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Enhanced DR5 binding capacity of nanovectorized TRAIL compared to its cytotoxic version by affinity chromatography
Albatoul Zakaria1, Fabien Picaud1, Yves Claude Guillaume1
1NanoMedicine, Imagery and Therapeutics Lab EA 4662, University of Franche-Comte, CHU Jean Minjoz, 25030, Besançon cedex, France.
Abstract:
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis of cancer cells when bound to its cognate receptors, TRAIL-R1 and TRAIL-R2 (DR4 and DR5), without being toxic to healthy cells. Nanovectorized TRAIL (abbreviated as NPT) is 10 to 20 times more efficient than one of the most potent soluble TRAIL used in preclinical studies (His-TRAIL). To determine whether differences in affinity may account for NPT superiority, a thermodynamic study was undertaken to evaluate NPT versus TRAIL binding affinity to DR5. Docking calculations showed that TRAIL in homotrimer configuration was more stable than in heterotrimer, because of the presence of one Zn ion in its structure. Indeed, TRAIL trimers can have head-to-tail orientations when Zn is missing. Altogether these data suggest that TRAIL homotrimer structures are predominant in solution and then are grafted on NPT. When docked to DR5, NPT carrying TRAIL homotrimer leads to a more stable complex than TRAIL monomer-based NPT. To comfort these observations, the extracellular domain of DR5 was immobilized on a chromatographic support using an "in situ" immobilization technique. The determination of the thermodynamic data (enthalpy ∆H° and entropy ∆S°*) of TRAIL and NPT binding to DR5 showed that the binding mechanism was pH dependent. The affinity of NPT to DR5 increased with pH, and the ionized energy was more important for NPT than for soluble TRAIL. Moreover, because of negative values of ∆H° and ∆S°* quantities, we demonstrated that van der Waals and hydrogen bonds governed the strong NPT-DR5 association for pH > 7.4 (as for TRAIL alone). Copyright © 2016 John Wiley & Sons, Ltd.
Insights
Nanovectorized TRAIL (NPT) exhibits superior cancer cell apoptosis induction compared to soluble TRAIL. Thermodynamic studies reveal NPT
Area of Science:
- Biochemistry
- Molecular Biology
- Nanotechnology
Background:
- Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces cancer cell apoptosis via TRAIL-R1/R2 (DR4/DR5) binding.
- Nanovectorized TRAIL (NPT) demonstrates significantly enhanced efficacy over soluble TRAIL in preclinical models.
- Understanding the thermodynamic basis of NPT's superior binding affinity to DR5 is crucial for its therapeutic development.
Purpose of the Study:
- To investigate the thermodynamic parameters governing the binding affinity of NPT to the DR5 receptor.
- To elucidate the structural basis for NPT's enhanced efficacy compared to soluble TRAIL.
- To determine the influence of pH on the binding kinetics and thermodynamics of NPT-DR5 interactions.
Main Methods:
- Molecular docking simulations to assess TRAIL homotrimer stability and NPT-DR5 complex formation.
- Immobilization of the DR5 extracellular domain on a chromatographic support for thermodynamic analysis.
- "In situ" immobilization technique for real-time binding studies.
- Determination of thermodynamic data (enthalpy and entropy) for TRAIL and NPT binding to DR5.
Main Results:
- TRAIL homotrimer configuration is more stable than heterotrimer, particularly with Zn ion presence.
- NPT-grafted TRAIL homotrimers form more stable complexes with DR5 than monomer-based NPT.
- NPT binding affinity to DR5 increases with pH, with greater ionized energy contribution compared to soluble TRAIL.
- Van der Waals and hydrogen bonds are key drivers of strong NPT-DR5 association at pH > 7.4.
Conclusions:
- NPT's enhanced efficacy is attributed to stable TRAIL homotrimer structures and favorable thermodynamic binding to DR5.
- The pH-dependent binding mechanism, driven by van der Waals and hydrogen bonds, underlies NPT's potent anticancer activity.
- These findings support NPT as a promising nanomedicine for cancer therapy, offering improved target engagement.
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