Selection of Aptamers Against FGF8b Oncoprotein for Diagnostic Application in Detecting Cancerous Cells by Finding Overexpression of Oncoprote

Taher Jamali
Target: mFGF8b
Pool: N71

Abstract:

Breast and prostate cancer(Jemal et al. 2009). Nasopharyngeal carcinoma. Structure of the midbrain and cerebellum. What do these three seemingly unrelated phenomena of the body have in common? Mammalian fibroblast growth factor 8b (mFGF8b). This unique protein has been found to be involved in the cell signaling pathway that can lead to uncontrolled growth in hormonal cancers, such as breast and prostate(Tanaka et al. 1998; Mattila and Harkonen, 2007), as well as nasopharyngeal cancer. mFGF8b is also interestingly involved with other isoforms and receptors to control the structure of midbrain and cerebellum during development(Olsen et al. 2006).

            An intricate approach to dealing with many of these possibly fatal cancerous diseases involves detection, and conceivably, inhibition of the mFGF8b protein using aptamers. Aptamers are small nucleic acid strands made up of DNA or RNA that can be selected against a protein of interest for utilization in a specific purpose. In this case, the aptamer selected against mFGF8b can attach to the protein of interest, and- with the proper detection signal such as radioactivity or fluorescence- help to identify overexpression of the oncoprotein. After finding overexpressed proteins in specific cells in the body, further action can be taken to remove the cells before the cancer can progress. The aptamer may also be utilized therapeutically if allowed to attach to mFGF8b in its cellular pathways, and hopefully, block any unnecessary effects of the protein in cells showing uncontrolled growth by inhibiting the entire protein. A representation of this therapeutic inhibition can be seen in Figure 1.

Figure 1: By preventing mFGF8b from binding to FGFR with the use of aptamers in cells such as prostate cells, we can prevent the oncoprotein’s effect in cellular pathways affecting uncontrolled cancerous growth.

Specific Aim 1: Selection of RNA aptamers against mFGF8b

The first major goal is to complete rounds of selection to find a specific aptamer that can bind to the mFGF8b protein.

Specific Aim 2: Perform Binding Assay to Test Binding Affinity of mFGF8b aptamers

The second major goal is to test the binding affinity of the selected aptamer to the mFGF8b protein so that it can be possible to see how effectively the aptamer will bind and inhibit the protein in its cellular pathways in the future.

Target Order Information:
Keatinge-Clay Lab
mFGF8b is currently available in the lab (Purified by Shawn Piasecki)
Lab Info is as follows-
Office: WEL 4.230B
Office Phone: (512) 471-2977
E-mail: adriankc@utexas.edu
Lab: WEL 4.234
Website: http://keatinge-clay.cm.utexas.edu/research/index.html
Target Cost: $0
Cost Per Round: $0

References:

1.     "His-Tagged Protein Isolation & Pulldown Using Dynabeads®." His-Tagged Protein Isolation & Pulldown Using Dynabeads. Life Technologies Corportation, 2013. Web. 21 Apr. 2013.

2.     Jemal A, Siegel R, Ward E, Yongping H, Xu Jiaquan, Thun, Michael. (2009). Cancer Statistics, 2009. A Cancer Journal for Clinicians. 59, 4. pp 215- 275

3.     Mattila MM, Harkonen PL. (2007). Role of fibroblast growth factor 8 in growth and progression of hormonal cancer. Cytokine Growth Factor Rev 18: 257–266.

4.     Olsen S, Li J, Bromleigh C, et al. (2006) Structural basis by which alternative splicing modulates the organizer activity of FGF8 in the brain.  Genes Dev. 20: 185-198.

5.     Tanaka A, Furuya A, Yamasaki M, Hanai N, Kuriki K, Kamiakito T et al. (1998). High frequency of fibroblast growth factor (FGF) 8 expression in clinical prostate cancers and breast tissues, immunohistochemically demonstrated by a newly established neutralizing monoclonal antibody against FGF 8. Cancer Res 58: 2053–2056.

6.     V W Y Lui, D M-S Yau, C S-F Cheung, S C C Wong, A K-C Chan, et al. (2010). FGF8b oncogene mediates proliferation and invasion of Espstein-Barr virus-associated nasopharyngeal carcinoma cells: implication for viral-mediated FGF8b upregulation. Nature Publishing Group. Oncogene (2011) 30, pp 1518-1530

Proposal:

https://docs.google.com/file/d/0B8agoD1PeGsSNHlCMWN6VUhWbG8/edit?usp=sharing
First Progress Report:
https://docs.google.com/file/d/0B8agoD1PeGsSY2pOUlpRYkNtZVU/edit?usp=sharing
Second Progress Report:
https://drive.google.com/file/d/0B8agoD1PeGsScmxSUnlLOFJiZUE/edit?usp=sharing
Final Report:
https://drive.google.com/file/d/0B8agoD1PeGsSMWxsa3Budk55djQ/edit?usp=sharing