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Genetic Engineering Searching for a target for the treatment of atherosclerosis

Genetic Engineering Searching for a target for the treatment of atherosclerosis. RISK FACTORS. Consumption of foods high in saturated fats and a sedentary lifestyle increase the risk of suffering from cardiovascular diseases such as atherosclerosis. WHAT IS ATHEROSCLEROSIS?.

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Genetic Engineering Searching for a target for the treatment of atherosclerosis

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  1. Genetic EngineeringSearching for a target for the treatment of atherosclerosis

  2. RISK FACTORS Consumption of foods high in saturated fats and a sedentary lifestyle increase the risk of suffering from cardiovascular diseases such as atherosclerosis.

  3. WHAT IS ATHEROSCLEROSIS? Atherosclerosis is a vascular disease caused by the accumulation of fats on the walls of the blood vessels. There are many different signs and degrees of severity. • Normal artery • Moderate • atherosclerosis • Severe atherosclerosis Formation of atheromatous plaque, which obstructs the blood flow.

  4. When there is too much cholesterol in the diet, it either ends up being stored as fat or continues circulating in the blood in the form of LDL (low-density lipoprotein) or what is more commonly known as “bad cholesterol”. • LDL is one of the components of the atheromatous plaque. “BAD” OR LDL CHOLESTEROL

  5. ACCORDING TO THE WORLD HEALTH ORGANISATION (WHO) • Although heart attacks and strokes are major killers in all parts of the world, 80% of premature deaths from these causes could be avoided by controlling the main risk factors • Every year, an estimated 17 million people globally die of cardiovascular diseases (CVD), particularly heart attacks and strokes. • CVDs occur almost equally in men and women. • Are the leading cause of death in developing countries, as well as developed ones.

  6. LDL Macrophage Oxidised LDL Oxidation of LDL THE MACROPHAGES, A “CLEAN-UP SYSTEM” • The macrophages act as a “cleaning” system to stop the cholesterol from being deposited on the walls. • This clean-up system is efficient if the increased cholesterol is not too excessive.

  7. Proliferation of endothelial cells Immune system activation Foam cell LDL Macrophage Oxidised LDL Oxidation of LDL HOW DOES AN ATHEROMATOUS PLAQUE FORM? If the quantities of cholesterol are very excessive: The macrophages continue to pick up the LDL But, once they have engulfed large amounts, they turn into what is known as “foam” cells. These induce inflammation and the proliferation of cells in the artery wall Formation of the atheromatous plaque

  8. RESEARCH INTO ATHEROSCLEROSIS One of the objectives is:To understand how macrophages are involved in the regulation of cholesterol levels and what role they play in the development of atherosclerosis. ?

  9. STUDY OF THE LDL RECEPTOR AND MYLIP We know that the macrophages recognise oxidised LDL thanks to some receptors. We know that a protein in the macrophages called MYLIP breaks down the LDL receptor. Macrophage LDL receptor Oxidised LDL If the macrophages produce MYLIP in large quantities, they ingest less cholesterol. Oxidation of LDL

  10. MYLIP, A POTENTIAL THERAPEUTIC TARGET Scientists are studying the MYLIP protein in depth because they think that it could be regulated by a drug and thus stop the macrophages from ingesting less cholesterol. Macrophage Therapeutic target Oxidised LDL Oxidation of LDL

  11. HOW CAN WE STUDY THE PROTEINS? • Large quantities of a protein are needed to study its function. • One of the tools in molecular biology which allows us to study the proteins is genetic engineering. • Genetic engineering is the technology of the manipulation and transfer of DNA from one organism to another.

  12. FIRST WE CLONE THE GENE OF THE PROTEIN OF INTEREST: MYLIP We insert it into a fragment of circular DNA called plasmid Human cell ADN MYLIP gene

  13. WE PERFORM A BACTERIAL TRANSFORMATION WITH THE GENE OF THE PROTEIN OF INTEREST: MYLIP 1.Bacterial transformation 2.Selection of transformed bacteria 3.Bacterial growth

  14. MYLIP THE GENE OF THE MYLIP PROTEIN IS THEN INTRODUCED INTO CELLS IN ORDER TO PRODUCE MORE Protein 3. Bacterial growth 4.We isolate the DNA 5.Production of the protein(introducing the DNA into eukaryotic cells)

  15. HOW DO WE DO A BACTERIAL TRANSFORMATION? ? With drastic changes in temperature and by adding cations, we help the entry of the DNA in the form of plasmid into the cells.

  16. HOW CAN WE MAKE SURE THAT TRANSFORMED BACTERIA WILL GROW? ? The plasmid has a gene that is resistant to an antibiotic.

  17. HOW CAN WE MAKE SURE THAT TRANSFORMED BACTERIA WILL GROW?

  18. HOW DO WE ISOLATE THE PLASMID DNA FROM THE BACTERIAL CULTURE? ? Doing a Mini-prep: Using various solvents and centrifugation cycles, we gradually discard the different components

  19. WITH THE DNA ISOLATED, WE CAN GET ON WITH THE RESEARCH! 5.Production of the protein: (introducing the DNA into eukaryotic cells). • 6.Once the scientists obtain the protein, they can then: • Study its role in cholesterol regulation • Search for new drugs for atherosclerosis.

  20. AUTHOR FUNDED BY: PROJECT PARTNERS: Put on a lab coat!!You too can be a researcher!! Researchers who have contributed to the writing of this presentation: Theresa León, Jonathan Matalonga, Barcelona University This work is under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported licence. To see a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

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