Target arm affinities determine preclinical efficacy and safety of anti-HER2/CD3 bispecific antibody

Staflin, Karin; Zafra, Christina L. Zuch de; Schutt, Leah; Clark, Vanessa; Zhong, Fiona; Hristopoulos, Maria; Clark, Robyn; Li, Ji; Mathieu, Mary; Chen, Xiaocheng; Johnston, Jennifer; Low, Justin; Ybarra, Ryan; Slaga, Dionysos; Yang, Jihong; Ovacik, Meric; Dybdal, Noël; Tótpál, Klára; Junttila, Melissa R.; Ellerman, Diego; Lee, Genee; Dennis, Mark S.; Prell, Rodney A.; Junttila, Teemu T.

doi:10.1172/jci.insight.133757

Cited by 69 publications

(55 citation statements)

References 39 publications

(40 reference statements)

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“…These data clearly demonstrate that, similar to the Fc/FcgRIIIa system, which could be potently modulated by Fc-engineering, high-affinity receptor engagement of NKp30 results in fundamentally improved cytotoxic response. Also, interestingly, variants displaying only a moderate affinity improvement resulted in significantly increased lytic activity, which is in line with other bispecific effector cell engagers and engineered Fc domains, for which it has been demonstrated that maximal affinity enhancement may not necessarily result in improved cytolytic activity (37,(51)(52)(53). Because the observed cytolytic activity of our novel molecules does not directly correlate with binding affinity of the receptor-ligand interaction, the mechanisms of enhanced recognition might be more complex.…”

Section: Discussionsupporting

confidence: 78%

Affinity Maturation of B7-H6 Translates into Enhanced NK Cell–Mediated Tumor Cell Lysis and Improved Proinflammatory Cytokine Release of Bispecific Immunoligands via NKp30 Engagement

Pekar

Klausz

Busch

et al. 2021

The Journal of Immunology

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Activating NK cell receptors represent promising target structures to elicit potent antitumor immune responses. In this study, novel immunoligands were generated that bridge the activating NK cell receptor NKp30 on NK cells with epidermal growth factor receptor (EGFR) on tumor cells in a bispecific IgG-like format based on affinity-optimized versions of B7-H6 and the Fab arm derived from cetuximab. To enhance NKp30 binding, the solitary N-terminal IgV domain of B7-H6 (DB7-H6) was affinity matured by an evolutionary library approach combined with yeast surface display. Biochemical and functional characterization of 36 of these novel DB7-H6-derived NK cell engagers revealed an up to 45-fold-enhanced affinity for NKp30 and significantly improved NK cell-mediated, EGFR-dependent killing of tumor cells compared with the NK cell engager based on the wild-type DB7-H6 domain. In this regard, potencies (EC 50 killing) of the best immunoligands were substantially improved by up to 87-fold. Moreover, release of IFN-g and TNF-a was significantly increased. Importantly, equipment of the DB7-H6-based NK cell engagers with a human IgG1 Fc part competent in Fc receptor binding resulted in an almost 10-fold superior killing of EGFR-overexpressing tumor cells compared with molecules either triggering FcgRIIIa or NKp30. Additionally, INF-g and TNF-a release was increased compared with molecules solely triggering FcgRIIIa, including the clinically approved Ab cetuximab. Thus, incorporating affinity-matured ligands for NK cell-activating receptors might represent an effective strategy for the generation of potent novel therapeutic agents with unique effector functions in cancer immunotherapy.

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Section: Discussionsupporting

confidence: 78%

Affinity Maturation of B7-H6 Translates into Enhanced NK Cell–Mediated Tumor Cell Lysis and Improved Proinflammatory Cytokine Release of Bispecific Immunoligands via NKp30 Engagement

Pekar

Klausz

Busch

et al. 2021

The Journal of Immunology

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“… 41 Though we did not evaluate the impact of TAA affinity in cytokine secretion, a recent publication on HER2/CD3 bispecific demonstrated that higher HER2 affinity leads to stronger cytotoxicity as well as lower tolerability. 42 One could argue that for a bispecific platform, it is possible to optimize the antigen-targeting epitope to achieve potent cytotoxic activity and relatively low cytokine release, if the antigen is large enough to offer epitopes with a range of distances to the cellular membrane. On the other hand, in the case when the antigen is small, certain bispecific platforms may be inherently suboptimal, for example, as demonstrated by the DbFc format in the T0 cell line, in which case cytotoxicity always goes hand in hand with cytokine release, regardless of the affinity of the CD3 binder chosen.…”

Section: Discussionmentioning

confidence: 99%

One size does not fit all: navigating the multi-dimensional space to optimize T-cell engaging protein therapeutics

Chen

Yang

Wang

et al. 2021

mAbs

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T-cell engaging biologics is a class of novel and promising immune-oncology compounds that leverage the immune system to eradicate cancer. Here, we compared and contrasted a bispecific diabody-Fc format, which displays a relatively short antigen-binding arm distance, with our bispecific IgG platform. By generating diverse panels of antigen-expressing cells where B cell maturation antigen is either tethered to the cell membrane or located to the juxtamembrane region and masked by elongated structural spacer units, we presented a systematic approach to investigate the role of antigen epitope location and molecular formats in immunological synapse formation and cytotoxicity. We demonstrated that diabody-Fc is more potent for antigen epitopes located in the membrane distal region, while bispecific IgG is more efficient for membrane-proximal epitopes. Additionally, we explored other parameters, including receptor density, antigen-binding affinity, and kinetics. Our results show that molecular format and antigen epitope location, which jointly determine the intermembrane distance between target cells and T cells, allow decoupling of cytotoxicity and cytokine release, while antigen-binding affinities appear to be positively correlated with both readouts. Our work offers new insight that could potentially lead to a wider therapeutic window for T-cell engaging biologics in general.

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“…Another approach to mitigate CRS is to reduce the binding affinity of a bsAb to CD3. Two previous studies showed that the CRS of anti-CLL-1 × anti-CD3 in cynomolgus monkeys [ 168 ] and anti-HER2 × anti-CD3 in mice [ 169 ] was dependent on the bsAbs’ binding affinity to CD3: the higher-affinity variants induced higher levels of cytokines released. A project of anti-CD38 × anti-CD3 bsAbs by Amgen also demonstrated the positive correlation between CD3 affinity and CRS [ 170 ].…”

Section: Prospectsmentioning

confidence: 99%

Development of bispecific antibodies in China: overview and prospects

Zhang

Zhou

2020

Antibody Therapeutics

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A bispecific antibody (bsAb) can simultaneously bind two different epitopes or antigens, allowing for multiple mechanistic functions with synergistic effects. BsAbs have attracted significant scientific attentions and efforts towards their development as drugs for cancers. There are 21 bsAbs currently undergoing clinical trials in China. Here, we review their platform technologies, expression and production, and biological activities and bioassay of these bsAbs, and summarize their structural formats and mechanisms of actions. T-cell redirection and checkpoint inhibition are two main mechanisms of the bsAbs that we discuss in detail. Furthermore, we provide our perspective on the future of bsAb development in China, including CD3-bsAbs for solid tumors and related cytokine release syndromes, expression and chemistry, manufacturing and controls, clinical development, and immunogenicity.

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Target arm affinities determine preclinical efficacy and safety of anti-HER2/CD3 bispecific antibody

Cited by 69 publications

References 39 publications

Affinity Maturation of B7-H6 Translates into Enhanced NK Cell–Mediated Tumor Cell Lysis and Improved Proinflammatory Cytokine Release of Bispecific Immunoligands via NKp30 Engagement

Affinity Maturation of B7-H6 Translates into Enhanced NK Cell–Mediated Tumor Cell Lysis and Improved Proinflammatory Cytokine Release of Bispecific Immunoligands via NKp30 Engagement

One size does not fit all: navigating the multi-dimensional space to optimize T-cell engaging protein therapeutics

Development of bispecific antibodies in China: overview and prospects

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