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Duokines: a novel class of dual-acting co-stimulatory molecules acting in cis or trans
Sina Fellermeier-Kopf1,2, Friederike Gieseke2, Ugur Sahin2
1Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany.
Abstract:
Co-stimulatory signals induced by ligands of the tumor necrosis factor superfamily (TNFSF) play a central role in T cell activation and have emerged as a promising strategy in cancer immunotherapy. Here, we established a novel class of bifunctional co-stimulatory fusion proteins with the aim to boost T cell activation at the level of T cell - antigen-presenting cell (APC) interaction. These novel dual-acting cytokine fusion proteins were created by connecting two different homotrimeric TNFSF ligands to form homotrimeric bifunctional molecules (Duokines) or by connecting single-chain derivatives of two different homotrimeric TNFSF with a single, flexible linker (single-chain Duokines, scDuokines). By linking the TNFSF ligands 4-1BBL, OX40L and CD27L in all possible combinations, cis-acting Duokines were generated that act on the same or adjacent T cells, while combining CD40L with 4-1BBL, OX40L and CD27L resulted in trans-acting Duokines acting simultaneously on APCs and T cells. In vitro, co-stimulation of T cells was seen for cis- and trans-acting Duokines and scDuokines in an antigen-independent as well as antigen-specific setting. Trans-acting molecules furthermore activated B cells, which represent a subclass of APCs. In a pilot experiment using the syngeneic B16-FAP mouse tumor model scDuokines displayed antitumoral activity in vivo in combination with a primary T cell-activating bispecific antibody, evident from reduced number of lung metastasis compared to the antibody-only treated group. Our data show that the bifunctional, co-stimulatory duokines are capable to enhance T cell-mediated anti-tumor immune responses, suggesting that they can serve as a new class of immuno-stimulatory molecules for use in cancer immunotherapy strategies.
Insights
Novel bifunctional fusion proteins, called Duokines and scDuokines, enhance T cell activation for cancer immunotherapy. These molecules boost anti-tumor immune responses by improving T cell-antigen-presenting cell interactions, showing promise in preclinical models.
Area of Science:
- Immunology
- Cancer Immunotherapy
- Molecular Engineering
Background:
- Co-stimulatory signals from tumor necrosis factor superfamily (TNFSF) ligands are crucial for T cell activation and cancer immunotherapy.
- Current strategies aim to enhance T cell responses, but novel approaches are needed to optimize T cell-antigen-presenting cell (APC) interactions.
Purpose of the Study:
- To establish a novel class of bifunctional co-stimulatory fusion proteins (Duokines and scDuokines) to enhance T cell activation.
- To investigate the potential of these engineered proteins in boosting T cell-mediated anti-tumor immunity.
Main Methods:
- Engineered dual-acting cytokine fusion proteins by linking TNFSF ligands (4-1BBL, OX40L, CD27L, CD40L) into homotrimeric bifunctional molecules (Duokines) or single-chain variants (scDuokines).
- Evaluated co-stimulatory activity of Duokines and scDuokines in vitro using T cells and APCs in antigen-dependent and -independent settings.
- Assessed in vivo anti-tumoral activity of scDuokines in a syngeneic B16-FAP mouse tumor model in combination with a bispecific antibody.
Main Results:
- Both cis-acting and trans-acting Duokines, as well as scDuokines, demonstrated potent T cell co-stimulation in vitro.
- Trans-acting molecules also activated B cells, a subset of APCs.
- In vivo studies showed that scDuokines, combined with a bispecific antibody, significantly reduced lung metastasis in the B16-FAP model, indicating anti-tumoral activity.
Conclusions:
- Bifunctional co-stimulatory Duokines and scDuokines represent a novel class of immuno-stimulatory molecules.
- These engineered proteins effectively enhance T cell-mediated anti-tumor immune responses.
- Duokines and scDuokines hold significant potential for development in cancer immunotherapy strategies.
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