EVALUATION OF HIGH DIMENTIONAL REDUCTION AND CLUSTERING IN THE PHENOTYPIC DICRIMINATION OF CD5+ B-CELL CHRONIC LYMPHOPROLIFERATIVE DISEASES

BAEZ, NATALIA; ARROYO, DANIELA; MANZONE-RODRIGUEZ, CLARISA; CECILIA M. RODRIGUEZ; PABLO IRIBARREN

Jornada; Reunión Anual de Sociedades de Biociencias (SAI); 2020

Introduction: Mature (peripheral) B-cell malignancies represent the malignant counterpart of normal mature B-cells that have differentiated into naive B cells or their progeny. Integration of a complex set of immunophenotypic, morphological, clinical, and cytogenetic information is essential for the subclassification of B-cell chronic lymphoproliferative diseases (B-CLPD).Phenotyping is essential for the diagnostic classification of many B-CLPD cases, but the current immunophenotyping strategies also face several difficulties. In view of these issues, new methods to facilitate the identification of abnormal B-cell populations in routine clinical flow cytometric data would be desirable.Methods: We used both 12 and 8 colour staining panels and we applied high dimensional reduction (viSNE) and clustering tools to discriminate between CD5+ B-CLPD cases. Samples from already diagnosed CLL and MCL were analyzed (n=6) and healthy patients? cells were used as controls. Results:High dimensional reduction (viSNE) revealed at least 6 individual clusters that corresponded to each CD5+ B-CLPD sample. In addition, the two dimensions spatial distribution of the populations showed segregation by disease type (n=6, p<0.05). Conversely, healthy control samples were separated in two clusters, but all the samples showed overlapping (p=NS). Clustering suggested the heterogeneity in markers expression in each disease sample. For instance, we detected clusters with higher expression of Ig-k and CD20, that corresponded to MCL samples, p<0.05 and p<0.01, respectively. Deeper analysis of these samples is currently under investigation. Conclusions:These preliminary results suggest that combination of high dimensional reduction and clustering might be an additional tool that can be used, at least, to distinguish between CD5+ B-CLPD. Further research is required to confirm these results and to evaluate the power of these tools in the classification of atypical forms of the B-CLPD.