Urine as a liquid biopsy for cancer detection

Abstract: This dissertation focused on exploiting the full potential of urine as liquid biopsy for cancer research. First this dissertations showed that urine is used for many cancer types and can be used as a multi-omics sample type. Next, we demonstrated that there is a gap in reporting of pre-analytical parameters since most publications (73.8%) reported no information on a pre-analytical parameter of their samples, and almost 25.0% did not report any pre-analytical parameter. To further elaborate on the clinical applicability of urine as liquid biopsy, there are some challenges to overcome, including but not limited to: degradation of interesting urinary analytes, collection of the correct fraction of urine using appropriate and user-friendly devices, and the low concentration of urinary analytes of interest. Therefore, we investigated the performance of a recently developed preservative, UAS, for the preservation of urinary analytes. Results clearly demonstrated that no microbial growth was observed in urine samples stored in UAS and cfDNA and cellular integrity were maintained for up to 14 days at RT. We also demonstrated that 4 cfDNA isolation methods performed similarly with respect to the cfDNA fragmentation profiles. And that three different Colli-Pee UAS device variants (FV-5010, collecting approx. 10 mL; FV-5020, collecting approx. 20 mL; FV-5040, collecting approx. 40 mL) performed similarly, indicating their potential in cancer research. Furthermore, it is important that devices developed for immediate preservation and collection of the correct fractions are easy, hygienic and user-friendly. Here, we showed that the vast majority was very positive about the urine collecting process with Colli-Pee UAS devices. Finally, we showed the design process of a potential new Colli-Pee variant for the enrichment of urinary analytes of interest, the URICH device, as case-example to demonstrate the inclusion of users and usability testing as early as possible in the design and development pathway. In conclusion, this dissertation identified the gap in reporting and standardization of pre-analytical workflows for urine as liquid biopsy and provided some initial solutions to this gap. Furthermore, the potential of Colli-Pee UAS, as a device for the volumetric collection of first-void urine and immediate preservation of urinary analytes for oncology applications, was clearly demonstrated as well as its user-friendliness and further innovative potential