Research Projects

Background knowledge gap: Lung cancer mortality is driven by metastasis and therapy resistance, fueled by cancer stem cells (CSCs) supported within a remodeled tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) are key architects of this pro-metastatic niche, orchestrating extracellular matrix (ECM) remodeling. Exosomes, particularly those carrying long non-coding RNAs (lncRNAs), are crucial mediators of tumor-stroma crosstalk. Oncogenic exosomal lncRNAs (e.g., H19, MALAT1, PVT1) are implicated in CAF activation, yet the precise mechanisms by which lung cancer-derived exosomal lncRNAs drive CAF-mediated ECM remodeling to sustain CSC plasticity and metastasis remain poorly defined, representing a critical knowledge gap. Hypothesis: Our central hypothesis is that exosomal lncRNAs secreted by lung cancer cells reprogram CAFs, inducing ECM remodeling that creates a permissive niche for CSC maintenance and metastatic dissemination. Furthermore, we propose that targeted disruption of this exosomal lncRNA-CAF-CSC axis using advanced nanotherapeutics can overcome therapy resistance. Overall aims: Aim-1) Elucidate the role of specific exosomal lncRNAs in CAF activation and ECM remodeling – Exosomes will be isolated from metastatic versus non-metastatic human lung cancer cell lines and patient-derived organoids. LncRNAs enriched in metastatic exosomes (with a focus on H19, MALAT1, PVT1) will be identified using RNA-seq and qRT-PCR. Functional validation will be performed by lentiviral knockdown/overexpression in donor cancer cells, and the impact of derived exosomes on recipient CAF activation markers (α-SMA, FAP), ECM gene/protein expression (Collagen I, Fibronectin, LOXL2, MMPs), contractility, and ECM stiffness will be assessed using 3D CAF-collagen gels and atomic force microscopy. Aim-2) Determine the impact of exosomal lncRNA-primed CAFs and remodeled ECM on CSC enrichment and metastasis – Lung CSCs will be co-cultured with CAFs pre-treated with metastatic cancer-derived exosomes or exosomes from lncRNA-knockdown cells. CSC stemness (sphere formation, ALDH activity, stemness gene expression - SOX2, OCT4, NANOG), chemoresistance, and invasive potential will be assessed. The direct influence of the CSC phenotype will be evaluated using decellularized ECM from these CAFs. The pro-metastatic capacity of this axis will be validated in orthotopic and tail vein metastasis mouse models using luciferase-tagged cancer cells, with CAF and CSC dynamics monitored by IHC/flow cytometry. Aim-3) Develop and evaluate targeted nanotherapy co-loaded with anti-lncRNA and chemo-drug to disrupt the CAF-CSC-ECM axis – Folic acid (FA) or hyaluronic acid (HA) decorated liposomal nanoparticles will be engineered for dual targeting (CAFs via folate receptor, CSCs via CD44). Nanoparticles will be co-loaded with anti-lncRNA(s) (identified in Aim 1) and a conventional chemotherapeutic (e.g., Cisplatin or Doxorubicin). Uptake, lncRNA knockdown efficiency, cytotoxicity in CAFs/CSCs, and impact on CAF activation/ECM deposition will be assessed in vitro. In vivo efficacy will be evaluated in metastatic lung cancer models measuring primary tumor growth, metastatic burden (via bioluminescence/IVIS, histology), CSC frequency, CAF activity, ECM composition, and overall survival. Restoration of chemosensitivity will be evaluated as a key endpoint. Impact: This project integrates molecular oncology, stromal biology, and nanomedicine to unravel a key mechanism of lung cancer lethality and translate it into a novel, rationally designed therapeutic strategy targeting the TME ecosystem, with high potential for clinical impact.