DSV: Thesis SL-DSV-16-0451 - instn

DSV: Thesis SL-DSV-16-0451
RESEARCH FIELD
Cellular biology, physiology and cellular imaging / Life Sciences
TITLE
Identification and study of molecular mechanisms regulating lineage specifications
ABSTRACT
Synchronization of self-renewal and cell differentiation is fundamentally connected to the regulation of cell cycle progression. The length of
cell cycle during the G1 phase is crucial for the cellular determination. The relationship between the molecular mechanisms that determine
the length of cell cycle and the activation of factors involved in the lineage choice remains widely unknown. It is the case for the erythroid
(red blood cell production) and megacaryocyte cell lines that are produced from the same precursor; as well as the white and brown fat
lineages. Our laboratory developed several models of studies that could help to understand this regulation:
- The slowdown of the G1/S progression is crucial for erythroid maturation. The transcription factor GATA-1, implicated in the expression
of all specific genes of these cells, is also involve in the control of the proliferating factor E2F-2 activity, and this, by direct association of
GATA-1 to the retinoblastoma protein (pRb) (Kadri et al, MCB 2005, PLoS Biol 2009, Genes & Dev 2015).
- Control of adipogenesis and the conversion of white fat to brown fat are operated by the association of the GATA co-factor FOG-2 with
pRb (Goupille et al, Nature Com in revision).
The length regulation of the G1/S transition by the binding of GATA-1 or FOG-2 on pRb could be, respectively, implicated in the choices of
the erythro/megakaryocyte lineages or of white/brown fat.
Methodology: GATA-1, FOG-2 and/or pRb genes will be mutated, by the strategy of CRISPR/Cas9, to decrease or increase
GATA/FOG/pRb complexes. Then, we will used the Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) technology to follow in vivo
and in real time the cellular cycle, to evaluate the length of each phase and to purified by cell sorter the cell at the same phase. These
strategies will allow the selection of mutations that modulate the length of G1/S phase. We shall focus our attention to the physiological
consequences of these modifications and we shall determine criteria that instruct the differentiation towards such or such lineages. For
this aim, we shall realize transcriptomique studies, analyses of phosphorylation modifications of various key proteins and we shall
characterize epigenetic and metabolic modifications. The ultimate goal of this research project will contribute to discover new strategies to
replace lost or destroyed cells (anemia, trombopenia…) or to correct damaged cells (obesity, genetic mutations, cancers…)
LOCATION
Institut des Maladies Emergentes et des Thérapies Innovantes
Service Thérapies Innovantes
Laboratoire d'Etude de la Prolifération et de la Diifférenciation Somatique
Place: Fontenay-aux-roses
Start date of the thesis: 01/10/2016
Commissariat à l'énergie atomique et aux énergies alternatives
Institut national des sciences et techniques nucléaires
www­instn.cea.fr
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CONTACT PERSON
Stany CHRETIEN
INSERM
DSV-IMTI
CEA Fontenay aux Roses
Email: [email protected]
UNIVERSITY / GRADUATE SCHOOL
Paris Sud
Cancérologie, Biologie, Médecine, Santé
THESIS SUPERVISOR
Stany CHRETIEN
INSERM
DSV-IMTI
CEA Fontenay aux Roses
Commissariat à l'énergie atomique et aux énergies alternatives
Institut national des sciences et techniques nucléaires
www­instn.cea.fr
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