Executive Summary : | Hepatocellular carcinoma (HCC) is the third most cause of cancer-related deaths worldwide and chemotherapy is the only treatment modality. Due to poor response with conventional chemotherapy, there is a dire need to develop natural anti-HCC drugs with therapeutic potential and less systemic toxicity. The ‘wonder tree’ Neem (Azadirachta Indica), well-known for its diverse medicinal uses, is one of the richest known sources of secondary metabolites, especially tetranortriterpenoids (limonoids). Gedunin, a limonoid, extracted from the neem seed oil, has anti-inflammatory, antipyretic, analgesic and antimalarial properties and recently, its anti-cancerous roles have been explored. However, anti-proliferative effects of Gedunin in liver cancer is yet to be explored. Naked Gedunin has limited bioavailability and pharmacokinetic profile due to poor solubility, making it less convenient for tumor targeting. Zinc oxide nanoparticle (ZnO NP), an FDA-approved pharmaceutical nanocarrier for delivery of drugs, is widely used due to their biocompatibility, stability and safety. Since naked Gedunin is poorly soluble in aqueous extracellular environment with limited bioavailability and pharmacokinetic profile, we hypothesize that nano-formulations of Gedunin embedded with ZnO may enhance its anti-proliferative effects. A major innovation of this proposal will be development of first of its kind, Gedunin-ZnO nanoparticles for efficient drug delivery in a 3D hepatosphere model. Poly-L-Lactic acid (PLLA), an FDA approved polymer, with excellent attributes such as biodegradability, biocompatibility and ease of fabricating nanofibrous scaffold, has been used extensively for efficient drug delivery during anti-cancer therapy. This proposal will explore the feasibility of using PLLA-based nanofibrous scaffold, incorporated with Gedunin-ZnO nanocomposite as a potent controlled drug-delivery vehicle, which has tremendous future therapeutic applications. Most of the current studies using nanofibrous scaffold involves conventional 2D culture, which is not exact representative of 3D tumor microenvironment. In this Proposal, we hypothesize development of 3D hepatosphere models generated from hepatocellular carcinoma cell line Huh7 and HepaRG (and co-cultured with HUVECs), which can be an appropriate in vivo mimic of HCC microenvironment. Such hepatospheres will be seeded on PLLA nanofibrous scaffolds incorporated with Gedunin-ZnO nanocomposite, for assessment of intracellular uptake, cytotoxicity and anti-proliferative activity on hepatocellular carcinoma cells in 3D condition, in a controlled release manner. The expected outcome of this project will be development of a novel, natural bioactive compound-based nano-formulation for effective anti-cancer treatment on a pathophysiologically relevant 3D model of HCC in a proof-of-concept style, which might have tremendous therapeutic application for HCC treatment in future. |