Research Projects

Project summary: Pancreatic neuroendocrine tumors (PanNETs) represent a significant clinical challenge, particularly in India, where the pancreas is the most common primary site of gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) among all. The rise in incidence and prevalence of PanNETs shows that there is an urgent need for improved diagnostic tools. Current diagnostic methods, including biomarkers like chromogranin-A and 5-HIAA, are not that specific and sensitive, while imaging techniques are expensive and not widely accessible. Exosomes (40–150 nm) secreted by all cell types, hold promise as a novel diagnostic tool. Tumor-derived exosomes carry proteomic and genomic signatures that reflect their cell of origin, make them reliable, non-invasive biomarker for early cancer detection. Despite their known role in other endocrine disorders and cancers, the potential of exosomes in PanNETs remains largely unexplored. This research aims to address this gap by identifying specific protein and miRNA signatures within plasma-derived exosomes from PanNET patients. Objectives: 1. Isolation, characterization, and validation of exosomes in blood and tissues of PanNETs. 2. To identify and validate exosome-derived protein and miRNA signatures unique to PanNETs. 3. Diagnostic Potential Assessment of the identified biomarkers in PanNET pathophysiology. Main Experiments to be Carried Out 1. Exosome Isolation and Characterization: exosomes will be isolated from PanNET patients and healthy controls plasma and tissues. Characterization and validation through nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), and western blotting for exosomal markers. 2. Proteomic Profiling: Mass spectrometry-based proteomic analysis will be performed to identify differentially expressed proteins in PanNET-derived exosomes compared to controls. Validation of candidate proteins will be conducted using ELISA or western blot. 3. miRNA Profiling: RNA will be extracted from exosomes for small RNA sequencing. Differentially expressed miRNAs will be validated using qRT-PCR. 4. Diagnostic Potential Assessment: ROC curve analysis will be used to assess the sensitivity and specificity of identified biomarkers in distinguishing PanNET patients from controls. Hypothesis of the study: We hypothesize that tissue and plasma-derived exosomes from PanNET patients carry unique proteomic and miRNA profiles distinct from healthy controls, which can be utilized for early detection and diagnosis of PanNETs. These signatures may also offer insights into the biological mechanisms underlying tumor progression and metastasis. Significance to the field of research: If successful, this research will establish a novel, non-invasive diagnostic tool for early detection of PanNETs. Additionally, the study will enhance our understanding of the molecular mechanisms underlying PanNET progression.