007;27:2297–303.

    CAS  PubMed  Google Scholar 

  19. Ma X, Guo W, Yang S, Zhu X, Xiang J, Li H. Serum GRP78 as a tumor marker and its prognostic significance in non-small cell lung cancers: a retrospective study. Dis Markers. 2015. https://doi.org/10.1155/2015/814670.

  20. Kwon D, Koh J, Kim S, Go H, Min HS, Kim YA, et al. Overexpression of endoplasmic reticulum stress-related proteins, XBP1s and GRP78, predicts poor prognosis in pulmonary adenocarcinoma. Lung Cancer. 2018;122:131–7.

    Article  Google Scholar 

  21. Imai H, Kaira K, Minato K. Clinical significance of post-progression survival in lung cancer. Thorac Cancer. 2017;8:379–86.

    Article  Google Scholar 

  22. Fu Y, Wey S, Wang M, Ye R, Liao CP, Roy-Burman P, et al. Pten null prostate tumorigenesis and AKT activation are blocked by targeted knockout of ER chaperone GRP78/BiP in prostate epithelium. Proc Natl Acad Sci USA. 2008;105:19444–9.

    Article  CAS  Google Scholar 

  23. Desai TJ, Brownfield DG, Krasnow MA. Alveolar progenitor and stem cells in lung development, renewal and cancer. Nature. 2014;507:190–4.

    Article  CAS  Google Scholar 

  24. Chapman HA, Li X, Alexander JP, Brumwell A, Lorizio W, Tan K, et al. Integrin alpha6beta4 identifies an adult distal lung epithelial population with regenerative potential in mice. J Clin Invest. 2011;121:2855–62.

    Article  CAS  Google Scholar 

  25. Wey S, Luo S, Tseng CC, Ni M, Zhou H, Fu Y, et al. Inducible knockout of GRP78/BiP in the hematopoietic system suppresses Pten-null leukemogenesis and AKT oncogenic signaling. Blood. 2012;119:817–25.

    Article  CAS  Google Scholar 

  26. Shen J, Ha DP, Zhu G, Rangel DF, Kobielak A, Gill PS, et al. GRP78 haploinsufficiency suppresses acinar-to-ductal metaplasia, signaling, and mutant Kras-driven pancreatic tumorigenesis in mice. Proc Natl Acad Sci USA. 2017;114:E4020–9.

    Article  CAS  Google Scholar 

  27. Du T, Li H, Fan Y, Yuan L, Guo X, Zhu Q, et al. The deubiquitylase OTUD3 stabilizes GRP78 and promotes lung tumorigenesis. Nat Commun. 2019;10:2914.

    Article  Google Scholar 

  28. Ye R, Jung DY, Jun JY, Li J, Luo S, Ko HJ, et al. Grp78 heterozygosity promotes adaptive unfolded protein response and attenuates diet-induced obesity and insulin resistance. Diabetes. 2010;59:6–16.

    Article  CAS  Google Scholar 

  29. Lee AS, Brandhorst S, Rangel DF, Navarrete G, Cohen P, Longo VD, et al. Effects of prolonged GRP78 haploinsufficiency on organ homeostasis, behavior, cancer and chemotoxic resistance in aged mice. Sci Rep. 2017;7:40919.

    Article  CAS  Google Scholar 

  30. Zhang X, He Z, Xiang L, Li L, Zhang H, Lin F, et al. Codelivery of GRP78 siRNA and docetaxel via RGD-PEG-DSPE/DOPA/CaP nanoparticles for the treatment of castration-resistant prostate cancer. Drug Des Devel Ther. 2019;13:1357–72.

    Article  CAS  Google Scholar 

  31. Cerezo M, Lehraiki A, Millet A, Rouaud F, Plaisant M, Jaune E, et al. Compounds triggering ER stress exert anti-melanoma effects and overcome BRAF inhibitor resistance. Cancer Cell. 2016;29:805–19.

    Article  CAS  Google Scholar 

  32. Bakewell SJ, Rangel DF, Ha DP, Sethuraman J, Crouse R, Hadley E, et al. Suppression of stress induction of the 78-kilodalton glucose regulated protein (GRP78) in cancer by IT-139, an anti-tumor ruthenium small molecule inhibitor. Oncotarget. 2018;9:29698–714.

    Article  Google Scholar 

  33. Burris HA, Bakewell S, Bendell JC, Infante J, Jones SF, Spigel DR, et al. Safety and activity of IT-139, a ruthenium-based compound, in patients with advanced solid tumours: a first-in-human, open-label, dose-escalation phase I study with expansion cohort. ESMO Open. 2016;1. https://doi.org/10.1136/esmoopen-2016-000154.

  34. Gifford JB, Huang W, Zeleniak AE, Hindoyan A, Wu H, Donahue TR, et al. Expression of GRP78, master regulator of the unfolded protein response, increases chemoresistance in pancreatic ductal adenocarcinoma. Mol Cancer Ther. 2016;15:1043–52.

    Article  CAS  Google Scholar 

  35. Lizardo MM, Morrow JJ, Miller TE, Hong ES, Ren L, Mendoza A, et al. Upregulation of glucose-regulated protein 78 in metastatic cancer cells is necessary for lung metastasis progression. Neoplasia. 2016;18:699–710.

    Article  CAS  Google Scholar 

  36. Denoyelle C, Abou-Rjaily G, Bezrookove V, Verhaegen M, Johnson TM, Fullen DR, et al. Anti-oncogenic role of the endoplasmic reticulum differentially activated by mutations in the MAPK pathway. Nat Cell Biol. 2006;8:1053–63.

    Article  CAS  Google Scholar 

  37. De Raedt T, Walton Z, Yecies JL, Li D, Chen Y, Malone CF, et al. Exploiting cancer cell vulnerabilities to develop a combination therapy for ras-driven tumors. Cancer Cell. 2011;3:400–13.

    Google Scholar 

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Acknowledgements

We thank Hal Chapman for the SPC-Cre mice, Peter Conti and Jennifer Choi for assistance with PET/CT, and Jorge Nieva and Robert Hsu for tumor samples. The work was supported by NIH grants R01 CA027607 and the Judy and Larry Freeman Chair (ASL), NIH Diversity Supplements (DFR), the Hastings Foundation (BZ, ZB), and NIH grant R35 HL135747 and Ralph Edgington Chair (ZB). We thank the USC Norris Comprehensive Cancer Translational Pathology Core and the USC Molecular Imaging Center (supported by P30 CA014089, 1S10OD012371 and 1S10OD18500) for technical assistance.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA

    Daisy Flores Rangel, Priscilla Chan, Dat P. Ha, Zea Borok, Ite A. Offringa & Amy S. Lee

  2. Norris Comprehensive Cancer Center, University of Southern California, Los Angeles, CA, USA

    Daisy Flores Rangel, Louis Dubeau, Priscilla Chan, Dat P. Ha, Mario A. Pulido, Daniel J. Mullen, Ite A. Offringa & Amy S. Lee

  3. Department of Pathology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA

    Louis Dubeau

  4. Department of Radiology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA

    Ryan Park & Ivetta Vorobyova

  5. Department of Surgery, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA

    Mario A. Pulido, Daniel J. Mullen & Ite A. Offringa

  6. Department of Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA

    Beiyun Zhou & Zea Borok

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Correspondence to Amy S. Lee.

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All protocols for animal use and euthanasia were reviewed and approved by the University of Southern California Institutional Animal Care and Use Committee. Patient lung tissues were obtained in accordance with a protocol approved by the Institutional Review Board of the University of Southern California. Confirmed consent was obtained from all participants.

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Rangel, D.F., Dubeau, L., Park, R. et al. Endoplasmic reticulum chaperone GRP78/BiP is critical for mutant Kras-driven lung tumorigenesis. Oncogene 40, 3624–3632 (2021). https://doi.org/10.1038/s41388-021-01791-9

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