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Open Access 01-12-2024 | Glioblastoma | Research

CD13 expression affects glioma patient survival and influences key functions of human glioblastoma cell lines in vitro

Authors: Wenying Zhang, Anne Blank, Irina Kremenetskaia, Anja Nitzsche, Güliz Acker, Peter Vajkoczy, Susan Brandenburg

Published in: BMC Cancer | Issue 1/2024

Abstract

CD13 (APN) is an Alanyl-Aminopeptidase with diverse functions. The role of CD13 for gliomas is still unknown. In this study, data of glioma patients obtained by TCGA and CGGA databases were used to evaluate the survival rate and prognostic value of CD13 expression level. Protein expression of CD13 was confirmed by immunofluorescence staining of fresh patient tissues. Eight human glioblastoma cell lines were studied by RT-PCR, Western Blot, immunofluorescence staining and flow cytometry to define CD13 expression. Cell lines with different CD13 expression status were treated with a CD13 inhibitor, bestatin, and examined by MTT, scratch and colony formation assaysas well as by apoptosis assay and Western Blots. Bioinformatics analysis indicated that patients with high expression of CD13 had poor survival and prognosis. Additionally, CD13 protein expression was positively associated with clinical malignant characteristics. Investigated glioblastoma cell lines showed distinct expression levels and subcellular localization of CD13 with intracellular enrichment. Bestatin treatment reduced proliferation, migration and colony formation of glioma cells in a CD13-dependent manner while apoptosis was increased. In summary, CD13 has an impact on glioma patient survival and is important for the main function of specific glioma cells.

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Louis DN, Perry A, Wesseling P, Brat DJ, Cree IA, Figarella-Branger D, Hawkins C, Ng HK, Pfister SM, Reifenberger G, et al. The 2021 WHO classification of tumors of the Central Nervous System: a summary. Neuro Oncol. 2021;23(8):1231–51. https://doi.org/10.1093/neuonc/noab106.CrossRefPubMedPubMedCentral

Schaff LR, Mellinghoff IK. Glioblastoma and other primary brain malignancies in adults: a review. JAMA. 2023;329(7):574–87. https://doi.org/10.1001/jama.2023.0023.CrossRefPubMed

Valentinis L, Tuniz F, Valent F, Mucchiut M, Little D, Skrap M, Bergonzi P, Zanchin G. Headache attributed to intracranial tumours: a prospective cohort study. Cephalalgia. 2010;30(4):389–98. https://doi.org/10.1111/j.1468-2982.2009.01970.x.CrossRefPubMed

Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352(10):987–96. https://doi.org/10.1056/NEJMoa043330.CrossRefPubMed

Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, Ludwin SK, Allgeier A, Fisher B, Belanger K, et al. Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol. 2009;10(5):459–66. https://doi.org/10.1016/S1470-2045(09)70025-7.CrossRefPubMed

Brown TJ, Brennan MC, Li M, Church EW, Brandmeir NJ, Rakszawski KL, Patel AS, Rizk EB, Suki D, Sawaya R, et al. Association of the extent of Resection with Survival in Glioblastoma: a systematic review and Meta-analysis. JAMA Oncol. 2016;2(11):1460–9. https://doi.org/10.1001/jamaoncol.2016.1373.CrossRefPubMedPubMedCentral

Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, Ohgaki H, Wiestler OD, Kleihues P, Ellison DW. The 2016 World Health Organization Classification of Tumors of the Central Nervous System: a summary. Acta Neuropathol. 2016;131(6):803–20. https://doi.org/10.1007/s00401-016-1545-1.CrossRefPubMed

Look AT, Ashmun RA, Shapiro LH, Peiper SC. Human myeloid plasma membrane glycoprotein CD13 (gp150) is identical to aminopeptidase N. J Clin Invest. 1989;83(4):1299–307. https://doi.org/10.1172/JCI114015.CrossRefPubMedPubMedCentral

Look AT, Peiper SC, Rebentisch MB, Ashmun RA, Roussel MF, Rettenmier CW, Sherr CJ. Transfer and expression of the gene encoding a human myeloid membrane antigen (gp150). J Clin Invest. 1985;75(2):569–79. https://doi.org/10.1172/JCI111733.CrossRefPubMedPubMedCentral

10.

Mina-Osorio P. The moonlighting enzyme CD13: old and new functions to target. Trends Mol Med. 2008;14(8):361–71. https://doi.org/10.1016/j.molmed.2008.06.003.CrossRefPubMedPubMedCentral

11.

Winnicka B, O’Conor C, Schacke W, Vernier K, Grant CL, Fenteany FH, Pereira FE, Liang B, Kaur A, Zhao R, et al. CD13 is dispensable for normal hematopoiesis and myeloid cell functions in the mouse. J Leukoc Biol. 2010;88(2):347–59. https://doi.org/10.1189/jlb.0210065.CrossRefPubMed

12.

Armulik A, Genove G, Mae M, Nisancioglu MH, Wallgard E, Niaudet C, He L, Norlin J, Lindblom P, Strittmatter K, et al. Pericytes regulate the blood-brain barrier. Nature. 2010;468(7323):557–61. https://doi.org/10.1038/nature09522.CrossRefPubMed

13.

Bhagwat SV, Lahdenranta J, Giordano R, Arap W, Pasqualini R, Shapiro LH. CD13/APN is activated by angiogenic signals and is essential for capillary tube formation. Blood. 2001;97(3):652–9. https://doi.org/10.1182/blood.v97.3.652.CrossRefPubMed

14.

Lendeckel U, Karimi F, Al Abdulla R, Wolke C. The role of the Ectopeptidase APN/CD13 in Cancer. Biomedicines. 2023;11(3). https://doi.org/10.3390/biomedicines11030724.

15.

Barnieh FM, Loadman PM, Falconer RA. Is tumour-expressed aminopeptidase N (APN/CD13) structurally and functionally unique? Biochim Biophys Acta Rev Cancer. 2021;1876(2):188641. https://doi.org/10.1016/j.bbcan.2021.188641.CrossRefPubMed

16.

Wickstrom M, Larsson R, Nygren P, Gullbo J, Aminopeptidase N. (CD13) as a target for cancer chemotherapy. Cancer Sci. 2011;102(3):501–8. https://doi.org/10.1111/j.1349-7006.2010.01826.x.CrossRefPubMed

17.

Sekine K, Fujii H, Abe F. Induction of apoptosis by bestatin (ubenimex) in human leukemic cell lines. Leukemia. 1999;13(5):729–34. https://doi.org/10.1038/sj.leu.2401388.CrossRefPubMed

18.

Terauchi M, Kajiyama H, Shibata K, Ino K, Nawa A, Mizutani S, Kikkawa F. Inhibition of APN/CD13 leads to suppressed progressive potential in ovarian carcinoma cells. BMC Cancer. 2007;7:140. https://doi.org/10.1186/1471-2407-7-140.CrossRefPubMedPubMedCentral

19.

Liu S, Xie F, Wang H, Liu Z, Liu X, Sun L, Niu Z. Ubenimex inhibits cell proliferation, migration and invasion in renal cell carcinoma: the effect is autophagy-associated. Oncol Rep. 2015;33(3):1372–80. https://doi.org/10.3892/or.2014.3693.CrossRefPubMed

20.

Wang X, Niu Z, Jia Y, Cui M, Han L, Zhang Y, Liu Z, Bi D, Liu S. Ubenimex inhibits cell proliferation, migration and invasion by inhibiting the expression of APN and inducing autophagic cell death in prostate cancer cells. Oncol Rep. 2016;35(4):2121–30. https://doi.org/10.3892/or.2016.4611.CrossRefPubMed

21.

Carl-McGrath S, Lendeckel U, Ebert M, Wolter AB, Roessner A, Rocken C. The ectopeptidases CD10, CD13, CD26, and CD143 are upregulated in gastric cancer. Int J Oncol. 2004;25(5):1223–32.PubMed

22.

Gros C, Giros B, Schwartz JC. Identification of aminopeptidase M as an enkephalin-inactivating enzyme in rat cerebral membranes. Biochemistry. 1985;24(9):2179–85. https://doi.org/10.1021/bi00330a011.CrossRefPubMed

23.

Noble F, Banisadr G, Jardinaud F, Popovici T, Lai-Kuen R, Chen H, Bischoff L, Parsadaniantz SM, Fournie-Zaluski MC, Roques BP. First discrete autoradiographic distribution of aminopeptidase N in various structures of rat brain and spinal cord using the selective iodinated inhibitor [125I]RB 129. Neuroscience. 2001;105(2):479–88. https://doi.org/10.1016/s0306-4522(01)00185-3.CrossRefPubMed

24.

Chen A, Zhao W, Li X, Sun G, Ma Z, Peng L, Shi Z, Li X, Yan J. Comprehensive Oncogenic features of Coronavirus receptors in Glioblastoma Multiforme. Front Immunol. 2022;13:840785. https://doi.org/10.3389/fimmu.2022.840785.CrossRefPubMedPubMedCentral

25.

Di Matteo P, Arrigoni GL, Alberici L, Corti A, Gallo-Stampino C, Traversari C, Doglioni C, Rizzardi GP. Enhanced expression of CD13 in vessels of inflammatory and neoplastic tissues. J Histochem Cytochem. 2011;59(1):47–59. https://doi.org/10.1369/jhc.2010.956644.CrossRefPubMedPubMedCentral

26.

Brandenburg S, Muller A, Turkowski K, Radev YT, Rot S, Schmidt C, Bungert AD, Acker G, Schorr A, Hippe A, et al. Resident microglia rather than peripheral macrophages promote vascularization in brain tumors and are source of alternative pro-angiogenic factors. Acta Neuropathol. 2016;131(3):365–78. https://doi.org/10.1007/s00401-015-1529-6.CrossRefPubMed

27.

Bungert AD, Urbantat RM, Jelgersma C, Bekele BM, Mueller S, Mueller A, Felsenstein M, Dusatko S, Blank A, Ghori A, et al. Myeloid cell subpopulations compensate each other for Ccr2-deficiency in glioblastoma. Neuropathol Appl Neurobiol. 2023;49(1):e12863. https://doi.org/10.1111/nan.12863.CrossRefPubMed

28.

Blank A, Kremenetskaia I, Urbantat RM, Acker G, Turkowski K, Radke J, Schneider UC, Vajkoczy P, Brandenburg S. Microglia/macrophages express alternative proangiogenic factors depending on granulocyte content in human glioblastoma. J Pathol. 2021;253(2):160–73. https://doi.org/10.1002/path.5569.CrossRefPubMed

29.

Ji S, Ma Y, Xing X, Ge B, Li Y, Xu X, Song J, Xiao M, Gao F, Jiang W, et al. Suppression of CD13 enhances the cytotoxic effect of chemotherapeutic drugs in Hepatocellular Carcinoma cells. Front Pharmacol. 2021;12:660377. https://doi.org/10.3389/fphar.2021.660377.CrossRefPubMedPubMedCentral

30.

Zhang M, Cai H, Du Y, Wang Y, Gong J, Xu J, Liu X. Enhancing the therapeutic efficacy of Gefitinib in Human Non-small-cell Lung Cancer through Drug Combination. Mol Pharm. 2021;18(3):1397–407. https://doi.org/10.1021/acs.molpharmaceut.0c01203.CrossRefPubMed

31.

Amin SA, Adhikari N, Jha T. Design of aminopeptidase N inhibitors as anti-cancer agents. J Med Chem. 2018;61(15):6468–90. https://doi.org/10.1021/acs.jmedchem.7b00782.CrossRefPubMed

32.

Tsukamoto H, Shibata K, Kajiyama H, Terauchi M, Nawa A, Kikkawa F. Aminopeptidase N (APN)/CD13 inhibitor, Ubenimex, enhances radiation sensitivity in human cervical cancer. BMC Cancer. 2008;8:74. https://doi.org/10.1186/1471-2407-8-74.CrossRefPubMedPubMedCentral

33.

Blomgren H, Naslund I, Esposti PL, Johansen L, Aaskoven O. Adjuvant bestatin immunotherapy in patients with transitional cell carcinoma of the bladder. Clinical results of a randomized trial. Cancer Immunol Immunother. 1987;25(1):41–6. https://doi.org/10.1007/BF00199299.CrossRefPubMed

34.

Ichinose Y, Genka K, Koike T, Kato H, Watanabe Y, Mori T, Iioka S, Sakuma A, Ohta M, Group NKLCS. Randomized double-blind placebo-controlled trial of bestatin in patients with resected stage I squamous-cell lung carcinoma. J Natl Cancer Inst. 2003;95(8):605–10. https://doi.org/10.1093/jnci/95.8.605.CrossRefPubMed

35.

Kobayashi T, Miyawaki S, Tanimoto M, Kuriyama K, Murakami H, Yoshida M, Minami S, Minato K, Tsubaki K, Ohmoto E, et al. Randomized trials between behenoyl cytarabine and cytarabine in combination induction and consolidation therapy, and with or without ubenimex after maintenance/intensification therapy in adult acute myeloid leukemia. The Japan Leukemia Study Group. J Clin Oncol. 1996;14(1):204–13. https://doi.org/10.1200/JCO.1996.14.1.204.CrossRefPubMed

36.

Urabe A, Mutoh Y, Mizoguchi H, Takaku F, Ogawa N. Ubenimex in the treatment of acute nonlymphocytic leukemia in adults. Ann Hematol. 1993;67(2):63–6. https://doi.org/10.1007/BF01788128.CrossRefPubMed

37.

Farsa O, Ballayova V, Zackova R, Kollar P, Kauerova T, Zubac P. Aminopeptidase N inhibitors as pointers for overcoming Antitumor Treatment Resistance. Int J Mol Sci. 2022;23(17). https://doi.org/10.3390/ijms23179813.

38.

Zhao SM, Yao K, Liu KD, Huang LM, Jiang YA, Li J, Dong ZM, Dong ZG. Bestatin cream impairs solar simulated light-driven skin inflammation and skin in mice. J Invest Dermatol. 2021;141(11):2699–. https://doi.org/10.1016/j.jid.2021.03.032.CrossRefPubMed

39.

Wan J, Ling XA, Wang J, Ding GG, Wang X. Inhibitory effect of Ubenimex combined with fluorouracil on multiple drug resistance and P-glycoprotein expression level in non-small lung cancer. J Cell Mol Med. 2020;24(21):12840–7. https://doi.org/10.1111/jcmm.15875.CrossRefPubMedPubMedCentral

40.

Mendez LR, Arrebola Y, Valdes-Tresanco ME, Diaz-Guevara L, Bergado G, Sanchez B, Charli JL, Alonso IP. Bestatin and bacitracin inhibit porcine kidney cortex dipeptidyl peptidase IV activity and reduce human melanoma MeWo cell viability. Int J Biol Macromol. 2020;164:2944–52. https://doi.org/10.1016/j.ijbiomac.2020.08.157.CrossRefPubMed

41.

Fuchs Y, Steller H. Programmed cell death in animal development and disease. Cell. 2011;147(4):742–58. https://doi.org/10.1016/j.cell.2011.10.033.CrossRefPubMedPubMedCentral

42.

Wang B, Wang Y, Zhang J, Hu C, Jiang J, Li Y, Peng Z. ROS-induced lipid peroxidation modulates cell death outcome: mechanisms behind apoptosis, autophagy, and ferroptosis. Arch Toxicol. 2023;97(6):1439–51. https://doi.org/10.1007/s00204-023-03476-6.CrossRefPubMed

43.

Priya R, Jain V, Akhtar J, Chauhan G, Sakhuja P, Goyal S, Agarwal AK, Javed A, Jain AP, Polisetty RV, et al. Plasma-derived candidate biomarkers for detection of gallbladder carcinoma. Sci Rep. 2021;11(1):23554. https://doi.org/10.1038/s41598-021-02923-7.CrossRefPubMedPubMedCentral

44.

Tran VT, Phan TT, Mac HP, Tran TT, Ho TT, Pho SP, Nguyen VN, Vo TM, Nguyen HT, Le TT, et al. The diagnostic power of CD117, CD13, CD56, CD64, and MPO in rapid screening acute promyelocytic leukemia. BMC Res Notes. 2020;13(1):394. https://doi.org/10.1186/s13104-020-05235-7.CrossRefPubMedPubMedCentral

45.

Zhao Y, Wu H, Xing X, Ma Y, Ji S, Xu X, Zhao X, Wang S, Jiang W, Fang C, et al. CD13 induces Autophagy to promote Hepatocellular Carcinoma Cell Chemoresistance through the P38/Hsp27/CREB/ATG7 pathway. J Pharmacol Exp Ther. 2020;374(3):512–20. https://doi.org/10.1124/jpet.120.265637.CrossRefPubMed

46.

Guo Q, Li X, Cui MN, Sun JL, Ji HY, Ni BB, Yan MX. CD13: a key player in Multidrug Resistance in Cancer Chemotherapy. Oncol Res. 2020;28(5):533–40. https://doi.org/10.3727/096504020X15919605976853.CrossRefPubMedPubMedCentral

47.

Mawrin C, Wolke C, Haase D, Kruger S, Firsching R, Keilhoff G, Paulus W, Gutmann DH, Lal A, Lendeckel U. Reduced activity of CD13/aminopeptidase N (APN) in aggressive meningiomas is associated with increased levels of SPARC. Brain Pathol. 2010;20(1):200–10. https://doi.org/10.1111/j.1750-3639.2009.00267.x.CrossRefPubMed

48.

Marletta S, Luchini C, Sperandio N, Torresani E, Sorio A, Girolami I, Scarpa A, Eccher A, Ghimenton C. CD13 is a useful tool in the differential diagnosis of meningiomas with potential biological and prognostic implications. Virchows Arch. 2022;480(6):1223–30. https://doi.org/10.1007/s00428-022-03304-9.CrossRefPubMedPubMedCentral

49.

Gao P, Ren G. Identification of potential target genes of non-small cell lung cancer in response to resveratrol treatment by bioinformatics analysis. Aging. 2021;13(19):23245–61. https://doi.org/10.18632/aging.203616.CrossRefPubMedPubMedCentral

50.

Orning L, Krivi G, Fitzpatrick FA. Leukotriene A4 hydrolase. Inhibition by bestatin and intrinsic aminopeptidase activity establish its functional resemblance to metallohydrolase enzymes. J Biol Chem. 1991;266(3):1375–8.CrossRefPubMed

51.

Tieku S, Hooper NM. Inhibition of aminopeptidases N, A and W. A re-evaluation of the actions of bestatin and inhibitors of angiotensin converting enzyme. Biochem Pharmacol. 1992;44(9):1725–30. https://doi.org/10.1016/0006-2952(92)90065-q.CrossRefPubMedPubMedCentral

52.

Strong MJ, Koduri S, Allison JA, Pesavento CM, Ogunsola S, Ogunsola O, Yee TJ, Khalsa SSS, Saadeh YS, Joseph JR, et al. Bone metastasis from glioblastoma: a systematic review. J Neurooncol. 2022;158(3):379–92. https://doi.org/10.1007/s11060-022-04025-4.CrossRefPubMed

53.

Toshiyama R, Konno M, Eguchi H, Takemoto H, Noda T, Asai A, Koseki J, Haraguchi N, Ueda Y, Matsushita K, et al. Poly(ethylene glycol)-poly(lysine) block copolymer-ubenimex conjugate targets aminopeptidase N and exerts an antitumor effect in hepatocellular carcinoma stem cells. Oncogene. 2019;38(2):244–60. https://doi.org/10.1038/s41388-018-0406-x.CrossRefPubMed

54.

Redza-Dutordoir M, Averill-Bates DA. Activation of apoptosis signalling pathways by reactive oxygen species. Biochim Biophys Acta. 2016;1863(12):2977–92. https://doi.org/10.1016/j.bbamcr.2016.09.012.CrossRefPubMed

Title: CD13 expression affects glioma patient survival and influences key functions of human glioblastoma cell lines in vitro
Authors: Wenying Zhang
Anne Blank
Irina Kremenetskaia
Anja Nitzsche
Güliz Acker
Peter Vajkoczy
Susan Brandenburg
Publication date: 01-12-2024
Publisher: BioMed Central
Keywords: Glioblastoma
Glioblastoma
Glioblastoma
Glioma
Glioma
Glioma
Published in: BMC Cancer / Issue 1/2024
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-024-12113-z

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