No abstract

Oxidative stress and hypoxia in the retinal pigment epithelium (RPE) have long been considered major risk factors in the pathophysiology of age-related macular degeneration (AMD), but systematic investigation of the interplay between these two risk factors was lacking. For this purpose, we treated a human RPE cell line (ARPE-19) with sodium iodate (SI), an oxidative stress agent, together with dimethyloxalylglycine (DMOG) which leads to stabilization of hypoxia-inducible factors (HIFs), key regulators of cellular adaptation to hypoxic conditions. We found that HIF stabilization aggravated oxidative stress-induced cell death by SI and iron-dependent ferroptosis was identified as the main cell death mechanism. Ferroptotic cell death depends on the Fenton reaction where H2O2 and iron react to generate hydroxyl radicals which trigger lipid peroxidation. Our findings clearly provide evidence for superoxide dismutase (SOD) driven H2O2 production fostering the Fenton reaction as indicated by triggered SOD activity upon DMOG + SI treatment as well as by reduced cell death levels upon SOD2 knockdown. In addition, iron transporters involved in non-transferrin-bound Fe2+ import as well as intracellular iron levels were also upregulated. Consequently, chelation of Fe2+ by 2’2-Bipyridyl completely rescued cells. Taken together, we show for the first time that HIF stabilization under oxidative stress conditions aggravates ferroptotic cell death in RPE cells. Thus, our study provides a novel link between hypoxia, oxidative stress and iron metabolism in AMD pathophysiology. Since iron accumulation and altered iron metabolism are characteristic features of AMD retinas and RPE cells, our cell culture model is suitable for high-throughput screening of new treatment approaches against AMD.

Radiotherapy plays a key role in the management of lung cancer patients in curative and palliative settings. Traditionally, radiotherapy was either given alone or in combination with surgery, classical cytotoxic chemotherapy, or both. Technical and physical innovations achieved during the last two decades have helped to enhance the accuracy of radiotherapy dose delivery and have facilitated geometric radiotherapy individualization. Furthermore, multimodal combinations with molecularly tailored drugs or immunotherapy yielded promising survival benefits in selected patients. Yet high locoregional failure rates and frequent development of metastases still limit the patient outcome. One major obstacle to successful treatment is the high molecular heterogeneity observed in lung cancer. So far, clinical radiotherapy does not routinely use the knowledge on molecular subtypes with regard to therapy individualization and predictive biomarkers are missing. Herein, altered cancer metabolism has attracted novel attention during recent years as it promotes tumor growth and progression as well as resistance to anticancer therapies. The present perspective will exemplarily highlight how clinically relevant molecular subtypes defined by co-occurring somatic mutations in KRAS-driven lung cancer impact the metabolic phenotype of cancer cells, how the metabolic phenotype supports intrinsic radioresistance by the improved antioxidant defense, and also discuss potential subtype-specific actionable metabolic vulnerabilities. Understanding metabolic phenotypes of radioresistance and metabolic bottlenecks of cancer cells undergoing radiotherapy in a cancer-specific context will offer largely unexploited future avenues for biological individualization and optimization of radiotherapy. Transcriptional profiles will provide additional benefit in defining metabolic phenotypes associated with radioresistance, particularly in cases, where such dependencies cannot be identified by specific somatic mutations.

Foaming process can be monitored under batch or continuous flows conditions. In the batch process, foaming is time-dependent and the foaming efficiency is controlled by the operator. On the other hand, in the continuous process, the foaming efficiency is only monitored by gas and liquid flow rates. The aim of this work is to compare the two technologies to perform porous scaffold biomaterial based on chitosan (a biocompatible polysaccharide) as well as calcium (Ca2+) and silica (SiO2) (two osteogenesis compounds). Diverse recipes using chitosan (CS) solution (2% (w/v)) in acetic acid (1% (v/v in distilled water)) mixed with whey protein isolate (WPI) (2% (w/v)) as natural surfactant were studied. They were supplemented or not by hydroxyapatite powder (HAp) and tetraethyl orthosilicate (TEOS). A jacketed narrow annular gap unit (NAGU) was used to perform the continuous foaming process. For all experimentations, the mixture flow rate was maintained at 30 mL min-1. The influence of operating conditions such as gas and liquid flow rates was studied to obtain foams and final scaffold material with different densities and porosities. Some other recipes followed foaming under batch conditions. Generally, the recipes were placed in a vessel under mixing allowing the gas phase to come from the roof of the vessel. In this case, it becomes very difficult to control the density and the size distribution of bubbles in the final product. In both cases, liquid foams were analysed (density, bubble size distribution) and then freeze-dried for mechanical and porosity investigations using the dynamic mechanical analysis (DMA) system and scanning electron microscopy (SEM). It has been shown that the controlled injected gas affected the continuous phase, resulting in a lighter and higher porous structure, a more homogeneous appearance, and a more uniform distribution of osteogenesis components compared to one obtained using batch operation. The obtained porous materials exhibited good properties (porosity, interconnectivity, and good HAp and silica distribution) and potential for future bone regeneration applications.