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Circulating tumour cell detection using isolation by size (ISET) and RT-qPCR in dogs with Melanoma

Grant number: 17/22374-0
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): May 01, 2018
Effective date (End): October 31, 2020
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Pathology
Principal Investigator:Cristina de Oliveira Massoco Salles Gomes
Grantee:Victor Nowosh
Home Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil


Canine melanoma is a highly metastatic malignant neoplasm. One approach to understanding metastasis mechanisms is liquid biopsy, wich consists on detecting Circulating Tumor Cells (CTCs) and other tumoral products in circulation, enabling assessment of tumor biology, prognosis and treatment response non-invasively and in real time. Isolation by size of epitelial tumor cells (ISET) method allows intact CTC detection and counting. Real-Time reverse-transcriptase PCR (RT-qPCR) method targets tumor-associated messenger RNA. Besides its extensive use in human patients, intact CTC detection was not used in dog patients, and RT-qPCR studies were already conducted with canine melanoma, but not to detect CTCs. The main goal of this study is develop and validate ISET and RT-qPCR protocols for detecting CTCs in dogs with oral and digit melanoma. In vitro study for essay validation and sensivity assessment will be conducted with blood samples from healthy dogs spiked with known quantities of tumor cells from a canine oral melanoma cell culture. Multiple samples of 30 canine patients with oral or digit melanoma will be assessed. This project main hypothesis is that liquid biopsy results will present prognostic correlation with overall survival of Melanoma canine patients, similarly as reported in human patients. (AU)