Fabrication of Multimodal Organic-Inorganic Hybrid Nanovesicles and Study on their Intracellular Fates in Cancer Cells by Single Particle Tracking

Abstract

Creation of magic bullets might be the dream of all scientists working on anticancer therapeutics. In reality, there are always pros and cons. Chemotherapeutics such as doxorubicin and paclitaxel can effectively inhibit growth of cancer cells; however, the drugs at a high dosage are not selective and can cause severe damage to normal tissues and/or organs. To minimize its side effects, anticancer therapeutics are often encapsulated using nanovesicles (NVs). Antibodies that target specific cancer cells may be conjugated with drug-carrying NVs to further improve the selectivity of drug delivery. In the design and fabrication of NV-based drug carriers, many structural and micro-environmental factors affect cellular uptake and internalization of drug-carrying NVs, drug release and distribution in tissues, and therapeutic efficiency. Two goals of my dissertation study is (1) to fabricate multimodal NVs for bio-imaging, selective targeting and drug delivery and (2) to unfold interplays between different factors and intracellular fates of these NVs in prostate cancer cells. This proposed work separates into four phases: (1) Novel organic-inorganic liposomal cerasomes will be used as drug carriers for delivery of potent anticancer doxorcubicin (DOX). Liposomes will be stabilized via an addition of inorganic polyorganosiloxane networks on their surfaces, creating liposomal cerasomes, and its potential to store and release DOX will be investigated. (2) Prostate cancer cells will be used as a model system to study endocytic pathway of cerasomes. Among various types of human cancer, prostate cancer is the second leading cause of death for man. It is anticipated that the proposed study will shed new insights into endocytosis drug-carrying NVs. Results from the study may facilitate the development of cerasome-based therapeutics for treating prostate cancer patients. (3) Since diagnostic, imaging and therapy are three major biomedical applications in nanotechnology, multifunctional NVs with combined bio-imaging, therapeutic efficiency and selective targeting modalities are essential as next-generation nano-carriers. Here, we proposed to incooperate fluorophores and prostate cancer targeted antibodies into cerasomes to achieve our multimodal NVs. (4) Single particle tracking (SPT) analysis will be used to study the intracellular fates of cerasome uptake in single cell level. SPT is an advanced imaging technique permits the real-time monitoring of cellular entry, intracellular transport and internalization of multiple NVs. Our main focus is to identify the different in cellular transportation mechanism between targeted and non-targeted vesicles. Moreover, SPT will be integrated into a microfluidic system, which not only minimizes the consumption of reagents/ materials but also allows us to precisely control biochemical environments around cells.