Title: Studying leukaemia and ageing-associated haematopoietic stem cell exhaustion in short-lived killifish
Funder: Cancer Research UK

Abstract: Background: Age is an often overlooked aspect of oncology, despite being the main risk factor for the majority of cancers. Research in this area is hampered by the shortcomings of established laboratory animal models. Flies and worms, the workhorses of ageing biology, are ill-suited for the study of cancer, as they lack the organs and adult stem cells that most frequently lead to cancer, while vertebrate models, such as mice and zebrafish, live for several years and thus drag out studies with ageing animals. We now want to initiate analyses of age-associated tumours in the short-lived African turquoise killifish. This fish has recently emerged as a potent vertebrate model system for ageing and associated complex diseases, facilitated by its rapid ageing and short lifespan of about 4 months. Aims: The overall aim is to establish the killifish model for the study of leukaemia in an ageing context. This new, potent model system, which accounts for ageing and allows for its experimental manipulation, will enable unique advances in our understanding of leukaemia and open up fresh opportunities for its prevention and treatment. In three objectives we want to: 1) generate killifish models for myelodysplastic syndrome/acute myeloid leukaemia (MDS/AML); 2) analyse the age-related decline of haematopoiesis; and 3) study the contribution of cell-autonomous and cell-extrinsic factors to the development of MDS/AML. Methods: We will generate MDS/AML models by 1) engineering a truncated TET2 protein variant using a CRISPR-Cas9-based approach, and 2) by expressing a mutated version of the splicing factor SRSF2 using a Tol2-based strategy. Both mutations are frequent in human MDS and AML. We will also use a Tol2-based approach to tag HSPC with a fluorescent reporter. Heterochronic transplants of HSPCs carrying Tet2 or SRSF2 mutations to recipients of different ages and subjected to caloric restriction will decouple cell-autonomous from cell-extrinsic factors contributing to cancer. To this end, we will adapt techniques of retro-orbital transplantation of HSPCs between individuals from routine zebrafish methods. How the results of this research will be used: This project will pioneer the use of killifish for cancer research. The MDS-AML killifish models and key assays developed, and the new insights into the contribution of ageing-related factors to MDS/AML, will empower future research on blood cancer. By demonstrating the value of killifish-based models of leukaemia, this project will act as a springboard for future analyses of cancer-related processes as a function of age, in leukaemia and beyond.

Lay abstract: Background: Age is an often overlooked aspect of oncology, despite being the main risk factor for the majority of cancers. Research in this area is hampered by the shortcomings of established laboratory animal models. Flies and worms, the workhorses of ageing biology, are ill-suited for the study of cancer, as they lack the organs and adult stem cells that most frequently lead to cancer, while vertebrate models, such as mice and zebrafish, live for several years and thus drag out studies with ageing animals. We now want to initiate analyses of age-associated tumours in the short-lived African turquoise killifish. This fish has recently emerged as a potent vertebrate model system for ageing and associated complex diseases, facilitated by its rapid ageing and short lifespan of about 4 months. Aims: The overall aim is to establish the killifish model for the study of leukaemia in an ageing context. This new, potent model system, which accounts for ageing and allows for its experimental manipulation, will enable unique advances in our understanding of leukaemia and open up fresh opportunities for its prevention and treatment. In three objectives we want to: 1) generate killifish models for myelodysplastic syndrome/acute myeloid leukaemia (MDS/AML); 2) analyse the age-related decline of haematopoiesis; and 3) study the contribution of cell-autonomous and cell-extrinsic factors to the development of MDS/AML. Methods: We will generate MDS/AML models by 1) engineering a truncated TET2 protein variant using a CRISPR-Cas9-based approach, and 2) by expressing a mutated version of the splicing factor SRSF2 using a Tol2-based strategy. Both mutations are frequent in human MDS and AML. We will also use a Tol2-based approach to tag HSPC with a fluorescent reporter. Heterochronic transplants of HSPCs carrying Tet2 or SRSF2 mutations to recipients of different ages and subjected to caloric restriction will decouple cell-autonomous from cell-extrinsic factors contributing to cancer. To this end, we will adapt techniques of retro-orbital transplantation of HSPCs between individuals from routine zebrafish methods. How the results of this research will be used: This project will pioneer the use of killifish for cancer research. The MDS-AML killifish models and key assays developed, and the new insights into the contribution of ageing-related factors to MDS/AML, will empower future research on blood cancer. By demonstrating the value of killifish-based models of leukaemia, this project will act as a springboard for future analyses of cancer-related processes as a function of age, in leukaemia and beyond.

Research activity codes: 2.1 BIOLOGICAL AND ENDOGENOUS FACTORS;2.2 FACTORS RELATING TO PHYSICAL ENVIRONMENT;2.6 RESOURCES AND INFRASTRUCTURE (AETIOLOGY)
Health categories: CANCER AND NEOPLASMS