Multi electrochemical sensor arrayed platform for screening biological markers of neurological lysosomal storage diseases and acute brain injuries – MECANO

Electrochemical Sensor Arrays (ESAs), in which each individual electrode is specifically tailored to sensitively detect a particular analyte, provide an easy access to the development of tools for the simultaneous detection of multiple species. By that way, ESAs offer potential applications for investigating the behaviour of cultured cells or slices of biological tissues coupled to the array surface in response to a locally delivered chemical or physical stimulus by collecting dynamic information about the desired specific analyte. The need to develop ESAs fits well with the desire to study the key roles of reactive oxygen and nitrogen species as superoxide (O2-), nitric oxide (NO) and their reaction product peroxynitrite (ONOO-) in cellular signal transduction or cellular dysfunction. This has been the subject of a considerable amount of research because the balance between NO, O2- and ONOO- controls essential cell functions including activation, proliferation and apoptosis. Inappropriate production of NO and O2- radicals and their metabolites leads to the development of various pathologies as (cardio)vascular and neurodegenerative diseases. Recent studies have described an important oxidative stress in several neurological lysosomal storage diseases. Also, several works are currently suggesting the consideration of NO and ONOO- as biomarkers for brain damages severity following traumatic brain injury or cerebral ischemia.Thus, several research groups in the world are now actively working to overcome the technical limitations for the detection of these species. The key step in achieving the demonstration that the ONOO-/NO balance can be a risk marker is tightly linked to the accessibility of an accurate analytical concept. The elaboration of ESAs devoted to the simultaneous sensitive and selective detection of these secreted analytes is a challenging area of research. The creation of easy-to-handle disposable analytical platforms for the electrochemical detection of relative NO/ONOO- quantities is the main innovation here. Our project addresses the mentioned issue by focusing on the following actions: 1. To develop and validate low cost single use electrode array-based platforms for pharmaceutical screening relying on electrochemical technology for simultaneous detection of extra-cellular metabolites. Microelectrode arrays will be manufactured by photolithographic techniques. Electrodes in arrays will be chemically modified to tune the specificity of electrodes for the above mentioned two analytes, i.e. NO and ONOO- and endow them with the desired sensitivity, selectivity and biocompatibility. These microelectrode arrays will then be integrated with cell culture wells and plates, resulting in biocompatible 'array-cultured cell plate' platform. 2. To design networks of transparent electrode array platform configuration allowing simultaneous optical and electrochemical measurements. This feature will allow multiparameter biological experiments using fluorescent and electrochemical techniques. 3. To develop an accurate disposable miniaturized arrayed implantable microsensors for the quantification of NO and ONOO- in biological tissues and slices (neuronal tissues, plasma) to mimic in vivo situations, easily applicable for neuronal testing, medical diagnostic and preclinical follow-up studies. 4. To apply the developed platforms to the screening of the effects of anti oxidative agents and the understanding of models such as traumatic brain injuries, ischemia and in a model of mucopolysaccharidose of the type IIIA (model MPS IIIA of the disease of Sanfilippo) in which the effects of antioxydant agents will be studied on cell culture, slices of biological tissues and in vivo animals. The main expected result is the development of devices for addressing important biological questions that remain unresolved due to the inadequacy of the current state of the art technology in fundamental, experimental and clinical research fields. Our project will create a basis for a new generation of analytical tools and drugs testing devices. This will be considered by focusing on openings towards in vivo applications such as anti oxidative pharmacology and elucidations of brain damage severity following traumatic brain injuries or in the MPS IIIA disease model.