H2020Индивидуална стипендия2016–2018

SKINFRET · The involvement of epidermal stem cells in spatiotemporal ERK activity propagation

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2016-06-27 → 2018-06-26
Финансиране от ЕС: 195 455 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Епидермалните стволови клетки и техните сигнали се наблюдават чрез специални флуоресцентни маркери в кожата на мишки и човешки клетки. Това помага да се разбере как промените в активността на протеина ERK управляват превръщането на стволовите клетки в специализирани клетки.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

The involvement of epidermal stem cells in spatiotemporal ERK activity propagation

In the project, we studied the spatial and temporal dynamics of ERK MAPK signal for the regulation of stem cell commitment to differentiation. As proposed in the Annex1 of Grant Agreement, ERK MAPK signal was observed at single-cell resolution by EKAREV, a FRET probe for ERK MAPK. It was applied to multiple experimental platforms from cultured human keratinocytes, 3D patterned substrates, and mouse in vivo skin. Together with ERK MAPK signal activity, we simultaneously monitored keratinocyte differentiation level by developing a reporter that expresses mCherry fluorescence under the regulation of Involucrin promoter. This revealed that stem cells and differentiated cells are characterized by stable high and low ERK MAPK activity, pattern, respectively. In contrast, cells that are committed to exit the stem cell compartment are characterized by pulsatile ERK MAPK activity. Differentiation was preceded by significant decrease in ERK MAPK pulses. Milestones of the projects were achieved as described below. (MS 1) in vivo imaging of stem cells and SPREAD : Achieved by the 6th month of Year 1 Pulse activation of ERK MAPK activity was successfully observed in both living mouse skin and cultured human epidermal stem cells. (MS 2) ERK activation pattern on engineered substrate: Achieved by the 3rd month of Year 2 On the 3D patterned substrate, cells with different ERK MAPK patterns were segregated. (MS 3) ERK activation pattern on 3D reconstituted skin: Still ongoing. live imaging of 3D reconstituted skin requires optimization of microscopy due to its fragile structure. (MS 4) Light-induced generation of SPREAD: Achieved by the 6th month of Year 2 Generation of ERK activation wave directed cells to migrate toward the origin of the waves. 1.1 Objectives In the project, we aimed to answer the following questions. Objective 1: Do epidermal stem cells operate as initiating cells of SPREAD? Objective 2: If so, what is the influence of stem cell microenvironment on SPREAD? Objective 3: Does SPREAD appear in human keratinocytes?

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

Stem cells are central to maintenance of mammalian epidermis. This proposal focuses on how stem cellscommunicate with their microenvironment by intercellular propagation of growth signal. I will investigate the involvement of epidermal stem cells in a novel growth signal propagation pattern I found in my PhD research. In this research, I visualized extracellular signal regulated kinase (ERK) activity in the skin of living mice by FRET-based observation of transgenic mouse expressing a sensitive biosensor for ERK. By this approach, I found a novel pattern of intercellular ERK activity propagation, which we named SPREAD. In SPREAD, bursts of localized ERK activation concentrically propagated to their surrounding 100 -300 cells. The timing and location of SPREAD emergence suggested the involvement of epidermal stem cells. To test this possibility I will combine my expertise of FRET imaging with in vivo and in vitro systems for mouse and human keratinocytes in the host laboratory. By crossing ERK biosensor mice with various mouse strains that marks Lgr5+, Lgr6+, or Lrig1+ stem cells, I will observe whether the centre cells of SPREADs are epidermal stem cells in vivo. In addition,I will culture keratinocytes expressing the ERK FRET biosensor on an artificially engineered substrate to recapitulate SPREAD in vitro. To study whether SPREADs are also seen in human skin, I will exploit a skin reconstruction system, called organotopic culture. The method allows culture of keratinocytes on collagen matrices embedded with various fibroblast populations with various densities. The significance of the features of SPREAD (shape, size, amplitude etc.) will be addressed by generating various types of ERK activity propagation by using light-induced ERK activation system. This approach will bring fresh insights into how stem cells control their local tissue structures via intercellular propagation of growth signal and how microenviromental factors effect on the propagation pattern.

Оригинален текст от CORDIS (на английски).

Участници

KING'S COLLEGE LONDON · LondonКоординаторОбединеното кралство