Single cell characterization of the biomaterial immune and stromal response

生物材料免疫和基质反应的单细胞表征

基本信息

批准号：
10431933
负责人：
JENNIFER H ELISSEEFF
金额：
$ 61.23万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-06 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10431933
关键词：
Adhesions Algorithms Antigen Presentation Atlases Autoimmunity Biocompatible Materials Biological Biological Models Biological Process Bladder Cell Communication Cell Separation Cells Chronic Cluster Analysis Complement Activation Data Development Disease Environment Extracellular Matrix Female Fibroblasts Fibrosis Flow Cytometry Foreign Bodies Foreign-Body Giant Cells Future Gender Genes Goals Human IL17 gene Immune Immune response Immune system Implant Individual Inflammation Inflammatory Innate Immune System Interleukin-17 Interleukin-4 Internal Breast Prosthesis Investigation Knowledge Lymphoid Malignant Neoplasms Methods Modeling Mus Muscle Myelogenous Operative Surgical Procedures PTPRC gene Pathology Phagocytosis Phenotype Population Process Proteins RNA Research Sorting - Cell Movement Surface T-Lymphocyte Techniques Time Tissue Model Tissues Trauma base capsule cell type cytokine differential expression first responder in vivo joint injury macrophage male medical implant migration neutrophil new therapeutic target novel polycaprolactone recruit repaired response scaffold senescence single cell analysis single cell sequencing single-cell RNA sequencing tissue degeneration tissue repair tool transcriptome tumor wound healing

项目摘要

Profiling single cells using single cell RNA sequencing (scRNAseq) is revolutionizing our understanding of development and disease. In this proposal, we will apply scRNAseq to create an atlas of cells that respond to biomaterials that induce divergent responses and serve as a model for tissue microenvironments of repair versus fibrosis. The proposed research aims to leverage single cell analysis to define key subpopulations in the lymphoid, myeloid and stromal fibroblasts response to biomaterial models of tissue fibrosis and repair. Minimally processed biological scaffolds induce a Type 2 immune response characterized by interleukin (IL)-4 and tissue repair, similar to muscle repair processes. Our preliminary data describes a Type 17 immune and senescent cell response to synthetic implants that induce fibrotic capsule formation in an IL-17-dependent manner. We also demonstrate the ability of scRNASeq to uncover new macrophage cell populations in biomaterial microenvironments. We hypothesize that by sorting cell subpopulations in the FBR in vivo, combined with single cell analysis, we will identify new and rare populations that will help elucidate mechanisms and provide new therapeutic targets to enhance tissue repair or reduce fibrosis. The following specific aims are proposed to accomplish this goal: Specific Aim 1. Identify and characterize lymphoid, myeloid, and fibroblast subpopulations isolated from synthetic and biological scaffold implants using single cell RNA sequencing analysis. Specific Aim 2. Computationally phenotype cell clusters both within and across cell types to define distinct subsets and interaction models using pseudotime analysis, RNA velocity, differential expression and gene set enrichment, cluster analysis to predict unique surface markers/combinations, and cell interactions analysis. Specific Aim 3. Define unique surface and intracellular markers from single cell analysis to identify subpopulations using standard experimental methods. Newly-identified immune and fibroblast subpopulations will be evaluated over time in male and female mice and results will be validated with diverse materials. The cell atlas created in the proposed research will enable future mechanistic studies and investigation into the potential broad applicability to wound healing, cancer and other tissue pathologies.

使用单细胞 RNA 测序 (scRNAseq) 分析单细胞正在彻底改变我们对单细胞的理解发育和疾病。在本提案中，我们将应用 scRNAseq 来创建响应的细胞图谱诱导不同反应并作为组织微环境修复模型的生物材料与纤维化。拟议的研究旨在利用单细胞分析来定义关键亚群淋巴、骨髓和基质成纤维细胞对组织纤维化和修复的生物材料模型的反应。经过最低限度加工的生物支架可诱导以白细胞介素 (IL)-4 为特征的 2 型免疫反应和组织修复，类似于肌肉修复过程。我们的初步数据描述了 17 型免疫和衰老细胞对合成植入物的反应，在 IL-17 依赖性细胞中诱导纤维化包膜形成方式。我们还证明了 scRNASeq 发现新巨噬细胞群的能力生物材料微环境。我们假设通过对体内 FBR 中的细胞亚群进行分类，结合单细胞分析，我们将识别新的和稀有的群体，这将有助于阐明机制并提供新的治疗靶点来增强组织修复或减少纤维化。下列为实现这一目标提出了具体目标：具体目标 1. 鉴定和表征从使用单细胞 RNA 测序分析合成和生物支架植入物。具体目标 2. 通过计算对细胞类型内部和跨细胞类型的细胞簇进行表型分析，以定义不同的细胞簇使用伪时间分析、RNA 速度、差异表达和基因集的子集和相互作用模型富集、预测独特表面标记/组合的聚类分析以及细胞相互作用分析。具体目标 3. 从单细胞分析中定义独特的表面和细胞内标记物以识别使用标准实验方法的亚群。新发现的免疫和成纤维细胞亚群随着时间的推移，将在雄性和雌性小鼠中进行评估，并用不同的材料验证结果。在拟议的研究中创建的细胞图谱将使未来的机制研究和调查成为可能其对伤口愈合、癌症和其他组织病理学的潜在广泛适用性。