Determining the Role of Antisense Transcripts in Regulating the Human Inflammatory Response

Determing the Role of Antisense Transcripts in Regulating the Human Inflammatory Response

Students

College

Project Summary

This summer, I worked under the guidance of Dr. Elliot Hersh in the Penn Dental’s Department of Oral Surgery and Pharmacology, and Drs. Katie Theken and Tilo Grosser in the Institute for Translational Medicine and Therapeutics (ITMAT).

 

Using data from a previous clinical study their group has published, I selected two antisense transcripts to investigate based on levels of expression following the extraction of impacted wisdom teeth. Antisense transcripts (asRNAs) are single-stranded RNAs complementary to messenger RNAs (mRNAs), which code for proteins. asRNAs, especially long non-coding RNAs (lncRNAs), are important in regulating gene expression.We focused on these asRNAs with high expression to determine their role in the inflammatory response in humans using the common model of third molar extraction. Subjects were given non-steroidal anti-inflammatory drugs (NSAIDs) for pain relief, and their inflammatory response was measured at different timepoints post-surgery by measuring prostaglandin content in their blood and urine. By measuring the subjects’ response to the medication and studying the activity of asRNAs during inflammation, we can potentially develop more effective ways to treat pain while minimizing cardiovascular and gastrointestinal risk, including individualizing analgesic regimens.

 

To begin, I first designed primers and conducted PCRs to verify that the product produced was within the region of interest. I then designed antisense oligonucleotides (ASOs) that can be used to knock down the genes in cell cultures via transfection. By knocking down the genes and then treating the cells with lipopolysaccharide (LPS), an inflammatory agent, we can determine if those genes have an active role in the inflammatory response by comparing the responses of the experimental group to the control group (no gene knockdown).

 

Over the course of the summer, I learned various techniques in the biological sciences, from qPCR to cDNA isolation from human monocytes. The learning curve was steep at first, especially since I did not have an extensive background in biology, but I familiarized myself with techniques and language in the field by practicing, reading papers, and asking questions about everything.

 

Although a lot of the knowledge I gained was specific to a field, the interdisciplinary nature of my research allowed me to apply what I learned to a variety of different fields. Additionally, many of the skills I acquired, though not specific to science or research, were useful nonetheless. I realized that patience is key, especially in research, where it can take years to achieve concrete, replicable results. I improved my presentation and communication skills, and I became far less afraid to ask questions or admit that I didn’t know something. I became an independent problem-solver, thinking creatively and applying what I had learned from reading papers, going to talks and conferences, and asking questions. I look forward to continuing my work in the future and expanding upon what I have learned so far.