Fast and Efficient Genome Editing of Human FOXP3+ Regulatory T Cells

Zeebroeck, Lauren Van; Hornero, Rebeca Arroyo; Côrte-Real, Beatriz F.; Hamad, Ibrahim; Meißner, Torsten; Kleinewietfeld, Markus

doi:10.3389/fimmu.2021.655122

Cited by 12 publications

(13 citation statements)

References 75 publications

(118 reference statements)

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“…Indeed, literature has shown IL-6R expression on human Tregs as well [43], which renders Tregs, and more specifically thymic Tregs [44], more susceptible to Th17 skewing [45,46]. Previous reports showed that IL-1 and IL-6 alone and together downregulate FOXP3 while inducing IL-17 expression on Tregs in a STAT3-dependent way [33,[47][48][49][50][51], also in EAE and rheumatoid arthritis [26,29,52]. As these cytokines are not BBB specific, these effects will not be limited to MS.…”

Section: Discussionmentioning

confidence: 99%

Rapamycin rescues loss-of-function in blood-brain barrier-interacting regulatory T cells

Baeten

Hamad

Hoeks

et al. 2022

Preprint

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In many autoimmune diseases, FOXP3+regulatory T cells (Tregs) skew towards a pro-inflammatory and non-suppressive phenotype and are therefore unable to control the exaggerated autoimmune response. This may largely impact the success of autologous Treg therapy which is currently under investigation for treatment of autoimmune diseases, including multiple sclerosis (MS). Thus, there is a need to ensurein vivostability of Tregs before successful Treg therapy can be applied. Using a murine genetic fate-mapping model, we demonstrate that inflammatory exFOXP3 T cells accumulate in the central nervous system (CNS) during experimental autoimmune encephalomyelitis (EAE). In a humanin vitroBBB model, we discovered that interaction with inflamed blood-brain barrier (BBB)-endothelial cells induces loss of suppressive function in Tregs. Transcriptome analysis further revealed that Tregs which migrated across inflamed BBB-endothelial cellsin vitrohave a pro-inflammatory Th1/17 signature and upregulate the mTORC1 signaling pathway compared to non-migrated Tregs. These findings suggest that interaction with BBB-endothelial cells is sufficient to affect Treg function, and that transmigration triggers an additive pro-inflammatory phenotype switch, which was also seen in CNS-derived exFOXP3 T cells of EAE mice.In vitrotreatment of migrated human Tregs with the clinically-approved mTORC1 inhibitor rapamycin completely restored the loss of suppressive function. Finally, flow cytometric analysis indicated an enrichment of inflammatory, less suppressive CD49d+Tregs in the cerebrospinal fluid of MS patients, thereby underscoring the relevance of our findings for human disease. In sum, our findings provide firm evidence that the inflamed BBB affects human Treg stability, which can be restored using a mTORC1 inhibitor. These insights can help in significantly improving the efficacy of autologous Treg therapy of MS.

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Section: Discussionmentioning

confidence: 99%

Rapamycin rescues loss-of-function in blood-brain barrier-interacting regulatory T cells

Baeten

Hamad

Hoeks

et al. 2022

Preprint

Self Cite

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“…Changes in IL2RA expression may affect immune and inflammatory signaling cascade responses, which in turn affect CD4 + T cell differentiation and TReg cell suppressive activity (Asouri et al, 2020). IL2 signaling is involved in the differentiation and homeostasis of regulatory T cells (Tregs), and IL2 signaling is involved in the induction of cell growth and effector T cell proliferation (Zeebroeck et al, 2021). Pre-activation of IL-12, IL-15, and IL-18 was shown to upregulate IL2RA (CD25) expression (Akman et al, 2021).…”

Section: Phaseol May Reduce Inflammatory Cell Activation and Inflamma...mentioning

confidence: 99%

Exploring the mechanism of action of licorice in the treatment of COVID-19 through bioinformatics analysis and molecular dynamics simulation

Cao

Gong²,

Wu³

et al. 2022

Front. Pharmacol.

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Purpose: The rapid worldwide spread of Corona Virus Disease 2019 (COVID-19) has become not only a global challenge, but also a lack of effective clinical treatments. Studies have shown that licorice can significantly improve clinical symptoms such as fever, dry cough and shortness of breath in COVID-19 patients with no significant adverse effects. However, there is still a lack of in-depth analysis of the specific active ingredients of licorice in the treatment of COVID-19 and its mechanism of action. Therefore, we used molecular docking and molecular dynamics to explore the mechanism of action of licorice in the treatment of COVID-19.Methods: We used bioinformatics to screen active pharmaceutical ingredients and potential targets, the disease-core gene target-drug network was established and molecular docking was used for verification. Molecular dynamics simulations were carried out to verify that active ingredients were stably combined with protein targets. The supercomputer platform was used to measure and analyze stability of protein targets at the residue level, solvent accessible surface area, number of hydrogen bonds, radius of gyration and binding free energy.Results: Licorice had 255 gene targets, COVID-19 had 4,628 gene targets, the intersection gene targets were 101. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene ontology (GO) analysis showed that licorice played an important role mainly through the signaling pathways of inflammatory factors and oxidative stress. Molecular docking showed that Glycyrol, Phaseol and Glyasperin F in licorice may playe a role in treating COVID-19 by acting on STAT3, IL2RA, MMP1, and CXCL8. Molecular dynamics were used to demonstrate and analyze the binding stability of active ingredients to protein targets.Conclusion: This study found that Phaseol in licorice may reduce inflammatory cell activation and inflammatory response by inhibiting the activation of CXCL8 and IL2RA; Glycyrol may regulate cell proliferation and survival by acting on STAT3. Glyasperin F may regulate cell growth by inhibiting the activation of MMP1, thus reducing tissue damage and cell death caused by excessive inflammatory response and promoting the growth of new tissues. Therefore, licorice is proposed as an effective candidate for the treatment of COVID-19 through STAT3, IL2RA, MMP1, and CXCL8.

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“…IL-2RA deletion resulted in the loss of STAT5 signaling and Treg suppressive function. Similarly, IL-6R alpha subunit showed reduction in STAT3 activation, suggesting the possibility of genetically engineered Tregs to be resistant to IL-6 mediated Treg instability (88). While transcriptional regulation is not a focus of this review, a paper by Schumann et al entitled "functional CRISPR dissection of gene networks controlling human regulatory T cell identity" pressured Tregs under inflammatory cytokines while using CRISPR/Cas9 to reveal important transcription factors regulating Treg instability in the inflammatory settings.…”

Section: Foxp3 Stability Through Functional and Phenotypic Screensmentioning

confidence: 99%

Section: Foxp3 Stability Through Functional and Phenotypic Screensmentioning

confidence: 99%

Tregs in Autoimmunity: Insights Into Intrinsic Brake Mechanism Driving Pathogenesis and Immune Homeostasis

2022

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CD4+CD25highFoxp3+ regulatory T-cells (Tregs) are functionally characterized for their ability to suppress the activation of multiple immune cell types and are indispensable for maintaining immune homeostasis and tolerance. Disruption of this intrinsic brake system assessed by loss of suppressive capacity, cell numbers, and Foxp3 expression, leads to uncontrolled immune responses and tissue damage. The conversion of Tregs to a pathogenic pro-inflammatory phenotype is widely observed in immune mediated diseases. However, the molecular mechanisms that underpin the control of Treg stability and suppressive capacity are incompletely understood. This review summarizes the concepts of Treg cell stability and Treg cell plasticity highlighting underlying mechanisms including translational and epigenetic regulators that may enable translation to new therapeutic strategies. Our enhanced understanding of molecular mechanism controlling Tregs will have important implications into immune homeostasis and therapeutic potential for the treatment of immune-mediated diseases.

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Fast and Efficient Genome Editing of Human FOXP3+ Regulatory T Cells

Cited by 12 publications

References 75 publications

Rapamycin rescues loss-of-function in blood-brain barrier-interacting regulatory T cells

Rapamycin rescues loss-of-function in blood-brain barrier-interacting regulatory T cells

Exploring the mechanism of action of licorice in the treatment of COVID-19 through bioinformatics analysis and molecular dynamics simulation

Tregs in Autoimmunity: Insights Into Intrinsic Brake Mechanism Driving Pathogenesis and Immune Homeostasis

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