Reference number : GC-2017-Q1-02774, amendment number : , agreement type : G, recipient legal name : Larjava, Hannu (The University of British Columbia), recipient country : CA, recipient province : nan, recipient city : British Columbia|Colombie-Britannique, recipient postal code : nan, program name english : Discovery Grants Program - Individual, program name french : Programme de subventions à la découverte - individuelles, program purpose english : Overall program goal
 
The overall goal of this research program is to unravel the mechanisms that regulate the niche and function of oral/gingival neural crest derived stromal stem cells (NCSSCs). The NCSSCs are retained in a few specific locations&mdash;such as bone marrow, hair follicles and oral mucosa&mdash;in adult tissue and their numbers generally decline with age. Using Wnt1-CRE/R26RYFP reporter mice, we have shown using tracing experiments that gingival NCSSCs are enriched in the gingival connective tissue. Interestingly, the NCSSCs are enriched at the connective tissue papilla and more specifically in close proximity to gingival oral epithelial cells (OECs). It is well known that connective tissue and epithelial cells engage in molecular crosstalk that can regulate the functions of both cell types. The crosstalk between OECs and NCSSCs is, therefore, likely to be involved in the regulation of NCSSC niche and function. There is no information available in the literature on how OECs could regulate NCSSC niche or fate. In the present proposal, we are proposing a new paradigm that OECs release extracellular vesicles that participate in the regulation of NCSSC fate.
 
 
 General hypothesis
 
The general hypothesis is that signals derived from adjacent OECs regulate the fate and niche of NCSSCs in adult gingival tissue. Because the signals need to be specific and protected from off-target effects, we further hypothesize that the OEC&ndash;NCSSC crosstalk is regulated by extracellular vesicles (EVs). The EVs, including exosomes (EXO) and microvesicles (MVs), provide safety and receptor-mediated specificity to this regulation.
 
 
 Objectives and methodology
 
The objective of the proposal is to find out how OEC-derived EXOs and MVs regulate the function and expression of differentiation-related genes and proteins in NCSSCs. Furthermore, we will investigate how these vesicles regulate the specific differentiation pathways, including osteogenic, adipogenic, chondrogenic and neuronal lineages. We will use a NCSSC cell line (named Ginestin, with over 90% nestin-positive cells) that we have isolated from adult human gingiva as our target cell line, expose it to OEC vesicles and measure the output with RT-qPCR, Western blotting, proteomics, immunostainings and other microscopic techniques.
 
 
 Expected outcomes and significance
 
This research program is expected to significantly increase our understanding of how neural crest derived cells can survive in certain microenvironments such as oral mucosa. The novel aspect is the way we expect the OECs to contribute to the formation of the niche and regulation of the gene expression profile that is conductive for cell survival., program purpose french : Overall program goal
 
The overall goal of this research program is to unravel the mechanisms that regulate the niche and function of oral/gingival neural crest derived stromal stem cells (NCSSCs). The NCSSCs are retained in a few specific locations&mdash;such as bone marrow, hair follicles and oral mucosa&mdash;in adult tissue and their numbers generally decline with age. Using Wnt1-CRE/R26RYFP reporter mice, we have shown using tracing experiments that gingival NCSSCs are enriched in the gingival connective tissue. Interestingly, the NCSSCs are enriched at the connective tissue papilla and more specifically in close proximity to gingival oral epithelial cells (OECs). It is well known that connective tissue and epithelial cells engage in molecular crosstalk that can regulate the functions of both cell types. The crosstalk between OECs and NCSSCs is, therefore, likely to be involved in the regulation of NCSSC niche and function. There is no information available in the literature on how OECs could regulate NCSSC niche or fate. In the present proposal, we are proposing a new paradigm that OECs release extracellular vesicles that participate in the regulation of NCSSC fate.
 
 
 General hypothesis
 
The general hypothesis is that signals derived from adjacent OECs regulate the fate and niche of NCSSCs in adult gingival tissue. Because the signals need to be specific and protected from off-target effects, we further hypothesize that the OEC&ndash;NCSSC crosstalk is regulated by extracellular vesicles (EVs). The EVs, including exosomes (EXO) and microvesicles (MVs), provide safety and receptor-mediated specificity to this regulation.
 
 
 Objectives and methodology
 
The objective of the proposal is to find out how OEC-derived EXOs and MVs regulate the function and expression of differentiation-related genes and proteins in NCSSCs. Furthermore, we will investigate how these vesicles regulate the specific differentiation pathways, including osteogenic, adipogenic, chondrogenic and neuronal lineages. We will use a NCSSC cell line (named Ginestin, with over 90% nestin-positive cells) that we have isolated from adult human gingiva as our target cell line, expose it to OEC vesicles and measure the output with RT-qPCR, Western blotting, proteomics, immunostainings and other microscopic techniques.
 
 
 Expected outcomes and significance
 
This research program is expected to significantly increase our understanding of how neural crest derived cells can survive in certain microenvironments such as oral mucosa. The novel aspect is the way we expect the OECs to contribute to the formation of the niche and regulation of the gene expression profile that is conductive for cell survival., agreement title english : Regulation of neural crest derived progenitor cell niche by epithelial cell derived extracellular vesicles, agreement title french : Regulation of neural crest derived progenitor cell niche by epithelial cell derived extracellular vesicles, agreement number : RGPIN, agreement value : 140000.0, agreement start date : 2017-05-10, agreement end date : nan, description english : nan, description french : nan, expected results english : nan, expected results french : nan, additional information english : Grant or Award spanning more than one fiscal year. (2017-2018 to 2022-2023), additional information french : Subvention ou bourse octroyée s'appliquant à plus d'un exercice financier. (2017-2018 à 2022-2023), owner organization : nserc-crsng, owner organization title : Natural Sciences and Engineering Research Council of Canada | Conseil de recherches en sciences naturelles et en génie du Canada