title: Induced Apoptosis Investigation in Wild-type and FLT3-ITD Acute Myeloid Leukemia Cells by Nanochannel Electroporation and Single-cell qRT-PCR.
abstract: Nanochannel electroporation (NEP) was applied to deliver precise dosages of myeloid cell leukemia-1 (Mcl-1)-specific siRNA and molecular beacons to two types of acute myeloid leukemia (AML) cells, FMS-like tyrosine kinase-3 wild-type (WT) and internal tandem duplications (ITD) type at the single-cell level. NEP, together with single-cell quantitative reverse transcription PCR, led to an observation showing nearly 20-folds more Mcl-1 siRNA than MCL1 mRNA were required to induce cell death for both cell lines and patient blasts, i.e., ~8,800 siRNAs for ~500 ± 50 mRNAs in ITD cells and ~6,000 siRNAs for ~300 ± 50 mRNAs in WT cells. A time-lapse study revealed that >75% MCL1 mRNA was downregulated within 1 hour after delivery of a small amount of siRNA. However, additional siRNA was required to inhibit the newly transcribed mRNA for >12 hours until the cell lost its ability of self-protection recovery. A multidelivery strategy of low doses and short delivery interval, which require 77% less siRNA and has the potential of lower side effects and clinical cost, was as effective as a single high-dose siRNA delivery. Our method provides a viable analytical tool to investigate gene silencing at the single-cell level for oligonucleotide-based therapy.
authors.affiliation: NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, Ohio, USA.|College of Pharmacy, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Keliang
authors.initials: K
authors.lastname: Gao
authors.affiliation: NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, Ohio, USA.|Molecular Cellular and Developmental Biology, The Ohio State University, Columbus, Ohio, USA.|Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Huang
authors.affiliation: NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, Ohio, USA.|Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Chi-Ling
authors.initials: CL
authors.lastname: Chiang
authors.affiliation: NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Xinmei
authors.initials: X
authors.lastname: Wang
authors.affiliation: NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Lingqian
authors.initials: L
authors.lastname: Chang
authors.affiliation: NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Pouyan
authors.initials: P
authors.lastname: Boukany
authors.affiliation: Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Guido
authors.initials: G
authors.lastname: Marcucci
authors.affiliation: College of Pharmacy, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Lee
authors.affiliation: NSF Nanoscale Science and Engineering Center for Affordable Nanoengineering of Polymeric Biomedical Devices, The Ohio State University, Columbus, Ohio, USA.|Molecular Cellular and Developmental Biology, The Ohio State University, Columbus, Ohio, USA.|Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Ly James
authors.initials: LJ
authors.lastname: Lee
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018274:Electroporation; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000628:therapy; D009154:Mutation; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000037:antagonists & inhibitors*; D034741:RNA, Small Interfering / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D059010:Single-Cell Analysis / Q000379:methods*; D020080:Tandem Repeat Sequences; D014162:Transfection; D051941:fms-Like Tyrosine Kinase 3 / Q000235:genetics*
chemical_list: C579095:MCL1 protein, human; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D034741:RNA, Small Interfering; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3