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de Figueiredo, D.

Publications and source records attributed to de Figueiredo, D..

2 recordsLinked to original sources

Tear fluid as noninvasive liquid biopsy reveals proteins associated with malignant transformation of oral lesions

Oral leukoplakias (OLs) are premalignant lesions that can progress into oral squamous cell carcinoma (OSCC). This study hypothesized that tear fluid, as a noninvasive biofluid, reflects proteomic alterations associated with malignant transformation. The tear proteome of 44 individuals, including healthy controls, OL/PVL (proliferative verrucous leukoplakia), and OSCC patients, was deeply profiled, revealing 828 protein groups clustered according to histopathological alterations. N-glycoproteome analysis identified immune-related proteins, while public RNA-seq integration indicated immune imbalance marked by increased B-cell and decreased macrophage signatures during disease progression. Several immune-associated proteins and epithelial markers, including desmoplakin, KRT14, and DSC1, emerged as potential indicators of malignant transformation. These findings demonstrate that tear fluid reflects oral carcinogenic processes, thereby serving as a noninvasive liquid biopsy for early detection and clinical monitoring.

biochemistry↗

Single-cell proteomics workflow for characterizing heterogeneous cell populations in saliva and tear fluid

Single-cell proteomics (SCP) has advanced considerably but still is largely limited to homogeneous populations and distant from clinical applicability. We present an SCP workflow for assessing the cellular heterogeneity in saliva and tear fluid. Initially, benchmarks were established using a standard HeLa digestion curve, resulting in more than 5,463 protein groups (PGs) at 50 pg. For single HeLa cells, the workflow was improved to minimize contamination and increase quantitative performance, reaching a maximum of 3,785 PGs per single cell. Following, SCP was benchmarked across heterogenous populations of saliva and tear fluid, collected from 10 healthy individuals. By improving cell isolation, contamination control, and DIA-based search and quantitation, single cells from saliva (n=110) and tear fluid (n=149), with average diameters of 8 and 11 {micro}m, respectively, yielded a maximum of 700 PGs per single cell. Downstream analysis indicated overrepresented protein functions, distinct cluster markers and twenty-three validated therapeutic targets identified from single-cell data. Taken together, this study demonstrates the robustness of our SCP workflow applied to biofluids, driving the discovery of biomarkers and therapeutic targets in complex microenvironments.

molecular biology↗