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Direct Epigenetic Reprogramming of T Cells

T 细胞的直接表观遗传重编程

基本信息

批准号：
8955075
负责人：
Artem Barski
金额：
$ 234万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-30 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8955075
关键词：
Affect Allergens Area Asthma Autoimmunity Back Beryllium Blood CD4 Positive T Lymphocytes Cell Lineage Cells Chimeric Proteins Chromatin Communicable Diseases DNA Binding Domain DNA Methylation Deposition Elements Engineering Enzymes Epigenetic Process Genes Helminths Helper-Inducer T-Lymphocyte Heterogeneity Human Hypersensitivity IL4 gene Immune Immune System Diseases Immune response Immunosuppressive Agents Immunotherapy Infection Inflammatory Knowledge Lead Learning Maintenance Memory Methylation Modeling Mus Mycoses Patients Phenotype Population Recording of previous events Regulatory Element Regulatory T-Lymphocyte Rest T memory cell T-Lymphocyte Technology Testing Th1 Cells Th2 Cells Tumor Antigens Work base cancer immunotherapy cytokine disease phenotype epigenetic regulation epigenome histone modification human disease interest memory CD4 T lymphocyte mouse model novel novel strategies pathogen public health relevance tool tumor

项目摘要

DESCRIPTION (provided by applicant): The heterogeneity within the CD4+ effector/memory T-cell compartment is critical for our ability to deal with diverse pathogens. For example, dedicated populations of CD4+ T helper cells are required for promoting immune defense against intracellular infections (Th1 cells), helminth infections (Th2 cells), and fungal infection (Th17 cells). On the other hand, each of these differentiated states is associated with human disease: Th1 and Th17 cells can promote autoimmunity, while Th2 cells can promote allergy and asthma. Thus, understanding and learning to exploit the mechanisms that underlie lineage choice is vital for understanding and treatment of immunological and infectious diseases. We are interested in epigenetic regulation of T-cell lineage commitment and T-cell memory. Here, we propose a novel strategy to target the Th cell epigenome in order to modify their phenotype and promote or inhibit inflammatory immune response. The cellular epigenome, represented by DNA methylation, histone modifications and ncRNA, is believed to reflect the differentiation history of the cell and determine its phenotype. Working with human helper T cells, we have found that the ability of memory CD4 T cells to quickly induce key cytokines (rapid recall ability) is correlated with epigenetic gene poising: the presence of positive histone modifications at several regulatory elements in the resting memory cells. Based on this observation, we hypothesize that memory T-cell lineage commitment is encoded in the epigenome. As a first step toward proving this hypothesis (and to T-cell reprogramming), we will attempt to modify T cell phenotype by changing chromatin marks at several key elements that we and others have previously identified. To do so, we are using TALEMs - fusion proteins of TAL-based engineered DNA binding domains (DBDs) with Epigenetic Modifier enzymes. We will use TALEMs to remove positive chromatin marks (e.g., H3K4 methylation) from the previously identified regulatory elements in the IL4/13 locus to test whether the presence of such marks is indeed required for the maintenance of Th2 phenotype and IL4/13 gene inducibility. We will also test whether deposition of negative marks (e.g., K9me2 and DNA methylation) there will be sufficient to reverse Th2 differentiation. Similarly, we will test whether deposition of the positie marks at these elements is sufficient to force Th2 cytokine inducibility in naïve T cells. Finallywe will test, whether such epigenetic reprogramming can affect disease phenotype in a mouse model of experimental asthma. If successful, this strategy may potentially lead to creation of therapies for immunological diseases and immunotherapy of cancer. In one hypothetical scenario, for asthma, allergen- specific inflammatory Th2 cells could be purified from patient's blood using tetramers, propagated and reprogrammed into immunosuppressive Tregs. These Treg cells could then be returned back to patient to restore tolerance and, potentially, to provide a cure. Importantly, the ability of allergen-specific Tregs to cure experimental asthma has already been demonstrated in a murine model. Similarly, tumor antigen-specific Treg cells could be purified from tumors and reprogrammed into Th1 effectors for immunotherapy. Knowledge gained through this study would enable similar applications in other biomedical areas.

描述（由申请人提供）：CD 4+效应/记忆T细胞区室内的异质性对于我们处理不同病原体的能力至关重要。例如，需要专门的CD 4 + T辅助细胞群来促进针对细胞内感染（Th 1细胞）、蠕虫感染（Th 2细胞）和真菌感染（Th 17细胞）的免疫防御。另一方面，这些分化状态中的每一种都与人类疾病有关：Th 1和Th 17细胞可以促进自身免疫，而Th 2细胞可以促进过敏和哮喘。因此，理解和学习利用谱系选择的机制对于理解和治疗免疫和传染病至关重要。我们对T细胞谱系定型和T细胞记忆的表观遗传调控感兴趣。在这里，我们提出了一种新的策略，以目标Th细胞表观基因组，以修改其表型和促进或抑制炎症免疫反应。以DNA甲基化、组蛋白修饰和ncRNA为代表的细胞表观基因组被认为反映了细胞的分化历史并决定了其表型。与人类辅助性T细胞合作，我们发现记忆性CD 4 T细胞快速诱导关键细胞因子的能力（快速回忆能力）与表观遗传基因平衡相关：在静息记忆细胞中的几个调节元件处存在阳性组蛋白修饰。基于这一观察结果，我们假设记忆T细胞谱系承诺编码在表观基因组中。作为证明这一假设（以及T细胞重编程）的第一步，我们将尝试通过改变我们和其他人先前已经确定的几个关键元件的染色质标记来修改T细胞表型。为此，我们正在使用TALEM-基于TAL的工程化DNA结合结构域（DBD）与表观遗传修饰酶的融合蛋白。我们将使用TALEM去除阳性染色质标记（例如，H3 K4甲基化），以测试这些标记的存在是否确实是维持Th 2表型和IL 4/13基因诱导所必需的。我们还将测试是否沉积负标记（例如，K9 me 2和DNA甲基化）将足以逆转Th 2分化。同样，我们将检测这些元件上的阳性标记是否足以在初始T细胞中诱导Th 2细胞因子。最后，我们将测试这种表观遗传重编程是否能影响实验性哮喘小鼠模型的疾病表型。如果成功，这一策略可能会导致免疫性疾病和癌症免疫疗法的产生。在一种假设的情况下，对于哮喘，过敏原特异性炎性Th 2细胞可以使用四聚体从患者的血液中纯化，增殖并重编程为免疫抑制性T细胞。然后，这些Treg细胞可以返回到患者体内以恢复耐受性，并可能提供解药了重要的是，过敏原特异性Tclase治愈实验性哮喘的能力已经在小鼠模型中得到证实。类似地，可以从肿瘤中纯化肿瘤抗原特异性Treg细胞，并将其重编程为用于免疫治疗的Th 1效应子。通过这项研究获得的知识将使类似的应用在其他生物医学领域。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Chromatin Preparation from Murine Eosinophils for Genome-Wide Analyses.

从鼠嗜酸性粒细胞中制备染色质用于全基因组分析。

DOI：
10.1007/978-1-4939-7896-0_20
发表时间：
2018
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Bouffi,Carine;Barski,Artem;Fulkerson,PatriciaC
通讯作者：
Fulkerson,PatriciaC

RNF8 and SCML2 cooperate to regulate ubiquitination and H3K27 acetylation for escape gene activation on the sex chromosomes.

DOI：
10.1371/journal.pgen.1007233
发表时间：
2018-03
期刊：
PLoS genetics
影响因子：
4.5
作者：
Adams SR;Maezawa S;Alavattam KG;Abe H;Sakashita A;Shroder M;Broering TJ;Sroga Rios J;Thomas MA;Lin X;Price CM;Barski A;Andreassen PR;Namekawa SH
通讯作者：
Namekawa SH

Polycomb directs timely activation of germline genes in spermatogenesis.

DOI：
10.1101/gad.302000.117
发表时间：
2017-08-15
期刊：
Genes & development
影响因子：
10.5
作者：
Maezawa S;Hasegawa K;Yukawa M;Sakashita A;Alavattam KG;Andreassen PR;Vidal M;Koseki H;Barski A;Namekawa SH
通讯作者：
Namekawa SH

Analysis of ChIP-Seq and RNA-Seq Data with BioWardrobe.

使用 BioWardrobe 分析 ChIP-Seq 和 RNA-Seq 数据。