LXRepair: Development of miniaturized DNA repair assays for animal models of ageing
i. Objective of research: To adapt and miniaturize a series of DNA repair assays for use in biological models of (accelerated) ageing.
ii. Current state of the art: Genome maintenance pathways represent pivotal defense mechanisms so that cells can maintain and faithfully transmit their genome intact to progeny. A number of studies have shown that DNA repair pathways are functionally impaired during ageing, reactive metabolites or environmental pollutants with deleterious consequences for overall cellular function and organismal survival. In line, a number of human syndromes and accompanied mouse models with defects in DNA repair manifest with the premature onset of age-related pathological features and diseases, including cancer.
iii. Research methodology and approach: We have recently established a multiplexed approach enabling the simultaneous screening of several enzymatic DNA Repair activities from a single sample on a microsupport. Here, we propose to use this system for multiple detection of various types of DNA lesions allowing the system vision and comparative quantification of different DNA repair pathways in C. elegans and primary cells derived from progeroid Cockayne syndrome (CS) patients and associated animal models (e.g. Csbm/m mice). Furthermore, our ESR will focus on to apply our developed technology to establish a DNA repair enzyme signature in cells or tissues derived from young adult and naturally aged mice as well as from animals exposed to low and high fat diet for a period of 6 months. The distinct DNA repair enzymes contained in these biological sample are expected to react with series of lesion-containing DNA on a biochip. The repair reaction occurring at each lesion site will then be measured by fluorescence analysis. The data will be coupled with cell cycle parameters as well as biomarkers for senescence, autophagy, endoplasmic reticulum stress and Golgi dispersal.
iv. Originality and innovative aspects of the ESR project: DNA damage is considered a major causal contributor of ageing. However, the exact nature of DNA lesions or DNA repair pathways involved remain currently unknown hindering the development of rationalized intervention strategies. Our project has the potential to provide us with invaluable insights as to the exact nature of the DNA lesions that accumulate during ageing (also as a consequence of chronic exposure to western diet) as well as the type of the DNA repair mechanism involved.
v. Integration of the ESR project to the overall research programme: Our ESR will work with the Schumacher group on cellular senescence, with the Garinis group to validate our system on NER progeroid animal models and with the ProtATonce for the monitoring of selected age-related biomarkers.