Ingenito, Francesco (2023) Development of innovative aptamers for cancer exosomes detection. [Tesi di dottorato]

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In recent years, the global incidence of cancer has witnessed a significant increase due to an increased life expectancy and the prevalence of unhealthy lifestyle habits . Unfortunately, cancer is frequently diagnosed when tumors have reached advanced stages, resulting in a reduced effectiveness of therapeutic interventions . Moreover, t raditional diagnostic techniques such as tissue biopsies present significant limitations due to their invasiveness and non repeatability. In response to th ese issues , liquid biopsy emerged as a promising opportunity for cancer diagnosis, particularly through the analysis of extracellular vesicles known as exosomes. Exosomes are small vesicles released by cells into biological fluids and contain a variety of molecules that could potentially provide information about patients’ pathological status. This study focuses on the development and characterization of aptamers able to identify exosomes originating from breast cancer and pancreatic ductal adenocarcinoma cells. These aptamers could provide a platform for the development of early cancer diagnosis methodologies based on tumoral exosomes detection . Here, we described the properties of ex 50.T aptamer, able to selectively recognize exosomes derived from breast cancer cells. Through a combination of in vitro experiments and computational analyses, such as proteomic analysis and in vitro modeling studies , we demonstrated that the aptamer's specificity for breast cancer derived exosomes arises from its interaction with a bone morphogenetic protein antagonist recently confirmed to be located on the exosome surface. Additionally, we introduced a novel methodology based on the use of a robotic system for the automated generation of aptamers able to target exosomes from pancreatic ductal adenocarcinoma cells. This approach can expedite the development of aptamers for the use in various applications , including the personalized medicine. Altogether, our findings highlight the considerable potential of aptamers in advancing the field of cancer diagnostics and liquid biopsy technology.

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