BRAINSTORM: NUCLEIC ACID APTAMER SELECTION AGAINST S100B

By: Theresa Deike

Full Proposal 
Progress Report 1
Progress Report 2
Final Report
Abstract

S100B is a Calcium binding peptide commonly associated with disorders and injuries sustained to the CNS. Its presence has been associated with medical conditions such as traumatic brain injury, Alzheimer’s, Down Syndrome, dementia, and other neurological disorders. Currently the levels of S100B mRNA found intra- and extra- cellular are used to determine levels of astrocyte cell activity and or death in the brain and spinal cord after injury (Figure 1). When the blood brain barrier ,BBB,  is functioning normally, increases in the concentration of the protein in the brain correlate with an increased immune response due to the creation of auto antibodies against it, a trait commonly seen in disorders such as Alzheimer’s and dementia.

An aptamer for S100B could not only serve as a diagnostic tool for these CNS disorders but also serve as a way to get in therapeutic drugs to help counteract the disruption of the blood brain barrier once it has occurred.  In cases such as Alzheimer’s, the binding of an aptamer to the protein could help to inhibit its interaction with glial cells by changing the shape so that it can no longer bind to calcium ,receptors, or its own antibody.

Specific Aim 1: Selection of RNA aptamers against S100B.                                                                                                

A high affinity RNA aptamer could serve the purpose of inhibiting or detecting S100B. By doing so, once levels of S100B reach certain levels the treatment, detection, and possible prevention of CNS disorders associated with antibodies against S100B could become more effective.

Figure 1 (Frances et al. 2003)                                                                                                                                                      Normally, S100B is used in glial activation and for the CNS system on the brain side of BBB in cells, such as the astrocyte cells pictured. Once the blood brain barrier has been penetrated, excess amounts of S100B are secreted into the blood allowing S100B to be used to test for the functionality of BBB. Levels on the brain side of the barrier can also be used to test for neurological disorders associated with high levels of S100B when the barrier is undisturbed.

S100B is currently in the lab so there are no additional costs associated with its acquisition. It was provided by RayBiotech Inc. and is listed as catalog number 230-00002, Recombinant Human S100 Calcium binding protein B. It can be purchased purchased for $326/100 ug with the estimated cost per round totaling $7.44.  RayBiotech Inc.’s contact information is http://www.raybiotech.com/ and (770) 729-2992. 

Works Cited                                                                                                                   

"Blood Brain Barrier Working Group." Blood Brain Barrier. John Hopkins University.

Verma, Neeraj, Madhumita Karmakar, Krishna P. Singh, and Suchi Smita. "Structural and Dynamic Insights into S100B Protein Activity Inhibition by Melittin for the Treatment of Epilepsy." International Journal of Computer Applications. International Journal of Computer Applications, 2013.

Wang, Doris D., and Angélique Bordey. "The Astrocyte Odyssey." PMC. U.S. National Institutes of Health's National Library of Medicine, 1 Dec. 2009.

Yardan, Turker, Ali Kemal Erenler, Ahmet Baydin, Cengiz Cokluk, and Keramettin Aydin. "Usefulness of S100B Protein in Neurological Disorders." Journal of Pakistan Medical Association, Mar. 2011.

Frances, Karen, Johan Van Beek, Cecile Canova, Jim W. Neal, and Philippe Gasque. "The Bloodöbrain Barrier (BBB)." Expert Reviews in Molecular Medicine. Cambridge University Press, 23 May 2003.

Marchi N, Bazarian JJ, Puvenna V, Janigro M, Ghosh C, et al.  “Consequences of Repeated Blood-Brain Barrier Disruption in Football Players” Plos One. Plos.org, Mar. 2013.

W, Pan, Xin P, Patrick S, Dean S, Keating C, and Clawson G. "Primer-free Aptamer Selection Using a Random DNA Library." PubMed. N.p., 26 July 2010.

Donato, Rosario, and Claus W. Heizmann. "S100B Protein in the Nervous System and Cardiovascular Apparatus in Normal and Pathological Conditions: Figure 2." Hinawi. Cardiovascular Psychiatry and Neurology, 2010.

Smith, S. P., and G. S. Shaw. "RCSB Protein Data Bank - RCSB PDB - 1UWO Structure Summary." RCSB Protein Data Bank - RCSB PDB - 1UWO Structure Summary. RCSB, 1998.

"S100B [Homo Sapiens]." National Center for Biotechnology Information. U.S. National Library of Medicine

Putnam, Chris. "Protein Calculator." Protein Calculator.