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

• 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
• 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

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）：现在在T细胞开发文献中已经确定了在胸腺壁基中以高度控制和顺序的方式呈现的两个主要信号在生成功能性T细胞中起着至关重要的作用：（i）Delta像配体这样的Delta，如配体 - Notch受体信号传导和（II）的（II）主要的组织相称复杂（MHC）-TCR（MHC） - TCR（MHC） - TCR（MHC） - TCR -TCR（TCR） - TCR（TCR） - TCR（TCR） - TCR。像控制缺口信号传导和直接胸腺中TCR信号传导的MHC分子的三角体一样，存在于基质细胞和抗原呈递细胞的表面上，而发育中的茎/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基质细胞在体外模仿缺口信号过程。最近，我们报道了具有表面毫米的凹口配体（Delta样配体4，dll4）的微粒可以有效地将造血祖细胞引导到早期T细胞谱系。此外，我们现在已经证明，使用表面毫米液的配体介导的信号传导和外源性，可溶性抗原载荷的MHC四聚体介导的TCR信号传导，小鼠胚胎干细胞（ES）细胞和小鼠诱导的多能干细胞（IPS）细胞都可以针对The+ to the+ to the+ to the+ to the+ to the+ to cd indy to the+ to the+ to cd。但是，迄今为止，包括我们自己的大多数努力都集中在模仿2D环境中这些特定于小众的信号。基于我们在这些2D系统中的有希望的数据，我们假设可以有效触发培养的ES或IPS细胞中有效触发Delta-Notch和MHC-TCR信号通路的3D组织样微环境将在T细胞产生的效率方面显着提高。我们的目标是设计这种合成的3D T细胞壁ni（人造胸腺样微环境），将多能干细胞引导到治疗性，抗原特异性T细胞中。这项探索性研究的具体目的是：目标1：开发和表征与T细胞分化必不可少的配体功能化功能化的蛋白石水凝胶支架。 AIM 2：使用Notch配体官能化的蛋白石水凝胶支架有效地将小鼠ES和IPS细胞分为早期的T细胞。 AIM 3：为了有效地产生抗原特异性，使用AIM 2中早期T细胞群的功能性CD8+ T细胞，使用抗原载荷，MHC官能化的逆蛋白质水凝胶并评估其抗原的特异性和对自我抗原的交叉反应性。 公共卫生相关性：这个为期两年的探索性项目的目标是开发三维胸腺类似微环境，以从小鼠胚胎或诱导的多能干细胞中产生功能性的抗原特异性T细胞。我们的方法是通过创建具有有效方式的特定T细胞分化信号的支架来模仿胸腺壁裂。如果成功，这可能会导致使用干细胞衍生细胞的产卵T细胞疗法。