The effect of caffeine and purinergic signaling in three-dimensional models of Alzheimer's Disease using bioprinting

• Grant number: 24/04690-6
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: May 01, 2024
• End date: March 31, 2025
• Field of knowledge: Biological Sciences - Biophysics - Cellular Biophysics
• Principal Investigator: Alexander Henning Ulrich
• Grantee: Fernanda Tibolla Viero
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 18/07366-4 - Purine and kinin receptors as targets of study and therapeutic interventions in neurological diseases, AP.TEM

Abstract

Alzheimer's disease (AD) is a progressive and disabling neurodegenerative condition, with an increasing prevalence, projected to affect approximately 100 million people by 2050. Initially, it manifests with memory impairment and language difficulties, progressing to total loss of communication and physical skills. Neuropathological features include neuritic plaques, neurofibrillary tangles, as well as neuronal loss in specific brain regions. Studies suggest that AD involves not only ²-amyloid plaque accumulation and tauopathy, but also inflammation and oxidative stress.Purinergic receptors, which recognize nucleotides and nucleosides, play a crucial role in AD. A2A receptors, activated by adenosine, have been associated with neuroprotection and the prevention of memory dysfunction in animal models. On the other hand, P2X7 and P2Y2 receptors, activated by ATP, have opposite roles in AD, with the first being considered pro-degenerative and the second neuroprotective. Therefore, this study aims to explore the molecular mechanisms underlying the neuroprotective effect of caffeine in a three-dimensional (3D) in vitro model of AD, using cell printing with bioink. Neurospheres from neural stem cells extracted from transgenic mice for the AD model, exposed to caffeine, will be used to evaluate their differentiation and response to the induction of neuronal death. The methodology involves techniques such as 3D bioprinting, neural stem cell cultivation, pharmacological treatments with caffeine and purinergic receptor antagonists, assessment of Tau, caspase-3/7, in addition to cell viability assays, A² quantification, ROS detection and molecular analyzes such as immunohistochemistry, Western blotting and real-time PCR. In summary, this study using caffeine and the possible modulation of purinergic receptors may offer a promising therapeutic approach for AD treatment, using an innovative 3D bioprinting model and detailed molecular analyses. (AU)