Cell cultures in assessing radioresistance of glioblastomas

Belyashova A.S.; Galkin M.V.; Antipina N.A.; Pavlova G.V.; Golanov A.V.

doi:https://doi.org/10.17116/neiro202286051126

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Abstract:

To date, no modern methods of treatment allow overcoming malignant potential of glial neoplasms and significant increase of survival. Analysis of glioblastoma radioresistance using cancer cell cultures is one of the perspective directions, as radiotherapy is standard and available treatment method for these neoplasms. This review summarizes current studies identifying many factors of radioresistance of glial tumors, such as hypoxia, microenvironment and metabolic features of tumor, stem cells, internal heterogeneity of tumor, microRNA, features of cell cycle, DNA damage and reparation. We obtained data on involvement of various molecular pathways in development of radioresistance such as MEK/ERK, c-MYC, PI3K/Akt, PTEN, Wnt, JAK/STAT, Notch, etc. Changes in activity of RAD51 APC, FZD1, LEF1, TCF4, WISP1, p53 and many others are determined in radioresistant cells. Further study of radioresistance pathways will allow development of specific target aptamers and inhibitors.

Keywords:

cancer cell culture

glioblastoma

radioresistance

Authors:

Belyashova A.S.

Burdenko Neurosurgical Center

Galkin M.V.

Burdenko National Medical Research Center for Neurosurgery

SPIN RSCI: 7745-6695
Scopus AuthorID: 36099071200
ResearcherID: AAH-4783-2019
ORCID: 0000-0002-1436-0010

Antipina N.A.

Burdenko Neurosurgical Center

Pavlova G.V.

Burdenko National Medical Research Center of Neurosurgery

SPIN RSCI: 4633-8339
Scopus AuthorID: 7005650683
ORCID: 0000-0002-6885-6601

Golanov A.V.

Burdenko Neurosurgical Center

ORCID: 0000-0002-0976-4547

Received:

08.08.2022

Accepted:

19.08.2022

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References:

Stupp R, Mason WP, van den Bent, Martin J, Weller M, Fisher B, Taphoorn MJB, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO. Radiotherapy plus Concomitant and Adjuvant Temozolomide for Glioblastoma. New England Journal of Medicine. 2005;352(10):987-996. https://doi.org/10.1056/nejmoa043330
Chinot OL, Wick W, Mason W, Henriksson R, Saran F, Nishikawa R, Carpentier AF, Hoang-Xuan K, Kavan P, Cernea D, Brandes AA, Hilton M, Abrey L, Cloughesy T. Bevacizumab plus radiotherapy-temozolomide for newly diagnosed glioblastoma. New England Journal of Medicine. 2014;370(8):709-722. https://doi.org/10.1056/NEJMoa1308345
Rapp M, Baernreuther J, Turowski B, Steiger H-J, Sabel M, Kamp MA. Recurrence Pattern Analysis of Primary Glioblastoma. World Neurosurgery. 2017;103:733-740. https://doi.org/10.1016/j.wneu.2017.04.053
McDonald MW, Shu H-KG, Curran WJ, Crocker IR. Pattern of failure after limited margin radiotherapy and temozolomide for glioblastoma. International Journal of Radiation Oncology. Biology. Physics. 2011;79(1):130-136. https://doi.org/10.1016/j.ijrobp.2009.10.048
Graf R, Hildebrandt B, Tilly W, Sreenivasa G, Ullrich R, Maier-Hauff K, Felix R, Wust P. Dose-Escalated Conformal Radiotherapy of Glioblastomas — Results of a Retrospective Comparison Applying Radiation Doses of 60 and 70 Gy. Oncology Research and Treatment. 2005;28(6-7):325-330. https://doi.org/10.1159/000085574
Coughlin C, Scott C, Langer C, Coia L, Curran W, Rubin P. Phase II, two-arm RTOG trial (94-11) of bischloroethyl-nitrosourea plus accelerated hyperfractionated radiotherapy (64.0 or 70.4 Gy) based on tumor volume (> 20 or ≤ 20 cm2, respectively) in the treatment of newly-diagnosed radiosurgery-ineligible glioblastoma multiforme patients. International Journal of Radiation Oncology, Biology, Physics. 2000;48(5):1351-1358. https://doi.org/10.1016/s0360-3016(00)01412-7
Zhang Y, Dube C, Gibert M, Cruickshanks N, Wang B, Coughlan M, Yang Y, Setiady I, Deveau C, Saoud K, Grello C, Oxford M, Yuan F, Abounader R. The p53 Pathway in Glioblastoma. Cancers. 2018;10(9):297. https://doi.org/10.3390/cancers10090297
Pesenti C, Navone SE, Guarnaccia L, Terrasi A, Costanza J, Silipigni R, Guarneri S, Fusco N, Fontana L, Locatelli M, Rampini P, Campanella R, Tabano S, Miozzo M, Marfia G. The Genetic Landscape of Human Glioblastoma and Matched Primary Cancer Stem Cells Reveals Intratumour Similarity and Intertumour Heterogeneity. Stem Cells International. 2019;20192617030. https://doi.org/10.1155/2019/2617030
LeBlanc VG, Trinh DL, Aslanpour S, Hughes M, Livingstone D, Jin D, Ahn BY, Blough MD, Cairncross JG, Chan JA, Kelly JJP, Marra MA. Single-cell landscapes of primary glioblastomas and matched explants and cell lines show variable retention of inter- and intratumor heterogeneity. Cancer Cell. 2022;40(4):379-392.e9. https://doi.org/10.1016/j.ccell.2022.02.016
Lee J, Kotliarova S, Kotliarov Y, Li A, Su Q, Donin NM, Pastorino S, Purow BW, Christopher N, Zhang W, Park JK, Fine HA. Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than do serum-cultured cell lines. Cancer Cell. 2006;9(5):391-403. https://doi.org/10.1016/j.ccr.2006.03.030
Gray LH, Conger AD, Ebert M, Hornsey S, Scott OC. The concentration of oxygen dissolved in tissues at the time of irradiation as a factor in radiotherapy. British Journal of Radiology. 1953;26(312):638-648. https://doi.org/10.1259/0007-1285-26-312-638
Marampon F, Gravina GL, Zani BM, Popov VM, Fratticci A, Cerasani M, Di Genova D, Mancini M, Ciccarelli C, Ficorella C, Di Cesare E, Festuccia C. Hypoxia sustains glioblastoma radioresistance through ERKs/DNA-PKcs/HIF-1α functional interplay. International Journal of Oncology. 2014;44(6):2121-2131. https://doi.org/10.3892/ijo.2014.2358
Li P, Zhou C, Xu L, Xiao H. Hypoxia enhances stemness of cancer stem cells in glioblastoma: An in vitro study. International Journal of Medical Sciences. 2013;10(4):399-407. https://doi.org/10.7150/ijms.5407
Zimmer M, Ebert BL, Neil C, Brenner K, Papaioannou I, Melas A, Tolliday N, Lamb J, Pantopoulos K, Golub T, Iliopoulos O. Small molecule inhibitors of HIF-2a translation link its 5’-UTR Iron-Responsive Element (IRE) to oxygen sensing. Molecular Cell. 2008;32(6):838-848. https://doi.org/10.1016/j.molcel.2008.12.004
Schiffer D, Annovazzi L, Casalone C, Corona C, Mellai M. Glioblastoma: Microenvironment and Niche Concept. Cancers. 2018;11(1):5. https://doi.org/10.3390/cancers11010005
Mannino M, Chalmers AJ. Radioresistance of glioma stem cells: Intrinsic characteristic or property of the ‘microenvironment‐stem cell unit’? Molecular Oncology. 2011;5(4):374-386. https://doi.org/10.1016/j.molonc.2011.05.001
Fidoamore A, Cristiano L, Antonosante A, d’Angelo M, Di Giacomo E, Astarita C, Giordano A, Ippoliti R, Benedetti E, Cimini A. Glioblastoma Stem Cells Microenvironment: The Paracrine Roles of the Niche in Drug and Radioresistance. Stem Cells International. 2016;2016(18):1-17. https://doi.org/10.1155/2016/6809105
Folkins C, Shaked Y, Man S, Tang T, Lee CR, Zhu Z, Hoffman RM, Kerbel RS. Glioma tumor stem-like cells promote tumor angiogenesis and vasculogenesis via vascular endothelial growth factor and stromal-derived factor 1. Cancer Research. 2009;69(18):7243-7251. https://doi.org/10.1158/0008-5472.CAN-09-0167
Hovinga KE, Shimizu F, Wang R, Panagiotakos G, van der Heijden M, Moayedpardazi H, Correia AS, Soulet D, Major T, Menon J, Tabar V. Inhibition of notch signaling in glioblastoma targets cancer stem cells via an endothelial cell intermediate. Stem Cells. 2010;28(6):1019-1029. https://doi.org/10.1002/stem.429
Farace C, Oliver JA, Melguizo C, Alvarez P, Bandiera P, Rama AR, Malaguarnera G, Ortiz R, Madeddu R, Prados J. Microenvironmental Modulation of Decorin and Lumican in Temozolomide-Resistant Glioblastoma and Neuroblastoma Cancer Stem-Like Cells. PLoS One. 2015;10(7):e0134111. https://doi.org/10.1371/journal.pone.0134111
Stanzani E, Martínez-Soler F, Mateos TM, Vidal N, Villanueva A, Pujana MA, Serra-Musach J, La Iglesia N de, Giménez-Bonafé P, Tortosa A. Radioresistance of mesenchymal glioblastoma initiating cells correlates with patient outcome and is associated with activation of inflammatory program. Oncotarget. 2017;8(43):73640-73653. https://doi.org/10.18632/oncotarget.18363
Zeppernick F, Ahmadi R, Campos B, Dictus C, Helmke BM, Becker N, Lichter P, Unterberg A, Radlwimmer B, Herold-Mende CC. Stem cell marker CD133 affects clinical outcome in glioma patients. Clinical Cancer Research. 2008;14(1):123-129. https://doi.org/10.1158/1078-0432.CCR-07-0932
Lopez-Bertoni H, Lal B, Li A, Caplan M, Guerrero-Cázares H, Eberhart CG, Quiñones-Hinojosa A, Glas M, Scheffler B, Laterra J, Li Y. DNMT-dependent suppression of microRNA regulates the induction of GBM tumor-propagating phenotype by Oct4 and Sox2. Oncogene. 2015;34(30):3994-4004. https://doi.org/10.1038/onc.2014.334
Ligon KL, Huillard E, Mehta S, Kesari S, Liu H, Alberta JA, Bachoo RM, Kane M, Louis DN, DePinho RA, Anderson DJ, Stiles CD, Rowitch DH. Olig2-regulated lineage-restricted pathway controls replication competence in neural stem cells and malignant glioma. Neuron. 2007;53(4):503-517. https://doi.org/10.1016/j.neuron.2007.01.009
Veselska R, Kuglik P, Cejpek P, Svachova H, Neradil J, Loja T, Relichova J. Nestin expression in the cell lines derived from glioblastoma multiforme. BMC Cancer. 2006;6:32. https://doi.org/10.1186/1471-2407-6-32
Liu G, Yuan X, Zeng Z, Tunici P, Ng H, Abdulkadir IR, Lu L, Irvin D, Black KL, Yu JS. Analysis of gene expression and chemoresistance of CD133+ cancer stem cells in glioblastoma. Molecular Cancer. 2006;5:67. https://doi.org/10.1186/1476-4598-5-67
Xu Y, Stamenkovic I, Yu Q. CD44 attenuates activation of the hippo signaling pathway and is a prime therapeutic target for glioblastoma. Cancer Research. 2010;70(6):2455-2464. https://doi.org/10.1158/0008-5472.CAN-09-2505
Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J, Dirks PB. Identification of a cancer stem cell in human brain tumors. Cancer Research. 2003;63(18):5821-5828.
Chang C-J, Hsu C-C, Yung M-C, Chen K-Y, Tzao C, Wu W-F, Chou H-Y, Lee Y-Y, Lu K-H, Chiou S-H, Ma H-I. Enhanced radiosensitivity and radiation-induced apoptosis in glioma CD133-positive cells by knockdown of SirT1 expression. Biochemical and Biophysical Research Communications. 2009;380(2):236-242. https://doi.org/10.1016/j.bbrc.2009.01.040
Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB, Dewhirst MW, Bigner DD, Rich JN. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature. 2006;444(7120): 756-760. https://doi.org/10.1038/nature05236
Kao GD, Jiang Z, Fernandes AM, Gupta AK, Maity A. Inhibition of phosphatidylinositol-3-OH kinase/Akt signaling impairs DNA repair in glioblastoma cells following ionizing radiation. The Journal of Biological Chemistry. 2007;282(29):21206-21212. https://doi.org/10.1074/jbc.M703042200
Li H-F, Kim J-S, Waldman T. Radiation-induced Akt activation modulates radioresistance in human glioblastoma cells. Radiation Oncology. 2009;4:43. https://doi.org/10.1186/1748-717X-4-43
Yaes RJ. Tumor heterogeneity, tumor size, and radioresistance. International Journal of Radiation Oncology - Biology - Physics. 1989;17(5):993-1005. https://doi.org/10.1016/0360-3016(89)90147-8
Yin N, Xie T, Zhang H, Chen J, Yu J, Liu F. IDH1-R132H mutation radiosensitizes U87MG glioma cells via epigenetic downregulation of TIGAR. Oncology Letters. 2020;19(2):1322-1330. https://doi.org/10.3892/ol.2019.11148
Wolf A, Agnihotri S, Micallef J, Mukherjee J, Sabha N, Cairns R, Hawkins C, Guha A. Hexokinase 2 is a key mediator of aerobic glycolysis and promotes tumor growth in human glioblastoma multiforme. Journal of Experimental Medicine. 2011;208(2):313-326. https://doi.org/10.1084/jem.20101470
Vartanian A, Agnihotri S, Wilson MR, Burrell KE, Tonge PD, Alamsahebpour A, Jalali S, Taccone MS, Mansouri S, Golbourn B, Aldape KD, Zadeh G. Targeting hexokinase 2 enhances response to radio-chemotherapy in glioblastoma. Oncotarget. 2016;7(43):69518-69535. https://doi.org/10.18632/oncotarget.11680
Calvert AE, Chalastanis A, Wu Y, Hurley LA, Kouri FM, Bi Y, Kachman M, May JL, Bartom E, Hua Y, Mishra RK, Schiltz GE, Dubrovskyi O, Mazar AP, Peter ME, Zheng H, James CD, Burant CF, Chandel NS, Davuluri RV, Horbinski C, Stegh AH. Cancer-Associated IDH1 Promotes Growth and Resistance to Targeted Therapies in the Absence of Mutation. Cell Reports. 2017;19(9):1858-1873. https://doi.org/10.1016/j.celrep.2017.05.014
You W-C, Chiou S-H, Huang C-Y, Chiang S-F, Yang C-L, Sudhakar JN, Lin T-Y, Chiang I-P, Shen C-C, Cheng W-Y, Lin J-C, Shieh S-H, Chow K-C. Mitochondrial protein ATPase family, AAA domain containing 3A correlates with radioresistance in glioblastoma. Neuro-Oncology. 2013;15(10):1342-1352. https://doi.org/10.1093/neuonc/not077
Sana J, Busek P, Fadrus P, Besse A, Radova L, Vecera M, Reguli S, Stollinova Sromova L, Hilser M, Lipina R, Lakomy R, Kren L, Smrcka M, Sedo A, Slaby O. Identification of microRNAs differentially expressed in glioblastoma stem-like cells and their association with patient survival. Scientific Reports. 2018;8(1):2836. https://doi.org/10.1038/s41598-018-20929-6
Møller HG, Rasmussen AP, Andersen HH, Johnsen KB, Henriksen M, Duroux M. A systematic review of microRNA in glioblastoma multiforme: micro-modulators in the mesenchymal mode of migration and invasion. Molecular Neurobiology. 2013;47(1):131-144. https://doi.org/10.1007/s12035-012-8349-7
Deng X, Ma L, Wu M, Zhang G, Jin C, Guo Y, Liu R. miR-124 radiosensitizes human glioma cells by targeting CDK4. Journal of Neuro-Oncology. 2013;114(3):263-274. https://doi.org/10.1007/s11060-013-1179-2
Toraih EA, El-Wazir A, Abdallah HY, Tantawy MA, Fawzy MS. Deregulated MicroRNA Signature Following Glioblastoma Irradiation. Cancer Control. 2019;26(1):1073274819847226. https://doi.org/10.1177/1073274819847226
Li W, Guo F, Wang P, Hong S, Zhang C. miR-221/222 confers radioresistance in glioblastoma cells through activating Akt independent of PTEN status. Current Molecular Medicine. 2014;14(1):185-195. https://doi.org/10.2174/1566524013666131203103147
Moskwa P, Zinn PO, Choi YE, Shukla SA, Fendler W, Chen CC, Lu J, Golub TR, Hjelmeland A, Chowdhury D. A functional screen identifies miRs that induce radioresistance in glioblastomas. Molecular Cancer Research. 2014;12(12):1767-1778. https://doi.org/10.1158/1541-7786.MCR-14-0268
Marampon F, Megiorni F, Camero S, Crescioli C, McDowell HP, Sferra R, Vetuschi A, Pompili S, Ventura L, Felice F de, Tombolini V, Dominici C, Maggio R, Festuccia C, Gravina GL. HDAC4 and HDAC6 sustain DNA double strand break repair and stem-like phenotype by promoting radioresistance in glioblastoma cells. Cancer Letters. 2017;3971-3911. https://doi.org/10.1016/j.canlet.2017.03.028
Kowalski-Chauvel A, Modesto A, Gouaze-Andersson V, Baricault L, Gilhodes J, Delmas C, Lemarie A, Toulas C, Cohen-Jonathan-Moyal E, Seva C. Alpha-6 integrin promotes radioresistance of glioblastoma by modulating DNA damage response and the transcription factor Zeb1. Cell Death and Disease. 2018;9(9):872. https://doi.org/10.1038/s41419-018-0853-x
Golding SE, Morgan RN, Adams BR, Hawkins AJ, Povirk LF, Valerie K. Pro-survival AKT and ERK signaling from EGFR and mutant EGFRvIII enhances DNA double-strand break repair in human glioma cells. Cancer Biology and Therapy. 2009;8(8):730-738. https://doi.org/10.4161/cbt.8.8.7927
Hitoshi S, Alexson T, Tropepe V, Donoviel D, Elia AJ, Nye JS, Conlon RA, Mak TW, Bernstein A, van der Kooy D. Notch pathway molecules are essential for the maintenance, but not the generation, of mammalian neural stem cells. Genes Development. 2002;16(7):846-858. https://doi.org/10.1101/gad.975202
Hitoshi S, Seaberg RM, Koscik C, Alexson T, Kusunoki S, Kanazawa I, Tsuji S, van der Kooy D. Primitive neural stem cells from the mammalian epiblast differentiate to definitive neural stem cells under the control of Notch signaling. Genes Development. 2004;18(15):1806-1811. https://doi.org/10.1101/gad.1208404
The Cancer Genome Atlas Research Network. Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature. 2008;455(7216):1061-1068. https://doi.org/10.1038/nature07385
Wu JK, Ye Z, Darras BT. Frequency of p53 tumor suppressor gene mutations in human primary brain tumors. Neurosurgery. 1993;33(5):824-830. https://doi.org/10.1227/00006123-199311000-00006
Shu H-KG, Kim MM, Chen P, Furman F, Julin CM, Israel MA. The intrinsic radioresistance of glioblastoma-derived cell lines is associated with a failure of p53 to induce p21BAX expression. Proceedings of the National Academy of Sciences of the United States of America. 1998;95(24):14453-14458.
Tompa M, Kalovits F, Nagy A, Kalman B. Contribution of the Wnt Pathway to Defining Biology of Glioblastoma. Neuromolecular Medicine. 2018;20(4): 437-451. https://doi.org/10.1007/s12017-018-8514-x
Lee Y, Lee J-K, Ahn SH, Lee J, Nam D-H. WNT signaling in glioblastoma and therapeutic opportunities. Laboratory Investigation. 2016;96(2):137-150. https://doi.org/10.1038/labinvest.2015.140
McCord M, Mukouyama Y-s, Gilbert MR, Jackson S. Targeting WNT Signaling for Multifaceted Glioblastoma Therapy. Frontiers in Cellular Neuroscience. 2017;11:318. https://doi.org/10.3389/fncel.2017.00318
Kim N, Kim SH, Kang S-G, Moon JH, Cho J, Suh C-O, Yoon H in, Chang JH. ATM mutations improve radio-sensitivity in wild-type isocitrate dehydrogenase-associated high-grade glioma: retrospective analysis using next-generation sequencing data. Radiation Oncology. 2020;15(1):184. https://doi.org/10.1186/s13014-020-01619-y
Balbous A, Cortes U, Guilloteau K, Rivet P, Pinel B, Duchesne M, Godet J, Boissonnade O, Wager M, Bensadoun RJ, Chomel J-C, Karayan-Tapon L. A radiosensitizing effect of RAD51 inhibition in glioblastoma stem-like cells. BMC Cancer. 2016;16:604. https://doi.org/10.1186/s12885-016-2647-9
Ventero MP, Fuentes-Baile M, Quereda C, Perez-Valeciano E, Alenda C, Garcia-Morales P, Esposito D, Dorado P, Manuel Barbera V, Saceda M. Radiotherapy resistance acquisition in Glioblastoma. Role of SOCS1 and SOCS3. PLoS One. 2019;14(2)e0212581. https://doi.org/10.1371/journal.pone.0212581
Tang Z, Dokic I, Knoll M, Ciamarone F, Schwager C, Klein C, Cebulla G, Hoffmann DC, Schlegel J, Seidel P, Rutenberg C, Brons S, Herold-Mende C, Wick W, Debus J, Lemke D, Abdollahi A. Radioresistance and Transcriptional Reprograming of Invasive Glioblastoma Cells. International Journal of Radiation Oncology, Biology, Physics. 2022;112(2):499-513. https://doi.org/10.1016/j.ijrobp.2021.09.017
Antipina NA, Smirnov GYu, Nikolaeva AA, Belyashova AS, Pavlova GV, Golanov AV. Physico-technical aspects of the experimental study of the effect of high radiation doses on human glioblastoma cell culture. Meditsinskaya fizika. 2019;(2):51-57. (In Russ).