DEVELOPMENT OF IPSC-BASED MODEL SYSTEM FROM PATIENTS WITH 22Q11.2 DUPLICATION SYNDROME FOR STUDYING NEURODEVELOPMENTAL DISORDERS

Abstract

22q11.2 Duplication Syndrome (22q11.2DupS) is associated with an elevated risk of developing neurodevelopmental disorders (NDDs), such as autism spectrum disorders (ASD) and attention-deficit/hyperactivity disorder. ASD is detected in 14-25% of patients, making 22q11.2DupS one of the genetic syndromes with the highest prevalence of ASD. To better understand the molecular mechanisms underlying the pathophysiology of NDDs, we have generated induced pluripotent stem cells (iPSCs) from patients with 22q11.2DupS. Mononuclear cells from 22q11.2DupS patients and healthy individuals were reprogrammed into iPSC lines using the CytoTuneTM-iPS 2.0 Sendai Reprogramming Kit. The established iPSC lines were characterized by examining the expression of stem cell markers through RT-PCR and their ability to differentiate into cells of three germ layers using the STEMdiffTM Trilineage Differentiation Kit. We differentiated the iPSCs into neurons using the dual SMAD inhibition method together with retinoic acid. The expression of neural and neuronal-specific markers in neural progenitors and mature neurons was analyzed using RT-PCR and immunocytochemistry. The established iPSC lines from three patients with an inherited form of 22q11.2DupS, their mothers who carry the microduplication, and three healthy individuals demonstrated the expression of stem cell markers and the ability to differentiate into the cells of all three germ layers. Both iPSCs from carriers of the 22q11.2 microduplication and healthy controls successfully differentiated into neurons, which was confirmed by the expression of neuronal markers. The generated 22q11.2DupS iPSC lines provide a valuable resource for understanding the molecular mechanisms underlying NDDs associated with 22q11.2 microduplication.Abstract book: FENS Regional Meeting 2025, Oslo, Norway, 16-19 June 202