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：
10230987
负责人：
JENNIFER H ELISSEEFF
金额：
$ 60.66万
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/10230987
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发展与疾病。在此提案中，我们将应用scrnaseq创建一个响应的单元格诱导反应并作为组织微环境的模型的生物材料与纤维化相对于纤维化。拟议的研究旨在利用单细胞分析来定义关键亚群淋巴样，髓样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。最小加工的生物支架诱导以白介素（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型免疫和对合成植入物的衰老细胞反应，这些植入物在IL-17依赖性中诱导纤维化胶囊形成方式。我们还证明了Scrnaseq在发现新的巨噬细胞种群中的能力生物材料微环境。我们假设通过对fbr in Vivo中的细胞亚群进行排序，结合单细胞分析，我们将确定新的和罕见的种群，以帮助阐明机制并提供新的治疗靶标，以增强组织修复或减少纤维化。下列提出了具体目标来实现这一目标：特定目标1。识别并表征从从中分离出来的淋巴样，髓样和成纤维细胞亚群使用单细胞RNA测序分析的合成和生物支架植入物。特定目的2。计算表型细胞簇在细胞类型内部和跨细胞类型中，以定义不同的使用伪分析，RNA速度，差异表达和基因集的子集和相互作用模型富集，聚类分析以预测独特的表面标记/组合和细胞相互作用分析。特定目标3。从单细胞分析中定义独特的表面和细胞内标记以识别使用标准实验方法的亚群。新鉴定的免疫和成纤维细胞亚群将随着时间的流逝评估男性和雌性小鼠，结果将通过各种材料进行验证。拟议的研究中创建的细胞地图集将使未来的机械研究和研究潜在的广泛适用于伤口愈合，癌症和其他组织病理。