Circumventing Traditional Bottlenecks In Glioblastoma Genetic Therapy Case Study

Case Study Assignment: RNA interference: what advances have been made in the last decade? Name: Ronghua Wei CID: 00740579 Supervisors:Professor Kurt Drikamer, Dr. Maureen Taylor MRes in Drug Discovery and Development (2015-2016) 1 Content Abstract.............................................................................................................................................. 3 Introduction ..................................................................................................................................... 3 Machinery.......................................................................................................................................... 4 Challenges ......................................................................................................................................... 8 Delivery ...................................................................................................................................................................

The company ‘Invivogen’ made a transfectable and simple-to-use plasmid, called psiRNA™, that allows the production of siRNAs within the cells. To transfect means to introduce foreign material into eukaryotic cells. psiRNA is an RNA polymerase III-based plasmid that produces ‘short hairpin’ RNAs (shRNAs). shRNAs are more stable than synthetic siRNAs and since they are continuously expressed within the cells rather than having to be injected into the cytoplasm, this method permits long-lasting silencing of the BLG gene.

siRNA shows the potential to selectively silence the expression of a disease gene based on the gene’s sequence. By down regulating cancerous transcripts and restoring the wild type function of a gene, siRNA has the potential to play an important role in cancer

In America today finding a cure for cancer is a huge priority with clinical trails being a big part in these goal there are some newly developed drugs and technology that are making a huge impact in the cancer world.

Nanocarriers (NCs) have emerged as a favored drug delivery approach towards improving the anticancer benefits of several bioactives for cancer therapy with recent reports showing the application of NC systems in clinical settings [18]. The NCs with size < 100 nm have been associated with enhanced permeation and retention effect (EPR) due to the presence of leaky vasculature in the tumor tissues which contributes to its enhanced efficacy. Also recently, NCs have shown to be effective in the treatment of malignant mesothelioma [19]. A recent study by Kanai et al. showed that the NC albumin-bound paclitaxel and carboplatin (nabPC) repetitively achieved tumor regression in malignant mesothelioma

The functions of telomeres includes protection of chromosomes from degradation and end to end fusions, maintains the stability of chromosomes, acts as a mitotic clock to understand the replicative history of a cell.Telomerase is a ribonucleoprotein DNA polymerase that helps in maintaining the telomeres by adding TTAGGG repeats at the ends of chromosomes (Greider and Blackburn,1989). Telomerase consists of two main subunits: telomerase RNA (hTR) and catalytic component human telomerase reverse transcriptase (hTERT). The enzymatic activity is usually absent in somatic cells but remains active in germ line cells,embryonic stem cells and tumor cells. Almost 80 to 90% of human cancers are characterized by the presence of telomerase activity.Studies on glioma shown that more than 50% of tumors have telomerase activity and its detection rate increases with the grades of malignancy (Tchirkov et al 2003). Therefore knockdown of hTERT expression may help in the development of novel therapeutic strategies. In the present study we evaluate the effect of siRNA to downregulate hTERT in LN18 cells and also investigated the effect on apoptosis and cell

Anthracyclines antitumour antibiotics are capable to inflict the action of topoisomerase involved in DNA replication which inhibits mitotic and disrupt cells proliferation. Doxorubicin (DOX, with the commercial name of Adriamycin) is one of anthracycline and is a potent FDA-approved chemotherapy drug with the great efficiency to combat cancerous cell dividing [1]. Doxorubicin has been acknowledged to treat various types of cancers as well as breast cancer, bone marrow cancer, osteosarcoma and others for several decades that its ability limited by adverse side effect and unpredictable toxicity on normal cells leads to reduce immune cells and also extremely cardiotoxicity. Patients become more susceptible to infections [2]. For this reason, nanoparticles

Cancer is one of the leading causes of death today. There are a variety of different treatments for cancer such as surgical removal, chemotherapy, hormone therapy, and radiation. Chemotherapy is the most common approach it kills the cancer cells by drug toxicity or preventing cell division (Jabir et al., 2012). The issue with Chemotherapy is that it is rarely successful. The radiation approach kills cancerous cells, but it also damages the surrounding healthy cells. However, newer techniques are being developed to just attack the cancerous cells. There is a need to find a cancer treatment approach that does not kill the healthy cells along with the cancerous cells. The answer to solving this challenge lies in the successful application of

Cancer is one of the leading causes of death worldwide as it can develop in almost any organ or tissue. Significant advances in understanding the cellular basis of cancer and the underlying biological mechanisms of tumour has been vastly improved in the recent years (Jiang et al. 1994). Cancer is a genetic disease which requires a series of mutation during mitosis to develop, its characteristics can be associated with their ability to grow and divide abnormal cells uncontrollable while in the mean time invade and cause nearby blood vessels to serve its need. Even though many people are affected by cancer today, the abilities which cancer cells have make it hard to find a single effective treatment for cancer. The focus of research now lies

Cancer has been a disease since the dawn of man and even since that moment, we as humans have strived to understand its’ lethality and discover ways to combat it. This proves to be a more difficult task than it seems. Cancer, by definition, is the uncontrolled division of abnormal cells in the body [1]. These divisions can spread into the surrounding tissues as well and cause damage to the regions they locate to. Many of these divisions can lead to solid growths in the body. The growths are usually referred to as tumors [1]. These tumors are masses of tissue that group together.

In addition, it was shown that macrophages can mediate the delivery of gold-shell NPs into multicellular human glioma spheroids 76. In the report, gold-shell NPs are composed of a silica core coated with a thin layer of gold which greatly absorbs near-infrared light to generate local heat around NPs. Gold nanoshells (AuNS) NPs were loaded in macrophages via phagocytosis, and it is interesting to observe no apparent toxicity of macrophages after their uptake of nanoparticles 77. When co-incubated with human glioma spheroids, nanoparticle-laden macrophages can migrate and filtrate in the tumor as usual 78. Furthermore, two-photon fluorescence microscopy of glioma spheroids showed that nanoparticle-laden macrophages can accumulate in necrotic sites. When irradiated with 810 nm laser light, the growth of glioma spheroid dramatically decreased compared with the control without of nanoparticle-loading. The efficacy of macrophage-AuNS-mediated photothermal therapy (PTT) on glioma spheroids was also tested, and the growth of spheroids was dramatically suppressed. The same group continued to examine the therapeutic efficacy of macrophage-mediated delivery of AuNS nanoparticles in rat glioma tumor model 79. C6 rat glioma cells were directly injected into brain to generate brain tumor, following the injection of macrophage-AuNS delivery system. After NIR laser irradiation, it was observed that the tissue irradiated by laser was damaged, suggesting the therapeutic effect of AuNS.

The most common approach in gene replacement therapy is through viral vectors. A piece of DNA is

Through Western blot analysis, gö6983 and AZD1480 have been shown to inhibit PKC (protein kinase C) and JAK2 (Janus kinase 2), respectively, in LN229P glioblastoma cells. To determine the levels of proteins PKC and JAK2, inhibition readouts of the downstream proteins were looked at. The focus of this experiment are the drugs gö6983, AZD1480, their combinations and their respective effects in cell line LN229P. The drugs' inhibition effect on the levels of protein is clear in in-vitro experiments against LN229P glioblastoma cells. These findings suggest new combinations of proteins to inhibit so that apoptosis can be induced in these tumor

As far as the structure guided chemical modification process the researchers began with HEK293 cells that were engineered to constitutively express GFP and spCAS-9. The reason they did this is the GFP protein was the target gene the researchers wanted to turn off since it is a simple way to observe editing effects and quantify levels of potency simply. After modifying various aspects of the sgRNA and exposing the HEK293 cells to each treatment they could simply take fluorescence readings and quantify how many cells were still expressing the GFP gene, and how many had been successfully edited. The modifications explored in this paper are modifications that had previously shown to decrease RNA’s susceptibility to nucleases in the blood,

Despite the promise of such approaches, a number of difficulties remain to be overcome, the most important of which is the need for more efficient systems of gene delivery. No gene transfer system is 100% efficient, unless germ-line therapy is contemplated. During the past two decades, there have been major advances in our understanding of how cancer develops, proving that cancer has a genetic basis (2). A series of genetic abnormalities that accumulate in one cell may result in a pattern of abnormal clonal proliferation. Our growing understanding of the genetic basis of cancer offers new opportunities for the molecular prevention and treatment of cancer. There has been a