Oxidative Stress and Erythrocyte Membrane Alterations in Autism Spectrum Disorder: on Search of a Biomarker and of New Therapeutic Prospects

Project location: Italy, Bologna
Project start date: June 2015 - Project end date: May 2016
Project number: 2015-014
Beneficiary: Associazione Nazionale Genitori Soggetti Autistici

The project is aimed at understanding why red blood cell membranes of autistic children are endowed of a number of anomalous features, as described in the above-mentioned scientific article published in PLoS One, 2013 Jun 19;8(6). In fact, red blood cell membranes of autistic children are more stiff, have an anomalous lipid composition and the activity of one of the most important membrane enzymes (Na+, K+-ATPase) is severely depressed. The project is aimed at identifying the source(s) of these anomalous features, which may be compatible with the presence of oxidative stress, as suggested also by an increase in urinary oxidative stress biomarkers. However, the origin of such redox unbalance is still unknown.

Twenty autistic children and twenty age- and sex-matched typically developing children will be studied. An accurate selection will be made at diagnosis in order to exclude from the study children with concomitant pathologies or taking supplements. The biochemical evaluations will be addressed at i) the activity of Na+, K+-ATPase, the expression of its different subunits and their possible oxidative modifications in both erythrocytes and leukocytes; ii) the composition of membrane lipids; iii) markers of oxidative and nitrosylative stress; iv)expression of erythrocyte enzymes involved in the control of oxidation, such as peroxyredoxin. Moreover, erythrocyte morphology will be evaluated by Trasmission and Scanner Electron Microscopy. A study will be carried out with hyperspectral microscopy, which has never been applied so far to the study of erythrocytes.
Autistic and typically developing children will be enrolled in the Italian Region Emilia Romagna. Samples will be collected by IRCCS Istituto delle Scienze Neurologiche, Ospedale Bellaria, Bologna, Italy. Most laboratory work will be carried out in Bologna, Dept. of Experimental, Diagnostic, and Specialty Medicine, School of Medicine, University of Bologna, Italy, by the research team headed by prof. M. Marini, who will also collaborate with researchers located in Ancona, Verona and Urbino (all in Italy).

The Nando Peretti Foundation has awarded a grant for this project. This study will hopefully lead to a better understanding of the biological basis of Autism Spectrum Disorder.
In turn, a better acquaintance with the biological alterations of membranes may in the future lead to therapeutic interventions, aimed at their phenotypic correction (e.g. nutraceutical supplementation of specific membrane lipids and antioxidants). In this way, we will be able to suggest a bottom-up approach that could hopefully lead to the relief of some ASD symptoms (such as hyperactivity), which were found to be statistically correlated with some of the above-mentioned anomalies.
Moreover, the marked decrease in Na+, K+-ATPase activity we found could also constitute a possible biomarker that, together with other biological features which we are going to explore, might help in making earlier diagnoses, thus anticipating the age when educational-behavioral interventions will be administered to ASD children.