Therapeutic investigation and modeling of resveratrol and siRNA targeted combination delivery to leukemia cells

Nearly 10 million deaths are caused by cancer around the globe every year. Blood cancer is responsible for roughly 86,000 deaths in the United States. Patients' lifespan is improving as new technologies and controlled drug delivery techniques emerge. Cancer drug resistance and high required drug dosage remain the major hurdles towards improving cancer treatment. Nutraceuticals, such as resveratrol, are proven to be safe, and chemo-preventive action on a wide array of cancer have been used as a method to improve the sensitivity of the other drugs in combination therapy. Nutraceuticals, however, are non-specific, sparingly soluble in aqueous solution, and bioavailability in vivo is very low, leading to limited therapeutic effectiveness even at higher doses. Clinical trials with 150 mg/day of resveratrol given to healthy volunteers for 30 days showed plasma levels in the nanomolar range. Hence, arises the need for drug delivery of combination therapy to reduce the dose required and drug resistance. In this project, resveratrol is combined with leukemia-targeted small interfering RNA (siRNA), and encapsulated in electrospun microfibers, and holo-transferrin PEG-liposomes, respectively. Experiments were per-formed using single K562 cell cultures, as well as K562 and HUVECs co-cultures. Resveratrol (40 uM) content was analyzed using HPLC and cell viability was assessed using Annexin V (Non-viable), and Propidium Iodide (PI) (Necrotic) based flow cytome-try. Electrospun microfibers with resveratrol were made using 1:1 PCL-GT blends. BCR-ABL siRNA (36 nM)-encapsulated holo-transferrin-conjugated PEG-liposomes were characterized using dynamic light scattering, and transmission electron microscopy. RT-qPCR was performed to assess silencing BCR-ABL gene. The combination therapy was additive, and controlled release in a timely manner resulted in 94.32 (+/-1.70)% K562 cells non-viability after 8-days of incubation. A computational fluid dynamics (CFD) model was developed in this project to assess the fluid flow impact on resveratrol release from electrospun microfibers. A custom-built bioreactor was designed to validate the model and study the stability of resveratrol in cell medium. Concentration profiles over 5 days were generated. The model suggested that perfusion velocity may not have a significant effect relative to cellular uptake rate and porosity of the surrounding tissue represented by that in the bone marrow microenvironment