Final Report Summary - CHEMIRNA (CHEMiRNA: Chemical-based Platforms for micro-RNA Detection. Towards Novel OncomiR Assays)
During the past decade microRNAs (a novel species of non-coding RNAs with gene regulatory function) have been linked virtually to all known physiological and pathological processes, including cancer. In the same way as certain key protein-coding genes, miRNAs can be deregulated in cancer. Understanding the role of miRNA in biological processes is thus key to investigate cancer phenotypes, diagnose and classified cancer types in a much stratified manner and also to develop new therapies which either target or mimics miRNAs. Components of the miRNA machinery and miRNAs themselves are involved in many cellular processes that are altered in cancer, such as differentiation, proliferation and apoptosis and also other pathologies such as cardio pathologies and liver injuries. In vitro studies have showed evidence implicating miRNAs such as miR-21 and miR-10 in cancer and miR-122 in drug induced liver injuries (DILI). To analyse miRNA scientist adapted tools which are used for the majority of DNA genotyping methods at present relying upon DNA polymerase chain reaction (PCR) for the exponential amplification of DNA target molecules. Therefore, most of the efforts to do miRNA analysis have been based on PCR technologies to create cDNA from miRNA. However this approach is particularly challenging to miRNA short nature (22-24 nucleotides). This means that standard protocols for miRNA analysis are long, still inefficient (to date a few reports comparing different platforms cannot find very good reproducibility between them) and lacking being truly quantitative. This means that current miRNA products do not offer any further innovation if compared with PCR based products. This lack of innovation will be a blocking point for them to deliver robust and cost-efficient products for diagnoses and prognosis of cancer based on miRNA analysis. However, this innovation is essential to facilitate Stratified Medicine Programmes which goal is to help doctors to decide the most suitable treatment for patients based on genetic tests, being miRNA profiling tests clearly a need. The drawbacks of the current technologies provide a real opportunity for technologies such as the presented in this proposal which are capable of developing and introducing innovative solutions. In recent years, we have created a game-changing technology for improved nucleic acid detection. This strategy applies dynamic chemistry to peptide nucleic acid (PNA) for the development of an entirely novel method for nucleic acid detection. This technology is able to identify nucleic acid sequences with very high specificity due to his unique chemical-based approach for nucleic acid “reading” which does not allow “mis-reading” and will provide solutions to obtain the needed sensitivity. In this research project we have developed new reagent sets to allow fluorescence based detection using a commercial bead-based platform. A lab-on-a chip formats has also been developed in order to reduce the cost of assays and increase the number of applications. This proposal has lead to the design, set up and validation of assays for profiling miRNAs in a direct manner which will be used in cancer diagnosis, prognosis and DILI. A robust and cost-effective miRNA profiling technology based on chemical-based reagents has been successfully developed.