Characterization of long non-coding RNAs in the regulatory T helper cell polarization
Kumpulainen, Venla (2017-03-13)
Characterization of long non-coding RNAs in the regulatory T helper cell polarization
Kumpulainen, Venla
(13.03.2017)
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Turun yliopisto
Kuvaus
Siirretty Doriasta
Tiivistelmä
Regulatory T helper cells (Treg) play an important role in controlling and suppressing other immune cells by secreting multiple different cytokines. Their differentiation process is controlled and mediated through various molecular mechanisms. Recently, the discovery of the emerging role of regulatory long non-coding RNAs (lncRNAs) in T helper (TH) cell differentiation added another layer of complexity to the differentiation process.
The aim of this project was to characterize and study the functional role of TH cell specific lncRNAs focusing on Treg. Here we used naïve CD4+ T lymphocytes isolated from human umbilical cord blood which were polarized by culturing under different cytokine conditions. Based on RNA sequencing (RNAseq) data, a number of Treg-specific and differentially regulated lncRNAs were selected for this study which included qRT-PCR-based lncRNA and miRNA expression, cellular localization, TGFβ titration and knockdown experiments.
Out of seven selected lncRNAs based on RNAseq data, five were validated with qRT-PCR (lncVK1, lncVK3, lncMV2, lncAH2, lncOD1). All others were upregulated in Treg cells compared to TH1, TH2, TH17 and non-polarized control cells, except lncAH2 which was downregulated both in Treg and TH17 cells. lncVK1 is a new un-annotated lncRNA residing mainly in the nucleus and is induced with TGFβ, cytokine participating in the differentiation of Treg cells. lncMV2, on the other hand, was mainly cytoplasmic and interacts possibly with miRX, a known Treg polarization regulator. These lncRNAs were shown to be promising regulators of Treg polarization, but the study needs to be continued with subsequent functional studies.
The aim of this project was to characterize and study the functional role of TH cell specific lncRNAs focusing on Treg. Here we used naïve CD4+ T lymphocytes isolated from human umbilical cord blood which were polarized by culturing under different cytokine conditions. Based on RNA sequencing (RNAseq) data, a number of Treg-specific and differentially regulated lncRNAs were selected for this study which included qRT-PCR-based lncRNA and miRNA expression, cellular localization, TGFβ titration and knockdown experiments.
Out of seven selected lncRNAs based on RNAseq data, five were validated with qRT-PCR (lncVK1, lncVK3, lncMV2, lncAH2, lncOD1). All others were upregulated in Treg cells compared to TH1, TH2, TH17 and non-polarized control cells, except lncAH2 which was downregulated both in Treg and TH17 cells. lncVK1 is a new un-annotated lncRNA residing mainly in the nucleus and is induced with TGFβ, cytokine participating in the differentiation of Treg cells. lncMV2, on the other hand, was mainly cytoplasmic and interacts possibly with miRX, a known Treg polarization regulator. These lncRNAs were shown to be promising regulators of Treg polarization, but the study needs to be continued with subsequent functional studies.