Menu
MapConnectionsTopicsBulgariaThe Night (K-TRIO)Marie Skłodowska-CurieFor studentsFor teachersMethodologySurprise me
H2020Individual fellowship2016–2018

IntestineUb · The role of Ubiquitin System on the homeostatic control of stem cell maintenance and differentiation in crypt stem cells

Horizon 2020 — Marie Skłodowska-Curie Actions

Duration
2016-01-01 → 2018-11-01
EU contribution
€195,455
Participants
1
Scheme
MSCA-IF-EF-ST

Lines connect the coordinator with its partners.

Results in brief

The role of Ubiquitin System on the homeostatic control of stem cell maintenance and differentiation in crypt stem cells

The epithelium of small intestine and skin is continuously exposed to environmental assault and therefore requires regular and fast self-renewal, which is fuelled by the adult stem cells (SCs). Tight regulation of the adult SC number is required to sustain an adequately sized population for optimal tissue homeostasis. Mis-regulation of homeostasis results in either hyperplasia or loss of SCs. Although transcriptional regulation of SC homeostasis regarding multipotency and differentiation has been extensively studied, only little is known about post-transcriptional and post-translational modifications. A pivotal mechanism of post-transcriptional modification is RNA methylation by RNA methyltransferase like the NSUN family; a key mechanism of post-translational modification is ubiquitination by the ubiquitin system (US) E1, E2, E3 ubiquitin ligase and deubiquitinase (DUBs). The role of NSUNs and DUBs in adult SC homeostasis control is poorly understood. The in vitro primary 3D intestinal organoid culture system recapitulates in vivo epithelium organization, is amenable to essentially all experimental technologies that have been developed for cell lines, and therefore serves as a novel, robust and powerful model system. Taking advantage of the intestinal organoids and the well-established human keratinocytes culture, this project aims to investigate the role of NSUNs and DUBs on the homeostatic control of stem cell maintenance and differentiation in intestine and skin adult stem cells by: 1. Screening of NSUNs/DUBs required for stem cell number regulation; 2. Characterization of the expression pattern and sub-cellular localization of candidate NSUNs/ DUBs; 3. Examining the effect of candidate NSUNs/DUBs in known signalling pathways involved in the homeostatic control; 4. Elucidating the physiological role of NSUNs/DUBs with conditional knockout mouse models.

Data: CORDIS, © European Union

Project objective

The epithelium of small intestine has a higher self-renewal rate than any other mammalian tissues. This is fuelled by the intestinal stem cells which reside near the bottom of crypt. Tight regulation of the adult stem cell number is required to sustain an adequately sized population for optimal tissue homeostasis. Mis-regulation of homeostasis results in either hyperplasia or loss of stem cells. Although transcriptional regulation of stem cell homeostasis regarding multipotency and differentiation has been extensively studied, only little is known about post-translational modifications. A key mechanism of post-translational modification is ubiquitination by the ubiquitin system (US).The in vitro primary intestinal organoid culture system recapitulates in vivo epithelium organization, is amenable to essentially all experimental technologies that have been developed for cell lines, and therefore serves as a novel, robust and powerful model system. Taking advantage of the intestinal organoids, this project is aimed to investigate the role of US on the homeostatic control of stem cell maintenance and differentiation in crypt stem cells by: 1. Screening of US required for stem cell number regulation by CRISPR/Cas9 technology; 2. Characterization of the expression pattern and sub-cellular localization of candidate US; 3. Examining the effect of candidate US in signalling pathways involved in the homeostatic control by reporter assay and target gene expression; 4. Identifying target proteins of candidate US using surface proteome analysis, IP-mass, BioID or in vitro ubiquitination of arrayed proteins; 5. Performing biochemical and cell biological analysis of candidate US with its identified target proteins and signalling pathway components in vitro; 6. Generation of conditional knockout mouse models to elucidate the function of candidate US in vivo; 7. Conducting a small molecule screening for inhibitors that attenuate hyperplasia phenotype caused by E3 ligases mutation.

Original text from CORDIS.

Participants

  • THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE · CAMBRIDGECoordinatorUnited Kingdom

Links

Data: CORDIS, © European Union