Biomod/2014/OhioMOD/project

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<h1>Novel microRNA antisense therapeutic delivery using DNA origami nanostructures (under construction)</h1> Here's the research we did <h2>Introduction</h2> <h2>Background: CANCER AND CLL</h2> <p1>Cancer is a group of diseases broadly characterized by the uncontrolled growth and spread of abnormal cells. In 2013, there were approximately 1.6 million new cases of cancer, and about 580 thousand people were expected to die of the disease. However, cancer isn’t necessarily fatal. The five year survival rate of all cancers diagnosed between 2002 and 2008 is 68%, up from 49% in 1977. (Why did this happen? Better drugs? Better diagnosis?) INSERT FIGURE HERE ABOUT CANCER SURVIVABILITY?</p1>

</br></br><p1>Leukemia is a type of cancer that develops when the bone marrow produces abnormal stem cells, which would normally differentiate into white blood cells. The abnormal cells don’t differentiate, reproduce continuously, and do not die. The unhealthy cells crowd out healthy cells in the blood, resulting in complications which can be fatal. Chronic lymphocytic leukemia (CLL), one specific type of the disease, is characterized by the buildup of abnormal, mature lymphoid progenitor cells. CLL typically grows slowly, affecting cells in the lymph nodes. It is the most common form of leukemia in the western hemisphere, accounting for nearly 38% of all leukemia cases. In 2013, around 16 thousand new cases were diagnosed, with about 4,500 estimated deaths. Over 60% of those diagnosed were in adults over 65 years of age. The outlook for CLL has improved dramatically over the last three decades. The 5-year survival rate is 82%, and most patients live for 10 years or more if the cancer is detected in stage A. CLL is clearly less of a threat than other types of cancer. However, it is ideal for studying potential cancer therapeutics for several reasons.</p1> </br></br><p1>The OSU-CLL cell line is an immortalized cell line created by Dr. John Byrd and his team at The Ohio State University to promote mechanistic in vitro and in vivo studies of CLL. OSU-CLL cells displace aberrant co-expression of CD5 protein, and trisomy in chromosomes 12 and 19, all of which are common indicators of CLL. Studies have also shown that OSU-CLL cells show similar motility and proliferation behaviors when compared to primary CLL cells. OSU-CLL is also readily transferable to mice for in vivo studies. (ADD A COUPLE EXAMPLES OF RESEARCH DONE WITH OSU-CLL?) (WHAT SHOULD I DO WITH THE “CAUSE” SECTION?)</p1>

</br></br><p1>Chemotherapy refers to the use of chemical therapeutics to kill or attack cells which divide rapidly. Along with radiation therapy and surgery, chemotherapy is one of the most commonly used methods of cancer treatment, prescribed to 22% of cancer patients. There are numerous common chemotherapeutics, but they have some common characteristics. Their mechanism of action typically involves disrupting the mechanisms of cellular reproduction. This makes them more toxic to cells which divide rapidly, perfect for treating cancer. However, this also causes some unfortunate side effects. Cells in the bone marrow also divide rapidly, and are responsible for producing the white blood cells which are central to the human immune system. Chemotherapy usually attacks these cells, making the body extremely vulnerable to infection. Hair follicles and digestive tissues also divide rapidly, so hair loss and severe digestive problems are other side effects. Other complications surround the application of chemotherapy, including drug resistance. The Blood-Brain Barrier also poses a huge challenge in the treatment of brain cancers. </p1> </br></br><p1>There are other complications surrounding the application of chemotherapy. In brain cancer, the blood brain barrier prevents most free drugs from reaching the tumor through the circulatory system. Nonspecific chemotherapy also suffers because the drug is distributed throughout the body. Biocompatibility can also be an issue. If the drug provokes an immune response from the body, or is broken down in the circulatory system, it won’t function as a chemotherapeutic agent, despite potentially attractive cytotoxic characteristics. Cancer cells are also capable of developing resistance to chemotherapeutic agents through a variety of methods, most of which stem from the idea of natural selection. For any tumor, a small amount of cells will probably be resistant to any given chemotherapeutic. If this drug is administered consistently, those cells will reproduce more and the whole tumor will become resistant.</p1> </br></br><p1>Clearly, chemotherapy is not a perfect system. However, nanotechnology may hold the key to solving a lot it’s problems. Most cancer cells show a modified protein expression profile, expressing some surface proteins at a much greater rate than normal cells. For instance, Beta Folate Receptors are significantly overexpressed in myeloid leukemia. This folate receptor has been targeted by liposomal drug carriers in the past to great effect (http://pubs.acs.org.proxy.lib.ohio-state.edu/doi/pdf/10.1021/bm0506142) and it should be possible to incorporater targetting into other nanoparticle therapeutics. Targeting allows for higher doses of chemotherapy with fewer side effects (CITATION). Encapsulation of therapeutics has also been shown to increase their viability in the circulatory system and increase efficacy of therapy in vitro (http://pubs.rsc.org/EN/content/articlelanding/2013/tb/c2tb00063f/unauth#!divAbstract). Encapsulation of chemotherapeutics can also be used to apply multiple drugs to one tumor, which significantly lowers the chance of resistance developing.</p1> One specific type of chemotherapy, antisense therapy, utilizes complementary base pairing to block RNA strands in cancerous cells. In all cells, messenger RNA is translated in the ribosomes to produce proteins, which regulate and catalyze all processes in the cell. In cancerous cells, proteins responsible for growth and division are overexpressed, while those responsible for inducing cell death (apoptosis) and regulating growth are underexpressed. Antisense therapy involves targeting a specific mRNA strand that is overexpressed in cancerous cells (the sense strand) and introducing a complementary strand (the antisense strand) to the cell to bind with it and block it’s translation. One study targeted the growth inducing protein Bcl-2. The authors found that, in live patients, Bcl-2 antisense therapy showed a positive tumor response and an improvement in symptoms. The drawback to antisense therapy lies in the structure of DNA. It is naturally degraded in the body, so it needs to be modified to make it a viable treatment </p1></br></br> <h2>Background: miRNA & PTEN</h2> <h2>Background: DNA Origami</h2> <h2>Structures</h2> <h2>miR 21 Sequestering</h2> <h2>Cellular Uptaking</h2> <h2>PTEN Expression</h2> <h2>Cellular Viability</h2> <h2>etc.</h2>

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