Development of a Novel Method for the Identification and Characterization of Intercellular Communication in the Cancer Niche

开发一种用于识别和表征癌症生态位中细胞间通讯的新方法

基本信息

批准号：
10426930
负责人：
PETER M GORDON
金额：
$ 7.75万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10426930
关键词：
Address Biological Models Cancer Biology Cell Adhesion Molecules Cell Communication Cell Separation Cell Surface Proteins Cell surface Cells Central Nervous System Leukemia Coculture Techniques Communication Complex Development Developmental Biology Diffuse Engineering Enzymes Flow Cytometry Glycine Homing Immune Immune system In Vitro Injections Integral Membrane Protein Knock-in Label Leukemic Cell Literature Malignant Neoplasms Measures Methods Microscopy Mus N-terminal Neighborhoods Neoplasm Metastasis Neuraxis Pattern Peptides Peptidyltransferase Proteins Public Health Radial Reaction Reagent Resistance Scheme Signal Transduction System T-Lymphocyte Technology Testing Time Transgenic Mice Transgenic Organisms Transplantation Wild Type Mouse Work cancer cell cancer therapy design extracellular fluorophore in vivo intercellular communication interest leukemia migration mouse model new technology novel novel strategies single cell analysis single-cell RNA sequencing sortase stem cell biology technology development uptake

项目摘要

ABSTRACT Intercellular interactions and communication between cancer cells the cells that comprise their niches are critical for many aspects of cancer development, progression, metastasis, and therapy resistance. These interactions are often dynamic, transient and complex. Moreover, intercellular communication occurs directly by contact dependent cell-cell interactions and indirectly by the secretion of soluble factors into the local microenvironment. Current limitations to proximity dependent labeling strategies designed to identify and characterize direct and indirect cellular interaction partners in vivo include reagents that are toxic and/or limit the isolation and subsequent characterization of interacting cells, non-specific or large radius labeling (i.e. direct vs. indirect contacts), or a requirement to pre-engineer both interacting cells which obviates the possibility of identifying novel cellular interaction partners. Thus, a more ideal in vivo proximity dependent labeling system would concurrently, but differentially, label both direct and indirect cellular contacts at multiple timepoints using technology that is compatible with an array of downstream analyses ranging from microscopy to single cell analyses. In this proposal we will address this shortcoming in current technologies for comprehensively identifying and characterizing direct and indirect intercellular interactions in vivo. We will build upon the literature and our preliminary work to develop a novel approach in which an engineered cancer cell, or other cell of interest, concurrently, differentially and temporally fluorescently labels both direct and indirect cellular contacts within the cancer niche (Aims 1 and 2). Importantly, labeled cells will be suitable for an array of downstream analyses such as microscopy, flow cytometry, and cell sorting followed by bulk or single cell analyses. Accordingly, as proof-of-concept we will then combine this niche labeling technology with single-cell RNA-seq to begin to define cellular contacts during early stages of central nervous system leukemia metastasis and niche development (Aim 3).

抽象的细胞间相互作用和癌细胞之间包含其壁ches的细胞是对于癌症发展，进展，转移和耐药性的许多方面至关重要。这些相互作用通常是动态的，瞬态和复杂的。此外，直接发生细胞间沟通通过接触依赖性细胞 - 细胞相互作用，并通过分泌可溶性因子向局部进行间接微环境。当前限制依赖性依赖性标签策略，旨在识别和在体内表征直接和间接的细胞相互作用伴侣包括有毒和/或极限的试剂相互作用细胞，非特异性或大半径标记的隔离和后续表征（即直接与间接触点），或要求预先工程师相互作用的细胞的要求识别新型细胞相互作用伴侣的可能性。因此，一个更理想的体内接近度依赖性标记系统将同时但差异地标记在多个的直接和间接细胞触点使用与显微镜的一系列下游分析兼容的技术的时间点进行单细胞分析。在此提案中，我们将解决当前技术中的这一缺点在体内全面识别和表征直接和间接的细胞间相互作用。我们将建造关于文献和我们的初步工作，以开发一种新型方法，其中一种设计的癌细胞，或其他感兴趣的单元，同时，差异和时间荧光标记直接和间接的标签癌症生态裂内的细胞接触（目标1和2）。重要的是，标记的细胞适合阵列下游分析，例如显微镜，流式细胞术和细胞分类，然后是散装或单细胞分析。因此，作为概念验证，我们将将这种利基标签技术与单细胞结合起来在中枢神经系统白血病的早期，RNA-Seq开始定义细胞接触转移和利基发展（AIM 3）。