Comparison of Single and Repeated Dosing of Anti-Inflammatory Human Umbilical Cord Mesenchymal Stromal Cells in a Mouse Model of Polymicrobial Sepsis

Abstract

Summary Mesenchymal stromal cells (MSCs) ameliorate pre-clinical sepsis and sepsis-associated acute kidney injury (SA-AKI) but clinical trials of single-dose MSCs have not indicated robust efficacy. This study investigated immunomodulatory effects of a novel MSC product (CD362-selected human umbilical cord-derived MSCs [hUC-MSCs]) in mouse endotoxemia and polymicrobial sepsis models. Initially, mice received intra-peritoneal (i.p.) lipopolysaccharide (LPS) followed by single i.p. doses of hUC-MSCs or vehicle. Next, mice underwent cecal ligation and puncture (CLP) followed by intravenous (i.v.) doses of hUC-MSCs at 4 h or 4 and 28 h. Analyses included serum/plasma assays of biochemical indices, inflammatory mediators and the AKI biomarker NGAL; multi-color flow cytometry of peritoneal macrophages (LPS) and intra-renal immune cell subpopulations (CLP) and histology/immunohistochemistry of kidney (CLP). At 72 h post-LPS injections, hUC-MSCs reduced serum inflammatory mediators and peritoneal macrophage M1/M2 ratio. Repeated, but not single, hUC-MSC doses administered at 48 h post-CLP resulted in lower serum concentrations of inflammatory mediators, lower plasma NGAL and reversal of sepsis-associated depletion of intra-renal T cell and myeloid cell subpopulations. Hierarchical clustering analysis of all 48-h serum/plasma analytes demonstrated partial co-clustering of repeated-dose hUC-MSC CLP animals with a Sham group but did not reveal a distinct signature of response to therapy. It was concluded that repeated doses of CD362-selected hUC-MSCs are required to modulate systemic and local immune/inflammatory events in polymicrobial sepsis and SA-AKI. Inter-individual variability and lack of effect of single dose MSC administration in the CLP model are consistent with observations to date from early-phase clinical trials. Graphical Abstract

Improving the immunomodulatory function of mesenchymal stem cells by defined chemical approach

“…Endotoxemia reactions are induced in BALB/c mice as previously described with minor adjustment by LPS (Sigma) ( 25 ). All mice were randomly divided into 3 groups: without MSC group, Ctrl-MSC group and SM-MSC group.…”

The Effects of Programmed Cell Death of Mesenchymal Stem Cells on the Development of Liver Fibrosis

“…A gene expression analysis also showed that after rats with LF were treated by MSC-EVs and MSCs, the expression levels of collagenases (e.g., MMP13 and MMP9), anti-inflammatory cytokines (e.g., IL-10 and TGF-β1), and antiapoptotic genes (e.g., BCL-2) were all upregulated, while the expression levels of proapoptotic genes (BAX), proinflammatory cytokines (e.g., TNF-α, IL-2), and the main factors contributing to fibrosis (Collα, α-SMA, and TIMP1) were downregulated. It was suggested that MSC-EVs could regulate the hepatic inflammatory microenvironment through the downregulation of immune cell infiltration and the regulation of both the expression and secretion of inflammatory factors, including anti-inflammatory and proinflammatory factors [3,39]. Other EVs from various sources of MSCs have shown similar effects on liver protection and regeneration, including BMSCs [40][41][42], human menstrual blood-derived MSCs [43], adipose-derived MSCs, and human liverderived MSCs [44,45] (Figure 1).…”

“…Mesenchymal stem cells (MSCs), a group of unique cells, have shown great potential for unlimited self-renewal [1]. They are found in various tissues, including the liver, bone marrow, intestine, connective tissues, spleen, and placenta [2,3]. MSCs can not only differentiate into mesenchymal cell lines (e.g., osteocytes, chondrocytes, skeletal muscle cells) but also develop into ectodermal and endodermal cells (e.g., hepatocytes and neurons) [4][5][6].…”

Lipopolysaccharide-Preconditioned Mesenchymal Stem Cell Transplantation Attenuates Critical Persistent Inflammation Immune Suppression and Catabolism Syndrome in Mice

“…Figure 1A presents a schematic of the timeline and the different treatments. The cell dose was selected based on a prior report of MSC treatment in mouse models (15). The mortality and weights were assessed daily for the next 3 days.…”