Travel Report: USC Visitation
Start: August 2, 2022
End: August 10, 2022
Location: USC/Norris Cancer Center
Laboratory: Dr. Ite Offringa
Attendee: Matthew D. Gregory
Lab Experience
August 2, 2022, served as a travel day, arrival in LA was later than expected due to delays. The following days (August 3-4th) were used to acclimate to the lab environment. During this time, I was given a tour of the research facilities by colleague, Pablo Puente. I also had the pleasure of shadowing USC graduate student Diego Velarde and Michigan State University med student Ryan Dingle on their current projects in which I assisted in the preservation of mouse organs. August 5th, Pablo and I designed and initiated a cell viability assay to treat cell lines developed by the Offringa lab with compounds synthesized by Dr. Lamango. The specific aim was to identify the sensitivity of various lung epithelial cell lines when treated with PCAIs. We continued working on the experiment from August 6-7th. After its conclusion on August 8th, we collected our data. On this same day, I attended Dr. Offringas’ weekly lab meeting gaining further insight on the software and projects currently in progress. On the following day, we analyzed our data and discussed with our PIs. Overall, visiting Dr. Offringas’ lab was a productive and educational experience. During my brief visit, I was introduced to cell counting with a hemocytometer, organoids, electroporation, and embedding of mouse organs. I feel that this trip has broadened my research knowledge and applicational skills.
Findings
Aim:
Identify and compare the sensitivity of various non-cancerous immortalized lung epithelial cell lines to KRAS driven cancer cells when treated with PCAIs.
Protocol:
To determine the potency of the PCAIs as potential anticancer agents, cells were plated at a density of 1.0 × 104 cells per well in 96-well tissue culture plates (Genesee Scientific, San Diego, CA, USA) and incubated for 24 h for the cells to adhere. The cells were then treated with varying concentrations of the PCAIs (0-50 µM) followed by the WST-8 reduction assay for cell viability. EC50 values were then calculated from nonlinear regression plots of fluorescence intensities against the concentrations of the respective PCAIs using GraphPad Prism version 9.0 for Windows (San Diego, CA).
Cells Cultured:
Cancer cell line: NCI-H23
Immortalized epithelial cell lines: hAEC-FT-C7, hAEC-ON-F9, hAEC-TN-B10
Results:
Work previously done in the Lamango lab
Work performed in the Offringa lab
Discussion:
After the initial experiments, we have found that when treated with PCAIs NCI-H23 cancer cells have a higher sensitivity and lower EC50 than the immortalized lung epithelial cells. Further replication of these results will be performed soon to validate this finding.
Extracurricular Activities
Upon my arrival at USC, I was immediately introduced to the familial cohesiveness of the Offringa lab personnel. Dr. Offringas’ hospitality reminded me of my own family! I was fortunate to have numerous home-cooked meals with Dr. Offringa and outings with the lab. Among my many outings, I would place surfing with Pablo and my visit to Santa Monica among the most memorable experiences. I also had the opportunity to attend the poster session of my previous summer research program, tour the medical school, and meet with other members of the CaRE2 program. I cannot express my gratitude enough to FAMU, Dr. Offringa, her husband Darryl, Pablo, Alex Hightower, and students Diego, Ryan, and Sarah for making my experience fun and educational.
