Abstract
Purpose
Multicolor flow cytometry (MFC) is widely available, fast and has an easy-to perform approach for finding neuroblastoma (NB) cells among normal bone marrow (BM) hematopoietic cells. Aim of the study was to investigate prognostic significance of initial MFC tumor cells’ detection in BM of children with NB.
Methods
51 patients (24 boys and 27 girls) aged from 6 days to 15 years (median age 1 year 3 months) with NB were included in the study. BM samples at the time of diagnosis were obtained from 2 to 5 aspiration sites per patient. CD45(−)CD56(+)CD81(+)GD2(+)-cells were evaluated by MFC.
Results
NB cells were detected in BM by FC more frequently compared to conventional cytomorphology (49.0% and 29.4% patients, respectively, р = 0.043). Patients with NB cells detected in BM by MFC had significantly worse event-free survival and cumulative incidence of relapse/progression [0.24(0.08) and 0.60(0.10), respectively] compared to children with negative result of immunophenotyping [0.85(0.07) and 0.12(0.06), respectively, p < 0.001 in both cases]. BM involvement detection by MFC maintained its prognostic significance in various patients groups. In multivariate analysis, immunophenotyping proved to be an independent prognostic factor when analyzed jointly with other NB risk factors. In 42 patients BM involvement was also studied by RQ-PCR for PHOX2B and TH genes expression. Within groups of patients divided by RQ-PCR positivity, MFC-positivity retained prognostic significance.
Conclusions
Thus flow cytometric BM involvement detection has very strong prognostic impact even stronger than RQ-PCR. It could be used in combination with other parameters for the treatment strategy choice in patients with NB.
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References
Beiske K et al (2009) Consensus criteria for sensitive detection of minimal neuroblastoma cells in bone marrow, blood and stem cell preparations by immunocytology and QRT-PCR: recommendations by the International Neuroblastoma Risk Group Task Force. Br J Cancer 100:1627–1637. https://doi.org/10.1038/sj.bjc.6605029
Berthold F, Spix C, Kaatsch P, Lampert F (2017) Incidence, survival, and treatment of localized and metastatic neuroblastoma in Germany 1979–2015. Paediatr Drugs 19:577–593. https://doi.org/10.1007/s40272-017-0251-3
Bomken SN, Redfern K, Wood KM, Reid MM, Tweddle DA (2006) Limitations in the ability of NB84 to detect metastatic neuroblastoma cells in bone marrow. J Clin Pathol 59:927–929. https://doi.org/10.1136/jcp.2005.035444
Bozzi F, Gambirasio F, Luksch R, Collini P, Brando B, Fossati-Bellani F (2006) Detecting CD56+/NB84+/CD45- immunophenotype in the bone marrow of patients with metastatic neuroblastoma using flow cytometry. Anticancer Res 26:3281–3287
Bozzi F et al (2008) Flow cytometric phenotype of rhabdomyosarcoma bone marrow metastatic cells and its implication in differential diagnosis with neuroblastoma. Anticancer Res 28:1565–1569
Burchill SA et al (2017) Recommendations for the standardization of bone marrow disease assessment and reporting in children with neuroblastoma on behalf of the International Neuroblastoma Response Criteria Bone Marrow Working Group. Cancer 123:1095–1105. https://doi.org/10.1002/cncr.30380
Cai JY et al (2012) Minimal residual disease is a prognostic marker for neuroblastoma with bone marrow infiltration. Am J Clin Oncol 35:275–278. https://doi.org/10.1097/COC.0b013e318210f51b
Cheung IY, Cheung NK (2001) Quantitation of marrow disease in neuroblastoma by real-time reverse transcription-PCR. Clin Cancer Res 7:1698–1705
Corrias MV et al (2008) Detection of GD2-positive cells in bone marrow samples and survival of patients with localised neuroblastoma. Br J Cancer 98:263–269. https://doi.org/10.1038/sj.bjc.6604179
Corrias MV et al (2012) Multiple target molecular monitoring of bone marrow and peripheral blood samples from patients with localized neuroblastoma and healthy donors. Pediatr Blood Cancer 58:43–49. https://doi.org/10.1002/pbc.22960
Cotterill SJ, Pearson AD, Pritchard J, Foot AB, Roald B, Kohler JA, Imeson J (2000) Clinical prognostic factors in 1277 patients with neuroblastoma: results of The European Neuroblastoma Study Group ‘Survey’ 1982–1992. Eur J Cancer 36:901–908
Druy AE et al (2018) Prospective investigation of applicability and the prognostic significance of bone marrow involvement in patients with neuroblastoma detected by quantitative reverse transcription. PCR Pediatr Blood Cancer. https://doi.org/10.1002/pbc.27354
Ferreira-Facio CS et al (2013) Contribution of multiparameter flow cytometry immunophenotyping to the diagnostic screening and classification of pediatric cancer. PLoS One 8:e55534. https://doi.org/10.1371/journal.pone.0055534
Gautam U, Srinivasan R, Rajwanshi A, Bansal D, Marwaha RK (2008) Comparative evaluation of flow-cytometric immunophenotyping and immunocytochemistry in the categorization of malignant small round cell tumors in fine-needle aspiration cytologic specimens. Cancer 114:494–503. https://doi.org/10.1002/cncr.23948
Hartmann O et al (1999) Prognostic factors in metastatic neuroblastoma in patients over 1 year of age treated with high-dose chemotherapy and stem cell transplantation: a multivariate analysis in 218 patients treated in a single institution. Bone Marrow Transpl 23:789–795. https://doi.org/10.1038/sj.bmt.1701737
Hedley BD, Keeney M (2013) Technical issues: flow cytometry and rare event analysis. Int J Lab Hematol 35:344–350. https://doi.org/10.1111/ijlh.12068
Ifversen MR, Kagedal B, Christensen LD, Rechnitzer C, Petersen BL, Heilmann C (2005) Comparison of immunocytochemistry, real-time quantitative RT-PCR and flow cytometry for detection of minimal residual disease in neuroblastoma. Int J Oncol 27:121–129
Monclair T et al (2009) The International Neuroblastoma Risk Group (INRG) staging system: an INRG Task Force report. J Clin Oncol 27:298–303. https://doi.org/10.1200/JCO.2008.16.6876
Okcu MF et al (2005) Flow cytometry and fluorescence in situ hybridization to detect residual neuroblastoma cells in bone marrow. Pediatr Blood Cancer 45:787–795. https://doi.org/10.1002/pbc.20428
Shen H, Tang Y, Xu X, Tang H (2013) Detection of the GD2+/CD56+/CD45− immunophenotype by flow cytometry in cerebrospinal fluids from a patient with retinoblastoma. Pediatr Hematol Oncol 30:30–32. https://doi.org/10.3109/08880018.2012.737094
Stutterheim J et al (2008) PHOX2B is a novel and specific marker for minimal residual disease testing in neuroblastoma. J Clin Oncol 26:5443–5449. https://doi.org/10.1200/JCO.2007.13.6531
Stutterheim J et al (2009) Detecting minimal residual disease in neuroblastoma: the superiority of a panel of real-time quantitative PCR Markers. Clin Chem 55:1316–1326. https://doi.org/10.1373/clinchem.2008.117945
Swerts K, De Moerloose B, Dhooge C, Brichard B, Benoit Y, Laureys G, Philippe J (2004) Detection of residual neuroblastoma cells in bone marrow: comparison of flow cytometry with immunocytochemistry. Cytomet B Clin Cytom 61:9–19. https://doi.org/10.1002/cyto.b.20019
Trager C, Kogner P, Lindskog M, Ponthan F, Kullman A, Kagedal B (2003) Quantitative analysis of tyrosine hydroxylase mRNA for sensitive detection of neuroblastoma cells in blood and bone marrow. Clin Chem 49:104–112
Tsang KS et al (2003) Detection of micrometastasis of neuroblastoma to bone marrow and tumor dissemination to hematopoietic autografts using flow cytometry and reverse transcriptase-polymerase chain reaction. Cancer 97:2887–2897. https://doi.org/10.1002/cncr.11389
van Wezel EM et al (2016) Neuroblastoma messenger RNA is frequently detected in bone marrow at diagnosis of localised neuroblastoma patients. Eur J Cancer 54:149–158. https://doi.org/10.1016/j.ejca.2015.11.007
Warzynski MJ, Graham DM, Axtell RA, Higgins JV, Hammers YA (2002) Flow cytometric immunophenotyping test for staging/monitoring neuroblastoma patients. Cytometry 50:298–304. https://doi.org/10.1002/cyto.10159
Warzynski MJ, Roys JL, Peterson JW, DeLa Vega H (2005) Availability of conjugated ganglioside GD2 monoclonal antibody. Cytomet B Clin Cytom 65:42–43. https://doi.org/10.1002/cyto.b.20054
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Popov, A., Druy, A., Shorikov, E. et al. Prognostic value of initial bone marrow disease detection by multiparameter flow cytometry in children with neuroblastoma. J Cancer Res Clin Oncol 145, 535–542 (2019). https://doi.org/10.1007/s00432-018-02831-w
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DOI: https://doi.org/10.1007/s00432-018-02831-w