Mesenchymal stem cells engineered to express selectin ligands and IL-10 exert enhanced therapeutic efficacy in murine experimental autoimmune encephalomyelitis

Abstract

Systemic administration of mesenchymal stem cells (MSCs) affords the potential to ameliorate the symptoms of Multiple Sclerosis (MS) in both preclinical and clinical studies. However, the efficacy of MSC-based therapy for MS likely depends on the number of cells that home to inflamed tissues and on the controlled production of paracrine and immunomodulatory factors. Previously, we reported that engineered MSCs expressing P-selectin glycoprotein ligand-1 (PSGL-1) and Sialyl-Lewisx (SLeX) via mRNA transfection facilitated the targeted delivery of anti-inflammatory cytokine interleukin-10 (IL-10) to inflamed ear. Here, we evaluated whether targeted delivery of MSCs with triple PSGL1/SLeX/IL-10 engineering improves therapeutic outcomes in mouse experimental autoimmune encephalomyelitis (EAE), a murine model for human MS. We found PSGL-1/SLeX mRNA transfection significantly enhanced MSC homing to the inflamed spinal cord. This is consistent with results from in vitro flow chamber assays in which PSGL-1/SleX mRNA transfection significantly increased the percentage of rolling and adherent cells on activated brain microvascular endothelial cells, which mimic the inflamed endothelium of blood brain/spinal cord barrier in EAE. In addition, IL-10-transfected MSCs show significant inhibitory activity on the proliferation of CD4+ T lymphocytes from EAE mice. In vivo treatment with MSCs engineered with PSGL-1/SLeX/IL-10 in EAE mice exhibited a superior therapeutic function over native (unmodified) MSCs, evidenced by significantly improved myelination and decreased lymphocytes infiltration into the white matter of the spinal cord. Our strategy of targeted delivery of performance-enhanced MSCs could potentially be utilized to increase the effectiveness of MSC-based therapy for MS and other central nervous system (CNS) disorders.

Elucidation of Exosome Migration Across the Blood–Brain Barrier Model In Vitro

“…For BMEC activation, cells were treated with TNF-α (50 ng/mL, BD Biosciences) for 6 hours 29 . BMEC monolayer permeability was studied using Fluorescein isothiocyanate (FITC)-dextran (1 mg/mL) (Sigma-Aldrich), which was added to the upper chamber (luminal) and aliquots from the lower chamber (abluminal) were measured for their fluorescence intensity using a Biotek Synergy HT microplate reader (excitation at 485 nm and emission at 520 nm).…”

Intranasal delivery of stem cell-based therapies for the treatment of brain malignancies

“…IND of stem-cell-based therapeutics takes advantage of stem cells’ natural tropism toward glioma and their capacity to administer multiple therapeutic modalities in a repeatable and minimally invasive course. Once the modality is fully understood, it may be used in the treatment of different brain malignancies or other brain disorders [2,15,31,124–126]. …”

Strategies to improve the migration of mesenchymal stromal cells in cell therapy

“…Potential repair mechanisms involve transdifferentiation to replace damaged neural cells and production of growth factors by MSCs. In addition, infused MSCs could modulate inflammation and stimulate endogenous neural stem cells to migrate to the target region [10,106,116,136,137] . The major obstacles for cell therapies are limited cell availability and engraftment, as well as reduced integration of grafted cells into the host tissue [138] .…”

“…In order to enhance the migration through vessel walls, MSCs were simultaneously transfected with mRNAs for P-selectin glycoprotein ligand-1 (PSGL-1) and Sialyl-Lewis(x) (SLeX). These cells produced functional ligands for P-selectins and E-selectins, resulting in significantly enhanced MSC homing to the inflamed spinal cord during experimental autoimmune encephalomyelitis (EAE) in mice [106] . Recently, microRNAs (miRNAs) have been described as regulators of MSC migration.…”