Project description:Adult neurogenesis occurs in mammals and provides a mechanism for continuous neural plasticity in the brain.However, little is known about the molecular mechanisms regulating hippocampal neural progenitor cells (NPCs) and whether their fate can be pharmacologically modulated to improve neural plasticity and regeneration. Here, we report the characterization of a unique small molecule (KHS101) that selectively induces a neuronal differentiation phenotype. Mechanism of action studies revealed a link of KHS101 to cell cycle exit and specific binding to the TACC3 protein, whose knockdown in NPCs recapitulates the KHS101-induced phenotype. Upon systemic administration, KHS101 distributed to the brainandresulted in a significant increase in neuronal differentiation in vivo. Our findings indicate that KHS101 accelerates neuronal differentiation by interaction with TACC3 and may provide a basis for pharmacological intervention.directed at endogenous NPCs. Compare expression profile of KHS101-treated hippocampal progenitor cells (2 concentrations) vs. DMSO (negative control), retinoic acid (positive control)
Project description:Cell cycle progression is linked to transcriptome dynamics and variations in the response of pluripotent cells to differentiation cues, through mostly unknown determinants. Here, we characterized the cell cycle–associated transcriptome and proteome of mouse embryonic stem cells (mESCs) in naïve ground state. We found that the thymine DNA glycosylase (TDG) is a cell cycle–regulated co-factor of the tumour suppressor p53. Further, TDG and p53 co-bind ESC-specific cis-regulatory elements and thereby control transcription of p53-dependent genes during self-renewal. We determined that the dynamic expression of TDG is required to promote the cell cycle–associated transcriptional heterogeneity. Moreover, we demonstrated that transient depletion of TDG influences cell fate decisions during the early differentiation of mESCs. Our findings reveal an unanticipated role of TDG in promoting molecular heterogeneity during the cell cycle, and highlight the central role of protein dynamics for the temporal control of cell fate during development.
Project description:Parasitic protists, including Plasmodium falciparum malaria parasites, evolved sophisticated biological features to adapt and survive in e.g. mosquito vectors and mammalian hosts. The parasite’s life cycle is extraordinarily controlled, oscillating between quiescent stages (e.g. sporozoites or gametocytes) and stages of intense proliferation. The atypical mode of asexual reproduction in erythrocytes (schizogony: asynchronous nuclear division in the absence of cytokinesis), is clearly divergent from cell division in higher eukaryotes but the mechanisms that control cell cycle progression are poorly understood. Here, depletion of exogenous factors was used to induce reversible cell cycle arrest and allowed transcriptomic investigations of cell cycle control mechanisms operating in the parasite. We show that in early stages of erythrocyte infection, parasites enter a quiescent, G0-like state due to implementation of a G1 restriction point prior to G1/S transition. This quiescence is reversible, with cells making a clear decision to re-engage the proliferation machinery in response to proliferation stimuli. Cell cycle arrest is an adaptive response rather than merely prolongation of the G1 phase, as demonstrated by a distinct transcriptome. The quiescence-proliferation decision-making process in malaria parasites has hallmarks of unique regulatory principles: quiescence is associated with a deregulation of the pre-replicative complex, underlined by transcriptional control of kinases, including PfPK5 as putative functional homologue of the master mammalian cell cycle regulator, CDK1. Cell cycle re-entry is highly coordinated and mediated by a set of early responsive transcripts involving NIMA and Aurora kinases, ApiAP2 transcription factors and calcium signaling. We therefore show that Apicomplexa are able to perform quiescence-proliferation decision-making in a highly coordinated fashion, using atypical cell cycle regulation machinery.
Project description:Dendritic cell (DC) maturation is a prerequisite for the induction of adaptive immune responses against pathogens and cancer. Transcription factor (TF) networks control differential aspects of early DC progenitor versus late stage DC cell fate decisions. Here, we identified the TF C/EBPβ as a key regulator for DC maturation and immunogenic functionality under homeostatic and lymphoma-transformed conditions. Gene expression profiles of splenic C/EBPβ-/- DCs showed a strong downregulation of E2F cell cycle target genes, whereas signatures of maturation were enriched. In accordance with E2F1 being a negative regulator of DC maturation, C/EBPβ-/- bone marrow-derived DCs matured much faster enabling them to strongly activate T cells. Conversely, the E2F transcriptional pathways were upregulated in lymphoma-exposed DCs and DC maturation was impaired. Pharmacological blockade of C/EBPβ/mTOR signaling in human DCs abrogated their pro-tumorigenic function in B-cell lymphoma co-cultures. Thus, C/EBPβ plays a unique role in DC maturation and functionality and emerges as a key factor of the microenvironment promoting lymphomagenesis.
Project description:Dendritic cell (DC) maturation is a prerequisite for the induction of adaptive immune responses against pathogens and cancer. Transcription factor (TF) networks control differential aspects of early DC progenitor versus late stage DC cell fate decisions. Here, we identified the TF C/EBPβ as a key regulator for DC maturation and immunogenic functionality under homeostatic and lymphoma-transformed conditions. Gene expression profiles of splenic C/EBPβ-/- DCs showed a strong downregulation of E2F cell cycle target genes, whereas signatures of maturation were enriched. In accordance with E2F1 being a negative regulator of DC maturation, C/EBPβ-/- bone marrow-derived DCs matured much faster enabling them to strongly activate T cells. Conversely, the E2F transcriptional pathways were upregulated in lymphoma-exposed DCs and DC maturation was impaired. Pharmacological blockade of C/EBPβ/mTOR signaling in human DCs abrogated their pro-tumorigenic function in B-cell lymphoma co-cultures. Thus, C/EBPβ plays a unique role in DC maturation and functionality and emerges as a key factor of the microenvironment promoting lymphomagenesis.
Project description:Protein kinase C (PKC) activation induces cellular reprogramming and differentiation in a variety of cellular models. Although many effectors of PKC physiological actions have been elucidated, the molecular mechanisms that regulate oligodendrocytes differentiation after PKC activation are still unclear. Here we applied a liquid chromatography - mass spectrometry (LC-MS/MS) approach to identify proteins, and biological processes, modulated after PKC activation in the established PKC-induced differentiation model, MO3.13 oligodendroglial cell line. Our analysis revealed that PKC signaling activation leads to the down-regulation of biological processes related to cell cycle and DNA replication and the up-regulation of a glial like developmental program. Our data suggested a role for ROCK in the modulation of MO3.13 phenotype, cytoskeletal and mechanotransduction dynamics but not in the modulation of cell cycle and calcium transport. Overall, our findings provides a resource for elucidating the PKC-mediated signaling network contributing to a more robust knowledge of the molecular dynamics leading to this cell fate transition.
Project description:Cell cycle progression is linked to transcriptome dynamics and variations in the response of pluripotent cells to differentiation cues, mostly through unknown determinants. Here, we characterized the cell-cycleassociated transcriptome and proteome of mouse embryonic stem cells (mESCs) in naive ground state. We found that the thymine DNA glycosylase (TDG) is a cell-cycle-regulated co-factor of the tumor suppressor p53. Furthermore, TDG and p53 co-bind ESC-specific cis-regulatory elements and thereby control transcription of p53-dependent genes during self-renewal. We determined that the dynamic expression of TDG is required to promote the cell-cycle-associated transcriptional heterogeneity. Moreover, we demonstrated that transient depletion of TDG influences cell fate decisions during the early differentiation of mESCs. Our findings reveal an unanticipated role of TDG in promoting molecular heterogeneity during the cell cycle and highlight the central role of protein dynamics for the temporal control of cell fate during development.
Project description:Epigenetic control of neural stem/progenitor cell fate is fundamental to achieve a fully brain architecture. Two intrinsic programs regulate neurogenesis, one by epigenetic-mediated gene transcription and another by cell cycle control. Whether and how these two are coordinated to determine temporally and spatially neural development remains unknown. Here we show that deletion of Trrap (Transcription translation associated protein), an essential cofactor for HAT (histone acetyltransferase), leads to severe brain atrophy due to a combination of cell death and a blockade of neuron production. Specifically, Trrap deletion forces differentiation of apical progenitor (AP) fate into basal progenitors (BP) and neurons thereby limiting the total neurogenic production. Despite TrrapM-bM-^@M-^Ys general role in transcriptional regulation, a genome-wide transcriptome analysis of neuroprogenitors identified the cell cycle regulators that are specifically affected by Trrap deletion. Furthermore, E2F-dependent recruitment of HAT and transcription factors to the promoter of cell cycle regulators is impaired in Trrap-deleted neuroprogenitors. Consistent with these molecular changes, Trrap deletion lengthens particularly G1 and S phases in APs in vivo. Therefore, our study reveals an essential and a distinct function of Trrap-HAT in regulation of cell cycle progression that is required for proper determination of neuroprogenitor fate. Determine gene transcriptions by comparing Trrap-deleted and wild type samples