Characterizing Schwann cell transcriptional states in peripheral nerve regeneration with single-cell RNA sequencing
Suresh Mohan, MD1, Alan Tenney, PhD2, Zhu Zhuo, PhD3, Brandon Pratt, BS2, Thomas Collins, BS2, Alon Gelber, BS2, Shannan J Ho Sui, PhD3, Tessa A Hadlock, MD4, Nate Jowett, MD4 and Elizabeth C Engle, MD2, (1)Mass. Eye and Ear/ Harvard Medical School, Boston, MA, (2)Boston Children's Hospital, Boston, MA, (3)Harvard Chan School of Public Health, Boston, MA, (4)Facial Nerve Center - Dept. of Otolaryngology, Harvard Medical School / Massachusetts Eye and Ear, Boston, MA
INTRODUCTION
Upregulation of transcription factor cJun is critical to the transdifferentiation of myelinating and non-myelinating Schwann cells (SCs) into repair SCs after peripheral nerve injury. However, the complex signaling that produces repair SCs requires further analysis to develop therapeutic tools to enhance regeneration through promotion and maintenance of the repair SC phenotype. Herein, we employ single-cell RNA sequencing (scRNAseq) to characterize the peripheral nerve cellulome and delineate transcriptional states of Schwann cells transitioning from mature to repair phenotypes.
MATERIALS & METHODS
Sox10-venus mice labeling Schwann cells underwent unilateral facial nerve transection. Five days post-injury, distal nerve segments were harvested and dissociated into a single-cell suspension. The suspension was processed through the 10X Chromium scRNAseq platform and sequencing of over 5,000 cells per condition was performed. Differential gene expression analysis was utilized to identify novel transcriptional signatures. Tissue expression of identified gene targets was assessed with in situ hybridization.
RESULTS
The peripheral nerve cellulome in healthy and injured murine facial nerve is characterized. Known and novel genes expressed by repair SCs and resident cells of the peripheral nerve were identified. In situ hybridization verified tissue expression of targets obtained in silico.
CONCLUSIONS
Characterization of the murine facial nerve cellulome with scRNAseq is described for the first time. Novel and established genes upregulated within repair SCs are presented, yielding new potential therapeutic targets to prolong the repair SC phenotype and enhance nerve regeneration. Forthcoming studies are utilizing human nerve samples to verify aforementioned target genes.
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