Abstract
Pediatric cancer is a relatively rare and heterogeneous group of hematological and non-hematological malignancies which require multiple procedures for its diagnostic screening and classification. Until now, flow cytometry (FC) has not been systematically applied to the diagnostic work-up of such malignancies, particularly for solid tumors. Here we evaluated a FC panel of markers for the diagnostic screening of pediatric cancer and further classification of pediatric solid tumors. The proposed strategy aims at the differential diagnosis between tumoral vs. reactive samples, and hematological vs. non-hematological malignancies, and the subclassification of solid tumors. In total, 52 samples from 40 patients suspicious of containing tumor cells were analyzed by FC in parallel to conventional diagnostic procedures. The overall concordance rate between both approaches was of 96% (50/52 diagnostic samples), with 100% agreement for all reactive/inflammatory and non-infiltrated samples as well as for those corresponding to solid tumors (n = 35), with only two false negative cases diagnosed with Hodgkin lymphoma and anaplastic lymphoma, respectively. Moreover, clear discrimination between samples infiltrated by hematopoietic vs. non-hematopoietic tumor cells was systematically achieved. Distinct subtypes of solid tumors showed different protein expression profiles, allowing for the differential diagnosis of neuroblastoma (CD56hi/GD2+/CD81hi), primitive neuroectodermal tumors (CD271hi/CD99+), Wilms tumors (>1 cell population), rhabdomyosarcoma (nuMYOD1+/numyogenin+), carcinomas (CD45−/EpCAM+), germ cell tumors (CD56+/CD45−/NG2+/CD10+) and eventually also hemangiopericytomas (CD45−/CD34+). In summary, our results show that multiparameter FC provides fast and useful complementary data to routine histopathology for the diagnostic screening and classification of pediatric cancer.
Highlights
More than 200,000 pediatric patients (,15 years old) are diagnosed with cancer every year [1]
Multiparameter flow cytometry (MFC) immunophenotyping has proven to be essential for rapid diagnosis, classification and monitoring of therapy of most hematological malignancies, including pediatric leukemias and lymphomas
While immunophenotypic studies are routinely applied in most pediatric lymphomas for the differential diagnosis between B- and T-cell precursor lymphoblastic lymphomas and Burkitt lymphoma [25,26,27,28], few studies have been reported so far, in which multiparameter flow cytometry (MFC) is systematically applied to the study of the phenotypic characteristics of pediatric solid tumors [29,30,31]
Summary
More than 200,000 pediatric patients (,15 years old) are diagnosed with cancer every year [1]. It remains a research tool for pediatric solid tumors [8,9,10,11,12,13,14,15] Such limited usage of MFC in the diagnosis of pediatric solid tumors probably relates to the need for single cell suspensions and the availability of relatively restricted panels of reliable and validated markers, among other factors [16,17,18]. Usage of flow cytometry in pediatric solid tumors has been almost restricted to the evaluation of tumor cell DNA contents in both paraffin-embedded and frozen tissue specimens, in combination or not with simultaneous staining for one or a few phenotypic markers [19,20,21,22,23,24]. The few reported studies have mainly focused on descriptive analyses of the staining patterns for one or a few markers, usually in a single diagnostic subtype of the disease
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