T cell invasion of the stem cell compartment during immune-mediated GI damage

免疫介导的胃肠道损伤过程中 T 细胞入侵干细胞区室

• 批准号: 10322754
• 负责人: Alan M Hanash
• 金额: $ 57.3万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322754
• 关键词: 3-Dimensional; Active immunity; Acute Graft Versus Host Disease; Address; Adoptive Cell Transfers; Affect; Allogeneic Bone Marrow Transplantation; Allogenic; Automobile Driving; Behavior; Biological; Biology; Bone Marrow Transplantation; Cell Adhesion Molecules; Cell Communication; Cell Compartmentation; Cells; Clinical; Coculture Techniques; Development; Endothelium; Enterocytes; Environment; Epithelial; Frequencies; Functional disorder; Gastrointestinal Diseases; Gastrointestinal tract structure; Gene Expression; Genetic Techniques; Genetic Transcription; Goals; Hematopoietic; Homeostasis; Homing; Homologous Transplantation; Human; Image; Immune; Immune Targeting; Immune response; Immune system; Immunologics; Immunology; Immunosuppression; In Situ; Individual; Infiltration; Inflammatory Bowel Diseases; Intervention; Intestinal Mucosa; Intestines; LGR5 gene; Ligands; Location; Lymphocyte; Lymphoid Cell; Maintenance; Measures; Mediating; Memorial Sloan-Kettering Cancer Center; Microscopy; Modeling; Molecular; Mucous Membrane; Natural regeneration; Organoids; Paneth Cells; Pathogenicity; Pathologic; Pathology; Pathway interactions; Phenotype; Physiological; Process; Publishing; Recovery; Regulation; Research Proposals; Resources; Role; Sampling; Science; Signal Transduction; Site; Source; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Thick; Three-Dimensional Imaging; Tissues; Toxic effect; Transplantation; Transplantation Conditioning; Tumor-infiltrating immune cells; Validation; base; cell injury; cell motility; chemokine; clinically significant; conditioning; crypt cell; dimer; epithelial injury; epithelial stem cell; gastrointestinal; graft vs host disease; hematopoietic transplantation; human model; immune imaging; immune system function; immunopathology; in vivo; in vivo Model; injury recovery; insight; interleukin-22; intestinal injury; intravital imaging; man; microscopic imaging; migration; mouse model; multidisciplinary; novel; novel therapeutics; post-transplant; protein expression; recruit; response; skills; stem cell biology; stem cells; targeted treatment; tissue regeneration; translational model; translational potential; transplant model; two-photon

Despite the importance of intestinal stem cells (ISCs) for epithelial maintenance in the gastrointestinal (GI) tract and the clinical significance of inflammatory intestinal disorders and immune-mediated GI damage, the potential of the immune system to directly engage with the ISC compartment remains poorly understood. GI graft vs. host disease (GVHD) is one of the greatest challenges for allogeneic hematopoietic/bone marrow transplantation (allo-BMT), causing severe toxicity, necessitating deeply immunosuppressive interventions, and limiting the wider usage of allo-BMT for potentially curable conditions. We have found that acute GVHD leads to substantial reduction of Lgr5+ ISCs and the Paneth cells providing their epithelial niche. While there is limited understanding of the precise mechanisms by which immune responses induce ISC compartment damage and regeneration, it is not possible to fully understand BMT-related complications in the GI tract or how immune cells may impact specific epithelial components without examining the three-dimensional (3-D) tissue environment of the intestines and the proximity of immune responses to the ISC compartment. We seek to understand the fundamental interactions occurring between the immune system and the ISC compartment, how these interactions are engaged in the context of hematopoietic transplantation, and how the underlying biology may be manipulated therapeutically for clinical intervention. To address these goals, we have developed a research proposal emphasizing 3-D imaging of immune effectors within GI tract, mechanistic ex vivo modeling of murine and human immune cell recruitment to the ISC compartment, and in vivo validation in experimental transplant models. We have also assembled a multi-disciplinary team of collaborators at the forefront of basic and translational immunology, ISC biology, and advanced microscopic imaging. Our preliminary findings indicate that the ISC compartment is the first site of allogeneic T cell invasion within the intestinal mucosa after BMT and thus the initial target of GVHD. Furthermore, we have found that co-cultures of immune cells and intestinal organoids can be used to model and mechanistically dissect these processes. Using 3-D confocal and 2-photon intravital imaging, we have developed the capacity to accurately quantify loss of ISCs, specific invasion of the ISC compartment, and localization of T cell homing ligands during homeostasis and during active immunity. We have also developed strategies for analysis of in vivo lineage- specific gene expression within the ISC compartment. This study will thus test the hypothesis that lymphocyte recruitment to and regulation of the ISC compartment are central features of GI damage post-transplant, impacting both physiologic and pathologic mucosal responses, and we will evaluate approaches to modulate lymphocyte homing and promote recovery of injured epithelium. This project will lead to a mechanistic understanding of fundamental interactions between the ISC compartment and the immune system, opening a new direction for treatment of GVHD and other GI disorders by protecting ISCs and augmenting regeneration.

尽管肠干细胞（ISC）对于胃肠道（GI）上皮的维持非常重要 以及炎症性肠道疾病和免疫介导的胃肠道损伤的临床意义， 免疫系统直接与 ISC 区室结合的潜力仍知之甚少。胃肠道 移植物抗宿主病（GVHD）是同种异体造血/骨髓面临的最大挑战之一 移植（allo-BMT），导致严重毒性，需要深度免疫抑制干预，以及 限制了异基因骨髓移植（allo-BMT）在潜在可治愈疾病中的更广泛使用。我们发现急性 GVHD 会导致 Lgr5+ ISC 和提供上皮生态位的潘氏细胞大幅减少。虽然数量有限 了解免疫反应诱导 ISC 区室损伤的精确机制 再生，不可能完全了解胃肠道中与 BMT 相关的并发症或免疫如何 细胞可能会影响特定的上皮成分，而无需检查三维（3-D）组织 肠道环境以及免疫反应与 ISC 隔室的接近程度。我们力求 了解免疫系统和 ISC 区室之间发生的基本相互作用， 这些相互作用如何在造血移植的背景下进行，以及潜在的作用如何 可以通过治疗手段操纵生物学以进行临床干预。为了实现这些目标，我们有 制定了一项研究提案，强调胃肠道内免疫效应器的 3D 成像，机械前 小鼠和人类免疫细胞招募至 ISC 隔室的体内建模，以及体内验证 实验移植模型。我们还组建了一支多学科合作团队 基础和转化免疫学、ISC 生物学和先进显微成像的前沿。我们的 初步研究结果表明，ISC区室是同种异体T细胞入侵的第一个部位。 BMT后的肠粘膜因此成为GVHD的初始目标。此外，我们发现共培养 免疫细胞和肠道类器官的研究可用于模拟和机械剖析这些过程。 使用 3-D 共焦和 2 光子活体成像，我们已经具备了准确量化损失的能力 ISC 的形成、ISC 区室的特异性侵袭以及 T 细胞归巢配体的定位 稳态和主动免疫期间。我们还开发了体内谱系分析的策略- ISC 区室内的特定基因表达。因此，本研究将检验淋巴细胞的假设 ISC 区室的募集和调节是移植后胃肠道损伤的核心特征， 影响生理和病理粘膜反应，我们将评估调节方法 淋巴细胞归巢并促进受损上皮的恢复。该项目将导致机械化 了解 ISC 区室和免疫系统之间的基本相互作用，开启了 通过保护 ISC 和增强再生来治疗 GVHD 和其他胃肠道疾病的新方向。