The role of micro-RNA (miRNA) in cardiovascular disease

  1. You lead a research project investigating the role of micro-RNA (miRNA) in cardiovascular disease. You have clinical collaborators within the Vascular Surgery Department at the local University Hospital who are providing carotid endarterectomy specimens, bloods, urines, as well as patient information (Age, gender, smoking status, BMI status, medication history). Your research team in the laboratory will be responsible for the extraction of miRNA and mRNA from patient samples, next generation sequencing analysis of miRNA and mRNA profiles within these samples, and subsequent completion of a data report for clinical colleagues. (a) Micro-RNA (miRNA) are a class of non-coding RNA. In your own words please explain how microRNA are synthesized in cells and how miRNA regulate gene expression when interacting with target mRNAs. Comment on the importance of nucleic acid complementarity here. (10 marks) You have extracted micro-RNA and mRNA from patient biopsy (Carotid endarterectomy) samples using a column extraction method and assessed the quantity and quality of the samples. You have chosen to perform Illumina next-generation sequencing on the micro-RNA and mRNA samples and following data analysis you identified a cluster of 17 miRNAs that are significantly correlated with disease status in these patients. You are now interested in exploring miRNA-mRNA interactions networks as well as the biomarker potential of these miRNA. (b) Describe the main advantages of next-generation sequencing technology over traditional microarray approaches when measuring gene/miRNA expression. (10 marks) (c) Outline how you would propose to investigate the biomarker potential of these miRNA using the clinical resources available to you in this specific project. (15 marks) Advances in sequencing technologies have led to the development of several highly innovative approaches. Regarding RNA sequencing, different approaches are now available that allow for the investigation of gene expression in bulk tissues, specific cell subtypes as well as expression localisation within tissue sections. (d) In your own words, define each of these approaches, including any major advantages or differences between each technology. Comment on which technologies are suitable for RNA sequencing analysis in patient blood samples. (15 marks) Your project leader on this project has asked you to present your interpretation of a specific research paper focused on micro-RNA and their role in kidney disease. You are tasked with reading this paper (https://www.kidney-international.org/article/S0085-2538(20)31553-2/fulltext) and providing a brief (1 page) summary of the study hypothesis, methods, results and conclusions. (e) In your own words, provide a 1-page summary report of the paper – including an overview of the study hypothesis, methods, results, conclusions. Also provide an opinion on the key strengths and limitations of this study that you feel the team need to consider prior to starting a miRNA-based project. (50 marks)