Role of NFKB2 on the early myeloid differentiation of CD34+ hematopoietic stem/progenitor cells.
To better understand the early events regulating lineage-specific hematopoietic differentiation, we analyzed the transcriptional profiles of CD34+ human hematopoietic stem and progenitor cells (HSPCs) subjected to differentiation stimulus. CD34+ cells were cultured for 12 and 40h in liquid cultures with supplemented media favoring myeloid or erythroid commitment. Serial analysis of gene expression (SAGE) was employed to generate four independent libraries. By analyzing the differentially expressed regulated transcripts between the un-stimulated and the stimulated CD34+ cells, we observed a set of genes that was initially up-regulated at 12h but were then down-regulated at 40h, exclusively after myeloid stimulus. Among those we found transcripts for NFKB2, RELB, IL1B, LTB, LTBR, TNFRSF4, TGFB1, and IKBKA. Also, the inhibitor NFKBIA (IKBA) was more expressed at 12h. All those transcripts code for signaling proteins of the nuclear factor kappa B pathway. NFKB2 is a subunit of the NF-κB transcription factor that with RELB mediates the non-canonical NF-κB pathway. Interference RNA (RNAi) against NFKB1, NFKB2 and control RNAi were transfected into bone marrow CD34+HSPC. The percentage and the size of the myeloid colonies derived from the CD34+ cells decreased after inhibition of NFKB2. Altogether, our results indicate that NFKB2 gene has a role in the early commitment of CD34+HSPC towards the myeloid lineage.
De Molfetta
Greice Andreotti
GA
Center for Cell Therapy, Blood Center of the University of Sao Paulo, Rua Tenente Catão Roxo 2501, 14051-140 Ribeirão Preto - SP, Brazil.
Lucíola Zanette
Dalila
D
Alexandre Panepucci
Rodrigo
R
Dos Santos
Anemarie Ramos Dinarte
AR
da Silva
Wilson Araújo
WA
Antonio Zago
Marco
M
D000328:Adult; D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism*; D001853:Bone Marrow / Q000378:metabolism; D002454:Cell Differentiation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D022423:Myeloid Cells / Q000166:cytology* / Q000378:metabolism; D052003:NF-kappa B p52 Subunit / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D018952:Antigens, CD34; D052003:NF-kappa B p52 Subunit; D034741:RNA, Small Interfering