Renal Cell Carcinoma (RCC) kills more than 14,800 people annually in the United States alone. While immune and targeted therapies have shown some promise, late stage RCC is an aggressive and lethal disease, and so, new therapies that can treat RCC are urgently needed. TRAIL (Tumor necrosis factor (TNF)-related apoptosis-inducing ligand) has been shown previously to induce apoptosis in cancer cells by binding to TRAIL receptors DR4 and DR5. Unfortunately, DR5 antibodies have failed to improve patient survival in phase II clinical trials due, in part, to the inability to cluster enough membrane receptors and kill RCC cells in the body. This project is investigating the effects that computationally designed protein nanocages bound to DR5 antibodies have on kidney cancer cells in culture. In collaboration with the Institute for Protein Design, we have recently shown that these nanoparticles induce significant activation of caspase 8 and caspase 3 in renal carcinoma TRAIL resistant cells, but not in healthy primary kidney tubular cells. To further investigate the nanocage-induced apoptosis specificity to DR5 and the involvement of other TRAIL receptors, we used CRISPR/Cas9 to generate knockouts of TRAIL receptors in RCC. We are also investigating the effects of these nanocages on other downstream pathways including the NFkB and cFLIP pathways. This project advances the knowledge of mechanisms involved in TRAIL receptor-mediated apoptosis, and our results suggest a potential new treatment for RCC.

GNE myopathy (GNEM) is an autosomal recessive inherited disease that causes progressive muscle weakness starting around ages 20 - 40, which leads to physical disability. GNE myopathy is caused by a mutation in the GNE gene, which encodes the enzyme glucosamine (UDP-N-acetyl)-2-epimerase/N-acetylmannosamine kinase. This enzyme is responsible for sialic acid (SA) production. SA is a sugar that is required by all cells, including muscle cells, to produce energy. SA is also an important component of the cell membrane. Thus, lacking SA causes muscle wasting. Interestingly, sialic acid produced in the liver could be used by skeletal muscle. Thus, our lab is developing new promoters for GNEM gene therapy that combine liver and muscle expression. By using PCR and Hifi-assembly, I cloned three control promoters--one muscle-specific enhancer/promoter (Ck8e), one liver-specific enhancer/promoter (ApoE-hAAT), and one constitutive promoter (CMV)--into three reporter plasmids ahead of firefly luciferase. These reporter plasmids also have CMV-renilla luciferase as an internal control. Now I am transfecting six tandem expression cassettes plasmids (already cloned) and the three plasmids that I cloned into the culture of liver cells (HepG2) to observe the expression of the genes based on the ratio of firefly and renilla luciferase. In this way, we can compare expression in liver cells from the novel promoters and the control promoters. Another member of the lab will be transfecting the plasmids into muscle cells (C2C12), so we will be able to compare our results. This research is important to figure out the most optimal promoter to express the GNE gene in both muscle and liver cells. Thus, after treatment, the cells could produce SA by themselves to maintain their functions and prevent muscle wasting for patients with GNEM.

In Winslow Homer’s 1892 Coast in Winter, a single skeleton tree stands upon a snow-bitten shore amid the crashing waves of the North Atlantic. After a celebrated career painting at the center of the American art scene, Homer would spend the last years his life on the remote coast of Maine. Captured by this environment, Homer’s paintings from this period became less and less populated with human figures until he removed them from the frame entirely. The paintings that come from this final chapter in Homer’s body of work have come to be known as his late marines and have been hailed by art critics and historians as his zenith. And yet, something about these works has remained elusive. Due to a seeming lack of human representation within the late marines, these paintings have escaped the grasp of description. However, there may be something deeply human about these oceanscapes after all. This essay proposes a new point of entry into Homer’s depiction of the crashing waves of Prouts Neck, Maine through insights offered by the study of Biochemistry, the study of life’s creation from inanimate matter. By tracing how the fledgling field of biochemistry emerged in tandem with Homer’s challenging late work, it contributes to ongoing art historical efforts to situate his art within the scientific developments of the time. Scientists at this moment were beginning to reckon with the understanding that the complex molecules that govern life’s mechanical systems operate according to the very same physical and chemical laws that define the nonliving universe. The natural forces of the North Atlantic and the mortal experience captured by Homer while standing upon its shores ebb and flow into one another as a swirling tide.