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01-05-2019 | Astrocytoma | Laboratory Investigation

Cytometric analysis of cell suspension generated by cavitron ultrasonic surgical aspirator in pediatric brain tumors

Authors: Martina Vaskova, Michal Tichy, Josef Zamecnik, Petr Liby, Daniela Kuzilkova, Ales Vicha, Jan Hrabeta, Tomas Kalina, Jan Stary, Ondrej Hrusak

Published in: Journal of Neuro-Oncology | Issue 1/2019

Abstract

Purpose

The aim of this study was to test the possibility of using specimens obtained by a cavitron ultrasonic surgical aspirator (CUSA) in flow and mass cytometry investigations of pediatric brain tumors.

Methods

CUSA specimens obtained from 19 pediatric patients with brain tumors were investigated. Flow and mass cytometry methods were applied to analyze the composition of material collected using the CUSA. Cell suspensions were prepared from CUSA aspirates. Then sample viability was assessed by conventional flow cytometry and subsequently stained with a panel of 31 metal-labeled antibodies.

Results

Viability assessment was performed using conventional flow cytometry. Viability of cells in the acquired samples was below 50% in 16 of 19 cases. A mass cytometry investigation and subsequent analysis enabled us to discriminate brain tumor cells from contaminating leukocytes, whose proportions varied across the specimens. The addition of the viability marker cisplatin directly into the mass cytometry panel gave the means to selecting viable cells only for subsequent analyses. The proportion of non-viable cells was higher among tumor cells compared leukocytes.

Conclusions

When the analysis of the tumor cell immunophenotype is performed with markers for determining viability, the expression of the investigated markers can be evaluated. Suitable markers can be selected by high-throughput methods, such as mass cytometry, and those that are diagnostically relevant can be investigated using flow cytometry, which is more flexible in terms of time.

Louis DN, Perry A, Reifenberger G et al. (2016) The 2016 World Health Organization classification of tumors of the central nervous system: a summary. Acta Neuropathol 131:803–820. https://doi.org/10.1007/s00401-016-1545-1 CrossRefPubMed

Akagi Y, Yoshimoto K, Hata N et al. (2018) Reclassification of 400 consecutive glioma cases based on the revised 2016WHO classification. Brain Tumor Pathol 35:81–89. https://doi.org/10.1007/s10014-018-0313-4 CrossRefPubMed

Epstein F (1983) The cavitron ultrasonic aspirator in tumor surgery. Clin Neurosurg 31:497–505CrossRefPubMed

Day BW, Stringer BW, Wilson J et al. (2013) Glioma surgical aspirate: a viable source of tumor tissue for experimental research. Cancers 5:357–371. https://doi.org/10.3390/cancers5020357 CrossRefPubMedPubMedCentral

Bandura DR, Baranov VI, Ornatsky OI et al. (2009) Mass cytometry: technique for real time single cell multitarget immunoassay based on inductively coupled plasma time-of-flight mass spectrometry. Anal Chem 81:6813–6822. https://doi.org/10.1021/ac901049w CrossRefPubMed

Bendall SC, Nolan GP, Roederer M, Chattopadhyay PK (2012) A deep profiler’s guide to cytometry. Trends Immunol 33:323–332. https://doi.org/10.1016/j.it.2012.02.010 CrossRefPubMedPubMedCentral

Hrušák O, Basso G, Ratei R et al. (2014) Flow diagnostics essential code: a simple and brief format for the summary of leukemia phenotyping. Cytometry B Clin Cytom 86:288–291CrossRefPubMed

Liu G, Yuan X, Zeng Z et al. (2006) Analysis of gene expression and chemoresistance of CD133+ cancer stem cells in glioblastoma. Mol Cancer 5:67. https://doi.org/10.1186/1476-4598-5-67 CrossRefPubMedPubMedCentral

Lange F, Kaemmerer D, Behnke-Mursch J et al. (2018) Differential somatostatin, CXCR4 chemokine and endothelin a receptor expression in WHO grade I-IV astrocytic brain tumors. J Cancer Res Clin Oncol 144:1227–1237. https://doi.org/10.1007/s00432-018-2645-1 CrossRefPubMed

10.

Terakawa Y, Agnihotri S, Golbourn B et al. (2013) The role of drebrin in glioma migration and invasion. Exp Cell Res 319:517–528. https://doi.org/10.1016/j.yexcr.2012.11.008 CrossRefPubMed

11.

Vaskova M, Kovac M, Volna P et al. (2011) High expression of cytoskeletal protein drebrin in TEL/AML1pos B-cell precursor acute lymphoblastic leukemia identified by a novel monoclonal antibody. Leuk Res 35:1111–1113. https://doi.org/10.1016/j.leukres.2011.03.028 CrossRefPubMed

12.

Flavahan WA, Wu Q, Hitomi M et al. (2013) Brain tumor initiating cells adapt to restricted nutrition through preferential glucose uptake. Nat Neurosci 16:1373–1382. https://doi.org/10.1038/nn.3510 CrossRefPubMedPubMedCentral

13.

Navale AM, Paranjape AN (2016) Glucose transporters: physiological and pathological roles. Biophys Rev 8:5–9. https://doi.org/10.1007/s12551-015-0186-2 CrossRefPubMedPubMedCentral

14.

Mansouri S, Nejad R, Karabork M et al. (2016) Sox2: regulation of expression and contribution to brain tumors. CNS Oncol 5:159–173. https://doi.org/10.2217/cns-2016-0001 CrossRefPubMedPubMedCentral

15.

Chumakova AP, Hitomi M, Sulman EP, Lathia JD (2019) High-throughput automated single-cell imaging analysis reveals dynamics of glioblastoma stem cell population during state transition. Cytometry A. https://doi.org/10.1002/cyto.a.23728 CrossRefPubMedPubMedCentral

16.

Hepburn AC, Steele RE, Veeratterapillay R et al. (2019) The induction of core pluripotency master regulators in cancers defines poor clinical outcomes and treatment resistance. Oncogene. https://doi.org/10.1038/s41388-019-0712-y CrossRefPubMedPubMedCentral

17.

Ludwig K, Kornblum HI (2017) Molecular markers in glioma. J Neurooncol 134:505–512. https://doi.org/10.1007/s11060-017-2379-y CrossRefPubMedPubMedCentral

18.

Kamamoto D, Saga I, Ohara K et al. (2018) Association between CD133, CD44, and nestin expression and prognostic factors in high-grade meningioma. World Neurosurg. https://doi.org/10.1016/j.wneu.2018.12.067 CrossRefPubMed

19.

Yamamuro S, Okamoto Y, Sano E et al. (2015) Characterization of glioma stem-like cells from human glioblastomas. Int J Oncol 47:91–96. https://doi.org/10.3892/ijo.2015.2992 CrossRefPubMedPubMedCentral

20.

Liang L, Aiken C, McClelland R et al. (2015) Characterization of novel biomarkers in selecting for subtype specific medulloblastoma phenotypes. Oncotarget 6:38881–38900. https://doi.org/10.18632/oncotarget.6195 CrossRefPubMedPubMedCentral

21.

Liang L, Coudière-Morrison L, Tatari N et al. (2018) CD271+ cells are diagnostic and prognostic and exhibit elevated MAPK activity in SHH medulloblastoma. Cancer Res 78:4745–4759. https://doi.org/10.1158/0008-5472.CAN-18-0027 CrossRefPubMed

22.

Gholamin S, Mitra SS, Feroze AH et al. (2017) Disrupting the CD47-SIRPα anti-phagocytic axis by a humanized anti-CD47 antibody is an efficacious treatment for malignant pediatric brain tumors. Sci Transl Med 9:eaaf2968. https://doi.org/10.1126/scitranslmed.aaf2968 CrossRefPubMed

23.

Li F, Lv B, Liu Y et al. (2018) Blocking the CD47-SIRPα axis by delivery of anti-CD47 antibody induces antitumor effects in glioma and glioma stem cells. Oncoimmunology 7:e1391973. https://doi.org/10.1080/2162402X.2017.1391973 CrossRefPubMed

24.

Amir ED, Davis KL, Tadmor MD et al. (2013) viSNE enables visualization of high dimensional single-cell data and reveals phenotypic heterogeneity of leukemia. Nat Biotechnol 31:545–552. https://doi.org/10.1038/nbt.2594 CrossRefPubMedCentral

25.

Mallawaaratchy DM, Hallal S, Russell B et al. (2017) Comprehensive proteome profiling of glioblastoma-derived extracellular vesicles identifies markers for more aggressive disease. J Neurooncol 131:233–244. https://doi.org/10.1007/s11060-016-2298-3 CrossRefPubMed

26.

Hutter G, Theruvath J, Graef CM et al. (2019) Microglia are effector cells of CD47-SIRPα antiphagocytic axis disruption against glioblastoma. Proc Natl Acad Sci USA 116:997–1006. https://doi.org/10.1073/pnas.1721434116 CrossRefPubMedPubMedCentral

27.

Majzner RG, Theruvath JL, Nellan A et al. (2019) CAR T cells targeting B7-H3, a pan-cancer antigen, Demonstrate potent preclinical activity against pediatric solid tumors and brain tumors. Clin Cancer Res. https://doi.org/10.1158/1078-0432.CCR-18-0432 CrossRefPubMedPubMedCentral

28.

Brown CE, Alizadeh D, Starr R et al (2016) Regression of glioblastoma after chimeric antigen receptor T-cell therapy. N Engl J Med 375:2561–2569. https://doi.org/10.1056/NEJMoa1610497 CrossRefPubMedPubMedCentral

29.

Choi BD, Maus MV, June CH, Sampson JH (2018) Immunotherapy for glioblastoma: adoptive T-cell strategies. Clin Cancer Res. https://doi.org/10.1158/1078-0432.CCR-18-1625 CrossRefPubMedPubMedCentral

Title: Cytometric analysis of cell suspension generated by cavitron ultrasonic surgical aspirator in pediatric brain tumors
Authors: Martina Vaskova
Michal Tichy
Josef Zamecnik
Petr Liby
Daniela Kuzilkova
Ales Vicha
Jan Hrabeta
Tomas Kalina
Jan Stary
Ondrej Hrusak
Publication date: 01-05-2019
Publisher: Springer US
Keywords: Astrocytoma
Medulloblastoma
Medulloblastoma
Astrocytoma
Medulloblastoma
Brain Tumor
Astrocytoma
Published in: Journal of Neuro-Oncology / Issue 1/2019
Print ISSN: 0167-594X
Electronic ISSN: 1573-7373
DOI: https://doi.org/10.1007/s11060-019-03135-w

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Cytometric analysis of cell suspension generated by cavitron ultrasonic surgical aspirator in pediatric brain tumors

Abstract

Purpose

Methods

Results

Conclusions

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

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