The interactions of amphiphilic antisense oligonucleotides with serum proteins and their effects on in vitro silencing activity
Source abstract
Abstract
Antisense oligonucleotides (AONs) are a class of compounds with high therapeutic potential. One of the challenges facing this platform is the development of effective techniques to achieve cellular delivery. AON conjugates, in which traditional AONs are attached to certain biomolecules, can exhibit improved intracellular bioavailability in the absence of delivery systems. In this study, the lipophilic moieties docosahexaenoic acid, cholesterol, and docosanoic acid (DSA) were conjugated to various phosphorothioated DNA and chemically-modified 2'-fluoro-arabinonucleic acid AONs via an amino-hexanol-linker added to the 5'-end of the molecule. The gene silencing potential of these compounds was evaluated in vitro in the absence or presence of a transfecting agent (polyion complex micelle). Incubation with sub-micromolar concentration of DSA-conjugates could, in the absence of serum proteins, downregulate more than 60% of the targeted mRNA under carrier-free and carrier-loaded delivery methods. Gene silencing activity of carrier-free DSA-conjugates was, however, decreased in a dose-dependent fashion by adding albumin in the transfection medium. Supplementing the medium with free fatty acid prevented the interaction of the DSA-conjugate with albumin, and restored its silencing activity. These findings suggest that strategies aiming at preventing the association of hydrophobized AONs to serum proteins at the site of action may improve their activity.
Source: europepmc · Quality: candidate unverified · Source record ↗
Research context
OpenAlex topics: RNA Interference and Gene Delivery · Advanced biosensing and bioanalysis techniques · DNA and Nucleic Acid Chemistry
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
References (63)
Show source-provided references
1. Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide10.1073/pnas.75.1.280 ↗
2. Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide10.1073/pnas.75.1.285 ↗
3. Interactions of antisense DNA oligonucleotide analogs with phospholipid membranes (liposomes)10.1093/nar/19.20.5551 ↗
4. Antisense oligonucleotides: promise and reality10.1146/annurev.pharmtox.41.1.403 ↗
5. Physicochemical properties and nuclease resistance of antisense-oligodeoxynucleotides entrapped in the core of polyion complex micelles composed of poly(ethylene glycol)-poly(L-lysine) block copolymers10.1016/S0928-0987(00)00205-0 ↗
6. Oligonucleotide conjugates as potential antisense drugs with improved uptake, biodistribution, targeted delivery, and mechanism of action10.1089/108729002760070849 ↗
7. A review of the issues in the pharmacokinetics and toxicology of phosphorothioate antisense oligonucleotides10.1016/S0167-4781(99)00140-2 ↗
8. Antisense oligonucleotides: from design to therapeutic application10.1111/j.1440-1681.2006.04403.x ↗
9. Hybrids of RNA and arabinonucleic acids (ANA and 2′F-ANA) are substrates of ribonuclease10.1021/ja982325+ ↗
10. 2′-Deoxy-2′-fluoro-β-d-arabinonucleic acid (2′F-ANA) modified oligonucleotides (ON) effect highly efficient, and persistent, gene silencing10.1093/nar/gkj455 ↗
11. Studies on the hydrolytic stability of 2′-fluoroarabinonucleic acid (2′F-ANA)10.1039/b900443b ↗
12. Potent gene-specific inhibitory properties of mixed-backbone antisense oligonucleotides comprised of 2′-deoxy-2′-fluoro-d-arabinose and 2′-deoxyribose nucleotides10.1021/bi0115075 ↗
13. RNA targeting therapeutics: molecular mechanisms of antisense oligonucleotides as a therapeutic platform10.1146/annurev.pharmtox.010909.105654 ↗
14. Potential roles of antisense oligonucleotides in cancer therapy. The example of Bcl-2 antisense oligonucleotides10.1016/S0939-6411(02)00060-7 ↗
15. Phosphorothioate antisense oligodeoxynucleotides: questions of specificity10.1016/0167-7799(96)20006-X ↗
16. Pharmacokinetics, biodistribution, and stability of oligodeoxynucleotide phosphorothioates in mice10.1073/pnas.88.17.7595 ↗
17. Biodistribution and metabolism of internally 3H-labeled oligonucleotides. I. Comparison of a phosphodiester and a phosphorothioate10.1016/S0026-895X(25)10208-3 ↗
18. Viral vector-mediated RNA interference10.1016/j.coph.2010.06.007 ↗
19. Intracellular distribution and mechanism of delivery of oligonucleotides mediated by cationic lipids10.1023/A:1016026101195 ↗
20. Polyethylenimine-based non-viral gene delivery systems10.1016/j.ejpb.2004.11.011 ↗
21. Characterization of polyion complex micelles designed to address the challenges of oligonucleotide delivery10.1007/s11095-008-9591-6 ↗
22. Delivery of nucleic acids through the controlled disassembly of multifunctional nanocomplexes10.1002/adfm.200901139 ↗
23. Block copolymer micelles for delivery of gene and related compounds10.1016/S0169-409X(02)00017-0 ↗
24. Polyalkylcyanoacrylate nanoparticles as polymeric carriers for antisense oligonucleotides10.1023/A:1015871809313 ↗
25. Toxicity of cationic lipids and cationic polymers in gene delivery10.1016/j.jconrel.2006.04.014 ↗
26. Drug delivery trends in clinical trials and translational medicine: challenges and opportunities in the delivery of nucleic acid-based therapeutics10.1002/jps.22243 ↗
27. Intracellular delivery of an antisense oligonucleotide via endocytosis of a G protein-coupled receptor10.1093/nar/gkq534 ↗
28. Antisense inhibition of P-glycoprotein expression using peptide–oligonucleotide conjugates10.1016/S0006-2952(00)00310-5 ↗
29. Pharmacokinetic properties of several novel oligonucleotide analogs in mice10.1016/S0022-3565(25)12951-0 ↗
30. Modification of antisense phosphodiester oligodeoxynucleotides by a 5′ cholesteryl moiety increases cellular association and improves efficacy10.1073/pnas.90.3.1048 ↗
31. Lipid conjugated oligonucleotides: a useful strategy for delivery10.1021/bc200422w ↗
32. Carrier-free cellular uptake and the gene-silencing activity of the lipophilic siRNAs is strongly affected by the length of the linker between siRNA and lipophilic group10.1093/nar/gkr1002 ↗
33. Mechanisms and optimization of in vivo delivery of lipophilic siRNAs10.1038/nbt1339 ↗
34. Tumor targeting by conjugation of DHA to paclitaxel10.1016/S0168-3659(01)00321-2 ↗
35. SiRNA nanocarriers based on methacrylic acid copolymers10.1016/j.jconrel.2010.12.012 ↗
36. Preparation of polyion complex micelles from poly(ethylene glycol)-block-polyions10.1016/j.jconrel.2011.07.027 ↗
37. 2′-Deoxy-2′-fluoro-β-d-arabinonucleosides and oligonucleotides (2′F-ANA): synthesis and physicochemical studies10.1093/nar/28.18.3625 ↗
38. Oligoribonucleotides with 2′-O-(tert-butyldimethylsilyl) groups
39. Off-target effects related to the phosphorothioate modification of nucleic acids10.1002/cmdc.201000156 ↗
40. Gene profiling study of G3139- and Bcl-2-targeting siRNAs identifies a unique G3139 molecular signature10.1038/sj.cgt.7700901 ↗
41. Oligoribonucleotide synthesis: the silyl-phosphoramidite method
42. Oligonucleotides comprised of alternating 2′-deoxy-2′-fluoro-β-d-arabinonucleosides and d-2′-deoxyribonucleosides (2′F-ANA/DNA ‘altimers’) induce efficient RNA cleavage mediated by RNase H10.1016/S0960-894X(02)00439-0 ↗
43. Locating high-affinity fatty acid-binding sites on albumin by x-ray crystallography and NMR spectroscopy10.1073/pnas.0506440102 ↗
44. Crystallographic analysis reveals common modes of binding of medium and long-chain fatty acids to human serum albumin10.1006/jmbi.2000.4158 ↗
45. Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs10.1038/nature03121 ↗
46. Synthesis and physical properties of anti-HIV antisense oligonucleotides bearing terminal lipophilic groups10.1093/nar/20.13.3411 ↗
47. Quantitative structure-property relationship (QSPR) study of n-octanol-water partition coefficients (logPo/w) of fatty acids using multiple linear regression (MLR)
48. Comparison of QSPR properties for cholesterol and cholestanol oligomeric association forms
49. Synthesis and preliminary antitumor activity evaluation of a DHA and doxorubicin conjugate10.1016/j.bmcl.2006.02.066 ↗
50. Docosahexaenoic acid enhances the efficacy of docetaxel in prostate cancer cells by modulation of apoptosis: the role of genes associated with the NF-κB pathway10.1002/pros.20830 ↗
51. A phase II open-label study of DHA-paclitaxel (taxoprexin) by 2-h intravenous infusion in previously untreated patients with locally advanced or metastatic gastric or oesophageal adenocarcinoma10.1007/s00280-007-0486-8 ↗
52. DHA-paclitaxel (taxoprexin) as first-line treatment in patients with stage IIIB or IV non-small cell lung cancer: report of a phase II open-label multicenter trial10.1097/01243894-200611000-00011 ↗
53. GPR120 is an omega-3 fatty acid receptor mediating potent anti-inflammatory and insulin-sensitizing effects10.1016/j.cell.2010.07.041 ↗
54. Enhanced extravasation, stability and in vivo cardiac gene silencing via in situ siRNA-albumin conjugation10.1021/mp2002522 ↗
55. A new concept of cellular uptake and intracellular trafficking of long-chain fatty acids10.1007/s11745-001-0809-2 ↗
56. Fatty acid transport across the cell membrane: regulation by fatty acid transporters10.1016/j.plefa.2010.02.029 ↗
57. The role of the protein-binding on the mode of drug action as well the interactions with other drugs10.1007/BF03190876 ↗
58. What is the true clinical significance of plasma protein binding displacement interactions?10.2165/00002018-199512040-00001 ↗
59. Influence of fatty acids on the binding of warfarin and phenprocoumon to human serum albumin with relation to anticoagulant therapy10.1111/j.2042-7158.1996.tb03990.x ↗
60. Multiple fatty acid binding to albumin in human blood plasma10.1111/j.1432-1033.1990.tb15495.x ↗
61. Fatty acid distribution in systems modeling the normal and diabetic human circulation. A 13C nuclear magnetic resonance study10.1172/JCI115149 ↗
62. Lipoprotein-inspired nanoparticles for cancer theranostics10.1021/ar200017e ↗
63. Design and development of polymers for gene delivery10.1038/nrd1775 ↗