Expanding RNAi therapeutics to extrahepatic tissues with lipophilic conjugates.
Journal
Nature biotechnology
ISSN: 1546-1696
Titre abrégé: Nat Biotechnol
Pays: United States
ID NLM: 9604648
Informations de publication
Date de publication:
10 2022
10 2022
Historique:
received:
28
09
2021
accepted:
22
04
2022
pubmed:
3
6
2022
medline:
12
10
2022
entrez:
2
6
2022
Statut:
ppublish
Résumé
Therapeutics based on short interfering RNAs (siRNAs) delivered to hepatocytes have been approved, but new delivery solutions are needed to target additional organs. Here we show that conjugation of 2'-O-hexadecyl (C16) to siRNAs enables safe, potent and durable silencing in the central nervous system (CNS), eye and lung in rodents and non-human primates with broad cell type specificity. We show that intrathecally or intracerebroventricularly delivered C16-siRNAs were active across CNS regions and cell types, with sustained RNA interference (RNAi) activity for at least 3 months. Similarly, intravitreal administration to the eye or intranasal administration to the lung resulted in a potent and durable knockdown. The preclinical efficacy of an siRNA targeting the amyloid precursor protein was evaluated through intracerebroventricular dosing in a mouse model of Alzheimer's disease, resulting in amelioration of physiological and behavioral deficits. Altogether, C16 conjugation of siRNAs has the potential for safe therapeutic silencing of target genes outside the liver with infrequent dosing.
Identifiants
pubmed: 35654979
doi: 10.1038/s41587-022-01334-x
pii: 10.1038/s41587-022-01334-x
doi:
Substances chimiques
Amyloid beta-Protein Precursor
0
RNA, Small Interfering
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1500-1508Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.
Références
Setten, R. L., Rossi, J. J. & Han, S. P. The current state and future directions of RNAi-based therapeutics. Nat. Rev. Drug Discov. 18, 421–446 (2019).
pubmed: 30846871
doi: 10.1038/s41573-019-0017-4
Adams, D. et al. Patisiran, an RNAi therapeutic, for hereditary transthyretin amyloidosis. N. Engl. J. Med. 379, 11–21 (2018).
pubmed: 29972753
doi: 10.1056/NEJMoa1716153
Kristen, A. V. et al. Patisiran, an RNAi therapeutic for the treatment of hereditary transthyretin-mediated amyloidosis. Neurodegener. Dis. Manag. 9, 5–23 (2019).
pubmed: 30480471
doi: 10.2217/nmt-2018-0033
Balwani, M. et al. Phase 3 trial of RNAi therapeutic givosiran for acute intermittent porphyria. N. Engl. J. Med. 382, 2289–2301 (2020).
pubmed: 32521132
doi: 10.1056/NEJMoa1913147
Garrelfs, S. F. et al. Lumasiran, an RNAi therapeutic for primary hyperoxaluria type 1. N. Engl. J. Med. 384, 1216–1226 (2021).
pubmed: 33789010
doi: 10.1056/NEJMoa2021712
Raal, F. J. et al. Inclisiran for the treatment of heterozygous familial hypercholesterolemia. N. Engl. J. Med. 382, 1520–1530 (2020).
pubmed: 32197277
doi: 10.1056/NEJMoa1913805
Ray, K. K. et al. Inclisiran in patients at high cardiovascular risk with elevated LDL cholesterol. N. Engl. J. Med. 376, 1430–1440 (2017).
pubmed: 28306389
doi: 10.1056/NEJMoa1615758
Ray, K. K. et al. Two phase 3 trials of inclisiran in patients with elevated LDL cholesterol. N. Engl. J. Med. 382, 1507–1519 (2020).
pubmed: 32187462
doi: 10.1056/NEJMoa1912387
Foster, D. J. et al. Advanced siRNA designs further improve in vivo performance of GalNAc-siRNA conjugates. Mol. Ther. 26, 708–717 (2018).
pubmed: 29456020
pmcid: 5910670
doi: 10.1016/j.ymthe.2017.12.021
Janas, M. M. et al. Selection of GalNAc-conjugated siRNAs with limited off-target-driven rat hepatotoxicity. Nat. Commun. 9, 723 (2018).
pubmed: 29459660
pmcid: 5818625
doi: 10.1038/s41467-018-02989-4
Nair, J. K. et al. Impact of enhanced metabolic stability on pharmacokinetics and pharmacodynamics of GalNAc-siRNA conjugates. Nucleic Acids Res. 45, 10969–10977 (2017).
pubmed: 28981809
pmcid: 5737438
doi: 10.1093/nar/gkx818
Nair, J. K. et al. Multivalent N-acetylgalactosamine-conjugated siRNA localizes in hepatocytes and elicits robust RNAi-mediated gene silencing. J. Am. Chem. Soc. 136, 16958–16961 (2014).
pubmed: 25434769
doi: 10.1021/ja505986a
Alterman, J. F. et al. A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system. Nat. Biotechnol. 37, 884–894 (2019).
pubmed: 31375812
pmcid: 6879195
doi: 10.1038/s41587-019-0205-0
Zhou, Y. et al. Blood–brain barrier-penetrating siRNA nanomedicine for Alzheimer’s disease therapy. Sci. Adv. 6, eabc7031 (2020).
pubmed: 33036977
pmcid: 7546706
doi: 10.1126/sciadv.abc7031
Gregory, J. V. et al. Systemic brain tumor delivery of synthetic protein nanoparticles for glioblastoma therapy. Nat. Commun. 11, 5687 (2020).
pubmed: 33173024
pmcid: 7655867
doi: 10.1038/s41467-020-19225-7
Eyford, B. A. et al. A nanomule peptide carrier delivers siRNA across the intact blood–brain barrier to attenuate ischemic stroke. Front. Mol. Biosci. 8, 611367 (2021).
pubmed: 33869275
pmcid: 8044710
doi: 10.3389/fmolb.2021.611367
Nagata, T. et al. Cholesterol-functionalized DNA/RNA heteroduplexes cross the blood–brain barrier and knock down genes in the rodent CNS. Nat. Biotechnol. 39, 1529–1536 (2021).
Gupta, A., Kafetzis, K. N., Tagalakis, A. D. & Yu-Wai-Man, C. RNA therapeutics in ophthalmology—translation to clinical trials. Exp. Eye Res. 205, 108482 (2021).
pubmed: 33548256
doi: 10.1016/j.exer.2021.108482
DeVincenzo, J. et al. A randomized, double-blind, placebo-controlled study of an RNAi-based therapy directed against respiratory syncytial virus. Proc. Natl Acad. Sci. USA 107, 8800–8805 (2010).
pubmed: 20421463
pmcid: 2889365
doi: 10.1073/pnas.0912186107
Chappell, A. E. et al. Mechanisms of palmitic acid-conjugated antisense oligonucleotide distribution in mice. Nucleic Acids Res. 48, 4382–4395 (2020).
pubmed: 32182359
pmcid: 7192618
doi: 10.1093/nar/gkaa164
Chen, Q. et al. Lipophilic siRNAs mediate efficient gene silencing in oligodendrocytes with direct CNS delivery. J. Control. Release 144, 227–232 (2010).
pubmed: 20170694
doi: 10.1016/j.jconrel.2010.02.011
Manoharan, M. Oligonucleotide conjugates as potential antisense drugs with improved uptake, biodistribution, targeted delivery, and mechanism of action. Antisense Nucleic Acid Drug Dev. 12, 103–128 (2002).
pubmed: 12074364
doi: 10.1089/108729002760070849
Wolfrum, C. et al. Mechanisms and optimization of in vivo delivery of lipophilic siRNAs. Nat. Biotechnol. 25, 1149–1157 (2007).
pubmed: 17873866
doi: 10.1038/nbt1339
Schlegel, M. K. et al. Chirality dependent potency enhancement and structural impact of glycol nucleic acid modification on siRNA. J. Am. Chem. Soc. 139, 8537–8546 (2017).
pubmed: 28570818
doi: 10.1021/jacs.7b02694
Parmar, R. et al. 5′-(E)-vinylphosphonate: a stable phosphate mimic can improve the RNAi activity of siRNA-GalNAc conjugates. ChemBioChem 17, 985–989 (2016).
pubmed: 27121751
doi: 10.1002/cbic.201600130
Sullivan, J. M. et al. Convective forces increase rostral delivery of intrathecal radiotracers and antisense oligonucleotides in the cynomolgus monkey nervous system. J. Transl. Med. 18, 309 (2020).
pubmed: 32771027
pmcid: 7414676
doi: 10.1186/s12967-020-02461-2
Wilcock, D. M. et al. Progression of amyloid pathology to Alzheimer’s disease pathology in an amyloid precursor protein transgenic mouse model by removal of nitric oxide synthase 2. J. Neurosci. 28, 1537–1545 (2008).
pubmed: 18272675
pmcid: 2621082
doi: 10.1523/JNEUROSCI.5066-07.2008
Colton, C. A. et al. mNos2 deletion and human NOS2 replacement in Alzheimer disease models. J. Neuropathol. Exp. Neurol. 73, 752–769 (2014).
pubmed: 25003233
doi: 10.1097/NEN.0000000000000094
Kan, M. J. et al. Arginine deprivation and immune suppression in a mouse model of Alzheimer’s disease. J. Neurosci. 35, 5969–5982 (2015).
pubmed: 25878270
pmcid: 4397598
doi: 10.1523/JNEUROSCI.4668-14.2015
Hall, H. et al. Magnetic resonance spectroscopic methods for the assessment of metabolic functions in the diseased brain. Curr. Top. Behav. Neurosci. 11, 169–198 (2012).
pubmed: 22076698
doi: 10.1007/7854_2011_166
Su, L. et al. Whole-brain patterns of
pubmed: 27576166
pmcid: 5022086
doi: 10.1038/tp.2016.140
Wang, H. et al. Magnetic resonance spectroscopy in Alzheimer’s disease: systematic review and meta-analysis. J. Alzheimers Dis. 46, 1049–1070 (2015).
pubmed: 26402632
doi: 10.3233/JAD-143225
Janas, M. M. et al. Safety evaluation of 2′-deoxy-2′-fluoro nucleotides in GalNAc-siRNA conjugates. Nucleic Acids Res. 47, 3306–3320 (2019).
pubmed: 30820542
pmcid: 6468299
doi: 10.1093/nar/gkz140
Osborn, M. F. & Khvorova, A. Improving siRNA delivery in vivo through lipid conjugation. Nucleic Acid Ther. 28, 128–136 (2018).
pubmed: 29746209
pmcid: 5994667
doi: 10.1089/nat.2018.0725
Biscans, A. et al. Diverse lipid conjugates for functional extra-hepatic siRNA delivery in vivo. Nucleic Acids Res. 47, 1082–1096 (2019).
pubmed: 30544191
doi: 10.1093/nar/gky1239
Raouane, M., Desmaele, D., Urbinati, G., Massaad-Massade, L. & Couvreur, P. Lipid conjugated oligonucleotides: a useful strategy for delivery. Bioconjug. Chem. 23, 1091–1104 (2012).
pubmed: 22372953
doi: 10.1021/bc200422w
Kubo, T. et al. Palmitic acid-conjugated 21-nucleotide siRNA enhances gene-silencing activity. Mol. Pharm. 8, 2193–2203 (2011).
pubmed: 21985606
doi: 10.1021/mp200250f
Byrne, M. et al. Novel hydrophobically modified asymmetric RNAi compounds (sd-rxRNA) demonstrate robust efficacy in the eye. J. Ocul. Pharmacol. Ther. 29, 855–864 (2013).
pubmed: 24180627
doi: 10.1089/jop.2013.0148
DiFiglia, M. et al. Therapeutic silencing of mutant huntingtin with siRNA attenuates striatal and cortical neuropathology and behavioral deficits. Proc. Natl Acad. Sci. USA 104, 17204–17209 (2007).
pubmed: 17940007
pmcid: 2040405
doi: 10.1073/pnas.0708285104
Alterman, J. F. et al. Hydrophobically modified siRNAs silence Huntingtin mRNA in primary neurons and mouse brain. Mol. Ther. Nucleic Acids 4, e266 (2015).
pubmed: 26623938
pmcid: 5014532
doi: 10.1038/mtna.2015.38
Nikan, M. et al. Docosahexaenoic acid conjugation enhances distribution and safety of siRNA upon local administration in mouse brain. Mol. Ther. Nucleic Acids 5, e344 (2016).
pubmed: 27504598
pmcid: 5023396
doi: 10.1038/mtna.2016.50
Beaucage, S. L. Solid-phase synthesis of siRNA oligonucleotides. Curr. Opin. Drug Discov. Dev. 11, 203–216 (2008).
Zhang, L., Peritz, A. & Meggers, E. A simple glycol nucleic acid. J. Am. Chem. Soc. 127, 4174–4175 (2005).
pubmed: 15783191
doi: 10.1021/ja042564z
Zhang, L., Peritz, A. E., Carroll, P. J. & Meggers, E. Synthesis of glycol nucleic acids. Synthesis 2006, 645–653 (2006).
doi: 10.1055/s-2006-926313
Schlegel, M. K. & Meggers, E. Improved phosphoramidite building blocks for the synthesis of the simplified nucleic acid GNA. J. Org. Chem. 74, 4615–4618 (2009).
pubmed: 19441799
doi: 10.1021/jo900365a
O’Shea, J. et al. An efficient deprotection method for 5′-[O,O-bis(pivaloyloxymethyl)]-(E)-vinylphosphonate containing oligonucleotides. Tetrahedron 74, 6182–6186 (2018).
doi: 10.1016/j.tet.2018.09.008
Kohonen, P. et al. A transcriptomics data-driven gene space accurately predicts liver cytopathology and drug-induced liver injury. Nat. Commun. 8, 15932 (2017).
pubmed: 28671182
pmcid: 5500850
doi: 10.1038/ncomms15932
Davis, J. et al. Early-onset and robust cerebral microvascular accumulation of amyloid β-protein in transgenic mice expressing low levels of a vasculotropic Dutch/Iowa mutant form of amyloid β-protein precursor. J. Biol. Chem. 279, 20296–20306 (2004).
pubmed: 14985348
doi: 10.1074/jbc.M312946200
Laubach, V. E., Foley, P. L., Shockey, K. S., Tribble, C. G. & Kron, I. L. Protective roles of nitric oxide and testosterone in endotoxemia: evidence from NOS-2-deficient mice. Am. J. Physiol. 275, H2211–H2218 (1998).
pubmed: 9843821
Li, J. et al. Discovery of a novel deaminated metabolite of a single-stranded oligonucleotide in vivo by mass spectrometry. Bioanalysis 11, 1955–1965 (2019).
pubmed: 31829055
doi: 10.4155/bio-2019-0118
Liu, J. et al. Oligonucleotide quantification and metabolite profiling by high-resolution and accurate mass spectrometry. Bioanalysis 11, 1967–1980 (2019).
pubmed: 31829056
doi: 10.4155/bio-2019-0137
Bolon, B. et al. STP position paper: recommended practices for sampling and processing the nervous system (brain, spinal cord, nerve, and eye) during nonclinical general toxicity studies. Toxicol. Pathol. 41, 1028–1048 (2013).
pubmed: 23475559
doi: 10.1177/0192623312474865