Translational Core (Expression Core)

摘要-核心B 调节性B细胞（Bcells）的发育、功能和调节机制， 炎性B效应细胞（Beff）仍然不清楚。到目前为止，对贝弗莉和贝夫的研究一直具有挑战性， 这是由于它们在临床样品中的数量少以及它们的异质性。但发展 基因组方法，例如来自低输入样品的大量群体的RNA-Seq和单细胞RNA-Seq （scRNA-Seq）以及CRISPR-Cas9基因组编辑工具，现在使这一目标触手可及。的 布罗德研究所的转录和微扰核心将与PPG的所有三个项目合作， 使用批量RNA-Seq、单细胞RNA-Seq和NanoString技术分析样品。我们将执行 计算分析，以生成电路模型，并确定潜在的监管机构和参与的途径， 布雷格和Beff调节。我们将通过使用CRISPR-Cas9基因组编辑来测试这些预测模型 原电池技术。核心将处理和分析所有项目的样本， PPG的所有研究者将共享数据。总的来说，这些结果将导致更好的 了解布雷格和Beff生物学、正常功能及其在疾病中的作用。这也可能导致 自身免疫、癌症和移植排斥的新疗法。

ABSTRACT — CORE B The mechanisms underlying the development, function, and regulation of regulatory B-cells (Bregs) and inflammatory B-effector (Beff) cells remain obscure. To date, studies of Bregs and Beff have been challenging due to their small numbers in clinical samples as well as their heterogeneity. However, the development of genomic approaches, such as RNA-Seq of bulk populations from low-input samples and single-cell RNA-Seq (scRNA-Seq) as well as CRISPR–Cas9 genome editing tools, now put this goal within reach. The Transcriptional and Perturbation Core at the Broad Institute will collaborate with all three projects of the PPG to profile samples using bulk RNA-Seq, single-cell RNA-Seq, and NanoString technology. We will perform computational analysis to generate circuit models and identify potential regulators and pathways involved in Breg and Beff regulation. We will test these predictive models by using CRISPR-Cas9 genome editing technology in primary cells. The Core will process and analyze samples across all projects and the resulting data will be shared among all the investigators of the PPG. Overall, these results will lead to better understanding of Breg and Beff biology, normal function, and their role in disease. It may also potentially lead to new avenues for therapy for autoimmunity, cancer, and transplant rejection.