Targeted Disruption of the <i>β</i>2-Microglobulin Gene Minimizes the Immunogenicity of Human Embryonic Stem Cells

Wang, Dachun; Yuan, Quan; Yan, Qing; Morales, John E.; Wetsel, Rick A.

doi:10.5966/sctm.2015-0049

Cited by 151 publications

(125 citation statements)

References 58 publications

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“…However, one potential limitation of recombinases is their immunogenicity due to their non-human origins (FlpO, for example, is derived from yeast). To mitigate transgene immunogenicity, one strategy is to leverage genome editing tools to eliminate the gene B2M, an important component for antigen display through class I human leukocyte antigen (HLA) 39,40 . This strategy has also been combined with HLA-E overexpression to reduce immunogenicity against pluripotent stem cells 41 .…”

Section: Discussionmentioning

confidence: 99%

Inducible gene switches with memory in human T cells for cellular immunotherapy

Chakravarti

Caraballo

Weinberg

et al. 2018

Preprint

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Cell-based therapies that employ engineered T cells-including the expression of chimeric antigen receptors (CARs)-to target cancer cells have demonstrated promising responses in clinical trials. However, engineered T cell responses must be regulated to prevent severe side effects such as cytokine storms and off-target responses. Here we present a class of recombinase-based gene circuits that will enable inducible switching between two states of adoptive T cell therapy using an FDA-approved drug, creating a generalizable platform that can be used to control when and how strongly a gene is expressed. These circuits exhibit memory such that induced T cells will maintain any changes made even when the drug inducer is removed. This memory feature avoids prolonged drug inducer exposure, thus reducing the complexity and potential side effect associated with the drug inducer. We have utilized these circuits to control the expression of an anti-Her2-CAR, demonstrating the ability of these circuits to regulate CAR expression and T cell activity. We envision this platform can be extended to regulate other genes in T cell behavior for various adoptive T cell therapies.

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Section: Discussionmentioning

confidence: 99%

Inducible gene switches with memory in human T cells for cellular immunotherapy

Chakravarti

Caraballo

Weinberg

et al. 2018

Preprint

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“…The cytolytic assay was performed as previously described. 6 The CD8 + T cells involved in the cytotoxicity experiments were from healthy volunteers with signed informed consent. Briefly, CD8 + T cells were isolated from human peripheral blood mononuclear cells by Ficoll density gradient centrifugation followed by further purification using the CD8 + T-Cell isolation kit (Stem Cell Technologies).…”

Section: Cytolytic Assaymentioning

confidence: 99%

“…Due to the polymorphic nature of the HLA-I genes, it is often difficult to identify a perfect match between donor and recipient prior to transplantation. 5,6 HLA-I is made up of a heavy chain and a light chain, which is also called β 2 -microglobulin (B2M). HLA-I structure is disrupted and non-functional when the B2M gene is deleted.…”

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confidence: 99%

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Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway

Zhang

Wang

Shao

et al. 2019

J Cellular Molecular Medi

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Generating universal human umbilical mesenchymal stem cells (UMSCs) without immune rejection is desirable for clinical application. Here we developed an innovative strategy using CRISPR/Cas9 to generate B2M‐UMSCs in which human leucocyte antigen (HLA) light chain β2‐microglobulin (B2M) was deleted. The therapeutic potential of B2M‐UMSCs was examined in a mouse ischaemic hindlimb model. We show that B2M‐UMSCs facilitated perfusion recovery and enhanced running capability, without inducing immune rejection. The beneficial effect was mediated by exosomes. Mechanistically, microRNA (miR) sequencing identified miR‐24 as a major component of the exosomes originating from B2M‐UMSCs. We identified Bim as a potential target of miR‐24 through bioinformatics analysis, which was further confirmed by loss‐of‐function and gain‐of‐function approaches. Taken together, our data revealed that knockout of B2M is a convenient and efficient strategy to prevent UMSCs‐induced immune rejection, and it provides a universal clinical‐scale cell source for tissue repair and regeneration without the need for HLA matching in the future.

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“…To rule out non-specific effects of As 2 O 3 and confirm the requirement for MHCI in IFNγ-dependent neurite outgrowth, we investigated the effect of blocking MHCI cell surface expression on IFNγ-induced morphological changes. To achieve this, we took advantage of the requirement for the B2M protein for cell surface expression of MHCI (37). Firstly, we sought to demonstrate that IFNγ-induced expression of B2M protein was required for MHCI expression in our NPCs.…”

Section: Ifnγ-mediated Increase In Mhci and Neurite Outgrowth Requirementioning

confidence: 99%

Antiviral signalling in human IPSC-derived neurons recapitulates neurodevelopmental disorder phenotypes

Warre-Cornish

Perfect

Nagy

et al. 2019

Preprint

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Maternal immune activation increases the risk of neurodevelopmental disorders.Elevated cytokines, such as interferon-gamma (IFNγ), in offspring's brains play a central role. IFNγ activates an antiviral cellular state, limiting viral entry and replication. In addition, IFNγ has been implicated in brain development. Here, we hypothesise that IFNγ-induced antiviral signalling contributes to molecular and cellular phenotypes associated with neurodevelopmental disorders. We find that transient IFNγ treatment of neural progenitors derived from human induced pluripotent stem cells (hIPSCs) persistently increases neurite outgrowth, phenocopying hIPSC-neurons from autistic individuals. IFNγ upregulates antiviral PML bodies and MHC class I (MHCI) genes, which persists through neuronal differentiation. Critically, IFNγ-induced neurite outgrowth requires both PML and MHCI. We also find that IFNγ disproportionately alters expression of autism and schizophrenia risk genes, suggesting convergence between these genetic and environmental risk factors. Together, these data indicate that IFNγ-induced antiviral signalling may contribute to neurodevelopmental disorder aetiology. PMLNormalised expression * *

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Targeted Disruption of the β2-Microglobulin Gene Minimizes the Immunogenicity of Human Embryonic Stem Cells

Cited by 151 publications

References 58 publications

Inducible gene switches with memory in human T cells for cellular immunotherapy

Inducible gene switches with memory in human T cells for cellular immunotherapy

Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway

Antiviral signalling in human IPSC-derived neurons recapitulates neurodevelopmental disorder phenotypes

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