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RNA-sequencing on sorted Nrn1-/- and Nrn1+/- effector Treg cells

ABSTRACT: RNA-sequencing on sorted Nrn1-/- and Nrn1+/- effector Treg cells

PROVIDER: PRJNA499046 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8127323_1.fastq.gz	Fastqsanger.gz
	SRR8127323_2.fastq.gz	Fastqsanger.gz
	SRR8127324_1.fastq.gz	Fastqsanger.gz
	SRR8127324_2.fastq.gz	Fastqsanger.gz
	SRR8127325_1.fastq.gz	Fastqsanger.gz

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RNA-sequencing on sorted Nrn1-/- and Nrn1+/- effector Treg and OTII cells

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Treg cells from human blood differentiate into non-lymphoid tissue-resident effector cells upon TNFR2 costimulation

Project description:Regulatory T (Treg) cells can facilitate transplant tolerance and attenuate autoimmune- and inflammatory diseases. Therefore, it is clinically relevant to stimulate Treg cell expansion and function in vivo and to create therapeutic Treg cell products in vitro. We report that TNF receptor 2 (TNFR2) is a unique costimulus for naïve, thymus-derived (t)Treg cells from human blood that promotes their differentiation into non-lymphoid tissue (NLT)-resident effector Treg cells, without Th-like polarization. In contrast, CD28 costimulation maintains a lymphoid tissue (LT)-resident Treg cell phenotype. We base this conclusion on transcriptome and proteome analysis of TNFR2- and CD28-costimulated CD4+ Treg cells and conventional T (Tconv) cells, followed by bioinformatic comparison with published transcriptomic Treg cell signatures from NLT and LT in health and disease, including autoimmunity and cancer. These analyses illuminate that TNFR2 costimulation promotes Treg cell capacity for survival, migration, immunosuppression and tissue regeneration. Functional studies confirmed improved migratory ability of TNFR2-costimulated tTreg cells. Flow cytometry validated the presence of the TNFR2-driven Treg cell signature in effector/memory Treg cells from the human placenta as opposed to blood. Thus, TNFR2 can be exploited as driver of NLT-resident Treg cell differentiation for adoptive cell therapy or antibody-based immunomodulation in human disease.

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Project description:Analysis of Treg cells isolated from WT and NCOR1-cKO mice indicated an important function of NCOR1 in regulating the transition from naïve to effector Treg cells. The aim of this NGS experiment was to determine the transcriptome profile of ex vivo isolated naïve and effector WT and NCOR1-cKO Treg cells under steady state condition.

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Project description:The inflammasome initiates innate defense and inflammatory response by activating caspase-1 and pyroptotic cell death in myeloid cells1,2. It is comprised of an innate immune receptor/effector, pro-caspase-1 and a common adaptor molecule, ASC (apoptotic speck-containing protein with a CARD). Consistent with their pro-inflammatory function, inflammasome components including caspase-1, ASC and NLRP3, are known to exacerbate autoimmunity during experimental autoimmune encephalomyelitis (EAE) by enhancing IL-1 and IL-18 secretion in myeloid cells3-6. Here we show an unexpected function of a DNA-binding inflammasome effector, AIM2 (Absent in Melanoma 2)7-10, in restraining autoimmunity by performing EAE in both whole body and Treg-specific deletion of Aim2–/– mice. AIM2 is highly expressed by human and mouse Treg cells and it is essential to attenuate EAE. RNA-seq, biochemical and metabolic analyses revealed that AIM2 attenuates mTOR, Myc and immune-metabolic functions in both Treg cells isolated in vivo and Treg cells induced in vitro with TGF-. Importantly, we found AIM2 physically interacted with RACK1 in Treg cells to facility the PP2A/RACK1/Akt-mTOR signaling, which is identified as a central component downstream of AIM2 that controls Treg cell function and stability. While AIM2 is generally accepted as an inflammasome effector in myeloid cells, this report reveals a previously unappreciated T cell-intrinsic role of AIM2 in maintaining Treg cell function to restrain autoimmunity. This is achieved by diminishing Akt-mTOR signaling to regulate Treg stability under inflammation, and altering immune-metabolism in Treg cells.

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Project description:Gene expression profiling of Bone Marrow FoxP3+ Treg cells. Glatman Zaretsky et al. revealed an unexpected role for Tregs in plasma cell biology. Here we determined the gene-expression profile of this new subset of FoxP3+ Treg cell, which express high levels of Treg effector molecules, similar to other non-lymphoid tissue Tregs. Gene-profiling of BM Tregs. Bone marrow Treg cells (30k) (gfp+CD25hiCD4+TCRÎ²+) (dump negative: CD19-CD8Î±-TCRÎ³Î´-CD11b-CD11c-NK1.1-Gr-1-Ter-119-) were triple-sorted from pools of two to three reporter mice (C57BL/6 Foxp3-IRES-gfp, 9 week-old males) into trizol per ImmGen SOP. RNA was amplified, labeled and hybridized to Affymetrix Mouse Gene 1.0 ST Arrays (Expression Analysis)

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