Balanced signaling cues to guide cell transitions in the blood lineage continuum

平衡的信号线索引导血统连续体中的细胞转变

基本信息

批准号：
9127318
负责人：
Arup K. Chakraborty
金额：
$ 76.09万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-15 至 2020-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9127318
关键词：
Acute T Cell Leukemia Alleles Antibodies Autoimmune Diseases Autoimmune Process Autoimmunity Automobile Driving Behavior Biochemical Biochemistry Biological Blood Blood Cells Bone Marrow Bone Marrow Cells CD4 Positive T Lymphocytes Cell Lineage Cell physiology Cell surface Cells Characteristics Childhood Classification Complex Computer Simulation Computer software Cues Cytokine Receptors Cytometry Data Data Collection Data Set Development Differential Equation Discipline Ensure Environment Equilibrium Extracellular Signal Regulated Kinases FRAP1 gene Future Generations Goals Health Helper-Inducer T-Lymphocyte Hematological Disease Hematopoiesis Hematopoietic Hematopoietic Neoplasms Human Immune system Immunologic Deficiency Syndromes Individual Investigation Knock-in Lead Link Lupus Lymphocyte Lymphocyte Subset Lymphoid MAP Kinase Modules Malignant Neoplasms Membrane Methods Mission Mitogen-Activated Protein Kinases Modeling Mus Mutate Mutation Nature Neurons Nuclear Oncogenic Oocytes Pathway interactions Patients Pattern Phenotype Play Population Process Proliferating Proteins Publications Publishing Regulation Reporting Research Research Personnel Resolution Role Sampling Second Messenger Systems Shapes Signal Transduction Signaling Protein Spleen Staging Stimulus Surface System T cell differentiation T cell regulation T-Cell Leukemia T-Lymphocyte Testing Thymocyte Selection Thymus Gland United States National Institutes of Health analog base biomarker panel digital dynamic system high throughput analysis human disease insight interest leukemia lupus-like lymph nodes mouse model novel overexpression receptor research study response second messenger self-renewal simulation

项目摘要

DESCRIPTION (provided by applicant): Blood cells need to self-renew, proliferate, and differentiate in a balanced fashion to enable self-sustaining blood systems such as the immune system. The biochemical signaling network that regulates this balance is complex, non-intuitive, and not well understood. Adding to the complexity, blood cells exist as diverse lineages with rare, but critical, subsets within each lineage. Traditional FACS analyses with limited cell surface marker panels have created the false notion of restricted subsets, with abrupt transitions in a lineage trajectory. Such limited subset classification (and analyses of signals herein) has obstructed a full understanding of the function of biochemical networks that regulate the cellular balance between proliferation and differentiation. Our recently pioneered single-cell mass cytometry (CyTOF) method broke this impasse and has revealed that hematopoiesis in the bone marrow is a continuum with over a hundred identifiable subsets. It is known that aberrant biochemical networks can form the basis for human diseases like cancer, autoimmune diseases, or immunodeficiency Our deterministic and stochastic computational models that explored the topology of Ras signaling predicted distinct patterns of Ras activation as a function of the Ras activator proteins Rasgrp and Sos. Testing these hypotheses, we found that analog Rasgrp1- Ras-ERK or bimodal Sos-Ras-ERK signals can occur in lymphocytes. Our new mouse models now indicate that different perturbation in Rasgrp1 lead to reshaping of the Ras signals and result in cancer, autoimmune diseases, or immunodeficiency. Here we hypothesize that blood cells develop through a continuum in a balanced manner as a function of the topology and character of the Ras signaling network. In Preliminary Results, we discuss our ordinary differential equation (ODE) and Stochastic simulation compile (SSC) computational models of Ras signaling, details of our CyTOF data collection and computational SPADE and ACCENSE analysis methods, as well as our biochemical phospho-flow analyses on defined subsets of lymphocytes. We also present several lines of evidence that the Ras activator Rasgrp1 shapes the character of the Ras network to balance proliferation and differentiation. Loss of Rasgrp1 leads to immunodeficiency. We present data from our recent 2013 publications on T cell leukemia caused by oncogenic Ras mutations or overexpression of the Ras activator Rasgrp1 as well as a lupus-like autoimmune phenotype in a mouse model with a point-mutated Rasgrp1Anaef allele. In this proposal we will combine computational hypothesis generation, high-resolution analytic approaches of high-dimensional CyTOF data, and high-throughput biochemical analyses of primary blood cells from mouse models with distinct Ras signals and human leukemia samples to understand the topology of the Ras signaling network in T lymphocytes properly transitioning through the normal continuum in the bone marrow (Aim 1) and thymus (Aim 2). We will also characterize how perturbations of the network's character can lead to immunodeficiency, autoimmunity, or T cell leukemia. Using reiterative loops between the three disciplines, we focus on the T cell lineage here to ensure a productive research plan but will also generate new insights relevant for all hematopoietic blood lineages to spur future investigations.

描述（由申请人提供）：血细胞需要以平衡的方式自我更新、增殖和分化，以实现自我维持的血液系统，例如免疫系统。调节这种平衡的生化信号网络是复杂的、非直观的，而且还没有被很好地理解。更复杂的是，血细胞以不同的谱系存在，每个谱系内都有稀有但重要的子集。传统的 FACS 分析使用有限的细胞表面标记物组，产生了受限子集的错误概念，并且谱系轨迹发生突然转变。这种有限的子集分类（以及本文中的信号分析）阻碍了对调节增殖和分化之间的细胞平衡的生化网络的功能的充分理解。我们最近首创的单细胞质谱流式细胞术 (CyTOF) 方法打破了这一僵局，并揭示了骨髓中的造血是一个具有一百多个可识别子集的连续体。众所周知，异常的生化网络可以构成癌症、自身免疫性疾病或免疫缺陷等人类疾病的基础。我们探索 Ras 信号拓扑的确定性和随机计算模型预测了 Ras 激活作为 Ras 激活蛋白 Rasgrp 函数的不同模式和索斯。测试这些假设，我们发现类似的 Rasgrp1-Ras-ERK 或双峰 Sos-Ras-ERK 信号可以出现在淋巴细胞中。我们的新小鼠模型现在表明，Rasgrp1 中的不同扰动会导致 Ras 信号重塑，并导致癌症、自身免疫性疾病或免疫缺陷。在这里，我们假设血细胞根据 Ras 信号网络的拓扑和特征以平衡的方式通过连续体发育。在初步结果中，我们讨论了 Ras 信号传导的常微分方程 (ODE) 和随机模拟编译 (SSC) 计算模型、CyTOF 数据收集和计算 SPADE 和 ACCENSE 分析方法的详细信息，以及我们的生化磷流分析定义的淋巴细胞亚群。我们还提供了一些证据表明 Ras 激活剂 Rasgrp1 塑造 Ras 网络的特征以平衡增殖和分化。 Rasgrp1 缺失会导致免疫缺陷。我们提供了 2013 年最近发表的关于由致癌 Ras 突变或 Ras 激活剂 Rasgrp1 过度表达引起的 T 细胞白血病的数据，以及具有点突变 Rasgrp1Anaef 等位基因的小鼠模型中的狼疮样自身免疫表型。在本提案中，我们将结合计算假设生成、高维 CyTOF 数据的高分辨率分析方法以及对具有不同 Ras 信号的小鼠模型和人类白血病样本的原代血细胞进行高通量生化分析，以了解 Ras 的拓扑结构T 淋巴细胞中的信号网络正确地通过骨髓（目标 1）和胸腺（目标 2）的正常连续体。我们还将描述网络特性的扰动如何导致免疫缺陷、自身免疫或 T 细胞白血病。利用三个学科之间的重复循环，我们在这里重点关注 T 细胞谱系，以确保研究计划富有成效，同时也将产生与所有造血血液谱系相关的新见解，以促进未来的研究。