title: Bolaamphiphiles as carriers for siRNA delivery: From chemical syntheses to practical applications.
abstract: In this study we have investigated a new class of cationic lipids--"bolaamphiphiles" or "bolas"--for their ability to efficiently deliver small interfering RNAs (siRNAs) to cancer cells. The bolas of this study consist of a hydrophobic chain with one or more positively charged head groups at each end. Recently, we reported that micelles of the bolas GLH-19 and GLH-20 (derived from vernonia oil) efficiently deliver siRNAs, while having relatively low toxicities in vitro and in vivo. Our previous studies validated that; bolaamphiphiles can be designed to vary the magnitude of siRNA shielding, its delivery, and its subsequent release. To further understand the structural features of bolas critical for siRNAs delivery, new structurally related bolas (GLH-58 and GLH-60) were designed and synthesized from jojoba oil. Both bolas have similar hydrophobic domains and contain either one, in GLH-58, or two, in GLH-60 positively charged head groups at each end of the hydrophobic core. We have computationally predicted and experimentally validated that GLH-58 formed more stable nano sized micelles than GLH-60 and performed significantly better in comparison to GLH-60 for siRNA delivery. GLH-58/siRNA complexes demonstrated better efficiency in silencing the expression of the GFP gene in human breast cancer cells at concentrations of 5μg/mL, well below the toxic dose. Moreover, delivery of multiple different siRNAs targeting the HIV genome demonstrated further inhibition of virus production.
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA; Department of Inorganic and Physical Chemistry, Indian Institute of Science, Bangalore, 560012, India.
authors.forename: Kshitij
authors.initials: K
authors.lastname: Gupta
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA; Department of Chemistry, University of North Carolina at Charlotte, 9201 University City Boulevard, Charlotte, NC 28223, USA.
authors.forename: Kirill A
authors.initials: KA
authors.lastname: Afonin
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA; Basic Science Program, Leidos Biomedical Research Inc., Frederick National Laboratory, Frederick, MD 21702, USA.
authors.forename: Mathias
authors.initials: M
authors.lastname: Viard
authors.affiliation: Department of Chemistry, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Virginia
authors.initials: V
authors.lastname: Herrero
authors.affiliation: Basic Science Program, Leidos Biomedical Research Inc., Frederick National Laboratory, Frederick, MD 21702, USA.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Kasprzak
authors.affiliation: HIV Drug Resistance Program, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Kagiampakis
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Taejin
authors.initials: T
authors.lastname: Kim
authors.affiliation: National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
authors.forename: Alexey Y
authors.initials: AY
authors.lastname: Koyfman
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Anu
authors.initials: A
authors.lastname: Puri
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Marissa
authors.initials: M
authors.lastname: Stepler
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Alison
authors.initials: A
authors.lastname: Sappe
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Vineet N
authors.initials: VN
authors.lastname: KewalRamani
authors.affiliation: Department of Chemistry, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Sarina
authors.initials: S
authors.lastname: Grinberg
authors.affiliation: Department of Biotechnology, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Charles
authors.initials: C
authors.lastname: Linder
authors.affiliation: Department of Clinical Biochemistry and Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer Sheva, Israel.
authors.forename: Eliahu
authors.initials: E
authors.lastname: Heldman
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Blumenthal
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA. Electronic address: shapirbr@mail.nih.gov.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Shapiro
mesh_terms: D002460:Cell Line; D045744:Cell Line, Tumor; D004337:Drug Carriers / Q000737:chemistry*; D005663:Furans / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006678:HIV / Q000235:genetics; D006801:Humans; D008823:Micelles; D056004:Molecular Dynamics Simulation; D011728:Pyridones / Q000737:chemistry*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection / Q000379:methods
chemical_list: D004337:Drug Carriers; D005663:Furans; D008823:Micelles; D011728:Pyridones; D034741:RNA, Small Interfering; C543575:bolaamphiphile; D049452:Green Fluorescent Proteins
keywords: Bolaamphiphiles; Cryo-EM; FRET; Molecular dynamics simulations; RNAi induced gene silencing; siRNA drug delivery