Advertisement

Targeting IgE polyadenylation signal with antisense oligonucleotides decreases IgE secretion and plasma cell viability

Published:November 02, 2021DOI:https://doi.org/10.1016/j.jaci.2021.09.039

      Background

      Allergy regroups numerous complex and various diseases classified as IgE-dependent or non–IgE-dependent hypersensitivities. IgEs are expressed as membrane and secreted forms by B cells and plasma cells, respectively. In IgE-mediated hypersensitivity, IgE secretion and binding to the high-affinity IgE receptor FcεRI on effector cells are responsible for the onset of allergic symptoms; in contrast, surface IgE expression as a B-cell receptor is barely detectable.

      Objective

      Our aim was to test an innovative antisense approach to reducing IgE secretion.

      Methods

      We designed an antisense oligonucleotide (ASO) targeting the polyadenylation signal of human secreted IgE to redirect IgE transcript polyadenylation from the secreted form to the membrane form. ASO treatments were performed on B cells from transgenic mice expressing humanized IgE (InEps mice), as well as on human primary B cells and myeloma cells. In vivo ASO delivery was tested by using an InEps mouse model.

      Results

      We demonstrated that treatment with a morpholino ASO targeting the secreted IgE polyadenylation signal drastically decreased IgE secretion and inversely increased membrane IgE mRNA expression. In addition, ASO treatment induced apoptosis of IgE-expressing U266 myeloma cells, and RNA sequencing revealed attenuation of their plasma cell phenotype. Remarkably, systemic administration of an ASO coupled with Pip6a as an arginine-rich cell-penetrating peptide decreased IgE secretion in vivo.

      Conclusion

      Altogether, this ASO strategy could be an effective way to decrease IgE secretion and allergic symptoms in patients with IgE-dependent allergies, and it could also promote allergen tolerance through apoptosis of IgE+ antibody-secreting cells.

      Graphical abstract

      Key words

      Abbreviations used:

      ASO (Antisense oligonucleotide), BCR (B-cell receptor), CHS (Constant heavy secretory), PAS (Polyadenylation signal), Pip6a (Peptide nucleic acids/Phosphorodiamidate Morholino Oligomer internalization peptide 6a)
      To read this article in full you will need to make a payment

      References

        • MacGlashan D.W.
        IgE-dependent signaling as a therapeutic target for allergies.
        Trends Pharmacol Sci. 2012; 33: 502-509
        • Kawakami T.
        • Blank U.
        From IgE to omalizumab.
        J Immunol. 2016; 197: 4187-4192
        • Wu L.C.
        • Zarrin A.A.
        The production and regulation of IgE by the immune system.
        Nat Rev Immunol. 2014; 14: 247-259
        • Laffleur B.
        • Duchez S.
        • Tarte K.
        • Denis-Lagache N.
        • Péron S.
        • Carrion C.
        • et al.
        Self-restrained B cells arise following membrane IgE expression.
        Cell Rep. 2015; 10: 900-909
        • Haniuda K.
        • Fukao S.
        • Kodama T.
        • Hasegawa H.
        • Kitamura D.
        Autonomous membrane IgE signaling prevents IgE-memory formation.
        Nat Immunol. 2016; 17: 1109
        • Yang Z.
        • Sullivan B.M.
        • Allen C.D.C.
        Fluorescent in vivo detection reveals that IgE+ B cells are restrained by an intrinsic cell fate predisposition.
        Immunity. 2012; 36: 857-872
        • Wade-Vallance A.K.
        • Allen C.D.C.
        Intrinsic and extrinsic regulation of IgE B cell responses.
        Curr Opin Immunol. 2021; 72: 221-229
        • Bennett C.F.
        Therapeutic antisense oligonucleotides are coming of age.
        Annu Rev Med. 2019; 70: 307-321
        • Smith C.I.E.
        • Zain R.
        Therapeutic oligonucleotides: state of the art.
        Annu Rev Pharmacol Toxicol. 2019; 59: 605-630
        • Levin A.A.
        Treating disease at the RNA level with oligonucleotides.
        N Engl J Med. 2019; 380: 57-70
        • Vickers T.A.
        • Wyatt J.R.
        • Burckin T.
        • Bennett C.F.
        • Freier S.M.
        Fully modified 2′ MOE oligonucleotides redirect polyadenylation.
        Nucleic Acids Res. 2001; 29: 1293-1299
        • Vorlová S.
        • Rocco G.
        • LeFave C.V.
        • Jodelka F.M.
        • Hess K.
        • Hastings M.L.
        • et al.
        Induction of antagonistic soluble decoy receptor tyrosine kinases by intronic polyA activation.
        Mol Cell. 2011; 43: 927-939
        • Elkon R.
        • Ugalde A.P.
        • Agami R.
        Alternative cleavage and polyadenylation: extent, regulation and function.
        Nat Rev Genet. 2013; 14: 496-506
        • Fu Y.
        • Foden J.A.
        • Khayter C.
        • Maeder M.L.
        • Reyon D.
        • Joung J.K.
        • et al.
        High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells.
        Nat Biotechnol. 2013; 31: 822-826
        • Grünewald J.
        • Zhou R.
        • Garcia S.P.
        • Iyer S.
        • Lareau C.A.
        • Aryee M.J.
        • et al.
        Transcriptome-wide off-target RNA editing induced by CRISPR-guided DNA base editors.
        Nature. 2019; 569: 433-437
        • Zuccarino-Catania G.V.
        • Sadanand S.
        • Weisel F.J.
        • Tomayko M.M.
        • Meng H.
        • Kleinstein S.H.
        • et al.
        CD80 and PD-L2 define functionally distinct memory B cell subsets that are independent of antibody isotype.
        Nat Immunol. 2014 Jul; 15: 631-637
        • Andreani V.
        • Ramamoorthy S.
        • Pandey A.
        • Lupar E.
        • Nutt S.L.
        • Lämmermann T.
        • et al.
        Cochaperone Mzb1 is a key effector of Blimp1 in plasma cell differentiation and β1-integrin function.
        Proc Natl Acad Sci. 2018; 115: E9630-E9639
        • Lehto T.
        • Castillo Alvarez A.
        • Gauck S.
        • Gait M.J.
        • Coursindel T.
        • Wood M.J.A.
        • et al.
        Cellular trafficking determines the exon skipping activity of Pip6a-PMO in mdx skeletal and cardiac muscle cells.
        Nucleic Acids Res. 2014; 42: 3207-3217
        • He J.-S.
        • Subramaniam S.
        • Narang V.
        • Srinivasan K.
        • Saunders S.P.
        • Carbajo D.
        • et al.
        IgG1 memory B cells keep the memory of IgE responses.
        Nat Commun. 2017 Sep 21; 8: 641
        • Laffleur B.
        • Debeaupuis O.
        • Dalloul Z.
        • Cogné M.
        B cell intrinsic mechanisms constraining IgE memory.
        Front Immunol. 2017; 8: 1277
        • Rosenbaum M.
        • Andreani V.
        • Kapoor T.
        • Herp S.
        • Flach H.
        • Duchniewicz M.
        • et al.
        MZB1 is a GRP94 cochaperone that enables proper immunoglobulin heavy chain biosynthesis upon ER stress.
        Genes Dev. 2014 Jan 6; 28: 1165-1178
        • Hoh R.A.
        • Joshi S.A.
        • Lee J.-Y.
        • Martin B.A.
        • Varma S.
        • Kwok S.
        • et al.
        Origins and clonal convergence of gastrointestinal IgE+ B cells in human peanut allergy.
        Sci Immunol. 2020 Mar 6; 5eaay4209
        • Cruse G.
        • Yin Y.
        • Fukuyama T.
        • Desai A.
        • Arthur G.K.
        • Bäumer W.
        • et al.
        Exon skipping of FcεRIβ eliminates expression of the high-affinity IgE receptor in mast cells with therapeutic potential for allergy.
        Proc Natl Acad Sci. 2016 Dec 6; 113: 14115-14120
        • Tang L.
        • Sandrasagra A.
        • Teng K.
        • Nyce J.W.
        • Li Y.
        • Mannion J.C.
        • et al.
        RASONs: a novel antisense oligonucleotide therapeutic approach for asthma.
        Expert Opin Biol Ther. 2001 Nov 1; 1: 979-983