A long-lived IL-2 mutein that selectively activates and expands regulatory T cells as a therapy for autoimmune disease

Laurence B Peterson; Charles J M Bell; Sarah K Howlett; Marcin L Pekalski; Kevin Brady; Heather Hinton; Denise Sauter; John A Todd; Pablo Umana; Oliver Ast; Inja Waldhauer; Anne Freimoser-Grundschober; Ekkehard Moessner; Christian Klein; Ralf J Hosse; Linda S Wicker

doi:10.1016/j.jaut.2018.10.017

A long-lived IL-2 mutein that selectively activates and expands regulatory T cells as a therapy for autoimmune disease

J Autoimmun. 2018 Dec:95:1-14. doi: 10.1016/j.jaut.2018.10.017. Epub 2018 Nov 13.

Authors

Laurence B Peterson¹, Charles J M Bell², Sarah K Howlett², Marcin L Pekalski³, Kevin Brady⁴, Heather Hinton⁵, Denise Sauter⁴, John A Todd³, Pablo Umana⁵, Oliver Ast⁵, Inja Waldhauer⁵, Anne Freimoser-Grundschober⁵, Ekkehard Moessner⁵, Christian Klein⁵, Ralf J Hosse⁵, Linda S Wicker⁶

Affiliations

¹ Roche Innovation Center Basel, Roche Pharma Research & Early Development (pRED), Grenzacherstrasse 124, 4070, Basel, Switzerland; JDRF/Wellcome Diabetes and Inflammation Laboratory, Department of Medical Genetics, Cambridge Institute for Medical Research, NIHR Cambridge Biomedical Research Centre, University of Cambridge, Cambridge, CB2 OXY, UK.
² JDRF/Wellcome Diabetes and Inflammation Laboratory, Department of Medical Genetics, Cambridge Institute for Medical Research, NIHR Cambridge Biomedical Research Centre, University of Cambridge, Cambridge, CB2 OXY, UK.
³ JDRF/Wellcome Diabetes and Inflammation Laboratory, Wellcome Centre for Human Genetics, Nuffield Department of Medicine, NIHR Oxford Biomedical Research Centre, University of Oxford, Oxford, UK; JDRF/Wellcome Diabetes and Inflammation Laboratory, Department of Medical Genetics, Cambridge Institute for Medical Research, NIHR Cambridge Biomedical Research Centre, University of Cambridge, Cambridge, CB2 OXY, UK.
⁴ Roche Innovation Center Basel, Roche Pharma Research & Early Development (pRED), Grenzacherstrasse 124, 4070, Basel, Switzerland.
⁵ Roche Innovation Center Zurich, Roche Pharma Research & Early Development (pRED), Wagistrasse 10, 8952, Schlieren, Switzerland.
⁶ JDRF/Wellcome Diabetes and Inflammation Laboratory, Wellcome Centre for Human Genetics, Nuffield Department of Medicine, NIHR Oxford Biomedical Research Centre, University of Oxford, Oxford, UK; JDRF/Wellcome Diabetes and Inflammation Laboratory, Department of Medical Genetics, Cambridge Institute for Medical Research, NIHR Cambridge Biomedical Research Centre, University of Cambridge, Cambridge, CB2 OXY, UK. Electronic address: [email protected].

Abstract

Susceptibility to multiple autoimmune diseases is associated with common gene polymorphisms influencing IL-2 signaling and T_reg function, making T_reg-specific expansion by IL-2 a compelling therapeutic approach to treatment. As an in vivo IL-2 half-life enhancer we used a non-targeted, effector-function-silent human IgG1 as a fusion protein. An IL-2 mutein (N88D) with reduced binding to the intermediate affinity IL-2Rβγ receptor was engineered with a stoichiometry of two IL-2N88D molecules per IgG, i.e. IgG-(IL-2N88D)₂. The reduced affinity of IgG-(IL-2N88D)₂ for the IL-2Rβγ receptor resulted in a T_reg-selective molecule in human whole blood pSTAT5 assays. Treatment of cynomolgus monkeys with single low doses of IgG-(IL-2N88D)₂ induced sustained preferential activation of T_regs accompanied by a corresponding 10-14-fold increase in CD4⁺ and CD8⁺ CD25⁺FOXP3⁺ T_regs; conditions that had no effect on CD4⁺ or CD8⁺ memory effector T cells. The expanded cynomolgus T_regs had demethylated FOXP3 and CTLA4 epigenetic signatures characteristic of functionally suppressive cells. Humanized mice had similar selective in vivo responses; IgG-(IL-2N88D)₂ increased T_regs while wild-type IgG-IL-2 increased NK cells in addition to T_regs. The expanded human T_regs had demethylated FOXP3 and CTLA4 signatures and were immunosuppressive. These results describe a next-generation immunotherapy using a long-lived and T_reg-selective IL-2 that activates and expands functional T_regsin vivo. Patients should benefit from restored immune homeostasis in a personalized fashion to the extent that their autoimmune disease condition dictates opening up the possibility for remissions and cures.

Keywords: Autoimmunity; Cytokine therapy; IL-2 mutein; Immunotherapy; T(reg) expansion.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Autoimmune Diseases / genetics
Autoimmune Diseases / immunology
Autoimmune Diseases / pathology
Autoimmune Diseases / therapy*
Binding Sites
CD8-Positive T-Lymphocytes / drug effects
CD8-Positive T-Lymphocytes / immunology
CD8-Positive T-Lymphocytes / pathology
CTLA-4 Antigen / genetics
CTLA-4 Antigen / immunology
Cell Proliferation
DNA Methylation / drug effects
Disease Models, Animal
Epigenesis, Genetic
Forkhead Transcription Factors / genetics
Forkhead Transcription Factors / immunology
Humans
Immunoglobulin G / administration & dosage
Immunoglobulin G / chemistry
Immunoglobulin G / genetics
Immunoglobulin G / immunology*
Immunotherapy / methods*
Interleukin-2 / administration & dosage
Interleukin-2 / chemistry
Interleukin-2 / genetics
Interleukin-2 / immunology*
Interleukin-2 Receptor beta Subunit / genetics
Interleukin-2 Receptor beta Subunit / immunology
Lymphocyte Activation / drug effects
Lymphotoxin-alpha / administration & dosage
Lymphotoxin-alpha / chemistry
Lymphotoxin-alpha / genetics
Lymphotoxin-alpha / immunology*
Macaca fascicularis
Male
Mice
Mice, Transgenic
Models, Molecular
Protein Binding
Protein Isoforms / genetics
Protein Isoforms / immunology
Protein Structure, Secondary
Recombinant Fusion Proteins / administration & dosage
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / immunology*
STAT5 Transcription Factor / genetics
STAT5 Transcription Factor / immunology
Signal Transduction
T-Lymphocytes, Regulatory / drug effects*
T-Lymphocytes, Regulatory / immunology
T-Lymphocytes, Regulatory / pathology

Substances

CTLA-4 Antigen
CTLA4 protein, human
FOXP3 protein, human
Forkhead Transcription Factors
IL2 protein, human
Immunoglobulin G
Interleukin-2
Interleukin-2 Receptor beta Subunit
Lymphotoxin-alpha
Protein Isoforms
Recombinant Fusion Proteins
STAT5 Transcription Factor

Abstract

Publication types

MeSH terms

Substances

Grants and funding