Exosomes : fighting cancer with cancer

Exosomes are nanovesicles secreted by many cells, including cancer cells. Extensive research has been carried out to validate potential applications of exosomes and to evaluate their efficiency in a wide range of diseases, including cancer. The current knowledge on the origin, biogenesis and composition of exosomes is described. This review then focuses on the use of exosomes in cancer diagnostics and therapeutics

Microfluidic manufacturing of different niosomes nanoparticles for curcumin encapsulation : physical characteristics, encapsulation efficacy, and drug release

Curcumin is a natural chemical compound found in Curcuma longa which has been used in several therapeutic applications such as an antitumor and anti-inflammation agent. However, curcumin has very limited water solubility and rapid in vivo degradation which limits its clinical application. To overcome these limitations, niosome nanoparticles were prepared by microfluidic mixing for curcumin encapsulation. Niosome nanoparticles are lipid-based, and composed of non-ionic surfactants with cholesterol orientated into a membrane bilayer structure. Two different non-ionic surfactants were used and the mixing parameters were varied to optimise the characteristics of the prepared niosomes. The prepared niosomes had an average particle size ranging from 70-230 nm depending on the type of non-ionic surfactant used and the mixing parameter. Moreover, all the prepared niosomes were monodisperse with an average polydispersity index ranging from 0.07-0.3. All the prepared niosomes were spherical in shape as demonstrated by transmission electron microscopy. Curcumin was encapsulated with a maximum encapsulation efficiency around 60% using Tween 85 as the non-ionic surfactant. Niosomes prepared by microfluidic mixing provided controlled release of curcumin, as indicated by the release profile of curcumin overtime, thereby improving its therapeutic capability. These results demonstrate that niosomes prepared by microfluidic mixing to encapsulate curcumin is a promising delivery system to reach target cells

Development of exosome-based anti-cancer therapeutics

Hepatocellular carcinoma (HCC) exosomes are found to be responsible for cancer progression, metastasis, and angiogenesis via cellular communications. However, studying cancer derived exosomes in vitro is limited, due to the way cell lines are grown in medium supplemented with foetal bovine serum (FBS) that contains naturally occurring exosomes. Bovine-derived exosomes can cause artefacts and interfere with interpretation of results. The aim of this study was to investigate the release of exosomes from human liver cancer cell line HepG2 (HepG2-Exo) under different conditioned media with modified FBS to deliver the best approach for exosome production. Thus, two media were developed for growing HepG2 cell line which are M1 and M2 using dulbecco's modified eagle medium (DMEM), where M1 supplemented with 10 % (v/v) FBS, and M2 with 10 % (v/v) exosome depleted FBS (Dep-FBS). However, after cells reached confluency, those two media were removed and replaced with serum free media (only DMEM), to create M3 and M4, respectively. This resulted in collecting four categories of media: M1, M2, M3, and M4. Consequently, four groups of exosomes were obtained (Exo(M1), Exo(M2), Exo(M3), and Exo(M4).;In regard to cell culture, findings confirmed that M2 was the best approach for cultivating HepG2 and collecting exosomes, as cell viability was enhanced and contamination with FBS-exosomes was minimal, compared to M1. However, analysis of the different HepG2-Exo groups showed significant difference in protein concentration, percentage of fluorescence, exosome marker detection, tetraspanins expression, particle count, metabolic and lipidomic profiling, RNA sequencing, and gene expression. This difference indicates that the effect of media composition is inevitable on cell-derived exosome which may cause misinterpretation of the effect of the exosomes of interest. Consequently, biological assessment and metabolomic profiling of HepG2-Exo effect on different cancer and normal cell lines was carried out: A375 (melanoma), A549 (lung cancer), and PNT2A (normal prostate epithelium). The biological assays revealed that HepG2-Exo induced the proliferation, migration, adhesion, and invasion of A549 at 50 µg/ml. While metabolome analysis showed that HepG2-Exo at 100 µg/ml, induced significant changes in the cell metabolome of A375.;The outcomes of this project provided an effective approach in developing successful cell culture for exosome collection without concern over contamination from FBS-derived exosomes and brought attention to the critical effect of media in exosome studies. Moreover, this project has highlighted the effect of HepG2-Exo on other cell lines and the potential of HepG2-Exo to be applied to the development of future lung cancer therapeutics.Hepatocellular carcinoma (HCC) exosomes are found to be responsible for cancer progression, metastasis, and angiogenesis via cellular communications. However, studying cancer derived exosomes in vitro is limited, due to the way cell lines are grown in medium supplemented with foetal bovine serum (FBS) that contains naturally occurring exosomes. Bovine-derived exosomes can cause artefacts and interfere with interpretation of results. The aim of this study was to investigate the release of exosomes from human liver cancer cell line HepG2 (HepG2-Exo) under different conditioned media with modified FBS to deliver the best approach for exosome production. Thus, two media were developed for growing HepG2 cell line which are M1 and M2 using dulbecco's modified eagle medium (DMEM), where M1 supplemented with 10 % (v/v) FBS, and M2 with 10 % (v/v) exosome depleted FBS (Dep-FBS). However, after cells reached confluency, those two media were removed and replaced with serum free media (only DMEM), to create M3 and M4, respectively. This resulted in collecting four categories of media: M1, M2, M3, and M4. Consequently, four groups of exosomes were obtained (Exo(M1), Exo(M2), Exo(M3), and Exo(M4).;In regard to cell culture, findings confirmed that M2 was the best approach for cultivating HepG2 and collecting exosomes, as cell viability was enhanced and contamination with FBS-exosomes was minimal, compared to M1. However, analysis of the different HepG2-Exo groups showed significant difference in protein concentration, percentage of fluorescence, exosome marker detection, tetraspanins expression, particle count, metabolic and lipidomic profiling, RNA sequencing, and gene expression. This difference indicates that the effect of media composition is inevitable on cell-derived exosome which may cause misinterpretation of the effect of the exosomes of interest. Consequently, biological assessment and metabolomic profiling of HepG2-Exo effect on different cancer and normal cell lines was carried out: A375 (melanoma), A549 (lung cancer), and PNT2A (normal prostate epithelium). The biological assays revealed that HepG2-Exo induced the proliferation, migration, adhesion, and invasion of A549 at 50 µg/ml. While metabolome analysis showed that HepG2-Exo at 100 µg/ml, induced significant changes in the cell metabolome of A375.;The outcomes of this project provided an effective approach in developing successful cell culture for exosome collection without concern over contamination from FBS-derived exosomes and brought attention to the critical effect of media in exosome studies. Moreover, this project has highlighted the effect of HepG2-Exo on other cell lines and the potential of HepG2-Exo to be applied to the development of future lung cancer therapeutics

siRNA delivery to melanoma cells with cationic niosomes

RNA interference (RNAi) is a posttranscriptional regulatory mechanism that employs siRNA. It typically results in the degradation of a target mRNA that encodes a particular protein. Treatment with siRNA therapeutics requires the use of an effective drug delivery system to assist in delivering these therapeutics into the cytoplasm of the transfected cells. Here we describe the transfection of melanoma cancer cells with siRNA using cationic niosome nanoparticles as a delivery system. The method of niosome preparation is first introduced and is followed by complex formation with siRNA and the transfection method

Use of nanoparticles in delivery of nucleic acids for melanoma treatment

Melanoma accounts for 4% of all skin cancer malignancies, with only 14% of diagnosed patients surviving for more than 5 years after diagnosis. Until now, there is no clear understanding of the detailed molecular contributors of melanoma pathogenesis. Accordingly, more research is needed to understand melanoma development and prognosis. All the treatment approaches that are currently applied have several significant limitations that prevent effective use in melanoma. One major limitation in the treatment of cancer is the acquisition of multidrug resistance (MDR). The MDR results in significant treatment failure and poor clinical outcomes in several cancers, including skin cancer. Treatment of melanoma is especially retarded by MDR. Despite the current advances in targeted and immune-mediated therapy, treatment arms of melanoma are severely limited and stand as a significant clinical challenge. Further, the poor pharmacokinetic profile of currently used chemotherapeutic agents is another reason for treatment failure. Therefore, more research is needed to develop novel drugs and carrier tools for more effective and targeted treatment. Nucleic acid therapy is based on nucleic acids or chemical compounds that are closely related, such as antisense oligonucleotides, aptamers, and small-interfering RNAs that are usually used in situations when a specific gene implicated in a disorder is deemed a therapeutically beneficial target for inhibition. However, the proper application for nucleic acid therapies is hampered by the development of an effective delivery system that can maintain their stability in the systemic circulation and enhance their uptake by the target cells. In this chapter, the prognosis of the different types of melanoma along with the currently used medications is highlighted, and the different types of nucleic acids along with the currently available nanoparticle systems for delivering these nucleic acids into melanoma cells are discussed. We also discuss recently conducted research on the use of different types of nanoparticles for nucleic acid delivery into melanoma cells and highlight the most significant outcomes