Synthetic ligands of death receptor 5 display a cell-selective agonistic effect at different oligomerization levels

Chekkat, Neila; Smulski, Cristian R.; Lombardo, Caterina Maria; Lechner, Marie-Charlotte; Seguin, Cendrine; Décossas, Marion; Spanedda, Maria Vittoria; Frisch, Benoı̂t; Guichard, Gilles; Fournel, Sylvie

doi:10.18632/oncotarget.10508

Cited by 12 publications

(11 citation statements)

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“…In accordance with results from earlier studies (19), the assembly of two scTRAIL moieties into a hexavalent molecule considerably increased activation of caspases and subsequent cell death compared with trivalent scTRAIL. Of note, recent data highlighted the importance not only of increased valency, but also of an appropriate spatial organization to allow multivalent receptor binding (33). Our results demonstrate that employing a single-chain version of the natural homotrimeric TRAIL ligand in the context of immunoglobulin-derived dimerization modules enables potent death receptor activation.…”

Section: Discussionmentioning

confidence: 60%

Superior Properties of Fc-comprising scTRAIL Fusion Proteins

Hutt

Marquardt

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et al. 2017

Molecular Cancer Therapeutics

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The TNF-related apoptosis-inducing ligand (TRAIL) has been considered as a promising molecule for cancer treatment. However, clinical studies with soluble TRAIL failed to show therapeutic activity, which resulted in subsequent development of more potent TRAIL-based therapeutics. In this study, we applied defined oligomerization and tumor targeting as strategies to further improve the activity of a single-chain version of TRAIL (scTRAIL). We compared three different formats of EGF receptor (EGFR)-targeting dimeric scTRAIL fusion proteins [Diabody (Db)-scTRAIL, scFv-IgE heavy chain domain 2 (EHD2)-scTRAIL, scFv-Fc-scTRAIL] as well as two nontargeted dimeric scTRAIL molecules (EHD2-scTRAIL, Fc-scTRAIL) to reveal the influence of targeting and protein format on antitumor activity. All EGFR-targeted dimeric scTRAIL molecules showed similar binding properties and comparable cell death induction , exceeding the activity of the respective nontargeted dimeric format and monomeric scTRAIL. Superior properties were observed for the Fc fusion proteins with respect to production and half-life. studies using a Colo205 xenograft model revealed potent antitumor activity of all EGFR-targeting formats and Fc-scTRAIL and furthermore highlighted the higher efficacy of fusion proteins comprising an Fc part. Despite enhanced cell death induction of targeted scTRAIL molecules, however, comparable antitumor activities were found for the EGFR-targeting scFv-Fc-scTRAIL and the nontargeting Fc-scTRAIL .

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Section: Discussionmentioning

confidence: 60%

Superior Properties of Fc-comprising scTRAIL Fusion Proteins

Hutt

Marquardt

Seifert

et al. 2017

Molecular Cancer Therapeutics

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“…We previously described two cyclic peptides, named 1m and 2m in their monovalent forms that only differ by the position of a lysine in their sequence (see Supplementary Materials ). Their divalent forms, known as 1d and 2d respectively, bound to TRAIL-R2 with high affinity as measured by SPR and induced apoptosis of various cell lines [ 27 , 28 ]. In the present study, we compared the pro-apoptotic activity of 1d and 2d on the human Burkitt lymphoma BJAB, T leukemia Jurkat and the colon carcinoma HCT116 cell lines.…”

Section: Resultsmentioning

confidence: 99%

“…We previously described synthetic peptides (1 and 2) targeting the TRAIL-R2 receptor and that induce TRAIL-R2-dependent apoptosis of BJAB cells, in divalent and trivalent forms [ 27 ]. SPR experiments indicated that both 1d and 2d formed stable complexes with recombinant TRAIL-R2 covalently immobilized on a sensor surface.…”

Section: Discussionmentioning

confidence: 99%

“…Finally, the mechanism of apoptosis resistance proposed to explain the poor efficiency of some mAbs, was an inefficient receptor clustering thus limiting DISC formation and apoptosis [ 25 , 26 ]. Indeed, it was shown that some anti-TRAIL-R2 antibodies did not induce a sufficient degree of TRAIL-R2 oligomerization to trigger apoptosis and that the cross-linking of these antibodies was required to induce optimal apoptosis in vitro [ 9 , 27 ]. To further investigate the mechanism of resistance, it seems crucial to characterize in detail the interaction between the various TRAIL-R2 binders and TRAIL-R2 at the membrane level.…”

Section: Introductionmentioning

confidence: 99%

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Relationship between the agonist activity of synthetic ligands of TRAIL-R2 and their cell surface binding modes

et al. 2018

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Tumor Necrosis Factor Receptor Apoptosis Inducing Ligand (TRAIL) appears as an interesting candidate for targeted cancer therapy as it induces apoptosis in cancer cells without toxicity to normal cells. TRAIL elicits apoptosis through agonist death receptor TRAIL-R1 and TRAIL-R2 engagement. Nevertheless, recombinant soluble TRAIL and monoclonal antibodies against these receptors demonstrated insufficient efficacy in clinical trials. This may be explained by the cell-type dependency of the apoptotic response, itself influenced by the effect on ligand binding mode of factors such as the level of receptor oligomerization or glycosylation. To investigate the relation between binding mode and signaling, we used previously described synthetic divalent and monovalent peptides specific for TRAIL-R2. We measured their pro-apoptotic activity on three cancer cell lines sensitive to rhTRAIL induced-apoptosis and monitored their cell-surface binding kinetics. The two divalent peptides bound with strong affinity to TRAIL-R2 expressed on B lymphoma BJAB cells and induced a high degree of apoptosis. By contrast, the same peptides bound weakly to TRAIL-R2 expressed at the surface of the human colon cancer HCT116 or T lymphoma Jurkat cell lines and did not induce their apoptosis. Cross-linking experiments suggest that these differences could be afforded by variations in the TRAIL-R2 oligomerization state at cell surface before ligand addition. Moreover divalent peptides showed a different efficiency in BJAB apoptosis induction, and kinetic distribution analysis of the BJAB binding curves suggested subtle differences in binding mechanisms. Thus our data support a relation between the cell-surface binding mode of the peptides and their pro-apoptotic activity. In this case the precise characterization of ligand binding to the surface of living cells would be predictive of the therapeutic potential of TRAIL-R2 synthetic ligands prior to clinical trials.

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“…Chemical shift assignment has been used to perform binding studies with TRAILmim peptidomimetics that bind to DR5 (Pavet V et al, 2010;Pulka-Ziach K et al, 2015;Beyrath J et al, 2016) and the results will be reported in a future publication.…”

Section: Biological Contextmentioning

confidence: 99%

1H, 13C, 15N NMR resonance assignments and secondary structure determination of the extra-cellular domain from the human proapoptotic TRAIL-R2 death receptor 5 (DR5-ECD)

et al. 2018

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Death receptors (DR) selectively drive cancer cells to apoptosis upon binding to the Tumor necrosis factor-a-Related Apoptosis-Inducing Ligand (TRAIL). Complex formation induces the oligomerization of the death receptors DR4 (TRAIL-R1) and DR5 (TRAIL-R2) and transduces the apoptogenic signal to their respective death domains, leading to Death Inducing Signaling Complex (DISC) formation, caspase activation and ultimately cell death. Several crystal structures of the ExtraCellular Domain from Death Receptor 5 (DR5-ECD) have been reported in complex with the TRAIL ligand or anti-DR5 antibodies, but none for the isolated protein. In order to fill this gap and to perform binding experiments with TRAIL peptidomimetics, we have produced isotopically labelled DR5-ECD and started a conformational analysis by using high-field 3D NMR spectroscopy. Herein, we present the first resonance assignment of a TRAIL receptor in solution and the determination of its secondary structure from NMR chemical shifts.

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Synthetic ligands of death receptor 5 display a cell-selective agonistic effect at different oligomerization levels

Cited by 12 publications

References 50 publications

Superior Properties of Fc-comprising scTRAIL Fusion Proteins

Superior Properties of Fc-comprising scTRAIL Fusion Proteins

Relationship between the agonist activity of synthetic ligands of TRAIL-R2 and their cell surface binding modes

1H, 13C, 15N NMR resonance assignments and secondary structure determination of the extra-cellular domain from the human proapoptotic TRAIL-R2 death receptor 5 (DR5-ECD)

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