CD166 Engagement Augments Mouse and Human Hematopoietic Progenitor Function via Activation of Stemness and Cell Cycle Pathways

Abstract

Hematopoietic stem (HSC) and progenitor (HPC) cells are regulated by interacting signals and cellular and noncellular elements of the hematopoietic niche. We previously showed that CD166 is a functional marker of murine and human HSC and of cellular components of the murine niche. Selection of murine CD166 + engrafting HSC enriched for marrow repopulating cells. Here, we demonstrate that CD166-CD166 homophilic interactions enhance generation of murine and human HPC in vitro and augment hematopoietic function of these cells. Interactions between cultured CD166 + Lineage − Sca-1 + c-Kit + (LSK) cells and CD166 + osteoblasts (OBs) significantly enhanced the expansion of colony-forming units (CFUs). Interactions between CD166 + LSK cells and immobilized CD166 protein generated more CFU in short-term cultures than between these cells and bovine serum albumin (BSA) or in cultures initiated with CD166 − LSK cells. Similar results were obtained when LSK cells from wildtype (WT) or CD166 knockout (KO) (CD166 −/− ) mice were used with immobilized CD166. Human cord blood CD34 + cells expressing CD166 produced significantly higher numbers of CFUs following interaction with immobilized CD166 than their CD166 − counterparts. These data demonstrate the positive effects of CD166 homophilic interactions involving CD166 on the surface of murine and human HPCs. Single-cell RNA-seq analysis of CD150 + CD48 − (signaling lymphocyte activation molecule (SLAM)) LSK cells from WT and CD166 −/− mice incubated with immobilized CD166 protein revealed that engagement of CD166 on these cells activates cytokine, growth factor and hormone signaling, epigenetic pathways, and other genes implicated in maintenance of stem cell pluripotency-related and mitochondria-related signaling pathways. These studies provide tangible evidence implicating CD166 engagement in the maintenance of stem/progenitor cell function. STEM CELLS 2019;37:1319-1330 SIGNIFICANCE STATEMENTThis study describes how homophilic interactions via CD166 that is expressed on hematopoietic cells augments the functional potential of these cells. This study also uses single-cell RNAseq to identify pathways activated by the engagement of CD166 on the surface of these cells to enhance the hematopoietic function of stem and progenitor cells.

Adipose-Derived Stem Cells from Fat Tissue of Breast Cancer Microenvironment Present Altered Adipogenic Differentiation Capabilities

“…It has been reported that the surface markers such as CD133, in combination with CD29/CD24 and CD44/CD166, may correlate with high Wnt activity and identify stem cells in various tumour types and their possible interaction with the microenvironment [51]. Interestingly, in BC ADSCs, the markers CD144, CD146, CD166, and KDR result significantly deregulated suggesting oncogenic alteration also in the tumour microenvironment with a possible role in favouring angiogenesis, tumour cell migration, and proliferation [52].…”

Osteomacs promote maintenance of murine hematopoiesis through megakaryocyte-induced upregulation of Embigin and CD166

“…Progenitor assays were performed to determine whether recombinant CD166 (rCD166) and Embigin (rEmb) could substitute for OM activity in vitro. BSA (control), rCD166, and rEmb were coated onto tissue culture plates ( Zhang et al., 2019 ), which were used to culture OB (from NCC). No significant differences in CFU fold change were observed between OM+OB, rCD166+OB, rEmb+OB, and rCD166+rEmb+OB, indicating that rCD166 and rEmb could partially substitute for OM-mediated enhancement of hematopoietic activity ( Figure 5 E).…”

“…CD166 studies were more expansive due to prior laboratory experience and availability of CD166 KO mice ( Cheng et al., 2011 ; Chitteti et al., 2010a , 2014 ; Xu et al., 2016 ). We previously demonstrated in multiple settings the importance of CD166 in maintaining HSC and progenitor cell function and the competence of the hematopoietic niche ( Cheng et al., 2011 ; Chitteti et al., 2010a , 2014 ; Zhang et al., 2019 ). Here, we demonstrated that OM function is also dependent on CD166 expression.…”

Cellular components of the hematopoietic niche and their regulation of hematopoietic stem cell function

“…This suggests that immature osteolineage cells are potent in expanding HSPC in vitro compared with mature osteoblasts. CD166, another protein expressed on immature osteoblasts, is also critical for maintenance of HSPC in vitro and homophilic engagement of CD166 expressed on osteoblasts and HSPC is required for HEA [51,52].…”