About me
Proteomic and transcriptomic exploration of Eph receptor forward signaling in vitro
Location: 29
Mentor: Dr. Daniel Palaez
Several studies have identified dysregulation of ephrin signaling in various neurodegenerative diseases. As the largest class of receptor tyrosine kinases (RTKs), Ephrin receptors (EphRs) and their ephrin ligands are widely known for their role in mammalian development, specifically concerning axon guidance and synaptic plasticity. However, the downstream mechanisms that result from Eph-ephrin binding are poorly characterized due to the complexity of their signaling. In previous in vivo studies, we identified hyperactivation of several Eph receptor tyrosine kinase domains during the early stages of optic neuropathy. Whether or not these forward signaling mechanisms contribute directly to neurodegeneration is of great interest to the field. To expand our in vivo observations, we examined the immediate proteomic and transcriptomic changes that are a direct result of Eph receptor activation. By computational analysis of several proteomic pathway databases, we identified 14 genes of interest related to canonical Eph receptor forward signaling. To experimentally test these gene targets, we stimulated EphRs with purified ephrin ligands in MCF7 and Sh-Sy5y cell lines. We conducted western blot analysis to verify the proteomic post-translational modifications, and qPCR to identify differential expression of several known downstream transcripts. By coupling this in vitro data with the proteomic and transcriptomic changes seen in the retinal tissue of our mice models, we can infer the pathways that are likely implicated in optic neuropathy. Understanding the role that Eph receptor hyperactivation plays during the onset of neurodegeneration will offer insight into better diagnostic and treatment strategies.
