Farzam Farahani's HIF-1a Aptamer Project

With more than 14 million cases and 8.2 million deaths just in 2012, cancer is one of the leading causes of death worldwide administered in order to lower the mortality rate such as chemotherapy but they are not always successful, particularly when the cancer is at a more aggressive stage.

Most solid tumors enter a state of hypoxia due to uneven rates of tumor growth

and angiogenesis. To be able to survive against the hypoxic environment, cancer cells have developed a set of responses that allows them to endure the stresses against the given environment. Hypoxia inducible factor­1 (HIF­1), a transcription factor, is one of the critical mediators of the response to low oxygen tension. Under hypoxic conditions, the protein aids in the transcription of over 40 genes that participate in angiogenesis, pH regulation, cell proliferation and survival including glucose transporters, glycolytic enzymes, vascular endothelial growth factor, and erythropoietin that binds specifically to HIF­1α (a subunit of HIF­1), tumors will less likely acquire a resistance to cancer therapy, resulting in less aggressive cancer cells and potentially a lower mortality rate. An aptamer is a single­stranded DNA or RNA oligonucleotide sequence, which can fold into specific tertiary structures and bind to proteins or molecules. When bound, the oligonucleotide acts as a form of inhibition as it changes the conformation of the protein.

Aptamers can be used for many different situations, some of which are diagnostics, therapeutics, disease treatment, and drug transfer. The aptamer for this project will serve as a therapeutic agent. There currently are antibodies and other molecular structures that can inhibit HIF­1α, but the cost to produce them are really expensive. Aptamers on the other hand are much cheaper and easier to make. The SELEX method (see Figure 2) using in vitro filter­based selection will be implemented to find an aptamer against HIF­1α. More specifically, the HIF­1α protein will be bound to a filter paper, and a pool of different RNA sequences of equal length suspended in buffer will run through the filter and bind to protein. After subsequent rounds of selection, there will be a few aptamer sequences that effectively bind to the protein and successfully inhibit it.

The goal is to find an aptamer that binds effectively to HIF­1α in order to inhibit the HIF­1 transcriptional pathway. Hopefully, this aptamer will be a more cost­effective drug in cancer therapy and can help lower the mortality rate of cancer in the future.

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References

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2. Ke, Q., & Costa, M. (2006, August 3). Hypoxia­Inducible Factor­1 (HIF­1).

Retrieved September 1, 2015, from http://molpharm.aspetjournals.org/content/70/5/1469.full

3. Koh, M., Spivak­Kroizman, T., & Prowis, G. (2010). HIF­1alpha and cancer

therapy. Retrieved September 2, 2015, from http://www.ncbi.nlm.nih.gov/pubmed/20033376

4. Patiar, S., & Harris, A. (2006, December 13). Role of hypoxia­inducible

factor­1alpha as a cancer therapy target. Retrieved September 1, 2015, from

http://www.ncbi.nlm.nih.gov/pubmed/17259560

5. UniProtKB ­ Q16665 (HIF1A_HUMAN). (n.d.). Retrieved September 1, 2015,

from http://www.uniprot.org/uniprot/Q16665