Last Minute Data
Bhuvi M -
Hello everyone! This past week has been extremely busy with last-minute data collection and finishing up my presentation. Additionally, during the editing process, Dr. Fleming and I decided to change the title of my project, altering the project’s outlook.
We started the week with a dot blot of the inhibition titration we had initiated. Below, you will find five different dot blot arrays. BMS777607 is a MET inhibitor but has many off-targets and a history of being in numerous clinical trials. One issue in the past with this inhibitor was the lack of patients with the MET biomarker. Our parental cell line had a MET biomarker, qualifying this patient for a MET inhibitor both before and after the occurrence of Afatinib-mediated resistance. TP-0903 is an Axl inhibitor, known to be extremely specific to the Axl receptor. However, one issue this inhibitor faced in clinical trials was its lack of efficacy in halting afatinib resistance.
In the figures above, we can observe that with the MET inhibitor in the resistant cell line, MET disappears while EGFR remains activated. In GD, the Axl inhibitor deactivates every tyrosine kinase receptor on the dot blot array, except with a lower dose, in which the ligand of MET is activated.
So, what conclusions can we draw from this?
Well, we cannot assign these results to a specific pathway due to concerns about off-targets with an inhibitor, but we can see how the deactivation of these tyrosine kinase receptors affects some of the properties we observed in the fall that differentiated the parental and resistant cell lines.
Due to time constraints, we opted for an invasion assay. I began this invasion assay on Thursday, following the protocol outlined in my invasion assay blog post.
Thus far, the results display reduced invasion with the inhibitors. I will harvest the other wells on Saturday and stain them Monday morning, ultimately adding the pictures to my paper.
Additionally, since we haven’t been able to test our transfection fully, we chose to change the title and focus on the genetic changes occurring and how they affect afatinib resistance.
I will present the invasion assay results on May 9th and explain the implications of my research in my field!
Lastly, I would like to thank Dr. Fleming, Ms. Sandor, and Ms. Bennett, who have all contributed significantly to my project and support system over the last year!
Thank you for reading, and I hope to see you all on May 9th!