Development of a high-throughput arrayed neural circuitry platform using human induced neurons for drug screening applications

Abstract

Development and functionality of a 96 well plate-based platform for high-throughput drug screening of compartmentalized neurocircuit models.

Advances in microfluidic in vitro systems for neurological disease modeling

“…Through precise control of microenvironments, MPS offer new opportunities to create in vitro physiologically and pathophysiologically relevant models. as axonal response to injury (Taylor et al, 2005), myelination (Park et al, 2009), synaptic formation and function (Shi et al, 2013), probing the direct and indirect response of neuronal cultures to chemical stimuli (Robertson et al, 2014), neurite growth (Frimat et al, 2010) (Nagendran et al, 2018), proof-of-concept pharmacology (MacKerron et al, 2017), and drug screening (Fantuzzo et al, 2020 (Holloway et al, 2019;Peyrin et al, 2011;Renault et al, 2016) (Figure 2a,b), and also more recently with induced pluripotent stem cell (iPSC)-derived neuronal cultures (Gribaudo et al, 2019).…”

“…The throughput of models for the lead discovery can range from 1000s for antibody screening to 100,000, even 1,000,000s (Fantuzzo et al, 2020), while Parrish et al, 2018, have taken a membrane-based dual perfusion chamber approach to provide a 96-well microplate platform amenable for BBB studies (Parrish et al, 2018). This last example and many other MPS models require complex bespoke perfusion systems and holders to operate the devices, which may limit up take or restrict the user to a single type of MPS, thus raising the issue of standardization within the field.…”

“…Based on the microfluidic design proposed in a landmark paper from 2003 (Taylor et al., 2003), several examples of how neuronal cell cultures can be grown, manipulated, and studied in microfluidic devices have been provided in the last 15 years, with a number of device options commercially available for neuroscientists to study axonal growth and connectivity, such as Merck's AXIS™ Axon Isolation Devices, Ananda's™ Neuro Devices, Ufluidic's Axon chips, eNUVIO OMEGA4 Neuronal Co‐Culture chips and Xona Microfluidics ® XonaChips (summarized in Table 1) . Various modifications of the device layout and the development of new methodologies based on the device topology have since been reported, expanding the breath of applications that miniaturized technologies can provide in neuroscience research, such as axonal response to injury (Taylor et al., 2005), myelination (Park et al., 2009), synaptic formation and function (Shi et al., 2013), probing the direct and indirect response of neuronal cultures to chemical stimuli (Robertson et al., 2014), neurite growth (Frimat et al., 2010) (Nagendran et al., 2018), proof‐of‐concept pharmacology (MacKerron et al., 2017), and drug screening (Fantuzzo et al., 2020). Microfluidic topologies have also shown the capability to promote the asymmetric spatial organization of dendritic and axonal subcellular components, leading to the formation of unidirectional connections where axons from one culture are free to grow in a permissive direction, but extension from a neighboring culture is impeded, thus forming asymmetric connections (Holloway et al., 2019).…”

“…Lee et al., 2019; S. Lee et al., 2019) (12 devices in a microscope slide footprint or 96 devices on a 96‐well plate format) or the platform presented by Phan et al (2017) (12 devices in a 96‐well plate format), all of which provide patterning of hydrogels for culture of multiple cell types with flanking perfusion channels supplied with flow by passive hydrostatic head. Fantuzzo et al have also taken the concept of axon guidance first presented by Taylor et al and developed a high‐throughput arrayed neural circuitry platform with 96 devices per plate (Fantuzzo et al., 2020), while Parrish et al., 2018, have taken a membrane‐based dual perfusion chamber approach to provide a 96‐well microplate platform amenable for BBB studies (Parrish et al., 2018). This last example and many other MPS models require complex bespoke perfusion systems and holders to operate the devices, which may limit up take or restrict the user to a single type of MPS, thus raising the issue of standardization within the field.…”

Drug Screening with Genetically Encoded Fluorescent Sensors: Today and Tomorrow

“…Increasing agonist concentration resulted in increasing cell activity in both chambers, indicating the presence of excitatory inputs through the chambers’ microchannels. The proposed platform provides a powerful means for automated HTS of therapeutic reagents in established neurocircuits carrying the genetic context of a patient and may be used for testing personalized drug therapies as well [ 130 ].…”