Abstract Details

Title Analyzing Normal and Multiple Sclerosis Brain Cells Using Single-Nucleus RNA Sequencing: Transcriptome Analyses of Normal and Diseased Cells Expressing S1P Receptors and Related Enzymatic Genes

Topic Multiple Sclerosis

Presentation(s) P2 - Poster Session 2 (5:30 PM-6:30 PM)

Poster/Presentation
Number 15-080

Objective Implement single-cell RNAseq analyses of astrocyte primary culture models and post-mortem human CNS to determine activated cellular subpopulations in multiple sclerosis with relevance to S1P receptor signaling.

Background Recent reports indicate that sphingosine 1-phosphate receptor (S1PR) modulating compounds used in the treatment of multiple sclerosis (MS) can have direct CNS effects, raising the question of how therapeutic benefits for these compounds are achieved and which cell types and gene pathways in the CNS are involved. One clear cell type affected are astrocytes which express multiple S1PR subtypes. Other cell types may also be relevant, in view of the extraordinarily diverse range of heterogeneous cell types present within the human brain.

Design/Methods

Recent advances in single-cell sequencing have begun to reveal the transcriptional diversity in the CNS, including cell types expressing a number of S1P receptors. Our approach pairs extraction of cellular nuclei with single-cell sequencing using 10X Genomics to assess differential activation of individual cellular subpopulations in primary human astrocytes and post-mortem human MS brain samples. Extraction of nuclei for single-cell sequencing provides 2 main advantages: 1) Nuclei provide an instantaneous “snapshot” of cellular transcriptional state, avoiding artifacts generated from creating whole-cell suspensions from the brain and 2) Nuclei isolation provides an unbiased method to sample all cell types within CNS tissue, independent of cellular morphology. Effects of S1P receptor modulators in primary human astrocytes are also being assessed.

Results This pipeline has been used to identify transcriptomic signatures of human brain cells at the level of single cells, which has identified multiple brain cell types relevant to S1PR modulation by virtue of their expressing a range of S1PR and related enzymatic genes.

Conclusions These data are being compared across human MS samples from the CNS and primary human astrocytes towards understanding the responses to S1P receptor modulators accessed through direct CNS activities.

Authors/Disclosures
PRESENTER	No disclosure on file
	No disclosure on file
	No disclosure on file
	No disclosure on file
	No disclosure on file
Jerold Chun	No disclosure on file

��ɫ��

��ɫ��