Off-Campus Research
Ferdinand Amanor, "Investigating a Potential Checkpoint in the Type-I Interferon Pathway Using Nanobody Technology"
The mechanism in which cells are able to block viral infection is not yet completely understood. Specifically, viral requirements for antiviral activity and secretion of Type-I Interferon, an antiviral cytokine, remain unknown. We aim to characterize the IFN pathway using nanobodies to gain insight into the innate immune response. Previous experiments conducted in a non-human cell line using Influenza virus specific nanobodies (NP2) have indicated that antiviral activity can be induced without IFN secretion from cells infected with a non-replicating virus. This would suggest that an uncharacterized checkpoint exists in the IFN pathway. This project aims to verify these results in a human cell line. By conducting a series of bioassays we were able to verify the inhibition of Influenza virus infection in 293T cells expressing NP2 while maintaining an induction of antiviral activity. Further experimentation will determine if antiviral induction occurs concomitant with IFN secretion.
Madeline Jones, "Net1A: Looking for Synergy"
Net1A is a RhoA specific guanine nucleotide exchange factor that promotes breast cancer cell motility when it relocates from the nucleus into the cytosol. We have recently found that its relocation is regulated by the Src tyrosine kinase, an oncoprotein, which stimulates phosphorylation of Net1A on Y373, leading to Net1A relocalization into the cytosol. We have also found that the nuclear exportin CRM1 is required for cytosolic relocalization of Net1A and that phosphorylation of Net1A on S52A by JNK also promotes Net1A accumulation in the cytosol. The purpose of this project was to determine whether inhibitors of Src, JNK, or CRM1 can synergistically block Net1A cytosolic relocalization and inhibit cancer cell motility. This was accomplished by transfecting MCF7 cells with Net1A and treating them with the inhibitors at various concentrations before exposure to the stimulant EGF. We used immunofluorescence to visualize and quantify the localization of Net1A, and cell migration assays to assess cell motility. Cells were pre-treated with the JNK inhibitor at 10 µM for 30 minutes and then stimulated with EGF for 15 minutes. Net1A cytosolic localization was significantly lower in cells treated with the JNK inhibitor. For the migration assay, there were no significant differences in cell motility with the JNK inhibitor. However, there was a trend towards reduced motility in cells treated with the JNK inhibitor. Overall, this work demonstrated that both JNK and CRM1 inhibition block Net1A cytosolic localization and laid the groundwork for testing for synergy between these inhibitors.
Nicholas Weirath, "Copper-Catalyzed Cyclopropanation of β-Nitrostyrenes with α-Diazoketoesters"
Diazo functionalities have gained interest among researchers for their broad use as building blocks in synthesis. Shaikh et al. have previously reported on the functionalization and transformation of diazo keto esters now seeks to explore further uses for such compounds. The increased reactivity of the carbon at the diazo site makes these compounds prime candidates for transformations that are traditionally restricted to other functional groups, which usually requires extra steps of protection and deprotection thus incurring a higher cost with more synthetic steps. Amino acids, specifically cyclopropyl amino acids, have also gained recent attention in their applicability towards pharmaceuticals. The demand for new ways of synthesizing these cyclopropyl compounds has been a noticeable focus in research lately and thus we provide a method to transform a synthesized diazo keto ester into a cyclopropyl βamino acid with nitro styrene and a copper catalyst, followed by a simple hydrogenation and hydrolysis.