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Tuning the potency and selectivity of ImmTAC molecules by affinity modulation.

Ian B Robertson¹, Rachel Mulvaney¹, Nele Dieckmann¹

¹Immunocore Limited, Drug Discovery and Protein Engineering, Abingdon, Oxon, UK.

Clinical and Experimental Immunology

|November 6, 2023

Summary

Optimizing T-cell-engaging bispecifics involves balancing binding affinities. Combining a high-affinity T-cell receptor with an intermediate-affinity anti-CD3 moiety enhances T-cell activation and selectivity for cancer therapy.

Keywords:

T-cell activation bispecific cluster of differentiation 3 cytokine release cytotoxicity

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Area of Science:

Immunology
Oncology
Biochemistry

Background:

T-cell-engaging bispecifics show promise for treating cancer and infectious diseases.
Understanding the relationship between binding affinity/kinetics and bispecific molecule potency/specificity is crucial but incomplete.

Purpose of the Study:

To investigate how altering target and CD3 binding affinities impacts the potency and specificity of redirected T-cell responses.
To model these relationships using immune mobilizing monoclonal TCRs against cancer (ImmTAC) molecules.

Main Methods:

Utilized ImmTAC molecules, which feature an affinity-enhanced T-cell receptor for target binding and an anti-CD3 effector moiety.
Systematically varied the binding affinities of both the targeting and CD3 domains.
Measured T-cell activation, cytokine release, and selectivity.

Main Results:

Optimal T-cell activation was achieved with a high-affinity T-cell receptor and an intermediate-affinity anti-CD3 domain.
High affinity in both targeting and effector domains reduced maximum cytokine release.
Optimizing affinities for both domains improved the molecule's selectivity.
Experimental observations were explained by a kinetic proofreading model with limited signaling.

Conclusions:

The affinity of both targeting and CD3 domains significantly influences bispecific molecule efficacy and specificity.
Excessively strong binding can lead to T-cell receptor inactivation via a signaling 'dark state'.
Findings provide critical insights for designing potent and selective anti-CD3-based bispecific therapies.