Heteroduplex oligonucleotide technology boosts oligonucleotide splice switching activity of morpholino oligomers in a Duchenne muscular dystrophy mouse model.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
26 Sep 2024
26 Sep 2024
Historique:
received:
02
03
2023
accepted:
23
04
2024
medline:
27
9
2024
pubmed:
27
9
2024
entrez:
26
9
2024
Statut:
epublish
Résumé
The approval of splice-switching oligonucleotides with phosphorodiamidate morpholino oligomers (PMOs) for treating Duchenne muscular dystrophy (DMD) has advanced the field of oligonucleotide therapy. Despite this progress, PMOs encounter challenges such as poor tissue uptake, particularly in the heart, diaphragm, and central nervous system (CNS), thereby affecting patient's prognosis and quality of life. To address these limitations, we have developed a PMOs-based heteroduplex oligonucleotide (HDO) technology. This innovation involves a lipid-ligand-conjugated complementary strand hybridized with PMOs, significantly enhancing delivery to key tissues in mdx mice, normalizing motor functions, muscle pathology, and serum creatine kinase by restoring internal deleted dystrophin expression. Additionally, PMOs-based HDOs normalized cardiac and CNS abnormalities without adverse effects. Our technology increases serum albumin binding to PMOs and improves blood retention and cellular uptake. Here we show that PMOs-based HDOs address the limitations in oligonucleotide therapy for DMD and offer a promising approach for diseases amenable to exon-skipping therapy.
Identifiants
pubmed: 39327422
doi: 10.1038/s41467-024-48204-5
pii: 10.1038/s41467-024-48204-5
doi:
Substances chimiques
Morpholinos
0
Dystrophin
0
Oligonucleotides
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
7530Subventions
Organisme : Japan Agency for Medical Research and Development (AMED)
ID : 23am0401006h0005
Organisme : MEXT | Japan Society for the Promotion of Science (JSPS)
ID : 19H01016
Organisme : MEXT | Japan Society for the Promotion of Science (JSPS)
ID : 19H01016
Organisme : MEXT | Japan Society for the Promotion of Science (JSPS)
ID : 22H00440
Organisme : MEXT | Japan Society for the Promotion of Science (JSPS)
ID : 16H05221
Informations de copyright
© 2024. The Author(s).
Références
Drousiotou, A. et al. Neonatal screening for Duchenne muscular dystrophy: a novel semiquantitative application of the bioluminescence test for creatine kinase in a pilot national program in Cyprus. Genet Test. 2, 55–60 (1998).
pubmed: 10464597
doi: 10.1089/gte.1998.2.55
Bushby, K. et al. Diagnosis and management of Duchenne muscular dystrophy, part 1: diagnosis, and pharmacological and psychosocial management. Lancet Neurol. 9, 77–93 (2010).
pubmed: 19945913
doi: 10.1016/S1474-4422(09)70271-6
Mah, J. K. Current and emerging treatment strategies for Duchenne muscular dystrophy. Neuropsychiatr. Dis. Treat. 12, 1795–1807 (2016).
pubmed: 27524897
pmcid: 4966503
doi: 10.2147/NDT.S93873
Boland, B. J., Silbert, P. L., Groover, R. V., Wollan, P. C. & Silverstein, M. D. Skeletal, cardiac, and smooth muscle failure in Duchenne muscular dystrophy. Pediatr. Neurol. 14, 7–12 (1996).
pubmed: 8652023
doi: 10.1016/0887-8994(95)00251-0
Blake, D. J., Weir, A., Newey, S. E. & Davies, K. E. Function and genetics of dystrophin and dystrophin-related proteins in muscle. Physiol. Rev. 82, 291–329 (2002).
pubmed: 11917091
doi: 10.1152/physrev.00028.2001
Hoffman, E. P. & McNally, E. M. Exon-skipping therapy: a roadblock, detour, or bump in the road? Sci. Transl. Med. 6, 230fs214–230fs214 (2014).
doi: 10.1126/scitranslmed.3008873
Watanabe, N. et al. NS-065/NCNP-01: an antisense oligonucleotide for potential treatment of exon 53 skipping in Duchenne muscular dystrophy. Mol. Ther. Nucleic Acids 13, 442–449 (2018).
pubmed: 30388618
pmcid: 6202794
doi: 10.1016/j.omtn.2018.09.017
Komaki, H. et al. Systemic administration of the antisense oligonucleotide NS-065/NCNP-01 for skipping of exon 53 in patients with Duchenne muscular dystrophy. Sci. Transl. Med. 10, eaan0713 (2018).
pubmed: 29669851
doi: 10.1126/scitranslmed.aan0713
Clemens, P. R. et al. Safety, tolerability, and efficacy of viltolarsen in boys With Duchenne muscular dystrophy amenable to exon 53 skipping: a phase 2 randomized clinical trial. JAMA Neurol. 77, 982–991 (2020).
pubmed: 32453377
doi: 10.1001/jamaneurol.2020.1264
Mendell, J. R. et al. Eteplirsen for the treatment of Duchenne muscular dystrophy. Ann. Neurol. 74, 637–647 (2013).
pubmed: 23907995
doi: 10.1002/ana.23982
Frank, D. E. et al. Increased dystrophin production with golodirsen in patients with Duchenne muscular dystrophy. Neurology 94, e2270–e2282 (2020).
pubmed: 32139505
pmcid: 7357297
doi: 10.1212/WNL.0000000000009233
Wagner, K. R. et al. Safety, tolerability, and pharmacokinetics of casimersen in patients with Duchenne muscular dystrophy amenable to exon 45 skipping: a randomized, double-blind, placebo-controlled, dose-titration trial. Muscle Nerve 64, 285–292 (2021).
pubmed: 34105177
pmcid: 9290993
doi: 10.1002/mus.27347
Komaki, H. et al. Viltolarsen in Japanese Duchenne muscular dystrophy patients: a phase 1/2 study. Ann. Clin. Transl. Neurol. 7, 2393–2408 (2020).
pubmed: 33285037
pmcid: 7732240
doi: 10.1002/acn3.51235
van Putten, M. et al. Low dystrophin levels increase survival and improve muscle pathology and function in dystrophin/utrophin double-knockout mice. FASEB J. 27, 2484–2495 (2013).
pubmed: 23460734
pmcid: 3659351
doi: 10.1096/fj.12-224170
Wu, B. et al. Dose-dependent restoration of dystrophin expression in cardiac muscle of dystrophic mice by systemically delivered morpholino. Gene Ther. 17, 132–140 (2010).
pubmed: 19759562
doi: 10.1038/gt.2009.120
Wu, B. et al. One-year treatment of morpholino antisense oligomer improves skeletal and cardiac muscle functions in dystrophic mdx mice. Mol. Ther. 19, 576–583 (2011).
pubmed: 21179007
doi: 10.1038/mt.2010.288
Goyenvalle, A. et al. Functional correction in mouse models of muscular dystrophy using exon-skipping tricyclo-DNA oligomers. Nat. Med. 21, 270–275 (2015).
pubmed: 25642938
doi: 10.1038/nm.3765
Rae, M. G. & O’Malley, D. Cognitive dysfunction in Duchenne muscular dystrophy: a possible role for neuromodulatory immune molecules. J. Neurophysiol. 116, 1304–1315 (2016).
pubmed: 27385793
pmcid: 5023417
doi: 10.1152/jn.00248.2016
Kandasamy, P. et al. Control of backbone chemistry and chirality boost oligonucleotide splice switching activity. Nucleic Acids Res. 50, 5443–5466 (2022).
pubmed: 35061895
pmcid: 9178015
doi: 10.1093/nar/gkac018
Relizani, K. et al. Palmitic acid conjugation enhances potency of tricyclo-DNA splice switching oligonucleotides. Nucleic Acids Res. 50, 17–34 (2021).
pmcid: 8754652
doi: 10.1093/nar/gkab1199
Mullard, A. Antibody-oligonucleotide conjugates enter the clinic. Nat. Rev. Drug Discov. 21, 6–8 (2022).
pubmed: 34903879
doi: 10.1038/d41573-021-00213-5
Desjardins, C. A. et al. Enhanced exon skipping and prolonged dystrophin restoration achieved by TfR1-targeted delivery of antisense oligonucleotide using FORCE conjugation in mdx mice. Nucleic Acids Res. 50, 11401–11414 (2022).
Gan, L. et al. A cell-penetrating peptide enhances delivery and efficacy of phosphorodiamidate morpholino oligomers in mdx mice. Mol. Ther. Nucleic Acids 30, 17–27 (2022).
pubmed: 36189424
pmcid: 9483789
doi: 10.1016/j.omtn.2022.08.019
Klein, A. F. et al. Peptide-conjugated oligonucleotides evoke long-lasting myotonic dystrophy correction in patient-derived cells and mice. J. Clin. Invest. 129, 4739–4744 (2019).
pubmed: 31479430
pmcid: 6819114
doi: 10.1172/JCI128205
Li, X. et al. The endosomal escape vehicle platform enhances delivery of oligonucleotides in preclinical models of neuromuscular disorders. Mol. Ther. Nucleic Acids 33, 273–285 (2023).
pubmed: 37538053
pmcid: 10393622
doi: 10.1016/j.omtn.2023.06.022
Nishina, K. et al. DNA/RNA heteroduplex oligonucleotide for highly efficient gene silencing. Nat. Commun. 6, 7969 (2015).
pubmed: 26258894
pmcid: 4918363
doi: 10.1038/ncomms8969
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).
pubmed: 34385691
doi: 10.1038/s41587-021-00972-x
Ohyagi, M. et al. DNA/RNA heteroduplex oligonucleotide technology for regulating lymphocytes in vivo. Nat. Commun. 12, 7344 (2021).
pubmed: 34937876
pmcid: 8695577
doi: 10.1038/s41467-021-26902-8
Burki, U. et al. Development and application of an ultrasensitive hybridization-based ELISA method for the determination of peptide-conjugated phosphorodiamidate morpholino oligonucleotides. Nucleic Acid Ther. 25, 275–284 (2015).
pubmed: 26176274
pmcid: 4576940
doi: 10.1089/nat.2014.0528
Zhuang, P. et al. Combined microRNA and mRNA detection in mammalian retinas by in situ hybridization chain reaction. Sci. Rep. 10, 351 (2020).
pubmed: 31942002
pmcid: 6962165
doi: 10.1038/s41598-019-57194-0
Dirks, R. M. & Pierce, N. A. Triggered amplification by hybridization chain reaction. Proc. Natl Acad. Sci. USA 101, 15275–15278 (2004).
pubmed: 15492210
pmcid: 524468
doi: 10.1073/pnas.0407024101
Glesby, M. J., Rosenmann, E., Nylen, E. G. & Wrogemann, K. Serum CK, calcium, magnesium, and oxidative phosphorylation in mdx mouse muscular dystrophy. Muscle Nerve 11, 852–856 (1988).
pubmed: 3173410
doi: 10.1002/mus.880110809
Wells, D. J. et al. Human dystrophin expression corrects the myopathic phenotype in transgenic mdx mice. Hum. Mol. Genet 1, 35–40 (1992).
pubmed: 1301134
doi: 10.1093/hmg/1.1.35
Yin, H. et al. Functional rescue of dystrophin-deficient mdx mice by a chimeric peptide-PMO. Mol. Ther. 18, 1822–1829 (2010).
pubmed: 20700113
pmcid: 2951563
doi: 10.1038/mt.2010.151
Tanihata, J. et al. Truncated dystrophin ameliorates the dystrophic phenotype of mdx mice by reducing sarcolipin-mediated SERCA inhibition. Biochem. Biophys. Res. Commun. 505, 51–59 (2018).
pubmed: 30236982
doi: 10.1016/j.bbrc.2018.09.039
Qiao, C. et al. Myostatin propeptide gene delivery by adeno-associated virus serotype 8 vectors enhances muscle growth and ameliorates dystrophic phenotypes in mdx mice. Hum. Gene Ther. 19, 241–254 (2008).
pubmed: 18288893
doi: 10.1089/hum.2007.159
Burdi, R. et al. Multiple pathological events in exercised dystrophic mdx mice are targeted by pentoxifylline: outcome of a large array of in vivo and ex vivo tests. J. Appl Physiol. (1985) 106, 1311–1324 (2009).
pubmed: 19131478
doi: 10.1152/japplphysiol.90985.2008
Chu, V. et al. Electrocardiographic findings in mdx mice: a cardiac phenotype of Duchenne muscular dystrophy. Muscle Nerve 26, 513–519 (2002).
pubmed: 12362417
doi: 10.1002/mus.10223
Sadeghi, A., Doyle, A. D. & Johnson, B. D. Regulation of the cardiac L-type Ca2+ channel by the actin-binding proteins alpha-actinin and dystrophin. Am. J. Physiol. Cell Physiol. 282, C1502–C1511 (2002).
pubmed: 11997265
doi: 10.1152/ajpcell.00435.2001
Markham, L. W., Spicer, R. L. & Cripe, L. H. The heart in muscular dystrophy. Pediatr. Ann. 34, 531–535 (2005).
pubmed: 16092627
doi: 10.3928/0090-4481-20050701-10
Duan D. Challenges and opportunities in dystrophin-deficient cardiomyopathy gene therapy. Hum. Mol. Genet. 15, R253–R261 (2006).
Quinlan, J. G. et al. Evolution of the mdx mouse cardiomyopathy: physiological and morphological findings. Neuromuscul. Disord. 14, 491–496 (2004).
pubmed: 15336690
doi: 10.1016/j.nmd.2004.04.007
Au, C. G. et al. Increased connective tissue growth factor associated with cardiac fibrosis in the mdx mouse model of dystrophic cardiomyopathy. Int J. Exp. Pathol. 92, 57–65 (2011).
pubmed: 21121985
pmcid: 3052757
doi: 10.1111/j.1365-2613.2010.00750.x
Bostick, B. et al. Adeno-associated virus serotype-9 microdystrophin gene therapy ameliorates electrocardiographic abnormalities in mdx mice. Hum. Gene Ther. 19, 851–856 (2008).
pubmed: 18666839
pmcid: 2888653
doi: 10.1089/hum.2008.058
Li, Y. et al. Blunted cardiac beta-adrenergic response as an early indication of cardiac dysfunction in Duchenne muscular dystrophy. Cardiovasc Res. 103, 60–71 (2014).
pubmed: 24812281
pmcid: 4133593
doi: 10.1093/cvr/cvu119
Bridges, L. R. The association of cardiac muscle necrosis and inflammation with the degenerative and persistent myopathy of MDX mice. J. Neurol. Sci. 72, 147–157 (1986).
pubmed: 3711930
doi: 10.1016/0022-510X(86)90003-1
Meyers, T. A. & Townsend, D. Early right ventricular fibrosis and reduction in biventricular cardiac reserve in the dystrophin-deficient mdx heart. Am. J. Physiol. Heart Circ. Physiol. 308, H303–H315 (2015).
pubmed: 25485898
doi: 10.1152/ajpheart.00485.2014
Morroni, J. et al. Accelerating the mdx heart histo-pathology through physical exercise. Life (Basel) 11, 706 (2021).
pubmed: 34357078
Sekiguchi, M. et al. A deficit of brain dystrophin impairs specific amygdala GABAergic transmission and enhances defensive behaviour in mice. Brain 132, 124–135 (2009).
pubmed: 18927146
doi: 10.1093/brain/awn253
McMillan, H. J., Gregas, M., Darras, B. T. & Kang, P. B. Serum transaminase levels in boys with Duchenne and becker muscular dystrophy. Pediatrics 127, e132–e136 (2011).
pubmed: 21149430
doi: 10.1542/peds.2010-0929
Wang, L. et al. Ratio of creatine kinase to alanine aminotransferase as a biomarker of acute liver injury in dystrophinopathy. Dis. Markers 2018, 6484610 (2018).
pubmed: 30018675
pmcid: 6029496
doi: 10.1155/2018/6484610
Lai, Y. et al. Dystrophins carrying spectrin-like repeats 16 and 17 anchor nNOS to the sarcolemma and enhance exercise performance in a mouse model of muscular dystrophy. J. Clin. Invest 119, 624–635 (2009).
pubmed: 19229108
pmcid: 2648692
doi: 10.1172/JCI36612
Percival, J. M., Anderson, K. N., Gregorevic, P., Chamberlain, J. S. & Froehner, S. C. Functional deficits in nNOSmu-deficient skeletal muscle: myopathy in nNOS knockout mice. PLoS One 3, e3387 (2008).
pubmed: 18852886
pmcid: 2559862
doi: 10.1371/journal.pone.0003387
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
Østergaard, M. E. et al. Conjugation of hydrophobic moieties enhances potency of antisense oligonucleotides in the muscle of rodents and non-human primates. Nucleic Acids Res. 47, 6045–6058 (2019).
pubmed: 31076766
pmcid: 6614849
Ezzat, K. et al. Self-assembly into nanoparticles is essential for receptor mediated uptake of therapeutic antisense oligonucleotides. Nano Lett. 15, 4364–4373 (2015).
pubmed: 26042553
pmcid: 6415796
doi: 10.1021/acs.nanolett.5b00490
Fletcher, S. et al. Morpholino oligomer-mediated exon skipping averts the onset of dystrophic pathology in the mdx mouse. Mol. Ther. 15, 1587–1592 (2007).
pubmed: 17579573
doi: 10.1038/sj.mt.6300245
Ahmad, A., Brinson, M., Hodges, B. L., Chamberlain, J. S. & Amalfitano, A. Mdx mice inducibly expressing dystrophin provide insights into the potential of gene therapy for Duchenne muscular dystrophy. Hum. Mol. Genet 9, 2507–2515 (2000).
pubmed: 11030755
doi: 10.1093/hmg/9.17.2507
Viola, H. M., Johnstone, V. P. A., Adams, A. M., Fletcher, S. & Hool, L. C. A morpholino oligomer therapy regime that restores mitochondrial function and prevents mdx cardiomyopathy. JACC Basic Transl. Sci. 3, 391–402 (2018).
pubmed: 30062225
pmcid: 6059013
doi: 10.1016/j.jacbts.2018.03.007
Sazani, P. et al. Repeat-dose toxicology evaluation in cynomolgus monkeys of AVI-4658, a phosphorodiamidate morpholino oligomer (PMO) drug for the treatment of Duchenne muscular dystrophy. Int J. Toxicol. 30, 313–321 (2011).
pubmed: 21540336
doi: 10.1177/1091581811403505
Lim, K. R. Q. et al. Efficacy of multi-exon skipping treatment in Duchenne muscular dystrophy dog model neonates. Mol. Ther. 27, 76–86 (2019).
pubmed: 30448197
doi: 10.1016/j.ymthe.2018.10.011
Kenjo, E. et al. Low immunogenicity of LNP allows repeated administrations of CRISPR-Cas9 mRNA into skeletal muscle in mice. Nat. Commun. 12, 7101 (2021).
pubmed: 34880218
pmcid: 8654819
doi: 10.1038/s41467-021-26714-w
Blanchard, D. C. & Blanchard, R. J. Innate and conditioned reactions to threat in rats with amygdaloid lesions. J. Comp. Physiol. Psychol. 81, 281–290 (1972).
pubmed: 5084445
doi: 10.1037/h0033521
Gaus, H. J. et al. Characterization of the interactions of chemically-modified therapeutic nucleic acids with plasma proteins using a fluorescence polarization assay. Nucleic Acids Res. 47, 1110–1122 (2019).
pubmed: 30566688
doi: 10.1093/nar/gky1260