Selection of Aptamer Against Myelin Basic Protein for the Promotion of Remyelination

Selection of Aptamer Against Myelin Basic Protein for the Promotion of Remyelination
Evan Roberts
Myelin basic protein (MPB) is a protein found in the central nervous system (CNS) of many species. MBP is responsible for the adhesion of cytosolic surfaces of multilayered compact myelin in the CNS and is essential for the formation of CNS myelin. The demyelinating disease multiple sclerosis (MS) occurs when an immune response is caused due to presence of myelin (J.M. Boggs, 2006). There is absence of treatment that prevents progression or induces repair to the myelin in the human body (Nastasijevic, 2012).
Recently it has been shown that a single stranded DNA aptamer 40-nucleotides long selected for affinity to murine myelin shows the property of remyelination (Smestad, 2013). Because this aptamer binds to many mice components of myelin such as MBP, it is reasonable to believe that an aptamer selected against bovine MBP could have the same properties and may also bind with human MBP. This aptamer selected for MBP could provide a base for the creation of a therapeutic for humans with MS.
Specific Aim 1: Selection of RNA aptamer against MBP.
An aptamer selected against mice MBP recently promoted remyelination in a study done with mice. The myelin-specific aptamer induced remyelination in 35% of the experimental CNS lesions, compared to 4% and 9% remyelination that took place in the negative control mice treated with other aptamers (Nastasijevic, 2012). This study provides evidence that an aptamer selected against MBP can provide regeneration of myelin sheath, thus potentially providing an effective therapeutic for MS patients around the world.

Figure 1. MS causes the immune system to destroy Myelin in the CNS. Specific aim one is part of finding a component that may be vital to induce proper regrowth of Myelin in patients with MS. 

The target MBP can be ordered at a reasonable price from United States Biological, telephone number (800)-520-3011. Their website is http://www.usbio.net/. The catalog number is M9758-10 and it cost $327.00 for 1 mg of MBP. If 400 pmol of MBP is used per round of selection, the cost per round will be $2.42. 
Boggs, J. M. (2006). Myelin basic protein: a multifunctional protein. Cell. Mol. Life Sci., 63, 1945-1961.
                http://dx.doi.org/10.1007/s00018-006-6094-7
Nastasijevic B, Wright BR, Smestad J, Warrington AE, Rodriguez M, et al. (2012) Remyelination Induced by a DNA Aptamer in a Mouse Model of MultipleSclerosis. PLoS ONE 7(6): e39595. doi:10.1371/journal.pone.0039595
Smestad, J., & Maher, J. L., III. (2013). Ion-dependent conformational switching by a DNA aptamer that induces remyelination in a mouse model of multiple sclerosis. Nucleic Acids Research, 41(2), 1329-1342. http://dx.doi.org/10.1093/nar/gks1093
Here is a link to my progress report number 1: https://www.dropbox.com/s/yzecdo0awberv3e/Roberts%2C%20Evan%2C%20MBP_Progress_Report_1.pdf 
Here is a link to my progress report number 2: https://www.dropbox.com/s/alddplzsckghtbd/Roberts%2C%20Evan%2C%20MBP%20Progress%20Report_2.pdf
Here is a link to my final report: https://www.dropbox.com/s/xrbxly8ojvn7gxw/Roberts%2C%20Evan%2C%20MBP%20Final%20Manuscript.pdf

Aptamer Selection Against IL21R for the Stimulation of Resistance to Type 1 Diabetes Mellitus



        Aptamer Selection Against IL21R for the Stimulation of Resistance to 
                                            Type 1 Diabetes Mellitus 
                                                 Lusaura Gutierrez 

           Type 1 diabetes mellitus is a disease brought about by the autoimmune attack and destruction of the beta cells located in the pancreas which causes an increase in blood glucose within the individual to occur. The autoimmune destruction of the pancreatic beta cells is in part provoked by the variation of CD8(+) T cells into Cytotoxic T Lymphocytes through the use of several cytokines, and transcription factors. Interleukin 21, a cytokine and growth factor for CD8(+) T cells, utilizes the T-bet transcription factor in order to promote Cytotoxic T Lymphocyte variation (Sutherland et al 2013). However, the Interleukin 21 receptor (Figure 1) functions as a controlling factor of antigen conveyance by dendritic cells in CD4(+) T cells which are required by CD8(+) T cells to effectively reach the pancreatic islets and induce diabetes (Van Belle et al 2012). Therefore, the hindering of the Interleukin 21 receptor (IL21R) would impair CD4(+)and CD8(+) T cells and consequently stimulate resistance to type 1 diabetes mellitus.
         A RNA aptamer, is a nucleic acid sequence that has been developed to recognize and bind with a high affinity and specificity to a selected target. Therefore, due to the inability of pancreatic infiltration to occur without the presence of IL21R signals created in CD4(+) T cells, a RNA aptamer against IL21R will prevent the IL21R signals from reaching auto reactive CD8(+) T cells and initiating pancreatic infiltration.
         Utilized as therapeutics, RNA aptamers are able to impede a specific function or pathway of a targeted protein. An aptamer against the IL21R will inhibit pancreatic infiltration and therefore, the overall development of type 1 diabetes mellitus.
         Specific Aim 1: Develop an anti-IL-21R aptamer capable of inhibiting the IL21R signal, and stimulating resistance to type 1 diabetes mellitus.
Figure 1:
Figure 1 demonstrates the signal producing IL-21 and IL-21R relationship in the CD4(+) T cells which are then transported to CD8(+) T cells for pancreatic infiltration, adapted from Rochman (2009).
Target order Information: 
Vendor: R & D Systems
Product catalog number: 991-R2-100
Vendor website: www.rndsystems.com
Vendor telephone number: (800) 343-7475
Cost per unit:  $319 per 100 ug
Cost per round: ~$32.98 ($16.49 per 100 pmol)

References: 
Sutherland AP, N. Joller, M. Michaud, SM Liu, VK Kuchroo, MJ Grusby. 2013. IL-21 Promote
            CD8(+) CTL Activity Via the Transcription Factor T-Bet.Journal of Immunology
            190(8):3977-3984.

Van Belle Tom, S.Nierkens, R. Arens, M. Von Herrath. 2012. Interleukin 21Receptor-Mediated
            Signals Control Autoreactive T Cell Infiltration in Pancreatic Islets. Immunity 

            36(6):1060-1072.

Click here for full proposal 
Click here for the first progress report 
Click here for the second progress report 
Click here for the final report 

Example of a pretend Blog Post (Fall, 2013)

An anti-dog aptamer:  Should we call it cat?  
Gwendolyn M. Stovall

[TEXT of my ABSTRACT]

[Figure]

[Budget - Include a budget, even if your target is free]

[Reference]


Aptamer Selection against Myelin Basic Protein for the Treatment of Multiple Sclerosis

Aptamer Selection against Myelin Basic Protein for the Treatment of Multiple Sclerosis
Su Min Cho

        Myelin membrane of nerve cells is composed of a protein called myelin basic protein (MBP).[1] Myelin basic protein forms and stabilizes the myelin membrane of nerves which allows the nerve cells to transmit signals with greater speed and efficiency.[2] An autoimmune disease known as multiple sclerosis is characterized by the active degradation of myelin membrane of the nerve cells which hinders the ability of the nerves cells to conduct signals to one another. 
  The main aim of this research is to find an RNA aptamer for myelin basic protein. Aptamers are single-stranded oligonucleic acid molecules that bind to their targets with high affinity and can cause changes to a target by binding to the target. The result is target inhibition which prevents the target from binding to other proteins and amino acids. 
        Finding an aptamer for myelin basic protein may lead to a treatment for multiple sclerosis. Figure 1 is a simple depiction of the role that aptamers may serve. If aptamers bind to myelin basic protein, the resultant change in the structure of the myelin basic protein may prevent the body’s immune system from attacking the myelin sheaths of the nerve cells. 

Specific Aim: Selection of RNA aptamers against Myelin Basic Protein that will change the target structure


Figure 1. As aptamers bind to the target (MBP), changes are made to the structure of MBP. This change results in target inhibition which may prevent the body’s immune system from attacking the myelin sheaths of nerve cells.

Target Order Information

  Vendor: United States Biological
  Product catalog number: M9758-10
  Vendor website:  http://www.usbio.net/
  Vendor telephone number: 800-520-3011
  Cost per unit: $327 per 1 mg
  Cost per round: $2.40 per round (400 pmol)

References
1.  Deber, C. M., Reynolds, S. J. (1991). “Central nervous system myelin: structure, function, and pathology.” 
    Clinical Biochemistry. 24:113-34. 
2.  Nye, S.H., Pelfrey, C.M., Burkwit, J.J., Voskhul, R.R., Lenardo, M.J., Mueller, J.P. (1995). “Purification of 
    immunologically active recombinant 21.5 kDa isoform of human myelin basic protein.” Molecular Immunology.32:1131-4


Target Full Proposal Link
1st Progress Report Link
2nd Progress Report Link
Final Progress Report Link