| Autor: |
Escalona-Noguero C; Fundación IMDEA Nanociencia, Campus Universitario de Cantoblanco, Madrid 28049, Spain., Alarcón-Iniesta H; Fundación IMDEA Nanociencia, Campus Universitario de Cantoblanco, Madrid 28049, Spain., López-Valls M; Fundación IMDEA Nanociencia, Campus Universitario de Cantoblanco, Madrid 28049, Spain., Del Carpio LP; Medical Oncology Department, Institut Català d'Oncologia─ICO, L'Hospitalet de Llobregat, IDIBELL, Barcelona 08908, Spain.; Cancer ImmunoTherapy (CIT) Group-iPROCURE, Bellvitge Biomedical Research Institute IDIBELL-OncoBell, L'Hospitalet de Llobregat, Barcelona 08908, Spain., Piulats JM; Medical Oncology Department, Institut Català d'Oncologia─ICO, L'Hospitalet de Llobregat, IDIBELL, Barcelona 08908, Spain.; Cancer ImmunoTherapy (CIT) Group-iPROCURE, Bellvitge Biomedical Research Institute IDIBELL-OncoBell, L'Hospitalet de Llobregat, Barcelona 08908, Spain., Somoza Á; Fundación IMDEA Nanociencia, Campus Universitario de Cantoblanco, Madrid 28049, Spain.; Unidad Asociada de Nanobiotecnología (CNB-CSIC e IMDEA Nanociencia), Madrid 28049, Spain., Sot B; Fundación IMDEA Nanociencia, Campus Universitario de Cantoblanco, Madrid 28049, Spain.; Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT), Unidad de Innovación Biomédica, Complutense 40, Madrid 28040, Spain.; Advanced Therapies Unit, Instituto de Investigación Sanitaria Fundación Jiménez Díaz (IIS-FJ UAM), Madrid 28040, Spain. |
| Abstrakt: |
Uveal melanoma (UM) is a rare ocular tumor characterized by high metastasis risk and poor prognosis. The in-depth characterization of UM's molecular profile is critical for better disease classification and prognosis. Furthermore, the development of detection tools to monitor UM evolution upon treatment is of great interest for designing optimal therapeutic strategies. However, commonly used techniques, such as ddPCR or NGS, are costly, and they involve sophisticated equipment and complex experimental design. The development of alternative sensing methods that are fast, simple, and inexpensive would be of great benefit to improve UM's diagnosis and management, especially when combined with liquid biopsy. Samples from liquid biopsy can be obtained with minimal invasiveness, and the detection of circulating tumor DNA (ctDNA) in UM patients' plasma has proven useful for the diagnosis of metastasis, prognosis prediction, and disease monitoring. In this context, CRISPR/Cas12a-derived molecular sensors, thanks to their high specificity and sensitivity and their potential for point of care diagnosis, are particularly interesting. Here, we developed a CRISPR/Cas12a-based approach for the specific detection of the UM-related mutation GNAQ Q209P that relies on the design of highly specific crRNAs. Coupled with allele-specific PCR, it constitutes a sensitive platform for liquid biopsy detection, capable of sensing GNAQ Q209P in plasma samples with a low ctDNA concentration and fractional abundance. Finally, our method was validated using plasma samples from metastatic UM patients. |
