Cergutuzumab amunaleukin (CEA-IL2v), a CEA-targeted IL-2 variant-based immunocytokine for combination cancer immunotherapy: Overcoming limitations of aldesleukin and conventional IL-2-based immunocytokines

Christian Klein, Inja Waldhauer, Valeria G. Nicolini, Anne Freimoser-Grundschober, Tapan Nayak, Danielle J. Vugts, Claire Dunn, Marije Bolijn, Jörg Benz, Martine Stihle, Sabine Lang, Michaele Roemmele, Thomas Hofer, Erwin van Puijenbroek, David Wittig, Samuel Moser, Oliver Ast, Peter Brünker, Ingo H. Gorr, Sebastian Neumann & 10 others Maria Cristina de Vera Mudry, Heather Hinton, Flavio Crameri, Jose Saro, Stefan Evers, Christian Gerdes, Marina Bacac, Guus van Dongen, Ekkehard Moessner, Pablo Umaña

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

We developed cergutuzumab amunaleukin (CEA-IL2v, RG7813), a novel monomeric CEA-targeted immunocytokine, that comprises a single IL-2 variant (IL2v) moiety with abolished CD25 binding, fused to the C-terminus of a high affinity, bivalent carcinoembryonic antigen (CEA)-specific antibody devoid of Fc-mediated effector functions. Its molecular design aims to (i) avoid preferential activation of regulatory T-cells vs. immune effector cells by removing CD25 binding; (ii) increase the therapeutic index of IL-2 therapy by (a) preferential retention at the tumor by having a lower dissociation rate from CEA-expressing cancer cells vs. IL-2R-expressing cells, (b) avoiding any FcγR-binding and Fc effector functions and (c) reduced binding to endothelial cells expressing CD25; and (iii) improve the pharmacokinetics, and thus convenience of administration, of IL-2. The crystal structure of the IL2v-IL-2Rβγ complex was determined and CEA-IL2v activity was assessed using human immune effector cells. Tumor targeting was investigated in tumor-bearing mice using 89Zr-labeled CEA-IL2v. Efficacy studies were performed in (a) syngeneic mouse models as monotherapy and combined with anti-PD-L1, and in (b) xenograft mouse models in combination with ADCC-mediating antibodies. CEA-IL2v binds to CEA with pM avidity but not to CD25, and consequently did not preferentially activate Tregs. In vivo, CEA-IL2v demonstrated superior pharmacokinetics and tumor targeting compared with a wild-type IL-2-based CEA immunocytokine (CEA-IL2wt). CEA-IL2v strongly expanded NK and CD8+ T cells, skewing the CD8+:CD4+ ratio toward CD8+ T cells both in the periphery and in the tumor, and mediated single agent efficacy in syngeneic MC38-CEA and PancO2-CEA models. Combination with trastuzumab, cetuximab and imgatuzumab, all of human IgG1 isotype, resulted in superior efficacy compared with the monotherapies alone. Combined with anti-PD-L1, CEA-IL2v mediated superior efficacy over the respective monotherapies, and over the combination with an untargeted control immunocytokine. These preclinical data support the ongoing clinical investigation of the cergutuzumab amunaleukin immunocytokine with abolished CD25 binding for the treatment of CEA-positive solid tumors in combination with PD-L1 checkpoint blockade and ADCC competent antibodies.

Original languageEnglish
Article numbere1277306
JournalOncoImmunology
Volume6
Issue number3
DOIs
Publication statusPublished - 4 Mar 2017

Cite this

Klein, Christian ; Waldhauer, Inja ; Nicolini, Valeria G. ; Freimoser-Grundschober, Anne ; Nayak, Tapan ; Vugts, Danielle J. ; Dunn, Claire ; Bolijn, Marije ; Benz, Jörg ; Stihle, Martine ; Lang, Sabine ; Roemmele, Michaele ; Hofer, Thomas ; van Puijenbroek, Erwin ; Wittig, David ; Moser, Samuel ; Ast, Oliver ; Brünker, Peter ; Gorr, Ingo H. ; Neumann, Sebastian ; de Vera Mudry, Maria Cristina ; Hinton, Heather ; Crameri, Flavio ; Saro, Jose ; Evers, Stefan ; Gerdes, Christian ; Bacac, Marina ; van Dongen, Guus ; Moessner, Ekkehard ; Umaña, Pablo. / Cergutuzumab amunaleukin (CEA-IL2v), a CEA-targeted IL-2 variant-based immunocytokine for combination cancer immunotherapy : Overcoming limitations of aldesleukin and conventional IL-2-based immunocytokines. In: OncoImmunology. 2017 ; Vol. 6, No. 3.
@article{c3698f177a304c34908d56ad5f91e5cf,
title = "Cergutuzumab amunaleukin (CEA-IL2v), a CEA-targeted IL-2 variant-based immunocytokine for combination cancer immunotherapy: Overcoming limitations of aldesleukin and conventional IL-2-based immunocytokines",
abstract = "We developed cergutuzumab amunaleukin (CEA-IL2v, RG7813), a novel monomeric CEA-targeted immunocytokine, that comprises a single IL-2 variant (IL2v) moiety with abolished CD25 binding, fused to the C-terminus of a high affinity, bivalent carcinoembryonic antigen (CEA)-specific antibody devoid of Fc-mediated effector functions. Its molecular design aims to (i) avoid preferential activation of regulatory T-cells vs. immune effector cells by removing CD25 binding; (ii) increase the therapeutic index of IL-2 therapy by (a) preferential retention at the tumor by having a lower dissociation rate from CEA-expressing cancer cells vs. IL-2R-expressing cells, (b) avoiding any FcγR-binding and Fc effector functions and (c) reduced binding to endothelial cells expressing CD25; and (iii) improve the pharmacokinetics, and thus convenience of administration, of IL-2. The crystal structure of the IL2v-IL-2Rβγ complex was determined and CEA-IL2v activity was assessed using human immune effector cells. Tumor targeting was investigated in tumor-bearing mice using 89Zr-labeled CEA-IL2v. Efficacy studies were performed in (a) syngeneic mouse models as monotherapy and combined with anti-PD-L1, and in (b) xenograft mouse models in combination with ADCC-mediating antibodies. CEA-IL2v binds to CEA with pM avidity but not to CD25, and consequently did not preferentially activate Tregs. In vivo, CEA-IL2v demonstrated superior pharmacokinetics and tumor targeting compared with a wild-type IL-2-based CEA immunocytokine (CEA-IL2wt). CEA-IL2v strongly expanded NK and CD8+ T cells, skewing the CD8+:CD4+ ratio toward CD8+ T cells both in the periphery and in the tumor, and mediated single agent efficacy in syngeneic MC38-CEA and PancO2-CEA models. Combination with trastuzumab, cetuximab and imgatuzumab, all of human IgG1 isotype, resulted in superior efficacy compared with the monotherapies alone. Combined with anti-PD-L1, CEA-IL2v mediated superior efficacy over the respective monotherapies, and over the combination with an untargeted control immunocytokine. These preclinical data support the ongoing clinical investigation of the cergutuzumab amunaleukin immunocytokine with abolished CD25 binding for the treatment of CEA-positive solid tumors in combination with PD-L1 checkpoint blockade and ADCC competent antibodies.",
keywords = "Antibody, CEA, CEACAM5, IL-2, IL2v, Immunocytokine",
author = "Christian Klein and Inja Waldhauer and Nicolini, {Valeria G.} and Anne Freimoser-Grundschober and Tapan Nayak and Vugts, {Danielle J.} and Claire Dunn and Marije Bolijn and J{\"o}rg Benz and Martine Stihle and Sabine Lang and Michaele Roemmele and Thomas Hofer and {van Puijenbroek}, Erwin and David Wittig and Samuel Moser and Oliver Ast and Peter Br{\"u}nker and Gorr, {Ingo H.} and Sebastian Neumann and {de Vera Mudry}, {Maria Cristina} and Heather Hinton and Flavio Crameri and Jose Saro and Stefan Evers and Christian Gerdes and Marina Bacac and {van Dongen}, Guus and Ekkehard Moessner and Pablo Uma{\~n}a",
year = "2017",
month = "3",
day = "4",
doi = "10.1080/2162402X.2016.1277306",
language = "English",
volume = "6",
journal = "OncoImmunology",
issn = "2162-4011",
publisher = "Landes Bioscience",
number = "3",

}

Klein, C, Waldhauer, I, Nicolini, VG, Freimoser-Grundschober, A, Nayak, T, Vugts, DJ, Dunn, C, Bolijn, M, Benz, J, Stihle, M, Lang, S, Roemmele, M, Hofer, T, van Puijenbroek, E, Wittig, D, Moser, S, Ast, O, Brünker, P, Gorr, IH, Neumann, S, de Vera Mudry, MC, Hinton, H, Crameri, F, Saro, J, Evers, S, Gerdes, C, Bacac, M, van Dongen, G, Moessner, E & Umaña, P 2017, 'Cergutuzumab amunaleukin (CEA-IL2v), a CEA-targeted IL-2 variant-based immunocytokine for combination cancer immunotherapy: Overcoming limitations of aldesleukin and conventional IL-2-based immunocytokines' OncoImmunology, vol. 6, no. 3, e1277306. https://doi.org/10.1080/2162402X.2016.1277306

Cergutuzumab amunaleukin (CEA-IL2v), a CEA-targeted IL-2 variant-based immunocytokine for combination cancer immunotherapy : Overcoming limitations of aldesleukin and conventional IL-2-based immunocytokines. / Klein, Christian; Waldhauer, Inja; Nicolini, Valeria G.; Freimoser-Grundschober, Anne; Nayak, Tapan; Vugts, Danielle J.; Dunn, Claire; Bolijn, Marije; Benz, Jörg; Stihle, Martine; Lang, Sabine; Roemmele, Michaele; Hofer, Thomas; van Puijenbroek, Erwin; Wittig, David; Moser, Samuel; Ast, Oliver; Brünker, Peter; Gorr, Ingo H.; Neumann, Sebastian; de Vera Mudry, Maria Cristina; Hinton, Heather; Crameri, Flavio; Saro, Jose; Evers, Stefan; Gerdes, Christian; Bacac, Marina; van Dongen, Guus; Moessner, Ekkehard; Umaña, Pablo.

In: OncoImmunology, Vol. 6, No. 3, e1277306, 04.03.2017.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Cergutuzumab amunaleukin (CEA-IL2v), a CEA-targeted IL-2 variant-based immunocytokine for combination cancer immunotherapy

T2 - Overcoming limitations of aldesleukin and conventional IL-2-based immunocytokines

AU - Klein, Christian

AU - Waldhauer, Inja

AU - Nicolini, Valeria G.

AU - Freimoser-Grundschober, Anne

AU - Nayak, Tapan

AU - Vugts, Danielle J.

AU - Dunn, Claire

AU - Bolijn, Marije

AU - Benz, Jörg

AU - Stihle, Martine

AU - Lang, Sabine

AU - Roemmele, Michaele

AU - Hofer, Thomas

AU - van Puijenbroek, Erwin

AU - Wittig, David

AU - Moser, Samuel

AU - Ast, Oliver

AU - Brünker, Peter

AU - Gorr, Ingo H.

AU - Neumann, Sebastian

AU - de Vera Mudry, Maria Cristina

AU - Hinton, Heather

AU - Crameri, Flavio

AU - Saro, Jose

AU - Evers, Stefan

AU - Gerdes, Christian

AU - Bacac, Marina

AU - van Dongen, Guus

AU - Moessner, Ekkehard

AU - Umaña, Pablo

PY - 2017/3/4

Y1 - 2017/3/4

N2 - We developed cergutuzumab amunaleukin (CEA-IL2v, RG7813), a novel monomeric CEA-targeted immunocytokine, that comprises a single IL-2 variant (IL2v) moiety with abolished CD25 binding, fused to the C-terminus of a high affinity, bivalent carcinoembryonic antigen (CEA)-specific antibody devoid of Fc-mediated effector functions. Its molecular design aims to (i) avoid preferential activation of regulatory T-cells vs. immune effector cells by removing CD25 binding; (ii) increase the therapeutic index of IL-2 therapy by (a) preferential retention at the tumor by having a lower dissociation rate from CEA-expressing cancer cells vs. IL-2R-expressing cells, (b) avoiding any FcγR-binding and Fc effector functions and (c) reduced binding to endothelial cells expressing CD25; and (iii) improve the pharmacokinetics, and thus convenience of administration, of IL-2. The crystal structure of the IL2v-IL-2Rβγ complex was determined and CEA-IL2v activity was assessed using human immune effector cells. Tumor targeting was investigated in tumor-bearing mice using 89Zr-labeled CEA-IL2v. Efficacy studies were performed in (a) syngeneic mouse models as monotherapy and combined with anti-PD-L1, and in (b) xenograft mouse models in combination with ADCC-mediating antibodies. CEA-IL2v binds to CEA with pM avidity but not to CD25, and consequently did not preferentially activate Tregs. In vivo, CEA-IL2v demonstrated superior pharmacokinetics and tumor targeting compared with a wild-type IL-2-based CEA immunocytokine (CEA-IL2wt). CEA-IL2v strongly expanded NK and CD8+ T cells, skewing the CD8+:CD4+ ratio toward CD8+ T cells both in the periphery and in the tumor, and mediated single agent efficacy in syngeneic MC38-CEA and PancO2-CEA models. Combination with trastuzumab, cetuximab and imgatuzumab, all of human IgG1 isotype, resulted in superior efficacy compared with the monotherapies alone. Combined with anti-PD-L1, CEA-IL2v mediated superior efficacy over the respective monotherapies, and over the combination with an untargeted control immunocytokine. These preclinical data support the ongoing clinical investigation of the cergutuzumab amunaleukin immunocytokine with abolished CD25 binding for the treatment of CEA-positive solid tumors in combination with PD-L1 checkpoint blockade and ADCC competent antibodies.

AB - We developed cergutuzumab amunaleukin (CEA-IL2v, RG7813), a novel monomeric CEA-targeted immunocytokine, that comprises a single IL-2 variant (IL2v) moiety with abolished CD25 binding, fused to the C-terminus of a high affinity, bivalent carcinoembryonic antigen (CEA)-specific antibody devoid of Fc-mediated effector functions. Its molecular design aims to (i) avoid preferential activation of regulatory T-cells vs. immune effector cells by removing CD25 binding; (ii) increase the therapeutic index of IL-2 therapy by (a) preferential retention at the tumor by having a lower dissociation rate from CEA-expressing cancer cells vs. IL-2R-expressing cells, (b) avoiding any FcγR-binding and Fc effector functions and (c) reduced binding to endothelial cells expressing CD25; and (iii) improve the pharmacokinetics, and thus convenience of administration, of IL-2. The crystal structure of the IL2v-IL-2Rβγ complex was determined and CEA-IL2v activity was assessed using human immune effector cells. Tumor targeting was investigated in tumor-bearing mice using 89Zr-labeled CEA-IL2v. Efficacy studies were performed in (a) syngeneic mouse models as monotherapy and combined with anti-PD-L1, and in (b) xenograft mouse models in combination with ADCC-mediating antibodies. CEA-IL2v binds to CEA with pM avidity but not to CD25, and consequently did not preferentially activate Tregs. In vivo, CEA-IL2v demonstrated superior pharmacokinetics and tumor targeting compared with a wild-type IL-2-based CEA immunocytokine (CEA-IL2wt). CEA-IL2v strongly expanded NK and CD8+ T cells, skewing the CD8+:CD4+ ratio toward CD8+ T cells both in the periphery and in the tumor, and mediated single agent efficacy in syngeneic MC38-CEA and PancO2-CEA models. Combination with trastuzumab, cetuximab and imgatuzumab, all of human IgG1 isotype, resulted in superior efficacy compared with the monotherapies alone. Combined with anti-PD-L1, CEA-IL2v mediated superior efficacy over the respective monotherapies, and over the combination with an untargeted control immunocytokine. These preclinical data support the ongoing clinical investigation of the cergutuzumab amunaleukin immunocytokine with abolished CD25 binding for the treatment of CEA-positive solid tumors in combination with PD-L1 checkpoint blockade and ADCC competent antibodies.

KW - Antibody

KW - CEA

KW - CEACAM5

KW - IL-2

KW - IL2v

KW - Immunocytokine

UR - http://www.scopus.com/inward/record.url?scp=85013488349&partnerID=8YFLogxK

U2 - 10.1080/2162402X.2016.1277306

DO - 10.1080/2162402X.2016.1277306

M3 - Article

VL - 6

JO - OncoImmunology

JF - OncoImmunology

SN - 2162-4011

IS - 3

M1 - e1277306

ER -