Generation 3 Polyline Dendron Gene Delivery Vector - Research Proposal Example

Proposal for a Research Paper on Generation 3 Polylysine Dendron Gene Delivery Vector Summary of the Proposed Research, Including Goals (Lay Readers): In the modern day, genetic medicine plays a crucial role in the treatment of a wide array of diseases such as cancer, Alzheimer’s and other chronics. Similarly gene therapy and gene delivery have a crucial role to play in the present day medical science. However, a lot of barriers exist in the gene delivery system, especially due to vulnerability of various agents used in the process DNA introduction into a host body. Thus, the scientific community has been researching the use of dendrons for various biological applications including gene delivery and gene therapy. These agents are known to possess a wide variety of applications, which can facilitate various non-viral systems for the introduction of DNA into the host body and enable easy and effective genetic therapy for the cure of dreaded diseases such as cancer. This study, attempts to identify and illustrate third generation polylysine dendrons as a potential source for the development of a non-viral platform for gene delivery. The study will delineate the characteristics of various dendrons to demonstrate their potential as a non-viral source of gene delivery. In addition, it will also illustrate the specific traits of third generation polylysine for potential use in gene delivery and gene therapy. To establish this, the study will comprise lab testing of mice to show that gene delivery can be efficiently be done using third generation polylysine dendrons. The goal of the research is to add value to the existing body of knowledge in this regard and to make it a more effective system for gene delivery as well as gene therapy. Summary of the Proposed Research, Including Goals (Scientific Readers): While the use of viral-vectors is an efficient method to introduce genes into host cells, they entail several cellular as well as extra cellular barriers. Though viral-vectors have both efficiency and high gene expression they ail from the major limitation of vulnerability to immunogenic reactions. Therefore, the medical fraternity’s interest rather slants to the alternative method of deploying non-viral methods for the delivery of genes into the host body as it entails lesser complexity. Thus, the use of dendrons or dendrimers has been gaining popularity in the field of genetic medicine, especially in its application for gene delivery and gene therapy. This is especially so because they provide a multifunctional platform by virtue of being synthetically capable of attaching tumour-targeting antibodies. Many studies confirm the validity of third generation polylysine as an effective monomer that has the potential to deliver bioactive drugs and genes. Thus, this dendron can be an instrumental non-viral platform in gene delivery as well as gene therapy. This study aims to illustrate the various characteristics of dendrons as a catalyst for the non-viral method of delivery of genes. For this purpose it will demonstrate the characteristic traits of various dendrons and also delineate their capabilities for convergent and divergent expansion as well as other features that make them potential candidates for consideration as non-viral vectors for the delivery of genes. Similarly, the study will also demonstrate various traits of third generation polylysine and show why it could be a useful agent for use as a non-viral vector for gene delivery. The study seeks will also evaluate the existing body on the topic and show what findings other researchers have endorsed in their studies. This study intends to add to the existing body of knowledge on the subject. Background to the Research and How This Proposal Relates to Other Studies in the Field: Background: The modern day is marked by a wide array of chronic and acute diseases that pester human life, especially with conditions such as cancer, cardio vascular diseases, Alzheimer’s etc. Genetic medicine offers a lot of advantages in the treatment of several diseases including cancer but the paucity of “safe and effective delivery systems” is a crucial limitation (Harris et al 2010: 998). The barriers in delivery systems include both “cellular barriers” and “extracellular barriers” (998). Gene delivery is the introduction of “foreign DNA particles” into a host cell through viral or non-viral methods (Pal, Banthia & Majumdar 2009: 27). The former process uses a virus to incorporate its “DNA into the host cells” (27). Viral vectors have both the kind of efficiency for transduction as well as “high gene expression” (27). However, this method suffers from the problem that viruses are prone to “immunogenic reactions” and the transfected cells can become subject to mutagenesis (27). Therefore, scientific interest leans towards the use of non-viral delivery system option that entails “less complexity” (27). On the other hand, dendrimers, because of their “multivalent and monodisperse” traits, enjoy great potential for both biological and chemical applications in gene delivery and gene therapy (Ajaykumar et al 2012: 1). Cancer is one disease that “epitomizes the challenges,” which the medical fraternity faces in the present day, especially in terms of gene delivery (1). Thus, the use o dendrimers in gene delivery can be instrumental in the treatment of cancer, Alzheimer’s as well as other such dreaded diseases. Similarly, dendrimers possess the synthetic capabilities of attaching tumor-targeting antibodies as well as a “potent payload” of drugs to the same molecule, which provides a multifunctional platform (4). On the other hand, evidence suggests that polylysine dendron can be of potential use in “biological applications” such as gene delivery (Patel et al 2010: 384). In addition, dendric fragments also have the capabilities to be extended both in “convergent or divergent direction” and thus enable the utility of both approaches (386). Accurate evaluation of polylysine dendrons can be undertaken using either “transmission electron microscopy (or) atomic force microscopy” (386). The properties of dendrimers make them efficient agents for the application in a wide range of applications such as: (1) “solubility enhancers” (2) effective gene delivery agents in non-viral platform (3) after modifications with other agents they can be used as nano-drugs (4) instrumental in drug delivery (5) therapy using photodynamic and (6) for gene transfections (386). Evidence further shows that dendrimers being macromolecules with “highly branched 3D structure” can facilitate high levels of surface functionality as well as versatility (Pandya et al 2012: 404). On the other hand, peptide-based dendrimers including polylysine possess potentials as a “promising vaccine” and it can also be used as an antibacterial agent after certain marginal modifications (412). Thus, these dendrimers have valuable pharmaceutical applications and they offer “new scaffolds” for the delivery of drugs, especially for cancer, as well as play a crucial role in non-viral gene delivery and therapy (418). In addition, studies also have validated the effectiveness of polylysine as the “microbicides for herpes simplex virus (HSV) types 1 and 2” (Loutsch, Ong & Hill 2003: 479). While in vivo studies reveal that application of solution BRI-2999 has significantly reduced the “occurrence of disease,” in virto indicate that the procedure blocks the infection in the “cyto-pathic effect (CPE) inhibition assays” (480). Similarly a study, conducted by Galli et al (2012) to investigate the effects of the application of phosphoserine-tethered poly(epsilon-lysine) dendrons as a “film to titanium surfaces” find that the process enhances the “differentiation of osteoblastic cells and activation of Wnt/β-catenin signalling” (1). The study further finds that titanium is the most popular choice in “dental and orthopaedic uses” and it improves the response to the cells in the bone, especially when it is enhanced with dendrons (4). On the other hand, evidence also indicates that dendrimers have significance implications on the study of biological events and their use can be quite promising in “gene therapy and drug delivery” (Cloninger 2002: 746). Gene therapy can be perceived as the transfer of “recombinant genetic material” such as DNA or RNA to a host cell for the purpose of changing its gene expression so as to attain better effect for any therapeutic interventions (Biceroglu & Memis 2005: 113). Evidence suggests that while gene therapy has been subjected to a wide range of research, the determination of common approach still “remains elusive” (Jones et al 2011: 416). Thus, the development of appropriate vectors for the delivery of DNA or RNA into host cells has become crucial for interventions such as gene therapy (416). On the other hand, evidence points to the fact that viral vectors can cause immuno-genicity as well as “adverse patient response” (416). Thus, the development of non-viral vectors has become the practical approach for yielding the best results through gene therapy. In this context, evidence suggests that second generation dendrons can “transfect in the absence” of chloroquine, apparently because of the presence of larger number of amines (426). On the other hand, studies emphasise the fact that Poly(L-lysine) (PLL) enjoys an “advantage for in vivo” due to its peptide structure as well a biodegradable nature (Passirani 2009: 3479). Similarly, another study by Tripathy and Das (2013) finds that multiple antigen peptide dendrimers, which is “dendron like” molecular construction on “polylysine skeleton,” can act as effective monomers for introducing a variety of branching points (144). The study concludes that such dendrimers are of potential use in the delivery “bioactive like drugs and gene” in the non-viral system (147). Hypotheses: H-1: Polylysine enjoys the edge of better functionality as a non-viral gene delivery vector over non-viral vectors. H-2: Third generation polylysine is an effective agent for use as a potential non-viral vector for the delivery of gene and in gene therapy. H-3: Third generation polylysine dendron has better capabilities to work efficiently as a non-viral delivery vector than other dendrons. Methods of Research: Overall Study Design: The study will comprise collection of both primary as well as secondary data to determine the validity of third generation polylysine as a potential non-viral vector for the delivery of gene and in gene therapy. The secondary data in this regard will be obtained through a review of the existing literature published on the topic. Information will be obtained through reliable sources including scientific journals, books and internet sources. Only recent publications that appearing in credible and reliable sources during the last ten years will be included in the literature review so as to glean the most relevant and genuine information. The university library, local library as well as online sources will be used for the purpose of the study. The determination of various aspects of the study to be covered during the primary data collection will be done after the completion of secondary data collection and arriving at relevant findings that will point to any gaps in the existing research. This will also enable the researcher to focus the primary data collection relevant to areas that are currently unexplored or questions that remain unanswered. Primary data collection will be based on conducting appropriate lab test using suitable methods so as to determine the validity of the use of dendrons as the best method for non-viral gene delivery. Details of Sample Population to be Used: The primary data collection will encompass lab tests on a sample population mouse to see the impact of the use of non-viral system of gene delivery. The researcher proposes to use 10 mice that will be tested within the lab facility. Clinical/Laboratory/Field Techniques: The tests will be conducted in an enclosed space within the university lab and access to the area will be restricted to only people who are involved with various processes of this study. Clinical tests on the rats will be done using third generation polylysine dendrons applied to them on specific conditions for the determination of various aspects envisaged in the study. Fibroblast cells will be target for the study and different dendrons will be administered on the samples at appropriate intervals to record the response to each delivery system. Operative Procedures and Data Analysis: The data gleaned at each stage of the lab tests will recorded and finally the data obtained will be consolidated. Once the entire perspective of data is evaluated, it will be subjected to an analysis, using appropriate statistical methods to draw the findings. Potential Impact of the Work: The main goal of the study is to establish whether third generation polylysine dendrons are the most appropriate and suitable non-viral vectors for the delivery of genes. Thus, the study will add to the existing body of knowledge on the topic. On the other hand, the determination of the effectiveness of this agent will help the use of this Dendron as an effective source of gene delivery. This can further be used for the effective treatment of cancer and other such diseases. Thus, the proposed study will be beneficial not only to the medical fraternity and scientific community, but will also be advantageous to the human race in general. Description of Any Difficulties Anticipated: As transpired from the wide range of journal articles, published in scientific journals, it appears that there is still a element of indecisiveness among the scientists and medical experts as to which is the best non-viral system for gene delivery and gene therapy. As such, the results of a study of this magnitude may not find acceptance by the established system. However, the study is expected to throw further light into the matter. Also, during the course of the literature review, the researcher has felt that there is a dearth of polylysine-specific studies in the existing material available through various sources. Brief Reference to Any Further Work Anticipated: Due to the paucity of other specific studies relating to polylysine as an agent for non-viral gene delivery, irrespective of the findings of the instant study the researcher will recommend further studies on the topic so that final confirmation of the views of the study can be obtained and validated through other studies. Key Milestones to the Achieved in the Work: The key milestones to be achieved in the research will include the determination of the validity of polylysine as an agent for non-viral delivery of genes. If appropriately validated in the lab tests, it will pave the way for advanced research on this specific topic. This will be of interest to the medical fraternity, healthcare professionals as well as the human society in general. Ethics Committee and HTA Considerations: The researcher will complete all formalities prior to the commencement of the study and all necessary permission will be obtained from the Ethics Committee and HTA. References Ajaykumar, B., Babu, N. N. V., Reddy, L. Ujwala, K. 2012. Applications of Dendrimers in Cancer Therapy. International Journal of Chemical and Pharmaceutical Sciences, 3/2: 1-7. Biceroglu, S. & Memis, A. 2005. Gene Therapy: Applications in Interventional Radiology. Diagnostic and Interventional Radiology, 11: 113-118. Galli, C. et al. 2012. Biomimetic Coating with Phosphoserine-tethered Poly(epsilon-lysine) Dendrons on Titanium Surfaces Enhances Wnt and Osteoblastic Differentiation. Clinical and Oral Implants Research, 0: 1-7. Jones, S. P. et al. 2011. Hydrophobically Modified Dendrons: Developing Structure-Activity Relationships for DNA Binding and Gene Transfection. Molecular Pharmaceutics, 8: 416-429. Loutsch, J. M., Ong, D. & Hill, J. M. 2003. Dendrimers: An Innovative and Ocular Drug Delivery System. New York: Marcel Dekker Inc. Pal, K., Banitha, A. K. & Majumdar, D. K. Polymeric Hydrogels: Characterization and Biomedical Application – A Mini Review. Designed Monomers and Polymers, 12: 197-220. Pandya, D. P., Chaudhari, M. J., Thakkar, P. P. Soni, A, M. & Bharadia, P. D. 2012. Dendrimer: A Novel Polymer. International Journal of Pharmaceutical Research Scholars, 1/2: 404-420. Passirani, M. M., Vonabourg, A., Clavreul, A. & Benoit, J. P. 2009. Progress in Developing Cationic Systems for Non-viral Vector Systemic Gene Therapy against Cancer. Biomaterials, 29/25: 3479-3496. Patel, S. N. et al. 2010. Dendrimers: A Novel Drug Delivery System. International Journal of Pharmacy & Life Sciences, 1/7: 382-388. Tripathy, S. & Das, M. K. 2013. Dendrimers and Their Applications as Novel Drug Deliver Carriers. Journal of Applied Pharmaceutical Science, 3/9: 142-149. Read More