Projects

Development of high-throughput in vitro assays for drug screening and validation

The DDD program is adopting an approach of utilizing inducible pluripotent stem cells (iPSCs) to develop cellular assays for in vitro drug candidate screening and validation. Currently, our in-house approach focuses on characterization of (1) neuronal morphological features and synaptic puncta, along with (2) neuronal network activity in human neurons generated from iPSC lines (patient-derived and in CRISP/Cas9 engineered) harboring ADNP, SHANK3, DDX3X and FOXP1 mutations and their respective control lines with the final aim of developing an in vitro human model for ASD that can be used as a platform for drug screening and validation.

Neuronal morphology and synaptic puncta characterization of induced neurons (iN) modeling monogenic forms of ASD – Our aim is to describe for the first time an in-depth high-content phenotypic characterization in human ASD in vitro models by establishing and optimizing a protocol for the characterization of neuronal morphology features and synaptic density. The methodology involves large-scale generation of mature neurons followed by an automated and simultaneous measurement of neuronal shape and synaptic density using the High-Content Screening (HCS) Platforms.

Neuronal network activity characterization of induced neurons (iN) derived from monogenic forms of ASD – To identify electrical network phenotypes, we exploit the multielectrode-array (MEA) technology (Axion BioSystems) technology and capture key features of neuronal network behavior, such as: activity, synchrony, and network oscillations or bursting. Our final aim is to define a robust electrophysiological feature with relevant translational medicine implication, which will result in an in vitro platform suitable for second-tier hit validation.

Development of the in vivo animal models for second tier screening and drug candidate validation
Our main aim is to define a battery of standard, translationally relevant animal models that will be used for further screening and validation of drug candidates identified in vitro. Taking advantage of the mouse models generated by the Seaver Autism Center, the DDD initiative includes performing (1) ex vivo electrophysiology assays and (2) in vivo electroencephalogram (EEG) measurements in mouse bearing clinically relevant Shank3, Ddx3x, Adnp and Foxp1 mutations.