Palma, Francesco (2023) Selection of RNA-Aptamers for detection of Rectal cancer-derived exosomes. [Tesi di dottorato]

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Abstract

Rectal adenocarcinoma ranks as one of the leading causes of global cancer-related mortality. The current diagnostic gold standard, colonoscopy, leaves much to be desired due to its invasiveness and sensitivity in detecting even minute residual disease. This deficiency results in an imperfect diagnostic process, characterized by invasiveness, non-reusability, and a lack of comprehensive insights into metastatic risk, disease progression, and treatment efficacy. The promising technology of liquid biopsies offers a non-invasive approach for disease monitoring, offering real-time insights into a patient's disease status. In our study, we employed RNA-based aptamers, known for their high specificity, to target exosomes derived from rectal cancer. We've performed a distinctive Systematic Evolution of Ligands by Exponential Enrichment (SELEX) strategy, this approach involved a cell-based SELEX, using different stages of rectal cancer cell lines (SW1463 - Stage III and HRA19 - Stage II) to select a panel of target-specific aptamers. This choice aimed to mimic the practices employed in the "watch and wait" approach, with the goal of selecting an aptamer capable of detecting residual disease. Real-time PCR binding studies have illuminated the exceptional affinity and selectivity of the selected aptamers for stage III rectal cancer when compared to stage II. Our data suggest that these aptamers can pinpoint specific blood-based biomarkers associated with rectal cancer, ushering in an innovative non-invasive tool for early disease detection and offering valuable prognostic insights. Furthermore, they hold the potential to guide tailored therapies. With the most promising aptamer candidates from our panel, we are poised to develop a novel ELONA aptamer-based sandwich assay, set to revolutionize rectal cancer diagnosis and monitoring.

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