Insulin resistance in dairy cattle and human beings:
Potential for extracellular vesicle based biomarker discovery
Non communicable diseases like Type 2 diabetes and insulin resistance (IR) have very high prevalence among humans and also emerging plethora of evidences suggest the same is happening in farm and companion animal species. In dairy cows, insulin resistance develops during late pregnancy and represents an important adaptation, switching of metabolic activity of the cow to use energy stored in adipose tissue (AT) and minimize glucose consumption in peripheral tissues, sparing it for milk synthesis. However; severe IR may lead to post-partum expansive AT fatty acids mobilization along with rapid body weight loss that predispose cows to a range of metabolic and infectious diseases/disorders, decreased milk production, compromised welfare, poor fertility and high mortality risk. Lack of early detection biomarkers and the inability to accurately predict the progress of the disease has for many years hindered the treatment and management of animal and human cases. Specially for dairy cows, developing IR detection tool using a non or less invasive sample-based technique could help to identify cows at potential risk and enable the application of preventive actions since all he current methods are based on very invasive sampling procedures. The EVs which are used in inter-cellular, inter-species and even in inter-kingdom level communication have attracted the attention as very potential candidates in devising early diagnosis platforms due to their protein and nucleic acids cargos. EVs are involved in regulating different innate immune responses, inflammation, and regeneration of the cells. Hence, they can be very good targets for theragnostic. A potential practical method for the diagnosis of IR in cattle should be non-invasive thus, milk is probably the best candidate that is readily available and easy to obtain from cows and is also relevant to humans (Figure 7). But urine is probably the easiest and most noninvasive biofluid with a high potential for biomarker discovery in human. Thus, in this project we will employ molecular biological “omics” tools with EVs present in milk and urine in identification of possible diagnostic and prognostic biomarkers for IR in cattle with the aim of translating the outcomes to the human diabetes type 2 diagnosis and potential complication of this disease in humans.
Figure 7. Characteristics of milk EVs as a tool in biomarker discovery
Project background
Type 2 diabetes can be considered as a remarkable disease condition worldwide and especially with a higher prevalence rate in developed regions. However, there are concerning trends of rising prevalence in lower-income countries as well. This disease is coupled with Insulin resistance (IR) which is a systemic disorder that affects many organs and insulin-regulated pathways. Insulin resistance can be defined as a state whereby a normal concentration of insulin induces a decreased biological response in insulin-sensitive tissues and is subdivided based on 2 distinct features; insulin sensitivity and insulin responsiveness. The disorder is characterized by a reduced action of insulin despite increased insulin concentrations (hyperinsulinemia). Not only among humans this is a massive problem in animal husbandry as well, especially related to the dairy industry. In dairy cows, insulin resistance develops during late pregnancy and represents an important adaptation, switching of metabolic activity of the cow to use energy stored in adipose tissue (AT) and minimize glucose consumption in peripheral tissues, sparing it for milk synthesis. However; severe IR may lead to post-partum expansive AT fatty acids mobilization along with rapid body weight loss, development of AT inflammatory status (which in turn amplify IR) that predispose cows to a range of metabolic and infectious diseases/disorders, decreased milk production, compromised welfare, poor fertility and high mortality risk. Relevant to humans, insulin resistance is a state where the body does not respond to insulin as well as it should. It's almost like a lack of communication between insulin and the cells of the body. Insulin resistance is a key feature of type 2 diabetes. Insulin resistance is a problem because it affects individuals in a number of ways. While genetics, aging, and ethnicity play roles in developing insulin sensitivity, the driving forces behind insulin resistance include excess body weight, too much belly fat, a lack of exercise, smoking, and even skimping on sleep. As insulin resistance develops, the human body fights back by producing more insulin. Over months and years, the beta cells in the pancreas which have been supplying insulin get degenerated and no longer keep pace with the demand for more and more insulin production. Then years after insulin resistance silently began, leading to the rise of blood sugar, type 2 diabetes, and the development of non-alcoholic fatty liver disease, a growing problem associated with insulin resistance that boosts the risk for liver damage and heart disease. The prognosis of people with type 2 diabetes varies. It depends on how well an individual modifies his or her risk of complications. Heart attack, stroke and kidney disease in these patients will result in premature death. Disability due to blindness, amputation, heart disease, stroke and nerve damage may also occur. It may be easy to diagnose type 2 diabetes but understanding how patients are reacting or which complications are in progress is not easy. A lack of biomarkers to accurately predict the progress of the disease has for many years hindered the treatment and management of patients with type 2 diabetes. An urgent need exists for the discovery of such biomarkers in humans. Similar to human beings, in cattle, early diagnosis of (possibly) developing IR could help to identify cows at potential risk and enable the application of preventive actions. In cattle, the gold standards for detection of IR are hyperinsulinaemic-euglycaemic clamp technique and intravenous glucose tolerance test. Unfortunately, both methods are invasive, laborious and time-consuming; therefore, unsuitable for practical use. Revised Quantitative Insulin Sensitivity Check Index (RQUICKI), which takes into account blood glucose, insulin and non-esterified fatty acids (NEFA) concentration, is suggested to be more practical; however, it is also invasive and appears to be not sensitive enough. At present, no good (non-invasive) biomarkers are available for the prognosis of IR in cattle.
EV are membrane-bound nano-scale particles (50nm to 1000nm in diameter) found in all biological fluids. Different types of EVs such as exosomes, micro-vesicles and apoptotic bodies have been described. All the studied species appear to produce EVs. EVs are used in inter-cellular, inter-species and even in inter-kingdom level communications making EV based communication the common language of life for the whole biome. The communications are involved in regulating different innate immune responses, inflammation, and regeneration of the cells. Hence, they can be very good targets for biomarker discovery as well as therapeutics. A potential practical method for the diagnosis of IR in cattle should be non-invasive. Milk is probably the best candidate for a body fluid that is readily available and easy to obtain from cows. From this point of view using milk EVs as a novel tool for IR prognosis may have high potential. On the other hand, in human beings, IR diagnoses may happen faster but the prognosis of further disease progression towards diabetes type 2 and in particular different complications such as nephropathy may develop and progress very silently. Hence, the availability of biomarkers to pinpoint the progress of the IR in humans and cattle will be of extreme importance for the management of patients in both species. However, bovine milk is probably the most noninvasive body fluid to obtain and is relevant to humans. On the other hand, urine is probably the easiest and most non-invasive biofluid with a high potential for biomarker discovery.
Type 2 diabetes can be considered as a remarkable disease condition worldwide and especially with a higher prevalence rate in developed regions. However, there are concerning trends of rising prevalence in lower-income countries as well. This disease is coupled with Insulin resistance (IR) which is a systemic disorder that affects many organs and insulin-regulated pathways. Insulin resistance can be defined as a state whereby a normal concentration of insulin induces a decreased biological response in insulin-sensitive tissues and is subdivided based on 2 distinct features; insulin sensitivity and insulin responsiveness. The disorder is characterized by a reduced action of insulin despite increased insulin concentrations (hyperinsulinemia). Not only among humans this is a massive problem in animal husbandry as well, especially related to the dairy industry. In dairy cows, insulin resistance develops during late pregnancy and represents an important adaptation, switching of metabolic activity of the cow to use energy stored in adipose tissue (AT) and minimize glucose consumption in peripheral tissues, sparing it for milk synthesis. However; severe IR may lead to post-partum expansive AT fatty acids mobilization along with rapid body weight loss, development of AT inflammatory status (which in turn amplify IR) that predispose cows to a range of metabolic and infectious diseases/disorders, decreased milk production, compromised welfare, poor fertility and high mortality risk. Relevant to humans, insulin resistance is a state where the body does not respond to insulin as well as it should. It's almost like a lack of communication between insulin and the cells of the body. Insulin resistance is a key feature of type 2 diabetes. Insulin resistance is a problem because it affects individuals in a number of ways. While genetics, aging, and ethnicity play roles in developing insulin sensitivity, the driving forces behind insulin resistance include excess body weight, too much belly fat, a lack of exercise, smoking, and even skimping on sleep. As insulin resistance develops, the human body fights back by producing more insulin. Over months and years, the beta cells in the pancreas which have been supplying insulin get degenerated and no longer keep pace with the demand for more and more insulin production. Then years after insulin resistance silently began, leading to the rise of blood sugar, type 2 diabetes, and the development of non-alcoholic fatty liver disease, a growing problem associated with insulin resistance that boosts the risk for liver damage and heart disease. The prognosis of people with type 2 diabetes varies. It depends on how well an individual modifies his or her risk of complications. Heart attack, stroke and kidney disease in these patients will result in premature death. Disability due to blindness, amputation, heart disease, stroke and nerve damage may also occur. It may be easy to diagnose type 2 diabetes but understanding how patients are reacting or which complications are in progress is not easy. A lack of biomarkers to accurately predict the progress of the disease has for many years hindered the treatment and management of patients with type 2 diabetes. An urgent need exists for the discovery of such biomarkers in humans. Similar to human beings, in cattle, early diagnosis of (possibly) developing IR could help to identify cows at potential risk and enable the application of preventive actions. In cattle, the gold standards for detection of IR are hyperinsulinaemic-euglycaemic clamp technique and intravenous glucose tolerance test. Unfortunately, both methods are invasive, laborious and time-consuming; therefore, unsuitable for practical use. Revised Quantitative Insulin Sensitivity Check Index (RQUICKI), which takes into account blood glucose, insulin and non-esterified fatty acids (NEFA) concentration, is suggested to be more practical; however, it is also invasive and appears to be not sensitive enough. At present, no good (non-invasive) biomarkers are available for the prognosis of IR in cattle.
EV are membrane-bound nano-scale particles (50nm to 1000nm in diameter) found in all biological fluids. Different types of EVs such as exosomes, micro-vesicles and apoptotic bodies have been described. All the studied species appear to produce EVs. EVs are used in inter-cellular, inter-species and even in inter-kingdom level communications making EV based communication the common language of life for the whole biome. The communications are involved in regulating different innate immune responses, inflammation, and regeneration of the cells. Hence, they can be very good targets for biomarker discovery as well as therapeutics. A potential practical method for the diagnosis of IR in cattle should be non-invasive. Milk is probably the best candidate for a body fluid that is readily available and easy to obtain from cows. From this point of view using milk EVs as a novel tool for IR prognosis may have high potential. On the other hand, in human beings, IR diagnoses may happen faster but the prognosis of further disease progression towards diabetes type 2 and in particular different complications such as nephropathy may develop and progress very silently. Hence, the availability of biomarkers to pinpoint the progress of the IR in humans and cattle will be of extreme importance for the management of patients in both species. However, bovine milk is probably the most noninvasive body fluid to obtain and is relevant to humans. On the other hand, urine is probably the easiest and most non-invasive biofluid with a high potential for biomarker discovery.
Project objectives
- To investigate the EV profile of bovine milk to develop novel biomarkers to identify IR
- To compare the EV profiles and EV genomic cargo obtained from bovine milk of high IR, low IR and healthy cows
- Investigating the EV profile of humans using urine to develop novel biomarkers to identify IR
- To compare the EV profiles and EV genomic cargo obtained from human urine of high IR patients, low IR patients and healthy people
Integration with EMU expertise, research strategy and development
This research theme combines different expertise in EMU and also take advantage of collaboration and cooperation between EMU and the University of Tartu (UT) Medical school. The project is well focused on an area of diseases that is relevant to human and animals. It tries to bring new dimensions to improve the health and welfare of animals and Humans.
In Emu, programmes in animal nutrition have been offered by the predecessors of the present University of Tartu and the Institute of Veterinary Medicine since the second half of the 19th century. The beginning of research on animal nutrition dates back to 1919, when the Faculty of Agriculture was established at the University of Tartu. In 1921, the Animal Breeding Experiment Station (Zootehnika Katsejaam) was established at Raadi that served as the main research facility for 50 years until a new unit was set up at Räni. In 1951, the Estonian Academy of Agriculture was founded. The Faculty of Zootechnics established at the Academy comprised a Chair of Animal Nutrition that primarily conducted basic research on nutrition, while the activities of the Institute of Animal Breeding and Veterinary Medicine (established in 1947) were mostly focused on applied research. In 1994, the two institutions were merged to form the Department of Animal Nutrition at the Estonian Agricultural University. There was a laboratory of feed analysis at the Department. Later on, the raw milk and metabolism research laboratory as well as the Department of Small Animal and Poultry Production were also merged with the Department. In September 2017, the Department was restructured into the Chair of Animal Nutrition. The current main focus of the Department of Animal Nutrition is on determining the nutrient content of feedstuff samples, which serve as basis for formulating proper feed rations. The major goal is to improve the yield and quality of animal produce through high-quality feedstuffs practices, which is also related to the overall health of a herd. In recent years, the institute has developed competence in organic livestock production. The research project “Insulin resistance in dairy cattle and human beings: potential for EVs based biomarker discovery” is bringing a new dimension in to the research strategy of the department. This research project involves the department in direct research collaboration with UT medical school. This is a very important and strategic development and will foster future collaborative research projects between Medical and Veterinary Medicine experts.
This research theme combines different expertise in EMU and also take advantage of collaboration and cooperation between EMU and the University of Tartu (UT) Medical school. The project is well focused on an area of diseases that is relevant to human and animals. It tries to bring new dimensions to improve the health and welfare of animals and Humans.
In Emu, programmes in animal nutrition have been offered by the predecessors of the present University of Tartu and the Institute of Veterinary Medicine since the second half of the 19th century. The beginning of research on animal nutrition dates back to 1919, when the Faculty of Agriculture was established at the University of Tartu. In 1921, the Animal Breeding Experiment Station (Zootehnika Katsejaam) was established at Raadi that served as the main research facility for 50 years until a new unit was set up at Räni. In 1951, the Estonian Academy of Agriculture was founded. The Faculty of Zootechnics established at the Academy comprised a Chair of Animal Nutrition that primarily conducted basic research on nutrition, while the activities of the Institute of Animal Breeding and Veterinary Medicine (established in 1947) were mostly focused on applied research. In 1994, the two institutions were merged to form the Department of Animal Nutrition at the Estonian Agricultural University. There was a laboratory of feed analysis at the Department. Later on, the raw milk and metabolism research laboratory as well as the Department of Small Animal and Poultry Production were also merged with the Department. In September 2017, the Department was restructured into the Chair of Animal Nutrition. The current main focus of the Department of Animal Nutrition is on determining the nutrient content of feedstuff samples, which serve as basis for formulating proper feed rations. The major goal is to improve the yield and quality of animal produce through high-quality feedstuffs practices, which is also related to the overall health of a herd. In recent years, the institute has developed competence in organic livestock production. The research project “Insulin resistance in dairy cattle and human beings: potential for EVs based biomarker discovery” is bringing a new dimension in to the research strategy of the department. This research project involves the department in direct research collaboration with UT medical school. This is a very important and strategic development and will foster future collaborative research projects between Medical and Veterinary Medicine experts.
Project team
|
Yasalath Gamage Madhusha Prasadani
Institute of Veterinary Medicine and Animal Sciences Student |
Alireza Fazeli
Institute of Veterinary Medicine and Animal Sciences Supervisor |
|
Suranga Kodithuwakku
Institute of Veterinary Medicine and Animal Sciences Supervisor |
Meelis Ots
Institute of Veterinary Medicine and Animal Sciences Supervisor |
|
Hanno Jaakson
Institute of Veterinary Medicine and Animal Sciences Supervisor |
Vallo Volke
Institute of Veterinary Medicine and Animal Sciences Supervisor |
