LuCamiR aims to develop and validate a minimally invasive microRNA-based approach for the early detection of non-small cell lung cancer (NSCLC). The project focuses on circulating miR-223 as a serum biomarker, its biological and cellular origin, and the evaluation of miR-21 and miR-205 to improve diagnostic accuracy and support histological subtyping.
The study was designed to enroll 300 subjects, including 150 consecutively recruited patients with suspected early-stage NSCLC and 150 controls. Controls underwent low-dose computed tomography to confirm the absence of lung cancer. Following histological and clinical assessment, patients with non-NSCLC diagnoses, advanced disease or pulmonary metastases were excluded. Serum miR-223 was quantified by droplet digital PCR (ddPCR), but its levels did not differ significantly between patients and controls. Thus, unlike our previous study, miR-223 was not confirmed as a standalone diagnostic biomarker in this cohort. Cross-cohort and re-extraction controls are being used to investigate whether pre-analytical, analytical or biological factors account for this discrepancy.
Circulating miR-21 also showed no significant difference between patients and controls. Conversely, in the samples available for this analysis, miR-205 levels were significantly higher in squamous cell carcinoma than in adenocarcinoma, supporting its potential role in NSCLC subtyping.
In lung cancer cell lines, miR-223 agomir transfection significantly reduced CLDN1 transcript and increased PUMA transcript. Selected direct targets are also being assessed at protein level.
Cell-of-origin studies showed detectable miR-223 signal in transfected NCI-H1299 cells, supporting the suitability of the optimized in situ hybridization protocol for FFPE tissues. Moreover, microdissected FFPE samples generally showed higher miR-223 levels in the peritumoral compartment than in the tumor fraction, suggesting a substantial contribution from inflammatory and stromal components of the tumor microenvironment.
The study was designed to enroll 300 subjects, including 150 consecutively recruited patients with suspected early-stage NSCLC and 150 controls. Controls underwent low-dose computed tomography to confirm the absence of lung cancer. Following histological and clinical assessment, patients with non-NSCLC diagnoses, advanced disease or pulmonary metastases were excluded. Serum miR-223 was quantified by droplet digital PCR (ddPCR), but its levels did not differ significantly between patients and controls. Thus, unlike our previous study, miR-223 was not confirmed as a standalone diagnostic biomarker in this cohort. Cross-cohort and re-extraction controls are being used to investigate whether pre-analytical, analytical or biological factors account for this discrepancy.
Circulating miR-21 also showed no significant difference between patients and controls. Conversely, in the samples available for this analysis, miR-205 levels were significantly higher in squamous cell carcinoma than in adenocarcinoma, supporting its potential role in NSCLC subtyping.
In lung cancer cell lines, miR-223 agomir transfection significantly reduced CLDN1 transcript and increased PUMA transcript. Selected direct targets are also being assessed at protein level.
Cell-of-origin studies showed detectable miR-223 signal in transfected NCI-H1299 cells, supporting the suitability of the optimized in situ hybridization protocol for FFPE tissues. Moreover, microdissected FFPE samples generally showed higher miR-223 levels in the peritumoral compartment than in the tumor fraction, suggesting a substantial contribution from inflammatory and stromal components of the tumor microenvironment.