Статья

Development and characterization of a DNA aptamer for MLL-AF9 expressing acute myeloid leukemia cells using whole cell-SELEX

Kaylin G. EarnestDepartment of Chemistry, University of Cincinnati, Cincinnati, OH, USAErin M. McConnellDepartment of Chemistry, Carleton University, Ottawa, ON, CanadaEman M. HassanDepartment of Chemistry, Carleton University, Ottawa, ON, CanadaMark WunderlichDivision of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USABahareh HosseinpourDepartment of Chemistry, Carleton University, Ottawa, ON, CanadaBianca S. BonoDepartment of Neuroscience, Carleton University, Ottawa, ON, CanadaMelissa J. CheeDepartment of Neuroscience, Carleton University, Ottawa, ON, CanadaJames C. MulloyDivision of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USAWilliam G. WillmoreDepartment of Chemistry, Carleton University, Ottawa, ON, CanadaMaria C. DeRosaDepartment of Chemistry, Carleton University, Ottawa, ON, Canada. [email protected]Edward J. MerinoDepartment of Chemistry, University of Cincinnati, Cincinnati, OH, USA. [email protected]

2021en

ABI

Аннотация

Current classes of cancer therapeutics have negative side effects stemming from off-target cytotoxicity. One way to avoid this would be to use a drug delivery system decorated with targeting moieties, such as an aptamer, if a targeted aptamer is available. In this study, aptamers were selected against acute myeloid leukemia (AML) cells expressing the MLL-AF9 oncogene through systematic evolution of ligands by exponential enrichment (SELEX). Twelve rounds of SELEX, including two counter selections against fibroblast cells, were completed. Aptamer pools were sequenced, and three candidate sequences were identified. These sequences consisted of two 23-base primer regions flanking a 30-base central domain. Binding studies were performed using flow cytometry, and the lead sequence had a binding constant of 37.5 + / - 2.5 nM to AML cells, while displaying no binding to fibroblast or umbilical cord blood cells at 200 nM. A truncation study of the lead sequence was done using nine shortened sequences, and showed the 5' primer was not important for binding. The lead sequence was tested against seven AML patient cultures, and five cultures showed binding at 200 nM. In summary, a DNA aptamer specific to AML cells was developed and characterized for future drug-aptamer conjugates.

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Идентификаторы

DOI: 10.1038/s41598-021-98676-4

Цитирования и источники

Цитирований: 2Использованных источников: 0

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