Next generation sequencing facilitates quantitative analysis of bulk hippocampal tissue in lupus mice.

Purpose: Next-generation sequencing (NGS) technology was used to map expression profile of hippocampal tissue in mouse model of Systemic Lupus Erythematosus (SLE). Methods: Total RNA was extracted from total hippocampal tissue using NucleoSpinRNA and mRNA libraries were generated using the Illumina TruSeq Sample Preparation kit. Single-end 100bp mRNA sequencing was performed on Illumina NextSeq500 platform. Quality of sequencing was assessed using FastQC software. Raw reads in fastq format were collected and aligned to the mouse genome (mm10 version) using STAR 2.6 algorithm. Gene quantification was performed using HTSeq and differential expression analysis was performed using edgeR package. Results: We defined hippocampal-specific molecular signatures of the murine lupus transcriptome. Conclusions: By the use of the mouse hippocampal-specific transcriptome and through characterization of hippocampal neurogenesis, we showed that inflammatory mediators induce neuropsychiatric changes in SLE as an early event via the disruption of hippocampal neurogenesis. These data underscore the role of brain inflammation in the pathogenesis of early disease and support the use of immunosuppressants for the management of diffuse NPSLE. New Zealand Black (NZB) female mice were crossed with New Zealand White (NZW) male mice. Female hybrids NZB/W-F1 (n=4-5 at each time-point) were sacrificed with perfusion with PBS at the pre-nephritic stage (3 month) and symptomatic nephritic stage (6 months old where proteinuria is more than 200 mg/dl for 3 consecutive days) of the disease. Bulk hippocampal tissue was extracted. Age-matched female C57BL/6 mice (n=5) were used as controls.