Počet záznamů: 1

High-resolution Antibody Array Analysis of Childhood Acute Leukemia Cells

SYSNO ASEP	0471866
Druh ASEP	J - Článek v odborném periodiku
Zařazení RIV	J - Článek v odborném periodiku
Poddruh J	Článek ve WOS
Název	High-resolution Antibody Array Analysis of Childhood Acute Leukemia Cells
Tvůrce(i)	Kanderová, V. (CZ) Kuzilkova, D. (CZ) Stuchlý, J. (CZ) Vašková, M. (CZ) Brdička, Tomáš (UMG-J)_RID Fišer, K. (CZ) Hrušák, O. (CZ) Lund-Johansen, F. (NO) Kalina, T. (CZ)
Celkový počet autorů	9
Zdroj.dok.	Molecular & Cellular Proteomics. - : Elsevier - ISSN 1535-9476 Roč. 15, č. 4 (2016), s. 1246-1261
Poč.str.	16 s.
Jazyk dok.	eng - angličtina
Země vyd.	US - Spojené státy americké
Klíč. slova	acute lymphoblastic-leukemia ; acute promyelocytic leukemia ; cytometric immunobead assay ; caspase-dependent cleavage ; acute myeloid-leukemia ; gene-expression ; fusion proteins ; flow-cytometry ; pcr data ; b-cells
Vědní obor RIV	EB - Genetika a molekulární biologie
CEP	2B06064 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy
Institucionální podpora	UMG-J - RVO:68378050
UT WOS	000373992600006
DOI	10.1074/mcp.M115.054593
Anotace	Acute leukemia is a disease pathologically manifested at both genomic and proteomic levels. Molecular genetic technologies are currently widely used in clinical research. In contrast, sensitive and high-throughput proteomic techniques for performing protein analyses in patient samples are still lacking. Here, we used a technology based on size exclusion chromatography followed by immunoprecipitation of target proteins with an antibody bead array (Size Exclusion Chromatography-Microsphere-based Affinity Proteomics, SEC-MAP) to detect hundreds of proteins from a single sample. In addition, we developed semi-automatic bioinformatics tools to adapt this technology for high-content proteomic screening of pediatric acute leukemia patients. To confirm the utility of SEC-MAP in leukemia immunophenotyping, we tested 31 leukemia diagnostic markers in parallel by SEC-MAP and flow cytometry. We identified 28 antibodies suitable for both techniques. Eighteen of them provided excellent quantitative correlation between SEC-MAP and flow cytometry (p < 0.05). Next, SEC-MAP was applied to examine 57 diagnostic samples from patients with acute leukemia. In this assay, we used 632 different antibodies and detected 501 targets. Of those, 47 targets were differentially expressed between at least two of the three acute leukemia subgroups. The CD markers correlated with immunophenotypic categories as expected. From non-CD markers, we found DBN1, PAX5, or PTK2 overexpressed in B-cell precursor acute lymphoblastic leukemias, LAT, SH2D1A, or STAT5A overexpressed in T-cell acute lymphoblastic leukemias, and HCK, GLUD1, or SYK overexpressed in acute myeloid leukemias. In addition, OPAL1 overexpression corresponded to ETV6-RUNX1 chromosomal translocation. In summary, we demonstrated that SEC-MAP technology is a powerful tool for detecting hundreds of proteins in clinical samples obtained from pediatric acute leukemia patients. It provides information about protein size and reveals differences in protein expression between particular leukemia subgroups. Forty-seven of SEC-MAP identified targets were validated by other conventional method in this study.
Pracoviště	Ústav molekulární genetiky
Kontakt	Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217
Rok sběru	2017

Počet záznamů: 1