Useful webinars and videos about ASO Safety and toxicology

Webinar on OTS website from Art Krieg (April 2018)

Sebastien Burel’s talk from OTS 2018

Art Krieg’s talk from OTS 2018

Sabine Sewing’s talk from OTS 2017

ASO Safety and toxicology useful references

Andersson P, den Besten C (2019) Preclinical and Clinical Drug-metabolism, Pharmacokinetics and Safety of Therapeutic Oligonucleotides. In Advances in Nucleic Acid Therapeutics, Agrawal S, Gait MJ (eds)


Barchet W, Wimmenauer V, Schlee M, Hartmann G (2008) Accessing the therapeutic potential of immunostimulatory nucleic acids. Curr Opin Immunol 20: 389-95


Braendli-Baiocco A, Festag M, Dumong Erichsen K, Persson R, Mihatsch MJ, Fisker N, Funk J, Mohr S, Constien R, Ploix C, Brady K, Berrera M, Altmann B, Lenz B, Albassam M, Schmitt G, Weiser T, Schuler F, Singer T, Tessier Y (2017) From the Cover: The Minipig is a Suitable Non-Rodent Model in the Safety Assessment of Single Stranded Oligonucleotides. Toxicol Sci 157: 112-128


Burel SA, Hart CE, Cauntay P, Hsiao J, Machemer T, Katz M, Watt A, Bui HH, Younis H, Sabripour M, Freier SM, Hung G, Dan A, Prakash TP, Seth PP, Swayze EE, Bennett CF, Crooke ST, Henry SP (2016) Hepatotoxicity of high affinity gapmer antisense oligonucleotides is mediated by RNase H1 dependent promiscuous reduction of very long pre-mRNA transcripts. Nucleic Acids Res 44: 2093-109


Carver MP, Charleston JS, Shanks C, Zhang J, Mense M, Sharma AK, Kaur H, Sazani P (2016) Toxicological Characterization of Exon Skipping Phosphorodiamidate Morpholino Oligomers (PMOs) in Non-human Primates. J Neuromuscul Dis 3: 381-393


Crooke ST, Baker BF, Pham NC, Hughes SG, Kwoh TJ, Cai D, Tsimikas S, Geary RS, Bhanot S (2018) The Effects of 2′-O-Methoxyethyl Oligonucleotides on Renal Function in Humans. Nucleic Acid Ther 28: 10-22


Dieckmann A, Hagedorn PH, Burki Y, Brugmann C, Berrera M, Ebeling M, Singer T, Schuler F (2018) A Sensitive In Vitro Approach to Assess the Hybridization-Dependent Toxic Potential of High Affinity Gapmer Oligonucleotides. Mol Ther Nucleic Acids 10: 45-54


Echevarria L, Aupy P, Relizani K, Bestetti T, Griffith G, Blandel F, Komisarski M, Haeberli A, Svinartchouk F, Garcia L, Goyenvalle A (2019) Evaluating the Impact of Variable Phosphorothioate Content in Tricyclo-DNA Antisense Oligonucleotides in a Duchenne Muscular Dystrophy Mouse Model. Nucleic Acid Ther 29: 148-160


Engelhardt JA, Fant P, Guionaud S, Henry SP, Leach MW, Louden C, Scicchitano MS, Weaver JL, Zabka TS, Frazier KS, Society of Toxicologic Pathology Vascular Injury Working G (2015) Scientific and Regulatory Policy Committee Points-to-consider Paper*: Drug-induced Vascular Injury Associated with Nonsmall Molecule Therapeutics in Preclinical Development: Part 2. Antisense Oligonucleotides. Toxicol Pathol 43: 935-44


Fedorov Y, Anderson EM, Birmingham A, Reynolds A, Karpilow J, Robinson K, Leake D, Marshall WS, Khvorova A (2006) Off-target effects by siRNA can induce toxic phenotype. RNA 12: 1188-96


Flierl U, Nero TL, Lim B, Arthur JF, Yao Y, Jung SM, Gitz E, Pollitt AY, Zaldivia MT, Jandrot-Perrus M, Schafer A, Nieswandt B, Andrews RK, Parker MW, Gardiner EE, Peter K (2015) Phosphorothioate backbone modifications of nucleotide-based drugs are potent platelet activators. J Exp Med 212: 129-37


Frazier KS (2015) Antisense oligonucleotide therapies: the promise and the challenges from a toxicologic pathologist’s perspective. Toxicol Pathol 43: 78-89


Frazier KS, Sobry C, Derr V, Adams MJ, Besten CD, De Kimpe S, Francis I, Gales TL, Haworth R, Maguire SR, Mirabile RC, Mullins D, Palate B, Doorten YP, Ridings JE, Scicchitano MS, Silvano J, Woodfine J (2014) Species-specific inflammatory responses as a primary component for the development of glomerular lesions in mice and monkeys following chronic administration of a second-generation antisense oligonucleotide. Toxicol Pathol 42: 923-35


Goemans NM, Tulinius M, van den Hauwe M, Kroksmark AK, Buyse G, Wilson RJ, van Deutekom JC, de Kimpe SJ, Lourbakos A, Campion G (2016) Long-Term Efficacy, Safety, and Pharmacokinetics of Drisapersen in Duchenne Muscular Dystrophy: Results from an Open-Label Extension Study. PLoS One 11: e0161955


Hagedorn PH, Hansen BR, Koch T, Lindow M (2017) Managing the sequence-specificity of antisense oligonucleotides in drug discovery. Nucleic Acids Res 45: 2262-2282


Hagedorn PH, Yakimov V, Ottosen S, Kammler S, Nielsen NF, Hog AM, Hedtjarn M, Meldgaard M, Moller MR, Orum H, Koch T, Lindow M (2013) Hepatotoxic potential of therapeutic oligonucleotides can be predicted from their sequence and modification pattern. Nucleic Acid Ther 23: 302-10


Hashemi N, Odze RD, McGowan MP, Santos RD, Stroes ES, Cohen DE (2014) Liver histology during Mipomersen therapy for severe hypercholesterolemia. J Clin Lipidol 8: 606-11


Henry SP, Beattie G, Yeh G, Chappel A, Giclas P, Mortari A, Jagels MA, Kornbrust DJ, Levin AA (2002) Complement activation is responsible for acute toxicities in rhesus monkeys treated with a phosphorothioate oligodeoxynucleotide. Int Immunopharmacol 2: 1657-66


Henry SP, Giclas PC, Leeds J, Pangburn M, Auletta C, Levin AA, Kornbrust DJ (1997a) Activation of the alternative pathway of complement by a phosphorothioate oligonucleotide: potential mechanism of action. J Pharmacol Exp Ther 281: 810-6


Henry SP, Narayanan P, Shen L, Bhanot S, Younis HS, Burel SA (2017) Assessment of the Effects of 2′-Methoxyethyl Antisense Oligonucleotides on Platelet Count in Cynomolgus Nonhuman Primates. Nucleic Acid Ther 27: 197-208


Henry SP, Novotny W, Leeds J, Auletta C, Kornbrust DJ (1997b) Inhibition of coagulation by a phosphorothioate oligonucleotide. Antisense Nucleic Acid Drug Dev 7: 503-10


Henry SP, Seguin R, Cavagnaro J, Berman C, Tepper J, Kornbrust D (2016) Considerations for the Characterization and Interpretation of Results Related to Alternative Complement Activation in Monkeys Associated with Oligonucleotide-Based Therapeutics. Nucleic Acid Ther 26: 210-5


Jaax ME, Krauel K, Marschall T, Brandt S, Gansler J, Furll B, Appel B, Fischer S, Block S, Helm CA, Muller S, Preissner KT, Greinacher A (2013) Complex formation with nucleic acids and aptamers alters the antigenic properties of platelet factor 4. Blood 122: 272-81


Janas MM, Schlegel MK, Harbison CE, Yilmaz VO, Jiang Y, Parmar R, Zlatev I, Castoreno A, Xu H, Shulga-Morskaya S, Rajeev KG, Manoharan M, Keirstead ND, Maier MA, Jadhav V (2018) Selection of GalNAc-conjugated siRNAs with limited off-target-driven rat hepatotoxicity. Nat Commun 9: 723


Kline JN, Krieg AM (2008) Toll-like receptor 9 activation with CpG oligodeoxynucleotides for asthma therapy. Drug News Perspect 21: 434-9


Krieg AM (1998) The CpG motif: implications for clinical immunology. BioDrugs 10: 341-6


Krieg AM (2006) Therapeutic potential of Toll-like receptor 9 activation. Nat Rev Drug Discov 5: 471-84


Lankveld DP, Van Loveren H, Baken KA, Vandebriel RJ (2010) In vitro testing for direct immunotoxicity: state of the art. Methods Mol Biol 598: 401-23


Levin AA, Henry SP (2008) Toxicology of Oligonucleotide Therapeutics and Understanding the Relevance of the Toxicities. In Preclinical Safety Evaluation of Biopharmaceuticals: A Science-based Approach to Facilitating Clinical Trials Cavagnaro JA (ed) pp 538-574. John Wiley & Sons, Inc.


Lindow M, Vornlocher HP, Riley D, Kornbrust DJ, Burchard J, Whiteley LO, Kamens J, Thompson JD, Nochur S, Younis H, Bartz S, Parry J, Ferrari N, Henry SP, Levin AA (2012) Assessing unintended hybridization-induced biological effects of oligonucleotides. Nat Biotechnol 30: 920-3


Marlowe JL, Akopian V, Karmali P, Kornbrust D, Lockridge J, Semple S (2017) Recommendations of the Oligonucleotide Safety Working Group’s Formulated Oligonucleotide Subcommittee for the Safety Assessment of Formulated Oligonucleotide-Based Therapeutics. Nucleic Acid Ther 27: 183-196


Moisan A, Gubler M, Zhang JD, Tessier Y, Dumong Erichsen K, Sewing S, Gerard R, Avignon B, Huber S, Benmansour F, Chen X, Villasenor R, Braendli-Baiocco A, Festag M, Maunz A, Singer T, Schuler F, Roth AB (2017) Inhibition of EGF Uptake by Nephrotoxic Antisense Drugs In Vitro and Implications for Preclinical Safety Profiling. Mol Ther Nucleic Acids 6: 89-105


Monteith DK, Henry SP, Howard RB, Flournoy S, Levin AA, Bennett CF, Crooke ST (1997) Immune stimulation–a class effect of phosphorothioate oligodeoxynucleotides in rodents. Anticancer Drug Des 12: 421-32


Narayanan P, Shen L, Curtis BR, Bourdon MA, Nolan JP, Gupta S, Hoffmaster C, Zhou F, Christian B, Schaubhut JL, Greenlee S, Burel SA, Witztum JL, Engelhardt JA, Henry SP (2018) Investigation into the Mechanism(s) That Leads to Platelet Decreases in Cynomolgus Monkeys During Administration of ISIS 104838, a 2′-MOE-Modified Antisense Oligonucleotide. Toxicol Sci 164: 613-626


Sazani P, Ness KP, Weller DL, Poage D, Nelson K, Shrewsbury AS (2011a) Chemical and mechanistic toxicology evaluation of exon skipping phosphorodiamidate morpholino oligomers in mdx mice. Int J Toxicol 30: 322-33


Sazani P, Ness KP, Weller DL, Poage DW, Palyada K, Shrewsbury SB (2011b) 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: 313-21


Sehgal A, Barros S, Ivanciu L, Cooley B, Qin J, Racie T, Hettinger J, Carioto M, Jiang Y, Brodsky J, Prabhala H, Zhang X, Attarwala H, Hutabarat R, Foster D, Milstein S, Charisse K, Kuchimanchi S, Maier MA, Nechev L et al. (2015) An RNAi therapeutic targeting antithrombin to rebalance the coagulation system and promote hemostasis in hemophilia. Nat Med 21: 492-7


Senn JJ, Burel S, Henry SP (2005) Non-CpG-containing antisense 2′-methoxyethyl oligonucleotides activate a proinflammatory response independent of Toll-like receptor 9 or myeloid differentiation factor 88. J Pharmacol Exp Ther 314: 972-9


Sewing S, Boess F, Moisan A, Bertinetti-Lapatki C, Minz T, Hedtjaern M, Tessier Y, Schuler F, Singer T, Roth AB (2016) Establishment of a Predictive In Vitro Assay for Assessment of the Hepatotoxic Potential of Oligonucleotide Drugs. PLoS One 11: e0159431


Sewing S, Roth AB, Winter M, Dieckmann A, Bertinetti-Lapatki C, Tessier Y, McGinnis C, Huber S, Koller E, Ploix C, Reed JC, Singer T, Rothfuss A (2017) Assessing single-stranded oligonucleotide drug-induced effects in vitro reveals key risk factors for thrombocytopenia. PLoS One 12: e0187574


Sheehan JP, Lan HC (1998) Phosphorothioate oligonucleotides inhibit the intrinsic tenase complex. Blood 92: 1617-25


Shen L, Engelhardt JA, Hung G, Yee J, Kikkawa R, Matson J, Tayefeh B, Machemer T, Giclas PC, Henry SP (2016) Effects of Repeated Complement Activation Associated with Chronic Treatment of Cynomolgus Monkeys with 2′-O-Methoxyethyl Modified Antisense Oligonucleotide. Nucleic Acid Ther 26: 236-49


Shen L, Frazer-Abel A, Reynolds PR, Giclas PC, Chappell A, Pangburn MK, Younis H, Henry SP (2014) Mechanistic understanding for the greater sensitivity of monkeys to antisense oligonucleotide-mediated complement activation compared with humans. J Pharmacol Exp Ther 351: 709-17


Shen W, De Hoyos CL, Migawa MT, Vickers TA, Sun H, Low A, Bell TA, 3rd, Rahdar M, Mukhopadhyay S, Hart CE, Bell M, Riney S, Murray SF, Greenlee S, Crooke RM, Liang XH, Seth PP, Crooke ST (2019) Chemical modification of PS-ASO therapeutics reduces cellular protein-binding and improves the therapeutic index. Nat Biotechnol 37: 640-650



Voit T, Topaloglu H, Straub V, Muntoni F, Deconinck N, Campion G, De Kimpe SJ, Eagle M, Guglieri M, Hood S, Liefaard L, Lourbakos A, Morgan A, Nakielny J, Quarcoo N, Ricotti V, Rolfe K, Servais L, Wardell C, Wilson R et al. (2014) Safety and efficacy of drisapersen for the treatment of Duchenne muscular dystrophy (DEMAND II): an exploratory, randomised, placebo-controlled phase 2 study. Lancet Neurol 13: 987-96


Vollmer J, Weeratna RD, Jurk M, Samulowitz U, McCluskie MJ, Payette P, Davis HL, Schetter C, Krieg AM (2004) Oligodeoxynucleotides lacking CpG dinucleotides mediate Toll-like receptor 9 dependent T helper type 2 biased immune stimulation. Immunology 113: 212-23


Younis HS, Vickers T, Levin AA, Henry SP (2006) CpG and Non-CpG oligodeoxynucleotides induce differential proinflammatory gene expression profiles in liver and peripheral blood leukocytes in mice. J Immunotoxicol 3: 57-68


Zanardi TA, Kim TW, Shen L, Serota D, Papagiannis C, Park SY, Kim Y, Henry SP (2018) Chronic Toxicity Assessment of 2′-O-Methoxyethyl Antisense Oligonucleotides in Mice. Nucleic Acid Ther 28: 233-241


Zhang A, Uaesoontrachoon K, Shaughnessy C, Das JR, Rayavarapu S, Brown KJ, Ray PE, Nagaraju K, van den Anker JN, Hoffman EP, Hathout Y (2015) The use of urinary and kidney SILAM proteomics to monitor kidney response to high dose morpholino oligonucleotides in the mdx mouse. Toxicol Rep 2: 838-849