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Tregs coculture with control or Capns1-knockdown SVEC4-10 cells

ABSTRACT: Tregs coculture with control or Capns1-knockdown SVEC4-10 cells

PROVIDER: PRJNA702836 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13741523_1.fastq.gz	Fastqsanger.gz
	SRR13741523_2.fastq.gz	Fastqsanger.gz
	SRR13741524_1.fastq.gz	Fastqsanger.gz
	SRR13741524_2.fastq.gz	Fastqsanger.gz

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Tregs coculture with control or Capns1-knockdown SVEC4-10 cells

Project description:Purpose: The goals of this study are to compare NGS-derived Treg transcriptome profiling (RNA-seq) to microarray methods and to evaluate phenotypic difference between Tregs cocultured with control and Capns1-knockdown SVEC4-10 cells. Methods: Isolated splenic CD4+CD25+ Tregs were cocultured with SVEC4-10 cells for 5 days. Then total RNA in cocultured Tregs were isolated using an RNeasy mini kit (Qiagen) with on-column DNaseI digestion. Three replicates per each groups were pooled, and were subjected to the RNA-seq analysis using a NovaSeq6000 (Illumina, San Diego, CA) to obtain ~40 million 150 bp paired-end reads by Eurofins genomics (Tokyo, Japan). Contlusions: There is no remarkable difference in gene profiels between Tregs cocultured with the control and Capns1-knockdown SVEC4-10 cells.

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Knockdown of the Prolactin Receptor Inhibits Recruitment of Tregs and their Promotion of EMT in a Syngeneic Breast Cancer Model

Project description:The clinical relevance and prognostic significance of tumor-infiltrating Tregs in different types of cancers makes them appealing therapeutic targets. However, while effective at inhibiting cancer progression, immunotherapeutics targeting Treg function have adverse effects in a substantial proportion of patients, ranging from mild inflammatory disorders to frank autoimmune disease. Here, we report that systemic knockdown of the long form of the prolactin receptor (LFPRLR) inhibits recruitment of Tregs to primary tumors and, importantly, metastasis-promoting activities of tumor Tregs, with no change in the production of TGFβ or IL-10 by tumor Tregs or the ability of peripheral Tregs to suppress anti-CD3/CD28-stimulated effector cell proliferation. Knockdown of the LFPRLR was accomplished using a derivatized DNA splice-modulating oligomer and outcome was assessed in the highly-metastatic, syngeneic, orthotopic 4T1-Balb/c immune-competent breast cancer model. We conclude that knockdown of the LFPRLR produces a more functionally-specific anti-cancer effect in Tregs, suggesting promise as a future therapeutic approach.

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Project description:The aim of the project is to do proteomic analysis of total cell lysate from rest and activated mouse Tregs. The proteomic data were mapped to the Gene Ontology Cellular Component (GOCC)database to obtain the list of membrane proteins on activated Tregs. The set of membrane proteins identified were compared with the potential Siglec-1 counter-receptors on activated Tregs.

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