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Comparative gene expression analyses of purified CD14+ monocytes from human umbilical cord blood, CD14+ monocytes from adult peripheral blood and the cell therapy product DUOC-01

ABSTRACT: DUOC-01 is a macrophage-like cell therapy product manufactured from human umbilical cord blood (CB) mononuclear cells (MNCs) and intended for use in treatment of demyelinating diseases. During the 21-day manufacturing process, most initiating MNCs die in culture, but a highly active, adherent population of cells that resemble hematopoietic lineage macrophage in morphology, phagocytic activity, and antigen expression emerges; this population comprises the final DUOC-01 product [Kurtzberg et al. 2015]. Using the cuprizone-mediated murine demyelination model, we have found that DUOC-01 cells are capable of accelerating the remyelination process. In the same demyelination model, human cord blood-derived CD14+ cells (CBCD14) were less active. In another cellular therapy project, we have found that CBCD14 protects brain cells from oxygen-glucose-deprivation (OGD), but adult peripheral blood-derived CD14+ monocytes (PBCD14) did not. To address the functional differences between these cell types, we have analyzed differential gene expression by these three cell types.

ORGANISM(S): Homo sapiens

PROVIDER: GSE76803 | GEO | 2016/01/13

SECONDARY ACCESSION(S): PRJNA308603

REPOSITORIES: GEO

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Comparative gene expression analyses of purified CD14+ monocytes from human umbilical cord blood, CD14+ monocytes from adult peripheral blood and the cell therapy product DUOC-01

Project description:DUOC-01 is a macrophage-like cell therapy product manufactured from human umbilical cord blood (CB) mononuclear cells (MNCs) and intended for use in treatment of demyelinating diseases. During the 21-day manufacturing process, most initiating MNCs die in culture, but a highly active, adherent population of cells that resemble hematopoietic lineage macrophage in morphology, phagocytic activity, and antigen expression emerges; this population comprises the final DUOC-01 product [Kurtzberg et al. 2015]. Using the cuprizone-mediated murine demyelination model, we have found that DUOC-01 cells are capable of accelerating the remyelination process. In the same demyelination model, human cord blood-derived CD14+ cells (CBCD14) were less active. In another cellular therapy project, we have found that CBCD14 protects brain cells from oxygen-glucose-deprivation (OGD), but adult peripheral blood-derived CD14+ monocytes (PBCD14) did not. To address the functional differences between these cell types, we have analyzed differential gene expression by these three cell types. We used four cord blood (CB) samples (from Carolinas Cord Blood Bank) that were collected within 24hrs of delivery. Then, highly purified CB CD14+ monocytes were isolated by flow sorting. We collected the peripheral blood (PB) from four healthy volunteers and isolated the purified PB CD14+ monocytes by cell sorting. Three separate batches of cultured DUOC-01 cells were collected from Robertson Cell and Translational Cell Therapy cGMP facility. Total RNAs were isolated at the same time and kept frozen. RNA samples were given to Duke Microarray core facility to analyse the gene expression profile using Affymetrix GeneChipÂ® Human Genome U133 Plus 2.0 Assay.

2016-01-13 | E-GEOD-76803 | biostudies-arrayexpress

Gene expression profiles of human blood classical monocytes (CD14++CD16-), CD16 positive monocytes (CD14+16++ and CD14++CD16+), and CD1c+ CD19- dendritic cells [human data]

Project description:In this study gene expression of human blood classical monocytes (CD14++CD16-), CD16 positive monocytes (consisting of non-classical CD14+16++ and intermediate CD14++CD16+ monocytes) and CD1c+ CD19- dendritic cells from healthy subjects were investigated. Keywords: expression profiling by array

2011-12-17 | GSE34515 | GEO

Expression data from primary human healthy donor-derived CD14- cDC2s, CD14+ cDC2s and monocytes

Project description:The human dendritic cell (DC) family has recently been expanded by CD1c+CD14+CD163+ DCs, introduced as DC3. DC3 are found in tumors and peripheral blood of cancer patients but which cells can serve as their pre-cursors remain unknown. CD1c+CD14+ cells share similarities with both CD1c+ DCs (cDC2s) and CD14+ monocytes on transcriptomic and phenotypic level. To investigate whether CD14+ cDC2s are closer related to CD14- cDC2s or monocytes on transcriptomic level, we analyzed their RNA.

2023-12-01 | GSE218218 | GEO

NanoString nCounter immune-related gene expression in blood sorted CD14+CD16- monocytes from sALS, fALS and HC subjects

Project description:We investigated the innate immune system in the SOD1 ALS model. We found that splenic Ly6CHi monocytes were activated and their progressive recruitment to the spinal cord, but not brain, correlated with neuronal loss. We found a decrease in resident microglia in the spinal cord with disease progression. Two months prior to disease onset, splenic Ly6CHi monocytes had an M1 signature which included increased CCR2. At one month prior to disease onset, microglia expressed increased CCL2 and other chemotaxis-associated molecules. Microglia derived from the spinal cord of SOD1 mice recruited Ly6C+ monocytes to the CNS. Treatment with anti-Ly6C mAb modulated the Ly6CHi monocyte cytokine profile, reduced monocyte recruitment to the spinal cord, diminished neuronal loss and extended survival. In humans with ALS, CD14+/CD16- monocytes (analogue of Ly6CHi monocytes) exhibited an ALS specific microRNA inflammatory signature similar to that observed in the SOD1 mouse providing a direct link between the animal model and the human disease. Thus, the SOD1-like profile of monocytes in ALS subjects may serve as a biomarker for disease stage or progression. Our results suggest that recruitment of inflammatory monocytes plays an important role in disease progression and that modulation of these cells is a potential therapeutic approach This study used the NanoString nCounter hybridization system and the Nanostring GX Human Immunology and Nanostring Human Inflammation assays to identify and quantitate immune-related genes in blood CD14+CD16- monocytes from ALS, MS and HC subjects Total RNA was isolated from FACS sorted CD14+CD16- blood-derived monocytes from sporadic sALS (n=10), fALS (n=4) and HC (n=10) subjects. RNA was profiled using the Nanostring GX Human Immunology and Nanostring Human Inflammation assays

2012-07-25 | E-GEOD-39642 | biostudies-arrayexpress

NanoString miRNA profiling of peripheral blood sorted CD14+CD16- monocytes from amyotrophic lateral sclerosis, multiple sclerosis and healthy control subjects

Project description:We investigated the innate immune system in the SOD1 ALS model. We found that splenic Ly6CHi monocytes were activated and their progressive recruitment to the spinal cord, but not brain, correlated with neuronal loss. We found a decrease in resident microglia in the spinal cord with disease progression. Two months prior to disease onset, splenic Ly6CHi monocytes had an M1 signature which included increased CCR2. At one month prior to disease onset, microglia expressed increased CCL2 and other chemotaxis-associated molecules. Microglia derived from the spinal cord of SOD1 mice recruited Ly6C+ monocytes to the CNS. Treatment with anti-Ly6C mAb modulated the Ly6CHi monocyte cytokine profile, reduced monocyte recruitment to the spinal cord, diminished neuronal loss and extended survival. In humans with ALS, CD14+/CD16- monocytes (analogue of Ly6CHi monocytes) exhibited an ALS specific microRNA inflammatory signature similar to that observed in the SOD1 mouse providing a direct link between the animal model and the human disease. Thus, the SOD1-like profile of monocytes in ALS subjects may serve as a biomarker for disease stage or progression. Our results suggest that recruitment of inflammatory monocytes plays an important role in disease progression and that modulation of these cells is a potential therapeutic approach. This study used the NanoString nCounter hybridization system and nCounter miRNA expression assays to identify and quantitate miRNAs in blood CD14+CD16- monocytes from ALS, MS and HC subjects Total RNA was isolated from FACS sorted CD14+CD16- blood-derived monocytes from sporadic ALS (n=8), MS (n=8) and HC (n=8) subjects. RNA was profiled using the NanoString nCounter miRNA expression assay

2012-07-25 | E-GEOD-39643 | biostudies-arrayexpress

Expression data from porcine monocytes sorted on expression of CD14 and CD163

Project description:Human and mouse monocytes can be divided into 2 different sub-populations, using CD14-CD16 and Ly6C-CX3CR1 respectively. We investigated the pig monocytes sub-populations and found that all porcine monocyte express CD16 and CD172M-NM-1 but can be divided into 2 subpopulation using CD14 and the scavenger receptor CD163. The CD14hi-CD163low population resemble to the inflammatory monocytes whereas the CD14low-CD163hi display more a resident monocyte type. Pig monocyte can be differentiated into macrophages when cultured with rhCSF-1 and show an increase in size, granularity and autofluorescence, and express the common macrophage markers CD14, CD16 and CD172M-NM-1. Gene expression in these 2 sub-populations was profiled using the newly-developed and annotated pig whole genome snowball microarray, showing a distinct pattern between inflammatory and resident monocytes but this difference would be more a maturation process instead of two separate subsets. Furthermore, the expression of certain genes such as CD36, CLEC4E or TREM-1 proved to share the same pattern as human monocytes, quite different from mouse monocytes. These results emphasize the potential role of the pigs as a model for human inflammatory disease and will improved our knowledge on the mononuclear phagocyte system development. Porcine PBMCs were isolated from the blood of three seperate pigs, FACS sorted on expression of CD14 and CD163 and RNA isolated from each sample, a total of 6 microarrays were hybridised

2013-02-14 | E-GEOD-43898 | biostudies-arrayexpress

Gene expression in CD14+ and CD16+ monocytes

Project description:The aim of our work was to evaluate what happens to the two most polar subpopulations of peripheral blood monocytes in women diagnosed preeclampsia in comparison with physiological pregnancy. We used microarrays for comparison of CD14+ and CD16+ monocytes from patients with preeclampsia and women with physiological pregnancy

2021-10-31 | GSE186819 | GEO

Single-cell RNA-seq analysis of human CD14+ monocytes

Project description:We performed single-cell RNA-seq on CD14+ monocytes isolated from the blood of healthy donors. Using the 10x chromium technology, we analyzed 425 and 431 cells from 2 individual donors.

2017-09-15 | GSE103544 | GEO

Transcriptome analysis of CD14+ monocytes from SSc patients

Project description:Purpose: To compare transcriptome profile of CD14+ blood monocytes from systemic sclerosis (SSc) patients and healthy controls. Methods: CD14+ monocytes were isolated from peripheral blood of lcSSc (n=5, age=54.4±6.7), dcSSc patients (n=5, age=51.8±7.2) and age- and sex-matched healthy controls (HC) (n=5, age=50.8±9.7). Total RNA was isolated and polyA libraries were prepared using TruSeq Stranded mRNA kit. Next Generation Sequencing was performed using Illumina HiSeq 4000 platform. Differentially expressed genes were computed using DESeq2 algorithm. Results: We detected 1440 differentially expressed genes between dcSSc vs HC and 225 between lcSSc and HC respectively (p≤0.01; log2 ratio≥0.5, Fig 1). Among those, in dcSSc 1076 were upregulated (e.g. MMP9, IL1R2, FLT3, MIF, TLR9) and 364 were downregulated (e.g. TGFBR1, CD44, CD244, HLA-DRA, HLA-G). In lcSSc 160 transcripts were upregulated (e.g. CCL2, WNT5B, MMP17) and 65 were downregulated (e.g. KLF11, IRAK2). We identified 123 commonly deregulated genes between SSc subgroups (e.g. CCL3, CD14, IL27, MMP17). Principal component analysis showed close clustering within SSc subgroups and clear separation from healthy controls. Pathway analysis revealed alterations in several biological processes important in fibrogenesis including antigen presentation, MIF-induced immune responses, TGF-β, NOTCH and WNT signalling pathways. qPCR analysis further confirmed differences in gene expression on mRNA level (n HC=8, n SSc=25, p≤0.05). Conclusions: To our knowledge, this is the first global transcriptome analysis of peripheral blood CD14+ monocytes in SSc. Our results suggest a primary activation of monocytes in peripheral blood, which might be further translated into novel cellular biomarker of the disease and potentially used for distinguishing between responders and non-responders to a novel treatment in future clinical trials.

2021-09-10 | GSE157840 | GEO

Transcription profiling by array of human serum-free fibrocytes, serum-containing fibrocytes, CD14++CD16- Monocytes, CD14++CD16+ Monocytes, CD14+CD16++ Monocytes and macrophages

Project description:Serum-free Fibrocytes, Serum-containing Fibrocytes, CD14++CD16- Monocytes, CD14++CD16+ Monocytes, CD14+CD16++ Monocytes, Macrophages were all generated from up to 3 biological replicates from each of 3 separate donors. RNA was extracted (Ambion RNAqueous), labelled with cy3, mixed with cy5 labelled human reference (Stratagene), and hybridised to slides printed with Human AROS v4.0 oligonucleotides (Operon). Slides were scanned using a Perkin Elmer GX plus, and the data then normalised with GEPAS v4.0 and collated. Final data analysis was carried out using TMEV 4.0. SAM was performed using a 0.1% FDR. PCA were plotted from this list, and interrogation carried out using DAVID to determine pathway enrichment.

2011-01-01 | E-MEXP-2811 | biostudies-arrayexpress

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