Periodic Reporting for period 1 - BESG (Bioengineered exosomes based approaches for the effective treatment of non-small cell lung cancer)
Periodo di rendicontazione: 2020-10-01 al 2022-09-30
The most prevalent type of lung cancers are non-small cell lung cancers (NSCLCs), which have a variety of pathological characteristics. NSCLCs have been identified as a collection of different diseases with cellular and genetic heterogeneity by detailed investigations of lung cancer genomes and signalling pathways during the past ten years (Chen Z. et al. 2014). 85% of lung cancer cases in the United States are NSCLCs. The epidemiology has identified smoking, air pollution, occupational exposure, and alcohol consumption as the main risk factors. At the time of diagnosis, it was discovered that 70% of patients had locally progressed or metastatic disease (Molina JR. et al. 2008). The progression of NSCLCs is influenced by a number of driver mutations, including ROS1, RET, and other translocations; EGFR mutations; KRAS mutations; BRAF mutations; MET splice site mutations (Hirsch, FR., et al 2016). The treatment of lung cancer is drastically altering as a result of targeted medicines. These treatments include pharmaceuticals or biologicals that target immune checkpoint inhibitors, anti-apoptotic molecules implicated in cancer cell survival and proliferation, and driver mutation-targeting agents. With the introduction of new drugs, targets, therapeutic combinations, and drug delivery methods, the field of targeted therapeutic research is advancing every day (Molina JR. et al. 2008).
"BESG" sought to create a therapeutic drug delivery system prototype with improved targeting capabilities for treating non-small cell lung cancer using exosomes modified with the a targeting peptide and loaded with small molecule and nucleic acid. Theranostic and therapeutic agent delivery methodologies were also combined by BESG in order to create advanced drug delivery systems that may one day be used to develop personalized medicines.
The overall objectives of "BESG" includes
a. Development of smart peptide functionalized exosome loaded with small molecule and nucleic acid and/or fluorescent dye
b. Characterization of developed products for their enhanced targeting capabilities, cytotoxicity with NSCLCs
c. Functional studies of developed products in the animal model of NSCLCs.
"BESG" sought to create a therapeutic drug delivery system prototype with improved targeting capabilities for treating non-small cell lung cancer using exosomes modified with the a targeting peptide and loaded with small molecule and nucleic acid. Theranostic and therapeutic agent delivery methodologies were also combined by BESG in order to create advanced drug delivery systems that may one day be used to develop personalized medicines.
The overall objectives of "BESG" includes
a. Development of smart peptide functionalized exosome loaded with small molecule and nucleic acid and/or fluorescent dye
b. Characterization of developed products for their enhanced targeting capabilities, cytotoxicity with NSCLCs
c. Functional studies of developed products in the animal model of NSCLCs.
Various strategies were used to isolate exosomes from cell supernatant. NSCLC targeting peptide was developed and characterized using various analytical technologies. Bioinformatics tools were used to develop stable nucleic acid with cell uptake and imaging capabilities. Peptide was functionalized with a molecule to make it directly usable for the click chemistry. Isolated exosomes were further modified with the chemical agents so that peptide can be attached using copper free click-chemistry reaction. The developed exosomes were further characterized and then used to develop florescent dye/active ingredients loaded exosomes. Various analytical technologies were used to characterize and successfully develop functionalized extracellular vesicles. 2D cell culture models of NSCLCs were developed and capabilities of nucleic acid to lower desired the protein levels and uptake was also evaluated. Methodologies were developed to study the cytotoxic effects of small molecules and nucleic acids with the cells. Imaging studies were also performed to evaluate the effect of small molecule and nucleic acid with the cells. The developed fluorescent dye loaded peptide functionalized exosomes were further studied for their uptake by the NSCLCs using various analytical technologies.
We have successfully developed technologies to isolate exosomes in good quantity, their surface modifications with peptide using copper free click chemistry and loading them with various fluorescent dyes/small molecules and nucleic acids. Our results showed that the nucleic acid construct have higher cell uptake capability in the NSCLCs and can effectively transfect the cells and lower the desired protein level. Further, peptide modified fluorescent dye loaded exosomes have higher targeting and cell uptake capabilities than unmodified exosomes and they can also deliver the higher amount of payload to the target site. Thus we were able to successfully develop a product prototype which can be further used to develop personalized therapies for treating NSCLC patients.