Engineered T cell therapy for central nervous system injury [CSF RNAseq + VDJ]
Traumatic injuries to the central nervous system (CNS) affect millions of people worldwide yet lack an effective treatment. These injuries contain infiltrating immune cells that can promote tissue repair and could be exploited for therapeutic benefit. Here, using single-cell RNA-sequencing of T cells infiltrating the injured CNS we demonstrate their clonal expansion and antigen specificity towards CNS derived self-peptides. We confirm the beneficial effect of these injury-associated autoimmune CD4+ T cells in murine models of optic nerve and spinal cord injury. Subsequently, using mRNA-based transient T cell receptor (TCR) reconstitution, we demonstrate a therapeutic T cell strategy to alleviate CNS injury. Treatment of CNS-injured mice with this therapy improved locomotion and alleviated histological signs of damage, through regulation of myeloid cells, without detrimental autoimmune side effects. This strategy provides a means of developing custom-designed T cell therapies for CNS injury, and possibly for other neurodegenerative disorders. Overall design: Human CSF was collected via a lumbar drain daily from SCI patients. 10 mL of CSF was used at each time point for cell isolation. CSF was transferred into a 15 mL centrifuge tube and centrifuged at 500 g for 5 minutes. The supernatant was removed and cell pellet was resuspended with PBS containing 0.04 % BSA and filtered with 40 µm cell strainer. Cell number and viability were determined using Viastain AOPI Staining Solution (CS2-0106, Nexcelom Bioscience) and the CellDrop FL Fluorescence Cell Counter (DeNovix). Cell samples were loaded onto a 10X Genomics Chromium platform for Gel Bead-in Emulsions (GEMs) and Human T cell Chromium Single Cell V(D)J Enrichment Kit and Chromium Single Cell 5' Library & Gel Bead Kit were used separately to generate a cDNA library targeting the TCR and for transcriptome expression.