Synthetic, Thymus-like 3D Niche for T Cell Generation from Stem Cells

用于从干细胞生成 T 细胞的合成类胸腺 3D 生态位

基本信息

批准号：
7874375
负责人：
KRISHNENDU ROY
金额：
$ 18.69万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7874375
关键词：
Address Adoptive Transfer Antigen-Presenting Cells Antigens Apoptosis Autoantigens Autologous Binding Biological Assay CD8B1 gene Cell Lineage Cell Separation Cell Size Cell Therapy Cell surface Cells Commit Complex Data Differentiation Antigens Embryo Engineering Ensure Environment Epithelial Cells Generations Goals Hematopoietic stem cells Hodgkin Disease Hydrogels Immunotherapy In Vitro Incubated Laboratories Lead Life Ligands Literature Lymphopoiesis MHC antigen Major Histocompatibility Complex Malignant neoplasm of nasopharynx Malignant neoplasm of prostate Manuscripts Mature T-Lymphocyte Mediating Methods Microspheres Mixed Lymphocyte Culture Test Morbidity - disease rate Multiple Myeloma Mus Pathway interactions Patient Education Patients Peptides Play Pluripotent Stem Cells Polymers Population Process Receptor Signaling Renal carcinoma Reporting Research Role Signal Transduction Source Specificity Staging Stem cells Stromal Cells Surface System T Cell Receptor Signaling Pathway T cell differentiation T-Cell Development T-Cell Receptor T-Lymphocyte Technology Testing Therapeutic Thymus Gland Time Tissues Training Transplantation advanced disease base computerized data processing cross reactivity density embryonic stem cell in vivo induced pluripotent stem cell leukemia melanoma notch protein nuclear transfer peripheral blood porous hydrogel public health relevance scaffold stem stem cell differentiation

项目摘要

DESCRIPTION (provided by applicant): It is now well established in the T cell development literature that two key signals, presented in the thymic niche, in a highly controlled and sequential manner, play crucial roles in generating functional T cells: (i) Delta like ligands - Notch receptor signaling and (ii) Major Histocompatibility Complex (MHC) - T cell receptor (TcR) signaling. The Delta like ligands that control Notch signaling and the MHC molecules that direct TcR signaling in the thymus are present on the surface of stromal cells and antigen presenting cells, with which the developing stem/progenitor cells directly interact. It has been shown that at least the Notch signaling process can be mimicked in vitro using retrovirally transfected stromal cells displaying Notch ligands. Recently, we have reported that microbeads with surface-immobilized Notch ligands (delta-like ligand 4, DLL4) can efficiently direct hematopoietic progenitor cells to the early T cell lineage. In addition, we have now demonstrated that, using a combination of surface-immobilized Notch ligand-mediated signaling and exogenous, soluble antigen-loaded MHC tetramer-mediated TcR signaling, both mouse embryonic stem (ES) cells and mouse induced pluripotent stem (iPS) cells can be directed to the T cell pathway and differentiated into functional, CD8+ T cells specific for the same antigen. However, most efforts to date, including our own has focused on mimicking these niche-specific signals in 2D environments. Based on our promising data in these 2D systems, we hypothesize that a 3D tissue-like microenvironment that can efficiently trigger Delta- Notch and MHC-TcR signaling pathways in cultured ES or iPS cells would provide significant improvement in the efficiency of T cell generation. Our goal here is to engineer such synthetic, 3D T cell niches (artificial thymus-like microenvironment) to direct pluripotent stem cells into therapeutic, antigen-specific T cells. The specific aims of this exploratory research are: Aim 1: To develop and characterize inverse opal hydrogel scaffolds functionalized with ligands essential for T cell differentiation. Aim 2: To efficiently differentiate mouse ES and iPS cells into early T cells using Notch ligand-functionalized inverse opal hydrogel scaffolds. Aim 3: To efficiently generate antigen-specific, functional CD8+ T cells from the early T cell population obtained in Aim 2, using antigen-loaded, MHC-functionalized inverse opal hydrogels and evaluate their antigen specificity and cross-reactivity to self antigens. PUBLIC HEALTH RELEVANCE: The goal of this two year exploratory project is to develop a three dimensional thymus- like microenvironment to generate functional, antigen-specific T cells from mouse embryonic or induced pluripotent stem cells. Our approach is to mimic the thymic niche by creating scaffolds that present specific T cell differentiation signals in an efficient manner. If successful this could lead to adoptive T cell therapy using stem cell derived cells.

描述（由申请人提供）：现在 T 细胞发育文献中已经明确，胸腺微环境中以高度受控和连续的方式呈现的两个关键信号在生成功能性 T 细胞中发挥着至关重要的作用：(i) Delta类似配体 - Notch 受体信号传导和 (ii) 主要组织相容性复合物 (MHC) - T 细胞受体 (TcR) 信号传导。控制Notch信号传导的Delta样配体和指导胸腺中TcR信号传导的MHC分子存在于基质细胞和抗原呈递细胞的表面，与发育中的干细胞/祖细胞直接相互作用。已经表明，至少可以使用逆转录病毒转染的展示Notch配体的基质细胞在体外模拟Notch信号传导过程。最近，我们报道了表面固定Notch配体（δ样配体4，DLL4）的微珠可以有效地将造血祖细胞引导至早期T细胞谱系。此外，我们现在已经证明，结合使用表面固定的Notch配体介导的信号传导和外源可溶性抗原负载的MHC四聚体介导的TcR信号传导，小鼠胚胎干（ES）细胞和小鼠诱导多能干细胞（iPS） ) 细胞可以定向至 T 细胞途径并分化为对相同抗原具有特异性的功能性 CD8+ T 细胞。然而，迄今为止，包括我们自己在内的大多数努力都集中在模仿 2D 环境中的这些特定于利基的信号。基于我们在这些 2D 系统中的有希望的数据，我们假设能够在培养的 ES 或 iPS 细胞中有效触发 Delta-Notch 和 MHC-TcR 信号通路的 3D 组织样微环境将显着提高 T 细胞的生成效率。我们的目标是设计这种合成的 3D T 细胞生态位（人工胸腺样微环境），以将多能干细胞引导为治疗性抗原特异性 T 细胞。这项探索性研究的具体目标是：目标 1：开发并表征用 T 细胞分化必需的配体功能化的反蛋白石水凝胶支架。目标 2：使用 Notch 配体功能化的反蛋白石水凝胶支架有效地将小鼠 ES 和 iPS 细胞分化为早期 T 细胞。目标 3：使用负载抗原的 MHC 功能化反蛋白石水凝胶，从目标 2 中获得的早期 T 细胞群中有效生成抗原特异性、功能性 CD8+ T 细胞，并评估其抗原特异性和与自身抗原的交叉反应性。公共健康相关性：这个为期两年的探索性项目的目标是开发三维胸腺样微环境，以从小鼠胚胎或诱导多能干细胞中产生功能性、抗原特异性 T 细胞。我们的方法是通过创建以有效方式呈现特定 T 细胞分化信号的支架来模拟胸腺生态位。如果成功的话，这可能会导致使用干细胞衍生细胞的过继性 T 细胞疗法。