Discipline: Biological Sciences
Subcategory: Cancer Research
Session: 1
Room: Virginia B
Ebony Nottingham - Florida A&M University
Co-Author(s): Sunil Napir, Florida Agricultural and Mechanical University, Florida; Stephen Safe , Texas A&M University, Texas; Arun K Rishi , Wayne State University, Michigan; and Mandip Singh, Florida Agricultural and Mechanical University, Florida
Hypothesis: Non-small cell lung cancer (NSCLC) accounts for 80% of all lung cancers, and is one of the most common malignant tumors worldwide. It remains difficult to treat due to late stage detection as well as its highly mutative nature that leads to the development of drug resistance. We have found that multiple growth factor signaling processes are involved in NSCLC drug resistance that are not easily mimicked in conventional 2D cell culture techniques, which can skew cell response measurements. It is necessary to expose cancer cells to the cell–cell and cell–matrix interactions they would experience in vivo to achieve more physiologically relevant results. We believe that a combinational approach of CFM 4.16 (CARP-1 functional mimetic 4.16) and Osimertinib in a 3D printed tumor environment will be able to improve response in a more realistic manner. Methods: H1975 wildtype and H1975-CL1 (Osimertinib resistant) cell lines were used in these studies. Cell viability assays were performed comparing monolayer cultured (m), spheroid cultured (s), and 3D printed (p) cells to compare their response to both single treatment Osimertinib and the combination of Osimertinib and CFM 4.16. Western blot analysis was performed measuring the expression of EGFR, MET, VEGFR, and PDGFR both phosphorylated and un-phosphorylated forms. Treatment response was also measured in the presence and absence of 10ng/ml TGF-B, VEGF, and EGF. Tumor invasion was modelled through growth factor exposure measuring the size and number of spheroids that showed the ability to pinch off and form new spheroids. 3D spheroid and printed models were also expanded to include co-culture with Human cancer associated fibroblasts. Results: Monolayer cultured cells showed increasing treatment sensitivity followed by spheroid culture and 3D printed cells respectively. Western blot analysis confirmed increased expression of epithelial markers E-cadherin and Vimentin which decreased in the presence of growth factors suggesting increased mesenchymal signaling. The presence of growth factors also decreased cell sensitivity in spheroid and printing cultures as well as provided increased sphere pinching to form new spheroids. Combination therapy provided increased sensitivity in each culture method although 3D printed cells still showed higher IC50 values. Co-culture models showed increased stemness when compared to single culture models. Conclusions: Based on the results of these studies, 3D culture methods have shown that increased cell-cell interactions lead to increased drug resistance. We have also shown that the combination of CFM 4.16 and Osimertinib was able to provide increased cell response.
Funder Acknowledgement(s): Funding was provided by CREST Center for Complex Designs for Multidimensional Printing award number 1735968
Faculty Advisor: Mandip Sachdeva, Mandip.sachdeva@famu.edu
Role: All experiments were performed by me with the guidance of my co-authors