The Role Of Fibins in Zebrafish Development, Gill Inflammation, & Remodelling
Lead Research Organisation:
Imperial College London
Department Name: Life Sciences
Abstract
The role of Fibins in zebrafish development, gill inflammation & remodelling
Chronic lung disease is characterised by persistent dysregulated inflammation. The mechanisms that contribute to this inflammation as well as those driving its resolution largely remain to be elucidated. Zebrafish (Danio rerio) has been shown to be a valuable model to study respiratory tissue inflammation. Previous research within the Dallman group (Imperial College London) demonstrated that zebrafish possess ability to efficiently resolve gill inflammation after cryoinjury and/or exposure to cigarette smoke (1). Importantly, the molecular mechanisms involved in the process allowed for a transient inflammatory stage followed by a scar-less resolution/tissue regeneration. Such outcome is in sharp contrast to the irreversible deposition of fibrotic tissue observed in mice and humans after repetitive injury to the alveolar epithelium (2).
In collaboration with Boehringer Ingelheim, analysis of gene expression in our zebrafish gill wounding model was carried out. Comparative analysis of regulated genes in the zebrafish model and those in mouse model of fibrosis identified several genes showing differential expression between the two models. Two genes in particular, the fin bud initiation factor (fibin) a and fibinb, were upregulated in the zebrafish gill during the inflammation resolution stage, but downregulated in the mouse model of fibrosis. To date, very little is known about the functions of the secreted Fibin proteins, apart from the putative role of zebrafish fibinb in pectoral fin formation (3). Understanding the role(s) that Fibins play in development and inflammation may identify new targets for the treatment of non-resolving inflammation and scarring present in human pulmonary fibrosis.
RESEARCH PLAN
AIM1: To characterise the expression pattern of fibin genes and the role of Fibin proteins during zebrafish development.
Firstly, the pattern of expression of both genes will be analysed during early stages of development in various tissues. Secondly, models of gene knockdown by morpholinos and gene knockout/knockdown using the CRISPR-Cas9 technology will be established to investigate the function of each protein individually, and in combination.
AIM2: To investigate the role of fibina and b in zebrafish gill during the stages of acute inflammation and resolution/tissue remodelling using the zebrafish model of gill injury developed in the lab.
Gene expression will be investigated by RT-PCR, q-PCR and in situ hybridisation. The expression of Fibin proteins will be studied using custom-made Fibina and Fibinb antibodies available to us.
The role of fibins in regeneration processes of injured gills will be assessed using knock-down and knock out tools developed in aim1.
AIM3: Investigate the molecular pathways involving the Fibin proteins during acute inflammation and resolution/tissue remodelling stages.
The results obtained in AIM1/2, in particular signs of strong phenotype in fibin knockdowns and differential tissue/cell-specific gene expression profiles, will guide the focus on specific molecular pathways
Chronic lung disease is characterised by persistent dysregulated inflammation. The mechanisms that contribute to this inflammation as well as those driving its resolution largely remain to be elucidated. Zebrafish (Danio rerio) has been shown to be a valuable model to study respiratory tissue inflammation. Previous research within the Dallman group (Imperial College London) demonstrated that zebrafish possess ability to efficiently resolve gill inflammation after cryoinjury and/or exposure to cigarette smoke (1). Importantly, the molecular mechanisms involved in the process allowed for a transient inflammatory stage followed by a scar-less resolution/tissue regeneration. Such outcome is in sharp contrast to the irreversible deposition of fibrotic tissue observed in mice and humans after repetitive injury to the alveolar epithelium (2).
In collaboration with Boehringer Ingelheim, analysis of gene expression in our zebrafish gill wounding model was carried out. Comparative analysis of regulated genes in the zebrafish model and those in mouse model of fibrosis identified several genes showing differential expression between the two models. Two genes in particular, the fin bud initiation factor (fibin) a and fibinb, were upregulated in the zebrafish gill during the inflammation resolution stage, but downregulated in the mouse model of fibrosis. To date, very little is known about the functions of the secreted Fibin proteins, apart from the putative role of zebrafish fibinb in pectoral fin formation (3). Understanding the role(s) that Fibins play in development and inflammation may identify new targets for the treatment of non-resolving inflammation and scarring present in human pulmonary fibrosis.
RESEARCH PLAN
AIM1: To characterise the expression pattern of fibin genes and the role of Fibin proteins during zebrafish development.
Firstly, the pattern of expression of both genes will be analysed during early stages of development in various tissues. Secondly, models of gene knockdown by morpholinos and gene knockout/knockdown using the CRISPR-Cas9 technology will be established to investigate the function of each protein individually, and in combination.
AIM2: To investigate the role of fibina and b in zebrafish gill during the stages of acute inflammation and resolution/tissue remodelling using the zebrafish model of gill injury developed in the lab.
Gene expression will be investigated by RT-PCR, q-PCR and in situ hybridisation. The expression of Fibin proteins will be studied using custom-made Fibina and Fibinb antibodies available to us.
The role of fibins in regeneration processes of injured gills will be assessed using knock-down and knock out tools developed in aim1.
AIM3: Investigate the molecular pathways involving the Fibin proteins during acute inflammation and resolution/tissue remodelling stages.
The results obtained in AIM1/2, in particular signs of strong phenotype in fibin knockdowns and differential tissue/cell-specific gene expression profiles, will guide the focus on specific molecular pathways
People |
ORCID iD |
| Margaret Dallman (Primary Supervisor) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/R50578X/1 | 01/10/2017 | 31/12/2021 | |||
| 2365164 | Studentship | BB/R50578X/1 | 29/09/2018 | 23/12/2022 |