Live imaging and genetic dissection of immunothrombosis in Drosophila
Lead Research Organisation:
King's College London
Department Name: Randall Div of Cell and Molecular Biophy
Abstract
Upon injury, a blood clot is immediately formed to stop bleeding and block infection. Subsequently, inflammatory cells are recruited to the wound to remove tissue debris and prevent infection, and finally the damaged tissue is repaired. The mechanisms controlling these wound healing responses are critical for survival and occur in nearly all animals. However, the challenge now is to understand how these seemingly disparate tissue repair processes are coordinated as it has become evident that clot formation cannot be disentangled from other aspects of the wound response.
Here we will exploit a simple organism, Drosophila melanogaster (fruit fly), which has been a powerhouse in terms of enhancing our understanding of wound healing and inflammation, to determine how clotting is regulated and subsequently controlling other aspects of tissue repair. In recent years we and others have revealed that the mechanisms behind each of these processes, clotting, inflammation, and tissue repair, can be elucidated with the help of fruit flies. For the first time we will holistically examine all of these processes simultaneously in the fly in order to examine how the process of coagulation is controlling - or controlled by - other aspects wound repair.
We will first exploit our ability to acquire movies of clot formation, inflammation, and wound healing in the living fruit fly to examine the dynamics of the coagulatory response throughout tissue repair. We will subsequently take advantage of our ability to rapidly analyse the function of specific genes in the fruit fly to determine the mechanisms controlling clot formation, including known signals emanating from damaged tissues or the inflammatory response. Finally we will test how the loss of clotting affects inflammatory cell recruitment and tissue healing.
Our goal is to holistically understand how the coagulatory cascade is intertwined with other aspects of the tissue repair process. As many of the wound response signals in the fruit fly are similar to higher organisms, the mechanisms identified here are likely to provide novel avenues that may allow us to intervene and modulate inflammation and tissue repair in humans.
Here we will exploit a simple organism, Drosophila melanogaster (fruit fly), which has been a powerhouse in terms of enhancing our understanding of wound healing and inflammation, to determine how clotting is regulated and subsequently controlling other aspects of tissue repair. In recent years we and others have revealed that the mechanisms behind each of these processes, clotting, inflammation, and tissue repair, can be elucidated with the help of fruit flies. For the first time we will holistically examine all of these processes simultaneously in the fly in order to examine how the process of coagulation is controlling - or controlled by - other aspects wound repair.
We will first exploit our ability to acquire movies of clot formation, inflammation, and wound healing in the living fruit fly to examine the dynamics of the coagulatory response throughout tissue repair. We will subsequently take advantage of our ability to rapidly analyse the function of specific genes in the fruit fly to determine the mechanisms controlling clot formation, including known signals emanating from damaged tissues or the inflammatory response. Finally we will test how the loss of clotting affects inflammatory cell recruitment and tissue healing.
Our goal is to holistically understand how the coagulatory cascade is intertwined with other aspects of the tissue repair process. As many of the wound response signals in the fruit fly are similar to higher organisms, the mechanisms identified here are likely to provide novel avenues that may allow us to intervene and modulate inflammation and tissue repair in humans.
Technical Summary
Drosophila has become a widely used model system to understand the mechanisms controlling blood coagulation, epithelial repair, and inflammation. However, each of these processes are normally studied in isolation. Here we will holistically investigate how coagulation, inflammation and tissue repair are mechanistically and functionally coordinated as it is increasingly evident that clotting reactions cannot be disentangled from other aspects of the repair process. We have recently revealed that the clot reaction can be live imaged in fruit fly embryos. As this embryonic repair model is also amenable to dissecting inflammatory cell recruitment and epithelial repair, this gives us the unique capacity to examine the coordination of these responses.
We will first characterise the dynamics of clotting using fluorescently tagged clot components, and determine the temporal relationship of coagulation with inflammation and epithelial repair. We will subsequently exploit the genetic tractability of fruit flies to determine the mechanisms controlling clot induction, including the role of the myriad of hypothesised clotting factors and enzymes, as well as the well-characterised repair response pathways. Finally, we will examine the ability of the coagulatory cascade to control inflammatory cell recruitment, epithelial repair, and systemic wound responses, by examining tissue repair when clot components are perturbed.
As many wound response signals and coagulatory factors are conserved in higher organisms, this work will provide an understanding of the unappreciated coordination between these processes with the future goal of intervening and modulating the wound repair process.
We will first characterise the dynamics of clotting using fluorescently tagged clot components, and determine the temporal relationship of coagulation with inflammation and epithelial repair. We will subsequently exploit the genetic tractability of fruit flies to determine the mechanisms controlling clot induction, including the role of the myriad of hypothesised clotting factors and enzymes, as well as the well-characterised repair response pathways. Finally, we will examine the ability of the coagulatory cascade to control inflammatory cell recruitment, epithelial repair, and systemic wound responses, by examining tissue repair when clot components are perturbed.
As many wound response signals and coagulatory factors are conserved in higher organisms, this work will provide an understanding of the unappreciated coordination between these processes with the future goal of intervening and modulating the wound repair process.