An Investigation to Study Morphology, Nanomechanical Attributes, Transcriptomics, and Proteomics of Tumor Associated Macrophages Derived Extracellular Vesicles in the Tumor Microenvironment.

INTRODUCTION: This study investigates the molecular and biophysical changes in Tumor-Associated Macrophage (TAM)-derived exosomes (TAME) compared to exosomes from non-tumor microenvironment (TE) macrophages (MO-E). TAMs are immune cells infiltrating tumors that promote cancer progression. MATERIALS AND METHODS: Characterizing the cargo of TAME via Next Generation RNA Sequencing protein microarray analysis and studying their morphological and nanomechanical properties using Atomic Force Microscopy (AFM). RESULTS: Key Findings regarding Morphology showed TAME are significantly smaller (~50.6 nm) than MO-E (~64.8 nm).TAME exhibit higher Young's modulus (~15.5 MPa) indicating increased stiffness. Surface roughness ot TAME Is slightly higher (~3.64 nm) than MO-E (~3.51 nm). RNA sequencing revealed differential expression of genes involved in drug metabolism, cell cycle regulation, survival pathways, and immune modulation. Pathways such as NF-KB, PI3K/Akt, IL-13 signaling, and metabolic reprogramming are influenced by TAME cargo.Proteomics studies showed several Top upregulated proteins include ORP150 (promotes VEGF secretion), CSRP1, C1qB, DNER, PEPD, with roles in tumor growth and metastasis.Downregulated proteins include tumor suppressors like GOLPH2 and VNN1.These protein profiles suggest TAME carry oncoproteins that support tumor proliferation, invasion, and immune evasion. CONCLUSION: This study demonstrates that TAM-derived exosomes are morphologically distinct and carry molecular cargo that modulates key cancer-related pathways, highlighting their potential as biomarkers and therapeutic targets. The significance lies in understanding TAME's role in cancer progression, which could lead to novel diagnostic biomarkers and therapeutic targets, especially considering current limitations in cancer diagnosis accuracy.