Mesothelin/CD3 Half-Life–Extended Bispecific T-Cell Engager Molecule Shows Specific Tumor Uptake and Distributes to Mesothelin and CD3-Expressing Tissues

Suurs, Frans V.; Lorenczewski, Grit; Bailis, Julie M.; Stienen, Sabine; Friedrich, Matthias; Lee, Fei; Vegt, Bert van der; Vries, Elisabeth G.E. de; Groot, Derk-Jan A de; Hooge, Marjolijn N Lub de

doi:10.2967/jnumed.120.259036

Cited by 11 publications

(4 citation statements)

References 43 publications

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“…Since T cell engagers (TCEs) are perhaps the most therapeutically relevant class of bsAbs, we chose to attempt the grafting of a Fab HER2 and a Fab CD3 on Fc CD20 , with the aim to produce an “IgG-like TCE”, able to recruit T cells (through CD3-binding) to HER2 + tumor cells. While incorporating an Fc moiety into these construct can be crucial for half-life extension, − the presence of an Fc with immune-effector function is not beneficial in the context of a TCE strategy, and is in fact detrimental due to the undesired depletion of engaged T cells and due to cytokine release syndrome (CRS) arising from immune overactivation through FcγR engagement . As in this case the goal was to generate a proof-of-concept bispecific SynAb rather than a therapeutically relevant moiety, we used Fc CD20 5 even though as a native Fc it has effector function which is suboptimal for a TCE.…”

Section: Resultsmentioning

confidence: 99%

Modular Chemical Construction of IgG-like Mono- and Bispecific Synthetic Antibodies (SynAbs)

et al. 2023

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In recent years there has been rising interest in the field of protein–protein conjugation, especially related to bispecific antibodies (bsAbs) and their therapeutic applications. These constructs contain two paratopes capable of binding two distinct epitopes on target molecules and are thus able to perform complex biological functions (mechanisms of action) not available to monospecific mAbs. Traditionally these bsAbs have been constructed through protein engineering, but recently chemical methods for their construction have started to (re)emerge. While these have been shown to offer increased modularity, speed, and for some methods even the inherent capacity for further functionalization (e.g., with small molecule cargo), most of these approaches lacked the ability to include a fragment crystallizable (Fc) modality. The Fc component of IgG antibodies offers effector function and increased half-life. Here we report a first-in-class disulfide rebridging and click-chemistry-based method for the generation of Fc-containing, IgG-like mono- and bispecific antibodies. These are in the FcZ-(FabX)-FabY format, i.e., two distinct Fabs and an Fc, potentially all from different antibodies, attached in a homogeneous and covalent manner. We have dubbed these molecules synthetic antibodies (SynAbs). We have constructed a T cell-engager (TCE) SynAb, FcCD20-(FabHER2)-FabCD3, and have confirmed that it exhibits the expected biological functions, including the ability to kill HER2+ target cells in a coculture assay with T cells.

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Section: Resultsmentioning

confidence: 99%

Modular Chemical Construction of IgG-like Mono- and Bispecific Synthetic Antibodies (SynAbs)

et al. 2023

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“…Especially, in a model that is strongly dependent on the immune response, this important factor cannot be dismissed. Prospectively, for in vivo experiments, the use of half-life-extended BiTE (HLE) [ 60 ] would be beneficial in order to ease animal stress during experiments; conventional BiTEs with a short half-life would require frequent application or continuous infusion. However, despite several studies, no HLEs have been clinically approved, therefore making the canonical platform the better model.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Bispecific T-Cell Engagers Targeting Murine Cytomegalovirus

Menschikowski,

Bednar,

Kübel

et al. 2024

Viruses

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Human cytomegalovirus is a ubiquitous herpesvirus that, while latent in most individuals, poses a great risk to immunocompromised patients. In contrast to directly acting traditional antiviral drugs, such as ganciclovir, we aim to emulate a physiological infection control using T cells. For this, we constructed several bispecific T-cell engager (BiTE) constructs targeting different viral glycoproteins of the murine cytomegalovirus and evaluated them in vitro for their efficacy. To isolate the target specific effect without viral immune evasion, we established stable reporter cell lines expressing the viral target glycoprotein B, and the glycoprotein complexes gN-gM and gH-gL, as well as nano-luciferase (nLuc). First, we evaluated binding capacities using flow cytometry and established killing assays, measuring nLuc-release upon cell lysis. All BiTE constructs proved to be functional mediators for T-cell recruitment and will allow a proof of concept for this treatment option. This might pave the way for strikingly safer immunosuppression in vulnerable patient groups.

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“…Protein modification of BiTEs to extend their half-life is a strategy that should be further investigated. Several current studies reported the benefit of the addition of partialor full-length Fc region to the BITE structure, which was shown to extend BiTE half-life 37,38 . The present in vitro study demonstrates the feasibility of BiTEs for minimizing immune suppression and promoting the anti-tumor activity of T cells; however, the persistency/sustainability of BiTE needs to be further developed to improve the therapeutic efficiency of this treatment strategy, especially in solid tumors like CCA.…”

Section: Discussionmentioning

confidence: 99%

Combination gemcitabine and PD-L1xCD3 bispecific T cell engager (BiTE) enhances T lymphocyte cytotoxicity against cholangiocarcinoma cells

Wathikthinnakon

Luangwattananun

Sawasdee

et al. 2022

Sci Rep

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Cholangiocarcinoma (CCA) is a lethal cancer with rapid progression and poor survival. Novel and more effective therapies than those currently available are, therefore, urgently needed. Our research group previously reported the combination of gemcitabine and cytotoxic T lymphocytes to be more effective than single-agent treatment for the elimination of CCA cells. However, gemcitabine treatment of CCA cells upregulates the expression of an immune checkpoint protein (programmed death-ligand 1 [PD-L1]) that consequently inhibits the cytotoxicity of T lymphocytes. To overcome this challenge and take advantage of PD-L1 upregulation upon gemcitabine treatment, we generated recombinant PD-L1xCD3 bispecific T cell engagers (BiTEs) to simultaneously block PD-1/PD-L1 signaling and recruit T lymphocytes to eliminate CCA cells. Two recombinant PD-L1xCD3 BiTEs (mBiTE and sBiTE contain anti-PD-L1 scFv region from atezolizumab and from a published sequence, respectively) were able to specifically bind to both CD3 on T lymphocytes, and to PD-L1 overexpressed after gemcitabine treatment on CCA (KKU213A, KKU055, and KKU100) cells. mBiTE and sBiTE significantly enhanced T lymphocyte cytotoxicity against CCA cells, especially after gemcitabine treatment, and their magnitudes of cytotoxicity were positively associated with the levels of PD-L1 expression. Our findings suggest combination gemcitabine and PD-L1xCD3 BiTE as a potential alternative therapy for CCA.

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Mesothelin/CD3 Half-Life–Extended Bispecific T-Cell Engager Molecule Shows Specific Tumor Uptake and Distributes to Mesothelin and CD3-Expressing Tissues

Cited by 11 publications

References 43 publications

Modular Chemical Construction of IgG-like Mono- and Bispecific Synthetic Antibodies (SynAbs)

Modular Chemical Construction of IgG-like Mono- and Bispecific Synthetic Antibodies (SynAbs)

Evaluation of Bispecific T-Cell Engagers Targeting Murine Cytomegalovirus

Combination gemcitabine and PD-L1xCD3 bispecific T cell engager (BiTE) enhances T lymphocyte cytotoxicity against cholangiocarcinoma cells

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