PROM2 overexpression induces metastatic potential through epithelial-to-mesenchymal transition and ferroptosis resistance in human cancers

Abstract

Introduction: Despite considerable therapeutic advances in the last 20 years, metastatic cancers remain a major cause of death. We previously identified prominin-2 (PROM2) as a biomarker predictive of distant metastases and decreased survival, thus providing a promising bio-target. In this translational study, we set out to decipher the biological roles of PROM2 during the metastatic process and resistance to cell death, in particular for metastatic melanoma.

Methods and results: Methods and results: We demonstrated that PROM2 overexpression was closely linked to an increased metastatic potential through the increase of epithelial-to-mesenchymal transition (EMT) marker expression and ferroptosis resistance. This was also found in renal cell carcinoma and triple negative breast cancer patient-derived xenograft models. Using an oligonucleotide anti-sense anti-PROM2, we efficaciously decreased PROM2 expression and prevented metastases in melanoma xenografts. We also demonstrated that PROM2 was implicated in an aggravation loop, contributing to increase the metastatic burden both in murine metastatic models and in patients with metastatic melanoma. The metastatic burden is closely linked to PROM2 expression through the expression of EMT markers and ferroptosis cell death resistance in a deterioration loop.

Conclusion: Our results open the way for further studies using PROM2 as a bio-target in resort situations in human metastatic melanoma and also in other cancer types.

Keywords

breast cancer / epithelial-to-mesenchymal transition / ferroptosis resistance / melanoma / metastases / prominin-2 / renal cancer

References

[1] Wolchok JD, Chiarion-Sileni V, Gonzalez R, et al. Long-term outcomes with nivolumab plus ipilimumab or nivolumab alone versus ipilimumab in patients with advanced melanoma. J Clin Oncol. 2022;40(2):127-137.

[2] Robert C, Grob JJ, Stroyakovskiy D, et al. Five-year outcomes with dabrafenib plus trametinib in metastatic melanoma. N Engl J Med. 2019;381(7):626-636.

[3] Serra-Bellver P, Versluis JM, Oberoi HK, et al. Real-world outcomes with ipilimumab and nivolumab in advanced melanoma: a multicentre retrospective study. Eur J Cancer. 2022;176:121-132.

[4] Nguyen TT, Gapihan G, Tetu P, et al. Increased risk of brain metastases among patients with melanoma and PROM2 expression in metastatic lymph nodes. Clin Transl Med. 2020;10(8):e198.

[5] Fargeas CA, Florek M, Huttner WB, Corbeil D. Characterization of prominin-2, a new member of the prominin family of pentaspan membrane glycoproteins. J Biol Chem. 2003;278(10):8586-8596.

[6] Florek M, Bauer N, Janich P, et al. Prominin-2 is a cholesterol-binding protein associated with apical and basolateral plasmalemmal protrusions in polarized epithelial cells and released into urine. Cell Tissue Res. 2007;328(1):31-47.

[7] Singh RD, Schroeder AS, Scheffer L, et al. Prominin-2 expression increases protrusions, decreases caveolae and inhibits Cdc42 dependent fluid phase endocytosis. Biochem Biophys Res Commun. 2013;434(3):466-472.

[8] Dowland SN, Madawala RJ, Poon CE, Lindsay LA, Murphy CR. Prominin-2 prevents the formation of caveolae in normal and ovarian hyperstimulated pregnancy. Reprod Sci. 2018;25(8):1231-1242.

[9] Brown CW, Amante JJ, Chhoy P, et al. Prominin2 drives ferroptosis resistance by stimulating iron export. Dev Cell. 2019;51(5):575-586e4.

[10] Tang D, Kroemer G. Ferroptosis. Curr Biol. 2020;30(21):R1292-R1297.

[11] Brown CW, Amante JJ, Goel HL, Mercurio AM. The alpha6beta4 integrin promotes resistance to ferroptosis. J Cell Biol. 2017;216(12):4287-4297.

[12] Brown CW, Amante JJ, Mercurio AM. Cell clustering mediated by the adhesion protein PVRL4 is necessary for alpha6beta4 integrin-promoted ferroptosis resistance in matrix-detached cells. J Biol Chem. 2018;293(33):12741-12748.

[13] Hanahan D, Weinberg RA. The hallmarks of cancer. Cell. 2000;100(1):57-70.

[14] Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646-674.

[15] Hanahan D. Hallmarks of cancer: new dimensions. Cancer Discov. 2022;12(1):31-46.

[16] Bousquet G, Feugeas JP, Ferreira I, et al. Individual xenograft as a personalized therapeutic resort for women with metastatic triple-negative breast carcinoma. Breast Cancer Res. 2014;16(1):401.

[17] Varna M, Gapihan G, Feugeas JP, et al. Stem cells increase in numbers in perinecrotic areas in human renal cancer. Clin Cancer Res. 2015;21(4):916-924.

[18] Bousquet G, Feugeas JP, Gu Y, et al. High expression of apoptosis protein (Api-5) in chemoresistant triple-negative breast cancers: an innovative target. Oncotarget. 2019;10(61):6577-6588.

[19] Bousquet G, El Bouchtaoui M, Leboeuf C, et al. Tracking sub-clonal TP53 mutated tumor cells in human metastatic renal cell carcinoma. Oncotarget. 2015;6(22):19279-19289.

[20] U.S, Department of Health and Human Services National Institutes of Health Office of Laboratory Animal Welfare. Public Health Service Policy on Humane Care and Use of Laboratory Animals. NIH Publication revised, 2015; p. 15-8013.

[21] Marangoni E, Vincent-Salomon A, Auger N, et al. A new model of patient tumor-derived breast cancer xenografts for preclinical assays. Clin Cancer Res. 2007;13(13):3989-3998.

[22] Varna M, Lehmann-Che J, Turpin E, et al. p53 dependent cell-cycle arrest triggered by chemotherapy in xenografted breast tumors. Int J Cancer. 2009;124(4):991-997.

[23] Falgarone G, Pamoukdjian F, Cailhol J, et al. Lung ultrasound is a reliable diagnostic technique to predict abnormal CT chest scan and to detect oxygen requirements in COVID-19 pneumonia. Aging (Albany NY). 2020;12(20):19945-19953.

[24] Bustin SA, Benes V, Garson JA, et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009;55(4):611-622.

[25] Liao Y, Jia X, Ren Y, et al. Suppressive role of microRNA-130b-3p in ferroptosis in melanoma cells correlates with DKK1 inhibition and Nrf2-HO-1 pathway activation. Hum Cell. 2021;34(5):1532-1544.

[26] Floros KV, Cai J, Jacob S, et al. MYCN-amplified neuroblastoma is addicted to iron and vulnerable to inhibition of the system Xc-/glutathione axis. Cancer Res. 2021;81(7):1896-1908.

[27] Elmallah MIY, Ortega-Deballon P, Hermite L, et al. Lipidomic profiling of exosomes from colorectal cancer cells and patients reveals potential biomarkers. Mol Oncol. 2022;16(14):2710-2718.

[28] Gardiner C, Ferreira YJ, Dragovic RA, Redman CW, Sargent IL. Extracellular vesicle sizing and enumeration by nanoparticle tracking analysis. J Extracell Vesicles. 2013;2.

[29] Saha SK, Islam SMR, Kwak KS, Rahman MS, Cho SG. PROM1 and PROM2 expression differentially modulates clinical prognosis of cancer: a multiomics analysis. Cancer Gene Ther. 2020;27(3-4):147-167.

[30] Li W, Zhu Y, Zhang K, et al. PROM2 promotes gemcitabine chemoresistance via activating the Akt signaling pathway in pancreatic cancer. Exp Mol Med. 2020;52(3):409-422.

[31] Zhang B, Chu W, Wen F, et al. Dysregulation of long non-coding RNAs and mRNAs in plasma of clear cell renal cell carcinoma patients using microarray and bioinformatic analysis. Front Oncol. 2020;10:559730.

[32] Liang W, Hao Z, Han JL, et al. CAV-1 contributes to bladder cancer progression by inducing epithelial-to-mesenchymal transition. Urol Oncol. 2014;32(6):855-863.

[33] Gai X, Lu Z, Tu K, Liang Z, Zheng X. Caveolin-1 is up-regulated by GLI1 and contributes to GLI1-driven EMT in hepatocellular carcinoma. PLoS One. 2014;9(1):e84551.

[34] Cokakli M, Erdal E, Nart D, et al. Differential expression of Caveolin-1 in hepatocellular carcinoma: correlation with differentiation state, motility and invasion. BMC Cancer. 2009;9:65.

[35] Bousquet G, Janin A. Patient-derived xenograft: an adjuvant technology for the treatment of metastatic disease. Pathobiology. 2016;83(4):170-176.

[36] Talty R, Bosenberg M. The role of ferroptosis in melanoma. Pigment Cell Melanoma Res. 2022;35(1):18-25.

[37] Han N, Li H, Wang H. MicroRNA-203 inhibits epithelial-mesenchymal transition, migration, and invasion of renal cell carcinoma cells via the inactivation of the PI3K/AKT signaling pathway by inhibiting CAV1. Cell Adh Migr. 2020;14(1):227-241.

[38] Zhu Q, Zhan D, Zhu P, Chong Y, Yang Y. CircAKT1 acts as a sponge of miR-338-3p to facilitate clear cell renal cell carcinoma progression by up-regulating CAV1. Biochem Biophys Res Commun. 2020;532(4):584-590.

[39] Lee S, Bae JS, Jung CK, Chung WY. Extensive lymphatic spread of papillary thyroid microcarcinoma is associated with an increase in expression of genes involved in epithelial-mesenchymal transition and cancer stem cell-like properties. Cancer Med. 2019;8(15):6528-6537.

[40] Zhu Z, Yu Z, Wang J, et al. Kruppel-like factor 4 inhibits pancreatic cancer epithelial-to-mesenchymal transition and metastasis by down-regulating caveolin-1 expression. Cell Physiol Biochem. 2018;46(1):238-252.

[41] Phiboonchaiyanan PP, Busaranon K, Ninsontia C, Chanvorachote P. Benzophenone-3 increases metastasis potential in lung cancer cells via epithelial to mesenchymal transition. Cell Biol Toxicol. 2017;33(3):251-261.

[42] Magtanong L, Ko PJ, To M, et al. Exogenous monounsaturated fatty acids promote a ferroptosis-resistant cell state. Cell Chem Biol. 2019;26(3):420-432e9.

[43] Ubellacker JM, Tasdogan A, Ramesh V, et al. Lymph protects metastasizing melanoma cells from ferroptosis. Nature. 2020;585(7823):113-118.

[44] Jaszai J, Janich P, Farkas LM, et al. Differential expression of Prominin-1 (CD133) and Prominin-2 in major cephalic exocrine glands of adult mice. Histochem Cell Biol. 2007;128(5):409-419.

[45] Jaszai J, Farkas LM, Fargeas CA, et al. Prominin-2 is a novel marker of distal tubules and collecting ducts of the human and murine kidney. Histochem Cell Biol. 2010;133(5):527-539.

[46] Santelli A, Sun IO, Eirin A, et al. Senescent kidney cells in hypertensive patients release urinary extracellular vesicles. J Am Heart Assoc. 2019;8(11):e012584.

[47] Luo W, Wang J, Xu W, et al. LncRNA RP11-89 facilitates tumorigenesis and ferroptosis resistance through PROM2-activated iron export by sponging miR-129-5p in bladder cancer. Cell Death Dis. 2021;12(11):1043.

[48] Raguraman P, Balachandran AA, Chen S, Diermeier SD, Veedu RN. Antisense oligonucleotide-mediated splice switching: potential therapeutic approach for cancer mitigation. Cancers (Basel). 2021;13(21).

[49] Kaczorowski M, Chlopek M, Kruczak A, et al. PRAME expression in cancer. a systematic immunohistochemical study of >5800 epithelial and nonepithelial tumors. Am J Surg Pathol. 2022;46(11):1467-1476.

RIGHTS & PERMISSIONS

2024 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.