Dottorato in SCIENZE DELLA VITA

Titolo della tesi: Targeting adenosine A2A receptor in FXS-patient derived human cortical culture for animal free drug discovery and repositioning.

Intellectual disabilities affect a significant percentage of the population, with Fragile X Syndrome (FXS) standing out as a prevalent neurodevelopmental disorder and a leading heritable cause of Autism Spectrum Disorders (ASDs). FXS is characterized by the loss of the fragile X mental retardation protein (FMRP), crucial for regulating mRNA associated with synaptic plasticity and neuronal development. Despite insights gained from animal models, translating findings into effective therapies for patients remains challenging due to species differences. The advent of human induced pluripotent stem cells (hiPSCs) provides a revolutionary platform, allowing for the modeling of FXS by incorporating patient-derived cells carrying the disease-causing mutation. This Ph.D. project aimed to investigate neurodevelopmental alterations in FXS at cellular and network levels, utilizing hiPSC-derived bidimensional cell cultures. These cultures, established from FXS patients and healthy controls, serve as reliable platforms for understanding the pathological mechanisms of FXS and facilitate drug discovery. The focus on a 2D culture model comprising neural progenitors, neurons, and glial cells revealed a persistent population of neural progenitor cells throughout neurodevelopment, highlighting FMRP's role in regulating progenitor proliferation and cortical specialization. Characterizing glutamatergic and GABAergic synapse development and assessing calcium activity within the neuronal network unveiled heightened activity and immaturity, consistent with FXS hallmarks. Leveraging this reliable cellular system, the study explored the therapeutic potential of istradefylline, an A2A receptor antagonist previously tested in FMRP-KO mice. Chronic treatment of FXS cell cultures demonstrated a significant rescue in cortical development and overall network activity. This study not only emphasizes the potential therapeutic value of istradefylline for FXS but also underscores the importance of humanized models in advancing treatments for neurodevelopmental disorders, including FXS.