title: TNF-alpha and Notch signaling regulates the expression of HOXB4 and GATA3 during early T lymphopoiesis.
abstract: During the early thymus colonization, Notch signaling activation on hematopoietic progenitor cells (HPCs) drives proliferation and T cell commitment. Although these processes are driven by transcription factors such as HOXB4 and GATA3, there is no evidence that Notch directly regulates their transcription. To evaluate the role of NOTCH and TNF signaling in this process, human CD34+HPCs were cocultured with OP9-DL1 cells, in the presence or absence of TNF. The use of a Notch signaling inhibitor and a protein synthesis inhibitor allowed us to distinguish primary effects, mediated by direct signaling downstream Notch and TNF, from secondary effects, mediated by de novo synthesized proteins. A low and physiologically relevant concentration of TNF promoted T lymphopoiesis in OP9-DL1 cocultures. TNF positively modulated the expression of both transcripts in a Notch-dependent manner; however, GATA3 induction was mediated by a direct mechanism, while HOXB4 induction was indirect. Induction of both transcripts was repressed by a GSK3β inhibitor, indicating that activation of canonical Wnt signaling inhibits rather than induces their expression. Our study provides novel evidences of the mechanisms integrating Notch and TNF-alpha signaling in the transcriptional induction of GATA3 and HOXB4. This mechanism has direct implications in the control of self-renewal, proliferation, commitment, and T cell differentiation.
authors.affiliation: Department of Genetics, Medical School of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil. josililian@gmail.com.|National Institute of Science and Technology in Stem Cell and for Cell Therapy (INCTC) Center for Cell Therapy (CTC) and Regional Blood Center, Ribeirão Preto, SP, Brazil. josililian@gmail.com.|Medical School of Ribeirão Preto, University of São Paulo , Ribeirão Preto, São Paulo, Brazil. josililian@gmail.com.
authors.forename: Josiane Lilian
authors.initials: JL
authors.lastname: Dos Santos Schiavinato
authors.affiliation: National Institute of Science and Technology in Stem Cell and for Cell Therapy (INCTC) Center for Cell Therapy (CTC) and Regional Blood Center, Ribeirão Preto, SP, Brazil.|Medical School of Ribeirão Preto, University of São Paulo , Ribeirão Preto, São Paulo, Brazil.
authors.forename: Lucila Habib Bourguignon
authors.initials: LH
authors.lastname: Oliveira
authors.affiliation: Medical School of Ribeirão Preto, University of São Paulo , Ribeirão Preto, São Paulo, Brazil.
authors.forename: Amélia Goes
authors.initials: AG
authors.lastname: Araujo
authors.affiliation: National Institute of Science and Technology in Stem Cell and for Cell Therapy (INCTC) Center for Cell Therapy (CTC) and Regional Blood Center, Ribeirão Preto, SP, Brazil.|Medical School of Ribeirão Preto, University of São Paulo , Ribeirão Preto, São Paulo, Brazil.
authors.forename: Maristela Delgado
authors.initials: MD
authors.lastname: Orellana
authors.affiliation: National Institute of Science and Technology in Stem Cell and for Cell Therapy (INCTC) Center for Cell Therapy (CTC) and Regional Blood Center, Ribeirão Preto, SP, Brazil.
authors.forename: Patrícia Viana Bonini
authors.initials: PV
authors.lastname: de Palma
authors.affiliation: National Institute of Science and Technology in Stem Cell and for Cell Therapy (INCTC) Center for Cell Therapy (CTC) and Regional Blood Center, Ribeirão Preto, SP, Brazil.|Medical School of Ribeirão Preto, University of São Paulo , Ribeirão Preto, São Paulo, Brazil.
authors.forename: Dimas Tadeu
authors.initials: DT
authors.lastname: Covas
authors.affiliation: National Institute of Science and Technology in Stem Cell and for Cell Therapy (INCTC) Center for Cell Therapy (CTC) and Regional Blood Center, Ribeirão Preto, SP, Brazil.|Medical School of Ribeirão Preto, University of São Paulo , Ribeirão Preto, São Paulo, Brazil.
authors.forename: Marco Antonio
authors.initials: MA
authors.lastname: Zago
authors.affiliation: Department of Genetics, Medical School of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.|National Institute of Science and Technology in Stem Cell and for Cell Therapy (INCTC) Center for Cell Therapy (CTC) and Regional Blood Center, Ribeirão Preto, SP, Brazil.
authors.forename: Rodrigo Alexandre
authors.initials: RA
authors.lastname: Panepucci
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D050990:GATA3 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D038041:Lymphopoiesis / Q000235:genetics; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D051880:Receptors, Notch / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism*; D000072056:Transcription Factor HES-1 / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D018797:Cell Cycle Proteins; D050990:GATA3 Transcription Factor; D018398:Homeodomain Proteins; D016328:NF-kappa B; D021122:Protein Subunits; D051880:Receptors, Notch; D000072056:Transcription Factor HES-1; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; C494749:GATA3 protein, human; C485631:HEY1 protein, human; C090460:HOXB4 protein, human; C078493:HES1 protein, human
keywords: CD34; HOXB4 and GATA3; Hematopoietic stem cells; Notch signaling