Description

Single cell transcriptomics of patient specific iPSC-derived motor neuron cultures for ALS disease modeling

Shila Mekhoubad¹, Dan Wagner², Alexander McCampbell¹, Chao Sun¹, Mei Liu¹, Norm Allaire¹, John Carulli¹, Allon Klein²

¹Biogen, 225 Binney St. Cambridge, MA 02142; ²Harvard Medical School, Department of Systems Biology, 200 Longwood Ave, Warren Alpert 536 Boston, MA 02115

Amyotrophic lateral sclerosis (ALS) is a late onset neurodegenerative disease that leads to the selective loss of upper and lower motor neurons in the brain and spinal cord. While the majority of ALS cases are sporadic, several mutations in familial cases have been identified to be directly causative of this disease. Patient derived induced pluripotent stem cells (iPSCs) and neuronal differentiations have provided a new platform to study ALS specific disease mechanisms in vitro. However, determining the consequences of the inherited genetic mutations on gene regulation and on phenotypic outcomes have been challenging due to the heterogeneity of the generated cells and the varying differentiation propensities between different lines.

Here we describe the use of a high-throughput single-cell RNA-Seq platform to profile motor neuron (MN) cultures derived from iPSCs with particular ALS mutations. Transcriptomic analyses of cell-type diversity revealed these cultures to be highly enriched for multiple subtypes of MNs as well as interneurons and glia, cell populations that are all found in vivo in the spinal cord have been implicated in ALS disease progression. We are currently working towards identifying cell-type specific "disease signatures" that could help empower in vitro disease modeling efforts and contribute to the development and validation of novel therapeutics.