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

Projektinformationen

IntestineUb

ID Finanzhilfevereinbarung: 660116

DOI 10.3030/660116

Abgeschlossenes Projekt

EK-Unterschriftsdatum 26 März 2015

Startdatum 1 Januar 2016

Enddatum 1 November 2018

Finanziert unter

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Gesamtkosten € 195 454,80

EU-Beitrag € 195 454,80

Koordiniert durch

THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE
United Kingdom

Ziel

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.

Wissenschaftliches Gebiet

• medical and health sciences › medical biotechnology › genetic engineering
• medical and health sciences › medical biotechnology › cells technologies › stem cells
• medical and health sciences › clinical medicine › oncology
• medical and health sciences › clinical medicine › embryology
• medical and health sciences › basic medicine › physiology › homeostasis

Programm/Programme

• H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme
• H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility

Thema/Themen

• MSCA-IF-2014-EF - Marie Skłodowska-Curie Individual Fellowships (IF-EF)

Aufforderung zur Vorschlagseinreichung

H2020-MSCA-IF-2014

Finanzierungsplan

MSCA-IF-EF-ST - Standard EF

Koordinator

THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE

• Netto-EU-Beitrag: € 195 454,80
• Adresse: TRINITY LANE THE OLD SCHOOLS ; CB2 1TN Cambridge ; Vereinigtes Königreich
• Region: East of England › East Anglia › Cambridgeshire CC
• Aktivitätstyp: Higher or Secondary Education Establishments
• Gesamtkosten: € 195 454,80