Dados do Resumo

Título

Detection of plasma CD26 by liquid biopsy for evaluation of disease progression in human colorectal cancer.

Introdução

Colorectal cancer (CRC) is a neoplasm with a high incidence and mortality rate among men and women according to recent estimates. Various biomarkers have been studied in the context of disease progression and prognosis, such as CD26, which has been shown to be a promising tumor marker. Thus, the early detection of CD26 in the peripheral blood of individuals diagnosed with CRC may indicate possible micrometastases and recurrences, as well as helping the clinical team to better manage the patient.

Objetivo

To detect serum CD26 through liquid biopsy using digital droplet polymerase chain reaction (ddPCR) to evaluate tumor progression and prognosis in patients diagnosed with colorectal cancer.

Métodos

Forty patients with a diagnosis of CRC, assisted at the Hospital da Baleia, in Belo Horizonte, MG, were included in the study (CAAE 02177612.0.3001.5091). Twenty milliliters (mL) of peripheral blood were collected intravenously during a surgical procedure to excise the tumor. Blood was centrifuged to separate the plasma and then subjected to cell-free DNA (cfDNA) isolation using magnetic beads, following the manufacturer's guidelines. At the end of the process, cfDNA was eluted and stored at -20 degrees for application in an ultrasensitive ddPCR absolute copy number detection assay. Oligonucleotides were designed according to the manufacturer’s instructions. In addition, the patients' clinical data was accessed through medical records and previous examinations. An additional blood sample will be taken no later than three years after the surgical procedure, and serum quantification of carcinoembryonic antigen (CEA) will be used to compare disease progression. Quantification of the data, as well as patient and tumor variables, will be correlated to define the role of serum CD26 in CRC progression.

Resultados

The yield and purity of the isolated cfDNAs were assessed using a spectrophotometer to obtain satisfactory values for inclusion in the ddPCR assay. In addition, the process of standardizing the concentrations of primers and nucleic acids began, with six samples being subjected to digital PCR. Drops were generated above the minimum amount, including the no-template control (NTC), and after setting the threshold, it was observed that positive copies of the gene of interest were found in the reaction. Next, the DNA concentrations were decreased to better establish the separation between the positive and negative populations of each sample.as

Conclusões

Based on the objectives established and the follow-up of the trials, this technique shows promise in the plasma detection of markers of disease progression, with the prospect of encompassing the liquid biopsy technique for the validation of disease and validation of CD26 as a prognostic marker. The inclusion of this target will guarantee an improvement in the patient's quality of life and progression-free survival, which will help lead to an assertive treatment, especially in the management of critically ill patients.

Financiador do resumo

CAPES e FAPEMIG

Palavras Chave

Colorectal cancer; biomarker; Liquid Biopsy