Defining malignant hematopoiesis via single-cell multi-omics

通过单细胞多组学定义恶性造血

基本信息

批准号：
10018257
负责人：
Seung Ha Nam
金额：
$ 42.38万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
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：
2020-09-15 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10018257
关键词：
Admixture Advisory Committees Affect Anatomy Appointment Biochemical Biological Assay Biomedical Research Blood Blood Cells Bone Marrow CD34 gene Cell Cycle Cell physiology Cell surface Cells Chromatin Clinical Communities Complex Core Facility DNA DNA Sequence Alteration Data Dependence Development Diagnostic Disease Environment Environmental Risk Factor Epigenetic Process Exhibits Faculty Funding Gene Mutation Genes Genetic Genetic Transcription Genome Genomic medicine Genomics Genotype Hematological Disease Hematopathology Hematopoiesis Hematopoietic Hematopoietic Neoplasms Hematopoietic stem cells Human Immune Immunologic Factors Individual Inorganic Chemistry Institutes Intrinsic factor Investigation Laboratories Lead Link MPL gene Malignant - descriptor Medicine Memorial Sloan-Kettering Cancer Center Mentors Modernization Molecular Molecular Genetics Morphology Mutate Mutation Myelogenous Myeloproliferative disease NF-kappa B Nature Neoplastic Processes New York Outcome Output Pathology Pathway interactions Patients Peer Review Physicians Population Process Protein Secretion Proteins Publications Research Research Personnel Research Training Resources Sampling Scientist Signal Transduction Somatic Mutation Specimen Supervision T-Lymphocyte Technology Testing Therapeutic Intervention Tissues Training Translational Research Trees United States National Institutes of Health Ursidae Family biobank calreticulin cell type clinical phenotype college cytokine disease phenotype doctoral student driver mutation epigenome epigenomics hematopoietic differentiation immunomodulatory therapies improved innovation insight medical schools medical specialties molecular array mouse model multidisciplinary multiple omics mutant neoplastic neoplastic cell next generation novel personalized medicine progenitor programs response single cell sequencing stem cells targeted treatment tool transcription factor transcriptome transcriptomics undergraduate student

项目摘要

PROJECT SUMMARY / ABSTRACT Clonal blood differentiation through the acquisition of somatic mutations result in abnormal accumulation of blood components and clinically manifest as myeloid disorders. The study of how these somatic mutations perturb the differentiation trajectories in human hematopoiesis is often challenged by the admixture of normal hematopoietic cells with the neoplastic cells that cannot be distinguished by cell surface markers. To overcome this limitation, we developed a novel single-cell multi-omics Genotyping of Transcriptomes (GoT) platform that directly links somatic genotypes with transcriptomes of thousands of single cells. Thus, GoT enabled the comparison of mutant and wildtype cells within the same sample in the context of progenitor identities, thereby turning the co-mingling of mutant and wildtype hematopoiesis from a limitation to an advantage. As proof of principle, GoT was applied to CD34+ progenitor cells from patients with calreticulin- mutated myeloproliferative neoplasms (MPN), revealing key pathways that were aberrantly activated in the mutant cells, such as a robust unfolded protein response in the megakaryocytic progenitors, on the one way, and NF-KB pathway in stem cell-enriched populations, on the other. Overall, GoT revealed that the transcriptional impact of calreticulin mutations is highly variable as a function of progenitor identity – which bears significant implications for therapy by enabling the discovery of targetable pathways specific to the earliest stem cells. Thus, to demonstrate the cell identity-dependency across other key driver mutations, as a fundamental concept in myeloid disorders, I will apply GoT to thrombopoietin receptor-mutated progenitor cells and to clonally-diverse cells from MPN samples (Aim 1). Next, in order to define cell extrinsic determinants of somatic mutation impact, I will determine the immune niche interactions with calreticulin-mutant and wildtype progenitor cells, as well as the impact of immunomodulatory therapy on these interactions (Aim 2). Finally, I will test the hypothesis that the cell’s epigenome precedes the cell identity-dependency of somatic mutation effects, by developing and applying a novel single-cell platform that integrates somatic genotyping with chromatin accessibility states of progenitor cells (Aim 3). Thus, I will define the genetic, epigenetic, transcriptional and environmental factors that culminate in the clinical output of somatic mutations in human hematopoiesis. These studies will, therefore, unveil not only fundamental concepts in clonal hematopoietic differentiation but also specific targets for therapeutic intervention.

项目摘要 /摘要克隆血液通过获得躯体突变的分化导致异常积累血液成分和临床表现为髓样疾病。这些体细胞突变的研究扰动人类造血的分化轨迹通常受到正常混合的挑战用细胞表面标记区分的肿瘤细胞的造血细胞。到克服了这一局限性，我们开发了一种新型的单细胞多态基因分型的转录组（GOT）直接将体细胞基因型与数千个单细胞的转录组联系起来的平台。那就得到了在祖细胞的背景下，可以比较同一样品中突变体和野生型细胞身份，从而将突变体和野生型造血的共汇合从限制到一个优势。作为原理证明，GOT应用于钙网蛋白患者的CD34+祖细胞突变的骨髓增生性肿瘤（MPN），揭示了在该关键途径中被异常激活的关键途径突变细胞，例如在巨核细胞祖细胞中的强大展开的蛋白质反应，另一方面，富含干细胞的人群中的NF-KB途径。总的来说，发现钙网蛋白突变的转录影响是祖先身份的函数高度可变的 - 通过发现特定的目标途径，对治疗产生重大影响最早的干细胞。这是为了证明其他关键驱动突变的细胞身份依赖性，如在髓样疾病中的基本概念，我将申请到血小板蛋白受体杂种祖细胞细胞和来自MPN样品的克隆多样性细胞（AIM 1）。接下来，为了定义细胞外部躯体突变影响的决定因素，我将确定与钙网蛋白突变剂的免疫利基相互作用和野生型提供商细胞，以及免疫调节疗法对这些相互作用的影响（AIM 2）。最后，我将检验以下假设：细胞的表观组在细胞身份依赖性之前通过开发和应用整合体细胞的新型单细胞平台，体细胞突变效应祖细胞的染色质可及性状态的基因分型（AIM 3）。那我将定义遗传，表观遗传，转录和环境因素，在体细胞突变的临床输出中达到顶峰在人类造血中。因此，这些研究将不仅揭示克隆的基本概念造血分化，也是治疗干预的特定靶标。