Skip to main content

Advertisement

SpringerLink
Log in

Considerations in the Immunogenicity Assessment Strategy for Oligonucleotide Therapeutics (ONTs)

  • Review Article
  • Published:
The AAPS Journal Aims and scope Submit manuscript

Abstract

Oligonucleotide therapeutics (ONTs) are a diverse group of short synthetic nucleic acid–based molecules that exploit innovative intracellular molecular strategies to create novel treatments for a variety of medical conditions. ONT molecules (~7–15 kDa) reside between traditional large and small molecules, and there has been debate regarding their immunogenicity risk. To date, 13 ON drugs have been approved, and as the field is relatively new, there are currently no specific regulatory guidelines to indicate how to develop, validate, and interpret the immunogenicity assays of ONTs. Some investigators do not test for immune responses to ONs while others test for antibodies (Abs) to components within the formulation, which may or may not include aspects of characterization such as domain mapping of ONT conjugates. Similar to other biopharmaceuticals, the immunogenic properties of ONTs could be influenced by sequence, route, dosage, target population, co-medications, etc. The current anti-drug antibody (ADA) data for different approved ONTs suggest that their administration poses a low immunogenicity risk without any significant impact on pharmacokinetics (PK), pharmacodynamics (PD), and safety; nevertheless, until the field matures with data from many more ON drugs, it remains prudent to assess immunogenicity. The emphasis of this article is to highlight how current ADA methodologies might be applied to the development of ONTs, discuss factors that may pose immunogenicity risks, and provide the authors’ current position on immunogenicity assessment strategies for ONTs. We also discuss assay parameters that may be appropriate for the detection and characterization of ADAs, including the evaluation of neutralizing ADAs, ADA isotyping, Abs to dsDNA, and pre-existing ADA. Immunogenicity risk assessments (IRAs) and early interactions with regulators will inform how to proceed in late stage/pivotal studies.

Graphical abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  1. Lundin KE, Gissberg O, CIE S. Oligonucleotide therapies: the past and the present. Hum Gene Ther. 2015;26:475–85.

  2. Bennett CF, Baker BF, Pham N, Swayze E, Geary RS. Pharmacology of antisense drugs [Internet]. Vol. 57, Annual Review of Pharmacology and Toxicology. Annual Reviews; 2017 [cited 2021 Nov 1]. p. 81–105. Available from: https://www.annualreviews.org/doi/abs/10.1146/annurev-pharmtox-010716-104846

  3. Stein CA, Castanotto D. FDA-approved oligonucleotide therapies in 2017. Mol Ther. 2017;25:1069–75.

  4. Bennett CF. Therapeutic antisense oligonucleotides are coming of age [Internet]. Vol. 70, Annual Review of Medicine. Annual Reviews; 2019 [cited 2021 Nov 1]. p. 307–21. Available from: https://www.annualreviews.org/doi/abs/10.1146/annurev-med-041217-010829

  5. Keefe AD, Pai S, Ellington A. Aptamers as therapeutics. Nat Rev Drug Discov. 2010;9:537–50.

  6. Khati M. The future of aptamers in medicine. J Clin Pathol. 2010;63:480–7.

  7. Crooke ST, Witztum JL, Bennett CF, Baker BF. RNA-targeted therapeutics. Cell Metab. 2018;27:714–39.

  8. Hu B, Weng Y, Xia XH, Jie LX, Huang Y. Clinical advances of siRNA therapeutics. J Gene Med. 2019;21.

  9. Bajan S, Hutvagner G. RNA-based therapeutics: from antisense oligonucleotides to miRNAs [Internet]. Vol. 9, Cells. Multidisciplinary Digital Publishing Institute; 2020 [cited 2021 Nov 1]. p. 137. Available from: https://www.mdpi.com/2073-4409/9/1/137/htm

  10. Yamakawa K, Nakano-Narusawa Y, Hashimoto N, Yokohira M, Matsuda Y. Development and clinical trials of nucleic acid medicines for pancreatic cancer treatment. Int J Mol Sci. 2019;20.

  11. Bennett CF, Swayze EE. RNA targeting therapeutics: molecular mechanisms of antisense oligonucleotides as a therapeutic platform [Internet]. Vol. 50, Annual Review of Pharmacology and Toxicology. Annu Rev Pharmacol Toxicol; 2010 [cited 2021 Nov 1]. p. 259–93. Available from: https://pubmed.ncbi.nlm.nih.gov/20055705/

  12. Yin W, Rogge M. Targeting RNA: a transformative therapeutic strategy. Clin Transl Sci. 2019;12:98–112.

  13. Bramsen JB, Kjems J. Development of therapeutic-grade small interfering RNAs by chemical engineering. Front Genet. 2012;3.

  14. Chernikov IV, Vlassov VV, Chernolovskaya EL. Current development of siRNA bioconjugates: from research to the clinic. Front Pharmacol. 2019;10.

  15. Wittrup A, Lieberman J. Knocking down disease: a progress report on siRNA therapeutics. Nat Rev Genet. 2015;16:543–52.

  16. Weng Y, Xiao H, Zhang J, Liang XJ, Huang Y. RNAi therapeutic and its innovative biotechnological evolution. Biotechnol Adv. 2019;37:801–25.

  17. Juliano RL. The delivery of therapeutic oligonucleotides. Nucleic Acids Res [Internet]. 2016 Aug 19 [cited 2021 Nov 2];44(14):6518–48. Available from: https://academic.oup.com/nar/article/44/14/6518/2468139

  18. Dahlman JE, Kauffman KJ, Langer R, Anderson DG. Nanotechnology for in vivo targeted siRNA delivery. Adv Genet. 2014;88:37–69.

    Article  CAS  Google Scholar 

  19. Leung AKK, Tam YYC, Cullis PR. Lipid nanoparticles for short interfering RNA delivery. In: Advances in Genetics; 2014. p. 71–110.

    Google Scholar 

  20. Perry CM, Balfour JAB. Fomivirsen. Drugs. 1999;57(3):375–80.

    Article  CAS  Google Scholar 

  21. Scott LJ. Givosiran: first approval. Drugs. 2020;80:335–9.

  22. Corey DR, Damha MJ, Manoharan M. Challenges and opportunities for nucleic acid therapeutics. Nucleic Acid Ther [Internet]. 2022 Feb 1 [cited 2022 Jul 22];32(1):8–13. Available from: https://pubmed.ncbi.nlm.nih.gov/34931905/

  23. Lange MJ, Burke DH, Chaput JC. Activation of innate immune responses by a CpG oligonucleotide sequence composed entirely of threose nucleic acid. Nucleic Acid Ther. 2019;29(1).

  24. Stebbins CC, Petrillo M, Stevenson LF. Immunogenicity for antisense oligonucleotides: a risk-based assessment. Bioanalysis. 2019;11:1913–6.

  25. Koren E, Smith HW, Shores E, Shankar G, Finco-Kent D, Rup B, et al. Recommendations on risk-based strategies for detection and characterization of antibodies against biotechnology products. J Immunol Methods. 2008;333:1–9.

  26. Shankar G, Devanarayan V, Amaravadi L, Barrett YC, Bowsher R, Finco-Kent D, et al. Recommendations for the validation of immunoassays used for detection of host antibodies against biotechnology products. J Pharm Biomed Anal. 2008;48:1267–81.

  27. Lorenz C, Gesell T, Zimmermann B, Schoeberl U, Bilusic I, Rajkowitsch L, et al. Genomic SELEX for Hfq-binding RNAs identifies genomic aptamers predominantly in antisense transcripts. Nucleic Acids Res. 2010;38(11):3794–808.

    Article  CAS  Google Scholar 

  28. Gupta S, Richards S, Amaravadi L, Piccoli S, Desilva B, Pillutla R, et al. 2017 White paper on recent issues in bioanalysis: A global perspective on immunogenicity guidelines & biomarker assay performance (part 3-LBA: immunogenicity, biomarkers and PK assays). In: Bioanalysis; 2017.

    Google Scholar 

  29. Gupta S, Devanarayan V, Finco D, Gunn GR, Kirshner S, Richards S, et al. Recommendations for the validation of cell-based assays used for the detection of neutralizing antibody immune responses elicited against biological therapeutics. J Pharm Biomed Anal. 2011;55:878–88.

  30. Gorovits B, Wakshull E, Pillutla R, Xu Y, Manning MS, Goyal J. Recommendations for the characterization of immunogenicity response to multiple domain biotherapeutics. J Immunol Methods. 2014;408:1–12.

  31. Kurki P. Compatibility of immunogenicity guidance by the EMA and the US FDA. Bioanalysis. 2019;11:1619–29.

  32. Patton A, Mullenix MC, Swanson SJ, Koren E. An acid dissociation bridging ELISA for detection of antibodies directed against therapeutic proteins in the presence of antigen. J Immunol Methods. 2005;304(1–2):189–95.

    Article  CAS  Google Scholar 

  33. Wadhwa M, Knezevic I, Kang HN, Thorpe R. Immunogenicity assessment of biotherapeutic products: an overview of assays and their utility. Biologicals. 2015;43:298–306.

  34. Pineda C, Castañeda Hernández G, Jacobs IA, Alvarez DF, Carini C. Assessing the immunogenicity of biopharmaceuticals. BioDrugs. 2016;30:195–206.

  35. Wang J, Lon HK, Lee SL, Burckart GJ, Pisetsky DS. Oligonucleotide-based drug development: considerations for clinical pharmacology and immunogenicity. Ther Innov Regul Sci. 2015;49:861–8.

  36. Clinical pharmacology considerations for the development of oligonucleotide therapeutics (draft guidance) [Internet]. 2022 [cited 2022 Jul 22]. Available from: https://www.fda.gov/regulatory-information/search-fda-guidance-documents/clinical-pharmacology-considerations-development-oligonucleotide-therapeutics

  37. FDA. Immunogenicity testing of therapeutic protein products — developing and validating assays for anti-drug antibody detection [Internet]. 2019. Available from: https://www.accessdata.fda.gov/drugsatfda_docs/nda/2018/210922Orig1s000OtherR.pdf

  38. EMA. Guideline on immunogenicity assessment of therapeutic proteins [Internet]. 2017. Available from: https://www.ema.europa.eu/en/documents/scientific-guideline/guideline-immunogenicity-assessment-therapeutic-proteins-revision-1_en.pdf

  39. Hershfield MS, Ganson NJ, Kelly SJ, Scarlett EL, Jaggers DA, Sundy JS. Induced and pre-existing anti-polyethylene glycol antibody in a trial of every 3-week dosing of pegloticase for refractory gout, including in organ transplant recipients. Arthritis Res Ther. 2014;16(2).

  40. Welink J, Xu Y, Yang E, Wilson A, Henderson N, Luo L, et al. 2018 White paper on recent issues in bioanalysis: “A global bioanalytical community perspective on last decade of incurred samples reanalysis (ISR)” (part 1-small molecule regulated bioanalysis, small molecule biomarkers, peptides & oligonucleotide bioanalysis). Bioanalysis. 2018;10(22):1781–801.

    Article  Google Scholar 

  41. Nieri P, Donadio D, Rossi S, Adinolfi B, Podesta A. Antibodies for therapeutic uses and the evolution of biotechniques. Curr Med Chem. 2009;16(6):753–79.

    Article  CAS  Google Scholar 

  42. Shankar G, Arkin S, Cocea L, Devanarayan V, Kirshner S, Kromminga A, et al. Assessment and reporting of the clinical immunogenicity of therapeutic proteins and peptides - harmonized terminology and tactical recommendations. AAPS J. 2014;16:658–73.

  43. Myler H, Gorovits B, Phillips K, Devanarayan V, Clements-Egan A, Gunn GR, et al. Report on the AAPS Immunogenicity Guidance Forum. AAPS J. 2019;21(4).

  44. Van Meer PJK, Kooijman M, Brinks V, Gispen-De Wied CC, Silva-Lima B, Moors EHM, et al. Immunogenicity of mAbs in non-human primates during nonclinical safety assessment. MAbs [Internet]. 2013 Sep [cited 2022 Jul 22];5(5):810–6. Available from: https://pubmed.ncbi.nlm.nih.gov/23924803/

  45. Mak IW, Evaniew N, Ghert M. Lost in translation: animal models and clinical trials in cancer treatment [Internet]. Vol. 6, Am J Transl Res. 2014. Available from: www.ajtr.org

  46. Peebles RS, Liu MC, Adkinson NF, Lichtenstein LM, Hamilton RG. Ragweed-specific antibodies in bronchoalveolar lavage fluids and serum before and after segmental lung challenge: IgE and IgA associated with eosinophil degranulation. J Allergy Clin Immunol. 1998;101(2 I):265–73.

  47. Stokes Peebles R, Liu MC, Lichtenstein LM, Hamilton RG. IgA, IgG and IgM quantification in bronchoalveolar lavage fluids from allergic rhinitics, allergic asthmatics, and normal subjects by monoclonal antibody-based immunoenzymetric assays. J Immunol Methods. 1995;179(1):77–86.

    Article  Google Scholar 

  48. Stokes Peebles R, Hamilton RG, Lichtenstein LM, Schlosberg M, Liu MC, Proud D, et al. Antigen-specific IgE and IgA antibodies in bronchoalveolar lavage fluid are associated with stronger antigen-induced late phase reactions. Clin Exp Allergy. 2001;31(2):239–48.

    Article  Google Scholar 

  49. Clements-Egan A, Gorovits B, Myler H. The increasing influx of biotherapeutics opens the door to designing new bioanalytical and validation strategies. AAPS Newsmagazine [Internet]. 2019; Available from: https://www.aapsnewsmagazine.org/aapsnewsmagazine/articles/2019/sep19/cover-story-sep19

  50. Wang X, Xia Y. Anti-double stranded DNA antibodies: origin, pathogenicity, and targeted therapies. Front Immunol. 2019;10(JULY).

  51. Yu RZ, Wang Y, Norris DA, Kim TW, Narayanan P, Geary RS, et al. Immunogenicity assessment of inotersen, a 2-O-(2-methoxyethyl) antisense oligonucleotide in animals and humans: effect on pharmacokinetics, pharmacodynamics, and safety. Nucleic Acid Ther. 2020;30(5).

  52. FDA Submission. Onpattro (patisiran) [Internet]. Available from: https://www.accessdata.fda.gov/drugsatfda_docs/nda/2018/210922Orig1s000OtherR.pdf

  53. Zhang X, Goel V, Attarwala H, Sweetser MT, Clausen VA, Robbie GJ. Patisiran pharmacokinetics, pharmacodynamics, and exposure-response analyses in the phase 3 APOLLO trial in patients with hereditary transthyretin-mediated (hATTR) amyloidosis. J Clin Pharmacol. 2020;60(1):37–49.

    Article  CAS  Google Scholar 

  54. Balwani M, Sardh E, Ventura P, Peiró PA, Rees DC, Stölzel U, et al. Phase 3 trial of RNAi therapeutic givosiran for acute intermittent porphyria. N Engl J Med [Internet]. 2020 Jun 10 [cited 2021 Nov 1];382(24):2289–301. Available from: https://www.nejm.org/doi/10.1056/NEJMoa1913147

  55. Agarwal S, Simon AR, Goel V, Habtemariam BA, Clausen VA, Kim JB, et al. Pharmacokinetics and pharmacodynamics of the small interfering ribonucleic acid, givosiran, in patients with acute hepatic porphyria. Clin Pharmacol Ther [Internet]. 2020 Jul 1 [cited 2021 Nov 1];108(1):63–72. Available from: https://onlinelibrary.wiley.com/doi/full/10.1002/cpt.1802

  56. SPINRAZA (nusinersen) [Internet]. FDA submission. Available from: https://www.accessdata.fda.gov/drugsatfda_docs/nda/2016/209531Orig1s000OtherR.pdf

  57. Pisetsky DS. Anti-DNA antibodies - quintessential biomarkers of SLE. Nat Rev Rheumatol. 2016;12:102–10.

  58. Lou H, Wojciak-Stothard B, Ruseva MM, Cook HT, Kelleher P, Pickering MC, et al. Autoantibody-dependent amplification of inflammation in SLE. Cell Death Dis. 2020;11(9).

Download references

Acknowledgements

The authors would like to thank Gopi Shankar and Stephen Ayers for reviewing the manuscript and providing valuable suggestions and discussion.

Author information

Authors and Affiliations

  1. Preclinical Sciences & Translational Safety, Janssen R&D LLC, Spring House, Pennsylvania, 19477, USA

    Nazneen Bano, Christopher Ehlinger, Tong-yuan Yang, Michael Swanson & Schantz Allen

Authors
  1. Nazneen Bano
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christopher Ehlinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tong-yuan Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michael Swanson
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Schantz Allen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nazneen Bano.

Ethics declarations

Conflict of Interest

All authors are employees of Janssen R&D, USA, a division of Johnson and Johnson Inc. The authors declare that there is no potential conflict of interests.

Additional information

Authorship

• I certified that each co-author listed above participated sufficiently in the work to take the responsibility for the content. All authors reviewed and approved the final version of manuscript.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bano, N., Ehlinger, C., Yang, Ty. et al. Considerations in the Immunogenicity Assessment Strategy for Oligonucleotide Therapeutics (ONTs). AAPS J 24, 93 (2022). https://doi.org/10.1208/s12248-022-00741-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1208/s12248-022-00741-x

Keywords

Search

Navigation