Hox genes are evolutionarily conserved transcriptional regulators that are essential for the proper patterning of the axial and appendicular skeleton. Posterior Hox genes (Hox 9-13) are critical for the patterning of the proximodistal axis of the limb. Loss of Hox11 function leads to severe disruption in muscle patterning, with muscle defects in Hox11 mutants independent of skeletal patterning. Recently, our lab demonstrated that Hox11 genes are continuously expressed from embryonic to adult mouse stages in skeletal stem cells. We also have evidence that Hox11 expression is maintained through postnatal and adult stages within muscle stromal cells. I plan to investigate the role of Hox11 and Hox11-expressing cells at postnatal and adult stages during homeostasis and in response to skeletal muscle injury.

Current gene therapies utilize virus vectors for treatment of disease, but they are expensive and have a limited shelf life. Polymer mediated gene therapies are a suitable alternative because they address these issues. Our group has previously used Raman spectroscopy to investigate quinine-based polymers to further understand the transfection mechanism of DNA. My project is investigating how quinine-based monomers intercalate with DNA to elucidate the effect different monomers will have on the transfection efficiency. Raman spectroscopy can monitor the local environment changes that occur when the polymer interacts with the DNA. We monitor the quinoline ring vibrational mode and its shift as a function of DNA concentration, for different quinine-based monomers. By comparing the trend of frequency shifts for each monomer with their respective transfection efficiency, we can understand the effect quinine-DNA interactions have on the overall efficiency of polymer gene delivery. These results will be used to further the development of new and efficient quinine-based polymers for gene delivery.

Kinases are critical enzymes within biological chemistry that are habitually involved in the phosphorylation of proteins using adenosine triphosphate (ATP). Phosphorylation is a reaction that adds a phosphate group to a protein. In doing so, kinases aid in cell signaling allowing a series of chemical reactions to relay messages for cell function. Due to their role in signaling, dysregulation of kinase activity is most often observed in cancer progression. Therefore, there is a need to find specific small molecule inhibitors that inhibit their activity and act as therapeutic targets to treat cancer. Wnt signaling has a major role in colon tissue growth and homeostasis. Aberrant activation of this pathway can lead to the over proliferation of cells, increasing oncogenic signaling and the development and progression of colorectal cancer. Previous studies in our lab revealed that inhibition of the CLK3 kinase disrupted Wnt signaling. Therefore, the present study conducts a drug screen utilizing a particular subset of kinases, specifically the CLK’s and DYRKs from the CMGC kinase family. Tests will be conducted regarding the specificity of the collection of pan-DYRK/pan-CLK inhibitors to each of the proteins; aiming to find inhibitors that solely bind to one of the aforementioned enzymes. Upon completion of this project, small molecules will be identified that target subsets of DYRKS and CLKS. If specified inhibitors for our families of kinases are found, then we may have the capacity to disrupt the Wnt pathway in cancer cells, further progressing cancer research.

There have been limited drug breakthroughs for the treatment of Alzheimer Disease (AD). Molecularly, a main characteristic of AD is the formation of toxic protein structures that harm neurons. These are made from the accumulation of the proteins tau and amyloid beta (Ab). To form such structures, tau tends to require other compounds that act as seeds to aid the growth of its toxic structures. As there has been evidence of Ab proteins seeding the formation of toxic tau structures, we hypothesize that the seeding of tau with Ab is a relevant step in the progression of AD. We are using biochemical methods such as electron microscopy and western blots with the objective of understanding the Ab and tau seeding interface. This will allow us to understand how this process happens and learn if it has biological relevance. This will deepen the scientific understanding of this elusive topic, laying the groundwork for the development of novel drugs to treat the disease.

Oxidative stress has been found to be an important factor in cell and tissue damage as well as severe health conditions including chronic inflammation, cardiovascular conditions, neurodegenerative diseases and cancer. This imbalance between the antioxidants and the free radicals in the body prevents the cells from the detoxification of oxygen reactive species (ROS). The purpose of this study is to simultaneously quantify the ratio of oxidized (GSSG) and reduced glutathione (GSH), biomarkers of oxidative stress. By modifying the glassy carbon electrode with multiwalled carbon nanotubes (MWCNT) we were able to detect the GSSG and GSH with cyclic voltammetry (CV). The simultaneous detection of both species is enabled by the electrocatalytic activity between the MWCNT functionalized with carboxylic groups and nanocomposite molecular material based on cobalt phthalocyanine (CoPc). The electrochemical behaviors of the electrodes were assessed by a conventional three-electrode cell having the glassy carbon electrode as the working electrode, and an Ag/AgCl reference electrode. The same quantification of glutathione is being studied with a carbon fiber microelectrode to compare and create the standards for the determination of oxidative stress that will later be used on biological fluids. The creation of the carbon fiber ultra-microelectrodes will provide less invasive technology that can later be implemented in other studies. When an organism is under oxidative stress the ratio of oxidized and reduced glutathione is not in equilibrium, consequently the determination of a method that can easily detect GSH/GSSG would be a great improvement in the development of clinical diagnosis.