Evaluation of Computationally Designed Peptides against TWEAK, a Cytokine of the Tumour Necrosis Factor Ligand Family.

The tumour necrosis factor-like weak inducer of apoptosis (TWEAK) is a member of the tumour necrosis factor ligand family and has been shown to be overexpressed in tumoral cells together with the fibroblast growth factor-inducible 14 (Fn14) receptor. TWEAK-Fn14 interaction triggers a set of intracellular pathways responsible for tumour cell invasion and migration, as well as proliferation and angiogenesis. Hence, modulation of the TWEAK-Fn14 interaction is an important therapeutic goal. The targeting of protein-protein interactions by external agents, e.g., drugs, remains a substantial challenge. Given their intrinsic features, as well as recent advances that improve their pharmacological profiles, peptides have arisen as promising agents in this regard. Here, we report, by in silico structural design validated by cell-based and in vitro assays, the discovery of four peptides able to target TWEAK. Our results show that, when added to TWEAK-dependent cellular cultures, peptides cause a down-regulation of genes that are part of TWEAK-Fn14 signalling pathway. The direct, physical interaction between the peptides and TWEAK was further elucidated in an in vitro assay which confirmed that the bioactivity shown in cell-based assays was due to the targeting of TWEAK. The results presented here are framed within early pre-clinical drug development and therefore these peptide hits represent a starting point for the development of novel therapeutic agents. Our approach exemplifies the powerful combination of in silico and experimental efforts to quickly identify peptides with desirable traits.

Synthesis of a superparamagnetic iron oxide based nano-complex for targeted cell death of glioblastoma cells.

In the last ten years, there has been little advancement in the treatment of the aggressive brain cancer Glioblastoma Multiforme (GBM). This research describes the synthesis of a superparamagnetic iron oxide (SPION)-based nanotheraputic complex for use in targeting and killing aggressive mesenchymal GBM cells. The average sizes and magnetic properties of the synthesized SPIONs are tailored via a novel time-controlled approach to a previously described electrochemical reaction. Through this synthetic method, the optimal particle size where maximal thermal energy is released upon stimulation with an external magnetic field was determined to be 21 nm. The nano-complex was further modified to selectively target GBM cells by adding a heterobifunctional poly(ethylene) glycol polymer crosslinked to TWEAK (a GBM targeting peptide). Preliminary investigation with FITC Annexin V/propidium iodide fluorescent probes and transmission electron microscopy revealed biochemical and morphological evidence of both SPION internalization and cytotoxic effects over the course of three hours. Thus, these nano-complexes hold promise as a potential treatment agent for an otherwise untreatable disease.

Serum soluble TWEAK levels in severe traumatic brain injury and its prognostic significance.

BACKGROUND: Severe traumatic brain injury (sTBI) is characterized by a high mortality. Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) participates in inflammation. We determined serum soluble TWEAK (sTWEAK) levels with respect to its prognostic ability. METHODS: This was a single-center prospective, observational study that was performed from December 2014 to December 2017. A total of 114 sTBI patients who met the inclusion criteria and 114 randomly selected healthy controls were included in the study. Serum sTWEAK levels were gauged. Patients were followed-up until death or completion of 6 months. Poor outcome was referred to as Glasgow outcome scale score of 1-3. RESULTS: In comparison with controls, patients displayed predominantly higher serum sTWEAK levels. Serum sTWEAK levels were strongly correlated with Glasgow coma scale scores and serum C-reactive protein levels. 32 patients (28.1%) died and 60 patients (52.6%) suffered from a poor outcome. Receiver operating characteristic curve analysis clearly showed that serum sTWEAK levels had substantially high predictive performance for 6-month mortality and poor outcome. Serum sTWEAK emerged as an independent predictor for 6-month mortality, overall survival and poor outcome. CONCLUSIONS: Raised serum sTWEAK levels are closely related to increasing inflammatory response, elevated trauma severity and worse clinical outcome after sTBI.