Abstract 1747: Selectivity of K-562-GFP-CLENs by CML: Development, characterization and in vitro analysis

Abstract

Abstract Background: Chronic myeloid leukemia is a blood cancer predominantly affecting older adults. Current treatments are initiated based on CML phase severity or progression. Although unable to cure, tyrosine kinase inhibitors are first-line due to their ability to increase survival and provide fewer side effects than other treatments. All therapies for CML present with limitations due to reasons including mutations, resistance, tolerability, and a lack of selective drug targeting. Liposomes are effective drug delivery systems, shown to increase drug half-life and allow for better targeting of drug substances at the intended site of drug action. The objective of the study was to evaluate the cellular uptake selectivity and specificity of CLENs (cell membrane lipid-extracted nanoliposomes) in CML cells. Methods: The CML cell line, K-562-GFP, was the chosen model for CML disease and drug selectivity studies. K-562-GFP cells were cultured and expanded In vitro for cellular extraction purposes. The K-562-GFP cell lipid extract (LE) was included to prepare K-562-GFP CLENs. Phase I of evaluation included physicochemical characterization and cellular uptake studies of preparations of CLENs including 0 to 40 mol% ofK-562-GFP LE content. Phase II of evaluation fixed the optimal LE content determined from the Phase I studies and varied the cholesterol content (0 to 40 mol%), to determine optimized K-562-GFP- CLENs for the target selectivity studies. The experimental cell lines used in the study represent other hematological diseases, RPMI 8226 (multiple myeloma), CCRF-CEM (acute lymphoblastic leukemia), and U937 (lymphoma). Conventional lipid ingredients used in the preparation of K-562-GFP-CLENs include DOPC, Cholesterol, and DPPE-Rhodamine (used as a fluorescence indicator) for fluorescence studies using a fluorescence microplate reader. Results: The results from the characterization studies revealed that the optimized composition and ratio for K-562-GFP CLENs was 85/10/5 (DOPC/Cholesterol/LE), the mean particle size was 292 +/-58 nm (n=5). The mean zeta potential was -15 +/- 9 mV (n=5). The average uptake for K-562-GFP-CLENs cells with 0% LE was 6822 +/- 50.5 and 21,767 +/- 3380 (p<0.05) with LE. The average uptake for ALL cells with 0% LE was 6,558 +/- 33 and with LE was 24,672 +/- 3107(p>0.05). The average uptake for CRL cells with 0% LE was 6780 +/- 73.67 and with LE was19312 +/- 1434 (p<0.05). The average uptake for MM cells with 0% LE was 6992 +/- 46.15 and with LE was 21,197 +/- 1910.0 (p<0.05). There was no statistically significant difference in the selective uptake of optimized preparations of K-562-GFP-CLENs by control ALL, CRL, and MM cells, when compared to the K-562-GFP (target) cell line (p>0.05). Conclusion: Further research comparing the uptake of various preparations of K-562-GFP-CLENs by normal, and other negative controls cell lines in local and external microenvironments, are currently underway. Citation Format: Sarah Boddie, Akrofi Akotiah, Dominique Walker, Robert Campbell. Selectivity of K-562-GFP-CLENs by CML: Development, characterization and in vitro analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1747.