Exosome miRNA sequencing for biomarker profiling using the Ion S5 System and Ion Total RNA-seq kit

Presented By:
Dr. Lesley Cheng - Bachelor of Medical Science, Hons, PhD

Speaker Biography:
Dr. Lesley Cheng is a Postdoctoral researcher. She was awarded a Bachelor of Medical Science with Honours and a PhD from Monash University, Australia, with a PhD Scholarship from Neurosciences Victoria. After her PhD, she undertook a position in Research and Development at the American Hospital Dubai, developing diagnostic tests to screen neonatal babies for genetic disorders. She returned to Australia to the Department of Biochemistry and Molecular Biology, University of Melbourne as a Postdoctoral researcher. In 2015, she moved to the La Trobe Institute for Molecular Science, La Trobe University. Her research expertise combines basic and clinical science through the use of novel cell and molecular biology approaches to identify biomarker candidates for diagnostic testing. Currently, her major focus is to develop a minimally invasive blood test for the early detection and monitoring of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and Prion disease. Her research involves detecting disease signatures of RNA molecules called microRNA which can be collected from small biological vesicles called 'exosomes' that travel in the bloodstream. She is also investigating the functional role of exosomes in neurodegenerative diseases as they have been found to cause the spread of disease in the body.

Webinar:
Exosome miRNA sequencing for biomarker profiling using the Ion S5 System and Ion Total RNA-seq kit

Webinar Abstract:
Learning Objectives
-How to maximise RNA yield and extraction from exosomes and biological fluids
-How to construct small RNA libraries from low RNA yields
-How to analyze your small RNA reads to select a list of biomarker candidates for validation

Alzheimer’s disease (AD) affects more than 55 million people worldwide and is expected to double every 20 years in the absence of disease-modifying drugs. Future therapeutic strategies aimed at limiting neurodegeneration require methods to diagnose the disease in preclinical patients. Several blood-based tests have been explored to detect AD however, evidence is required to determine whether blood sampling is an appropriate specimen to diagnose brain diseases. Previously we isolated serum exosomes from AD patients which displayed an abnormal composition of 16 specific microRNA (miRNA) biomarkers compared to controls. To provide evidence that our serum exosomal miRNA biomarkers are suitable for the detection of a brain condition, we also profiled exosomes isolated from post-mortem human AD and control brain tissues. Exosomes were extensively characterised to meet the minimal experiments requirements set out by The International Society for Extracellular Vesicles to be defined as exosomes and small RNA profiling was performed by next-generation sequencing.
The Ion Torrent Ion S5 and Ion Chef system was used to perform small RNA deep sequencing of the brain and serum exosome samples. Brain derived exosomes (BDEs) were found to contain a unique profile of small RNA, including miRNA, compared to whole tissue samples. Furthermore, all 16 serum biomarkers, identified in our previous study, were detected in BDEs including a panel of BDE specific miRNA that target genes involved in AD pathology. This work has identified a highly specific panel of miRNA that is both present in the brain and blood of AD patient samples. In the future, the miRNA candidates could be used to develop a blood-based diagnostic test highly relevant to a brain disease, equivalent to non-invasive brain biopsy. Furthermore, this biomarker discovery pipeline could be used to identify exosomal miRNA biomarkers for other diseases and conditions.

Sponsored By:
Thermo Fisher Scientific Ion Torrent

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