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Gene expression changes in human cord blood-derived hematopoietic stem and progenitor cells upon Centrifugation enhanced Nanostraw Transfection or Conventional Electroporation

ABSTRACT: Gene expression changes in human cord blood-derived hematopoietic stem and progenitor cells upon Centrifugation enhanced Nanostraw Transfection or Conventional Electroporation

PROVIDER: PRJNA634387 | ENA |

REPOSITORIES: ENA

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Gene expression changes in human cord blood-derived hematopoietic stem and progenitor cells upon Centrifugation enhanced Nanostraw Transfection or Conventional Electroporation

Project description:CD34+ human cord blood-derived cells were subjected to GFP mRNA delivery or mock treatment using Centrifugation enhanced Nanostraw Transfection (CeNT) or conventional electroporation.

2020-08-17 | GSE151027 | GEO

Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection

Project description:Chimeric antigen receptor (CAR)-T cell therapy is an innovative treatment for CD19-expressing lymphomas. CAR-T cells are primarily manufactured via lentivirus transfection or transposon electroporation. While anti-tumor efficacy comparisons between the two methods have been conducted, there is a current dearth of studies investigating the phenotype and transcriptome alterations induced in T cells by the two distinct manufacturing methods. Here, we established CAR-T signatures using fluorescent imaging, flow cytometry, and RNA-sequencing. A small fraction of CAR-T cells that were produced using the PiggyBac transposon (PB CAR-T cells) exhibited much higher expression of CAR than those produced using a lentivirus (Lenti CAR-T cells). PB and Lenti CAR-T cells contained a more cytotoxic T cell subsets than control T cells, and Lenti CAR-T cells presented a more pronounced memory phenotype. RNA-sequencing further revealed vast disparities between the two CAR-T cell groups, with PB CAR-T cells exhibiting greater upregulation of cytokines, chemokines, and their receptors. Intriguingly, PB CAR-T cells singularly expressed IL-9 and fewer cytokine release syndrome-associated cytokines when activated by target cells. In addition, PB CAR-T cells exerted faster in vitro cytotoxicity against CD19-expressing K562 cells but similar in vivo anti-tumor efficacy with Lenti CAR-T. Taken together, these data provide insights into the phenotypic alterations induced by lentiviral transfection or transposon electroporation and will attract more attention to the clinical influence of different manufacturing procedures.

2022-09-01 | GSE212072 | GEO

Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection

Project description:Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection

| PRJNA873765 | ENA

Gene expression profile at single cell level of murine neural stem cells (NSCs) subjected to bulk electroporation (BEP) and localized electroporation (LEP)

Project description:Localized Electroporation (LEP) has emerged as an effective tool for intracellular delivery and non-destructive analysis of cells. However, little is known about the effects of LEP on cellular gene expression which may adversely impact their downstream phenotype. Here we use single cell RNA sequencing to dissect the effects of LEP on murine neural stem cell (NSC) gene expression and compare it to Bulk Electroporation (BEP).

2023-03-10 | GSE205507 | GEO

Liposome transfection of miR-145 induces immune response via RIG-I in mesenchymal stem cells

Project description:Liposomal transfection reagents are frequenly used in gene deliverytransfer experiments. The validity of these experiments depends on the absence of changes in genes other than the target gene. Here we report that transfection of synthetic microRNA-145 using liposomes, but not electroporation, induced the upregulation of a range of immune-related genes in human mesenchymal stem cells. The immune response was dependent on the binding of microRNA-145 to retinoic acid inducible gene-I, and was independent on endosome functionality and toll-like receptors. Immune response was not observed after transfection of any of nine other small RNAs, suggesting a sequence restricted response. One of the upregulated mRNAs encode interferon β, which presumably in turn induced the upregulation of another group of immune associated genes. Interestingly, exposure of liposomes alone led to the upregulation of some immune genes, one of them retinoic acid inducible gene-I mRNA.

2012-10-01 | GSE40387 | GEO

Skin Transfection Patterns and Expression Kinetics of Electroporation-Enhanced Plasmid Delivery Using the CELLECTRA-3P, a Portable Next-Generation Dermal Electroporation Device.

Project description:The CELLECTRA-3P dermal electroporation device (Inovio Pharmaceuticals, Plymouth Meeting, PA) has been evaluated in the clinic and shown to enhance the delivery of an influenza DNA vaccine. To understand the mechanism by which this device aids in enhancing the host immune response to DNA vaccines we investigated the expression kinetics and localization of a reporter plasmid (pGFP) delivered via the CELLECTRA-3P. Histological analysis revealed green fluorescent protein (GFP) expression as early as 1?hr posttreatment in the epidermal and dermal layers, and as early as 2?hr posttreatment in the subdermal layers. Immunofluorescence techniques identified keratinocytes, fibrocytes, dendritic-like cells, adipocytes, and myocytes as the principal cell populations transfected. We proceeded to demonstrate elicitation of robust host immune responses after plasmid DNA (pDNA) vaccination. In guinea pigs equivalent humoral (antibody binding titers) immune responses were observed between protocols using either CELLECTRA-3P or intramuscular electroporation to deliver the DNA vaccine. In nonhuman primates, robust interferon-? enzyme-linked immunospot and protective levels of hemagglutination inhibition titers after pDNA vaccination were observed in groups treated with the CELLECTRA-3P. In conclusion, these findings may assist in the future to design efficient, tolerable DNA vaccination strategies for the clinic.

| S-EPMC5206769 | biostudies-literature

Generation of diffuse intrinsic pontine glioma mouse models by brainstem targeted in utero electroporation

Project description:Diffuse intrinsic pontine gliomas (DIPGs) are highly lethal childhood brain tumors. Their unique genetic makeup, pathological heterogeneity, and brainstem location all present challenges to treatment. Developing mouse models that accurately reflect each of these distinct features will be critical to advance our understanding of DIPG development, progression, and therapeutic resistance. The aim of this study was to generate new mouse models of DIPG, and characterize the role of specific oncogenic combinations in DIPG pathogenesis. We used in utero electroporation (IUE) to transfect neural stem cells in the developing brainstem with PiggyBac DNA transposon plasmids. Combinations of PDGFB or PdgfraD842V, dominant negative Trp53 (DNp53), and H3.3K27M expression induced fully penetrant brainstem gliomas. IUE enabled the targeted transfection of brainstem neural stem cells. PDGFB + DNp53 induced the rapid development of grade-IV gliomas. PdgfraD842V + DNp53 produced slower forming grade-III gliomas. Addition of H3.3K27M only significantly accelerated PdgfraD842V DIPG development. Glioma subgroup molecular signatures were associated with differences in bulk PDGFB and PdgfraD842V tumor composition. H3.3K27M induced both overlapping and unique gene expression changes in PDGFB and PdgfraD842V tumors. Paracrine effects of PDGFB promote disruption of pericyte-endothelial interactions and angiogenesis in PDGFB DIPG mouse models. Brainstem targeted in utero electroporation provides a rapid and flexible system to generate diverse DIPG mouse models. Using IUE to investigate mutation and pathohistological heterogeneity of DIPG will provide a valuable tool for future genetic and preclinical studies.

2019-11-14 | GSE128807 | GEO

Liposome transfection of miR-145 induces immune response via RIG-I in mesenchymal stem cells

Project description:Liposomal transfection reagents are frequenly used in gene deliverytransfer experiments. The validity of these experiments depends on the absence of changes in genes other than the target gene. Here we report that transfection of synthetic microRNA-145 using liposomes, but not electroporation, induced the upregulation of a range of immune-related genes in human mesenchymal stem cells. The immune response was dependent on the binding of microRNA-145 to retinoic acid inducible gene-I, and was independent on endosome functionality and toll-like receptors. Immune response was not observed after transfection of any of nine other small RNAs, suggesting a sequence restricted response. One of the upregulated mRNAs encode interferon Î², which presumably in turn induced the upregulation of another group of immune associated genes. Interestingly, exposure of liposomes alone led to the upregulation of some immune genes, one of them retinoic acid inducible gene-I mRNA. Comparison of immune gene expression patterns in BM-MSC donors after transfection of synthetic miR-145.

2012-10-01 | E-GEOD-40387 | biostudies-arrayexpress

Nr2f6 electroporation in tibialis anterior promotes muscle weakness

Project description:Nr2f6 electroporation in tibialis anterior promotes muscle weakness

| PRJNA948669 | ENA

Expression data from human cord blood CD161++/CD161+/CD161- CD8+ T cell subsets

Project description:We used microarray to compare gene expression between CD161++/CD161+/CD161-CD8+ T cells from human cord blood. Lymphocytes from freshly obtained human cord blood samples were isolated by Ficoll density centrifugation. CD8+ T cells were purified by negative selection using magnetic beads and subsequently labelled with fluorescent antibodies prior to sorting using MoFlo MLS cell sorter (Dako).

2012-01-25 | E-GEOD-33424 | biostudies-arrayexpress

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