Pulmonary Macrophage Transplantation for Pulmonary Alveolar Proteinosis

肺巨噬细胞移植治疗肺泡蛋白沉积症

• 批准号: 10213109
• 负责人: Bruce Colston Trapnell
• 金额: $ 45.97万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213109
• 关键词: ATAC-seq; Adopted; Alveolar; Alveolar Macrophages; Architecture; Binding; Biological Markers; Bone Marrow; Bronchoalveolar Lavage; CSF2RA gene; Catabolism; Cell Therapy; Cells; Child; Cholesterol; Cholesterol Homeostasis; Chromatin; Clinical Data; Clinical Research; Clonal Expansion; Computer software; DNA; Data; Data Reporting; Development; Diagnosis; Disease; Elements; Engraftment; Evaluation; Exposure to; Fatty Acids; Fetal Liver; Foundations; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor Receptors; Growth; Health; Hematopoietic stem cells; Histopathology; Homeostasis; Host Defense; Human; Inherited; Kinetics; Knowledge; Lecithin; Link; Lung; Lung diseases; Macrophage Colony-Stimulating Factor; Macrophage Colony-Stimulating Factor Receptor; Measurement; Measures; Messenger RNA; Methods; Mission; Molecular; Mus; Mutation; Myelogenous; Myeloid Cells; Nature; Outcome; PPAR gamma; Pathogenesis; Pathologic; Patients; Pharmacologic Substance; Pharmacology; Pharmacotherapy; Phenotype; Phospholipids; Population Dynamics; Process; Proto-Oncogene Protein c-kit; Public Health; Publishing; Pulmonary Alveolar Proteinosis; Receptor Signaling; Regulator Genes; Regulatory Element; Replacement Therapy; Reporting; Research; Resolution; Role; Safety; Severity of illness; Signal Transduction; Stem Cell Factor; Survanta; Techniques; Testing; Therapeutic; Therapeutic Effect; Time; To specify; Translations; Transplantation; Treatment Efficacy; United States National Institutes of Health; Withdrawal; base; cell type; chromatin remodeling; clinical center; cohort; combinatorial; comparative efficacy; donor stem cell; effective therapy; experience; granulocyte; indexing; innovation; macrophage; monocyte; novel; novel therapeutics; pre-clinical; progenitor; programs; protein expression; pulmonary function; self-renewal; single-cell RNA sequencing; surfactant; tool; transcription factor; transcriptome; transcriptome sequencing; transplantation therapy

ABSTRACT There is a significant gap in understanding how GM-CSF directs macrophage specification following pulmonary macrophage transplantation (PMT). This is an important problem, because, without this crucial information, the preclinical data will not adequately demonstrate PMT safety sufficient to gain approval to test PMT in humans. The long-term goal is to develop PMT as therapy for patients with hereditary pulmonary alveolar proteinosis (hPAP) aimed at restoring alveolar macrophage (AM) function. The objective here is to identify mechanism(s) directing AM specification during PMT. The central Hypothesis is that AM specification is determined primarily by factors in the alveolar microenvironment including 1) GM-CSF, which renders AM-specific DNA regulatory elements accessible; and 2) surfactant phospholipid-derived fatty acids, which activate PPARγ to bind newly accessible DNA and switch AMs from LXR-driven to PPARγ-driven specification (and cholesterol metabolism). Our rationale is that by demonstrating GM-CSF directs macrophages to adopt a normal AM transcription profile at single cell resolution (which `bulk' studies failed to show) will be crucial to establish PMT is safe. Guided by our strong preliminary data, this hypothesis will be tested by pursuing three specific aims to determine 1) the relationship between donor cell plasticity and therapeutic efficacy; 2) temporal dynamics of macrophage engraftment, specification, and fate after PMT; and 3) cis-regulatory architecture governing AM specification. In aim 1, myeloid cells of various developmental stages will be administered by PMT to Csf2raKO mice to deter- mine the plasticity of cells capable of conferring therapeutic benefit. In aim 2, PMT of normal hematopoietic stem/progenitor cell (HSPC) donors in Csf2raKO (or normal) mice with/without M-CSF inhibition will be done to determine donor survival, proliferation, clonal expansion, phenotype and function. Cell population dynamics will be tracked temporally by single-cell RNA-seq using Monocle. In aim 3, mouse or human HSPCs will be cultured with/without GM-CSF and surfactant or DPPC and gene regulatory dynamics will be measured by single-cell RNA-seq using Monocle to determine differentiation `trajectories'. DNA elements regulating cell population dynamics will be identified by measuring chromatin accessibility by sci-ATAC-seq and using Cicero to link DNA regulatory elements to the target genes they regulate. The proposed research is innovative because it supports the development of a new therapy for hPAP based on restoring AM function rather than physically removing surfactant (the current approach), and because it uses novel methods, software and statistical tools to identify elements directing AM specification as `upstream regulators' or `downstream' targets, which has not been possible with methods available previously. The proposed research is significant because it will identify the alveolar determinants and transcriptional mechanism(s) regulating AM specification during PMT, support the development of PMT as an effective, disease-specific cell therapy for children with hPAP, and provide a foundation for developing novel macrophage-based therapies for other lung diseases.

抽象的 在理解 GM-CSF 如何指导肺部感染后巨噬细胞的规范方面存在显着差距。 巨噬细胞移植（PMT）。这是一个重要的问题，因为如果没有这些关键信息， 临床前数据不足以证明 PMT 的安全性足以获得批准在人体中测试 PMT。 长期目标是开发 PMT 作为遗传性肺泡蛋白沉积症患者的治疗方法 （hPAP）旨在恢复肺泡巨噬细胞（AM）功能。这里的目标是确定机制 在 PMT 期间指导 AM 规范。中心假设是 AM 规格主要由 受肺泡微环境中的因素影响，包括 1) GM-CSF，它使 AM 特异性 DNA 调节 可访问的元素； 2) 表面活性剂磷脂衍生的脂肪酸，可激活 PPARγ 新结合 可访问的 DNA 并将 AM 从 LXR 驱动的规范转换为 PPARγ 驱动的规范（和胆固醇代谢）。 我们的理由是，通过证明 GM-CSF 指导巨噬细胞采用正常的 AM 转录谱 单细胞分辨率（“批量”研究未能证明）对于确定 PMT 的安全性至关重要。指导者 我们强有力的初步数据，将通过追求三个具体目标来测试这一假设，以确定 1) 供体细胞可塑性与治疗效果之间的关系； 2) 巨噬细胞的时间动态 PMT 后的植入、规格和命运； 3) 管理 AM 规范的顺式监管架构。在 目标1，通过PMT将不同发育阶段的骨髓细胞给予Csf2raKO小鼠，以阻止 挖掘能够赋予治疗效果的细胞的可塑性。目标2、正常造血功能的PMT 具有/不具有 M-CSF 抑制作用的 Csf2raKO（或正常）小鼠的干细胞/祖细胞 (HSPC) 供体将被用于 确定供体的存活、增殖、克隆扩增、表型和功能。细胞群动态将 使用 Monocle 通过单细胞 RNA-seq 进行时间跟踪。在目标 3 中，小鼠或人类 HSPC 将被 使用/不使用 GM-CSF 和表面活性剂或 DPPC 进行培养，并且基因调控动态将通过以下方式测量 使用 Monocle 进行单细胞 RNA-seq 来确定分化“轨迹”。调节细胞的DNA元件 将通过 sci-ATAC-seq 并使用 Cicero 测量染色质可及性来确定种群动态 将 DNA 调控元件与其调控的靶基因连接起来。所提出的研究具有创新性 因为它支持基于恢复 AM 功能而不是基于恢复 AM 功能的 hPAP 新疗法的开发 物理去除表面活性剂（当前的方法），并且因为它使用新颖的方法、软件和 统计工具，用于将指导增材制造规范的要素识别为“上游监管机构”或“下游”目标， 这是以前可用的方法不可能实现的。拟议的研究意义重大，因为 它将识别调节 AM 规范的肺泡决定因素和转录机制 PMT，支持将 PMT 开发为针对 hPAP 儿童的有效的疾病特异性细胞疗法， 并为开发针对其他肺部疾病的新型巨噬细胞疗法奠定了基础。

项目成果

Inferring Causal Gene Regulatory Networks from Coupled Single-Cell Expression Dynamics Using Scribe

• DOI: 10.1016/j.cels.2020.02.003
• 发表时间: 2020-03-25
• 期刊: CELL SYSTEMS
• 影响因子: 9.3
• 作者: Qiu, Xiaojie;Rahimzamani, Arman;Kannan, Sreeram
• 通讯作者: Kannan, Sreeram

Accelerating Scientific Advancement for Pediatric Rare Lung Disease Research. Report from a National Institutes of Health-NHLBI Workshop, September 3 and 4, 2015.

加速儿科罕见肺部疾病研究的科学进步。

• DOI: 10.1513/annalsats.201605-402ot
• 发表时间: 2016
• 期刊: Annals of the American Thoracic Society
• 影响因子: 8.3
• 作者: Young,LisaR;Trapnell,BruceC;Mandl,KennethD;Swarr,DanielT;Wambach,JenniferA;Blaisdell,CarolJ
• 通讯作者: Blaisdell,CarolJ

Mavrilimumab in patients with severe COVID-19 pneumonia and systemic hyperinflammation (MASH-COVID): an investigator initiated, multicentre, double-blind, randomised, placebo-controlled trial.

• DOI: 10.1016/s2665-9913(21)00070-9
• 发表时间: 2021-06
• 期刊: The Lancet. Rheumatology
• 作者: Cremer PC;Abbate A;Hudock K;McWilliams C;Mehta J;Chang SY;Sheng CC;Van Tassell B;Bonaventura A;Vecchié A;Carey B;Wang Q;Wolski KE;Rajendram P;Duggal A;Wang TS;Paolini JF;Trapnell BC;MASH-COVID study group
• 通讯作者: MASH-COVID study group