Poster #043, UC Davis Health System
Measurement of Cortical Organoid Growth as a Disease Model Phenotype for CNKSR2-related Neurodevelopmental Disorders
Mentors: Casiana Gonzalez, PhD; PI: Julian Halmai, PhD
CNKSR2-related neurodevelopmental and epilepsy disorder is a type of neurodevelopmental disorder marked by intellectual disabilities, early-onset seizures, and language delays. It is caused by de novo mutations in the CNKSR2 gene located on the X chromosome, which when mutated causes a loss of function of the CNKSR2 protein. Under healthy conditions, CNKSR2 is a scaffolding protein found in the synapses of neurons. Although information about the function of CNKSR2 in neuronal cells along with its connection to neurodevelopmental disorders is apparent, its role in neural cell development is not fully clear. To investigate CNKSR2’s influence on neuronal development, cortical organoids were used, as they are useful for modeling complex systems. These are produced by aggregating neural stem cells into a sphere and allowing them to mature further. The neural stem cells used are hemizygous to represent the population most affected, males. I hypothesized that the organoids with a loss of function of CNKSR2 would appear smaller due to a decrease in the overall cell size caused by CNKSR2’s role in promoting neurite outgrowth. To investigate the relationship between CNKSR2 and neuronal cells, we cultured cortical organoids that contained either the mutation c.2145+1G>A, c.2303G>A, or a healthy copy of CNKSR2. In 7 day increments, data on the organoids’ area were collected using ImageJ. The data showed that in organoids with the mutated copy, the cortical organoid size was around the same as or greater than that of the organoids with the healthy copy in a non time specific manner, indicating that phenotype changes in organoid size may not be caused by CNKSR2 but by technical differences in imaging. These results will help develop a greater understanding of CNKSR2 and its influence on neuronal development, helping bridge the gap in knowledge regarding rare neurodevelopmental disorders.