Long noncoding RNAs in innate lymphoid cell biology

项目总结： 先天淋巴样细胞(ILCs)是最近被描述的一组先天淋巴细胞组，对于防御A 病原体种类繁多，它们的功能障碍与多种病理有关。因此， 了解先天淋巴样细胞在体内是如何调节的，是开发新的治疗方法的关键。 各种疾病。最近已经清楚的是，尽管大多数人类基因组并不是 对蛋白质进行编码，但非编码区仍被转录。事实上，长长的基因间非编码RNA (LincRNA)转录本已被证明在各种环境中具有关键的调节功能，例如 胚胎发育。然而，lincRNAs在体内免疫系统中的作用并不是很好。 明白了。重要的是，在小鼠和人类中编码lincRNAs的基因座通常是保守的。此外， LincRNAs的表达通常以细胞类型特有的方式进行调节，而不是蛋白质编码 基因。这意味着lincRNAs在调节小鼠和人类的特定细胞方面发挥着重要作用， 它们可能会成为未来治疗的有用靶点。因此，我假设特定类型的细胞 ILCs中lincRNA的表达对其动态平衡起着关键作用。为此，我们发现了一种特定的lincRNA， Ak083360，它在第1组ILC中表达，在Ak083360缺失的情况下，我发现两个 在多种组织中，第1组ILC的数量和功能均显著减少。此外，我发现， Ak083360-/-NK细胞中Id2转录本的表达显著降低。因此，我假设Ak083360 通过调节第1组ILC的发育，对第1组ILC的动态平衡至关重要，而且确实如此 这是通过对Id2基因的特定调控来实现的。为了验证这一假设，我将确定两个发展阶段 Ak083360-/-中失调的阶段和生物学过程，如细胞增殖或凋亡 老鼠。我还将确定炎症性环境中第1组ILC反应是否需要Ak083360。 此外，我还发现在Ak083360缺陷的第1组ILCs中，Id2基因的表达发生了变化，并将 确定Id2表达失调是否与这种表型有关。最后，我预测 Ak083360调节Id2基因座染色质的可及性。为了测试这一点，我将确定染色质的状态 在Ak083360-/-使用atac-seq的第1组ILC中的可访问性，并将其与RNA下拉和 Western blotting以确定Ak083360是否与特定的蛋白质伙伴相互作用。总的来说，成功了 上述实验的完成将进一步加深我们对lincRNAs在 控制ILC人口的一般因素和特殊因素。

Project Summary: Innate lymphoid cells (ILCs) are recently described groups of innate lymphocytes critical for defense against a variety of pathogens, and their dysfunction has been associated with multiple pathologies. Therefore, understanding how innate lymphoid cells are regulated in vivo is key to developing novel treatments for a variety of diseases. It has become clear recently that although the majority of the human genome does not encode for proteins, noncoding regions are transcribed nonetheless. Indeed, long intergenic noncoding RNA (lincRNA) transcripts have been shown to have key regulatory function in a variety of contexts, such as embryonic development. However, the roles of lincRNAs in the immune system in vivo are not well understood. Importantly, the loci encoding lincRNAs in mice and humans are often conserved. Furthermore, the expression of lincRNAs is often regulated in a cell type-specific manner, more so than protein coding genes. This implies both that lincRNAs play important roles in regulating specific cells in mice and humans, and that they may be useful targets in future therapeutics. Therefore, I hypothesize that cell type-specific lincRNA expression in ILCs is critical for their homeostasis. To this end, we found a specific lincRNA, Ak083360, that is expressed in group 1 ILCs, and in the absence of Ak083360 I have found that both the numbers and function of group 1 ILCs are significantly reduced in multiple tissues. Furthermore, I found that expression of the Id2 transcript is greatly reduced in Ak083360-/- NK cells. Thus, I hypothesize that Ak083360 is critical for the homeostasis of group 1 ILCs through regulation of group 1 ILC development, and that it does this through specific regulation of the Id2 gene. To test this hypothesis, I will determine both the developmental stages and biological process, such as cell proliferation or apoptosis, that are dysregulated in Ak083360-/- mice. I will also determine whether Ak083360 is required for group 1 ILC responses in inflammatory settings. Furthermore, I found that the expression of the Id2 gene is altered in Ak083360-deficient group 1 ILCs, and will determine whether dysregulation of Id2 expression is responsible for this phenotype. Finally, I predict that Ak083360 regulates chromatin accessibility at the Id2 locus. To test this, I will determine the state of chromatin accessibility in Ak083360-/- group 1 ILCs using ATAC-seq, and will combine this with RNA pulldown and Western blotting to determine if Ak083360 interacts with specific protein partners. Altogether, successful completion of the described experiments will further our understanding of both gene regulation by lincRNAs in general and specific factors regulating ILC populations.