Identifying Inflammatory Mediators of Clonal Hematopoiesis

识别克隆造血的炎症介质

• 批准号: 10743540
• 负责人: Ines Fernandez Maestre
• 金额: $ 4.92万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10743540
• 关键词: Acute Myelocytic Leukemia; Age; Aging; Alleles; Anti-Inflammatory Agents; Attenuated; Automobile Driving; Bone Marrow; CRISPR screen; Candidate Disease Gene; Cardiovascular Diseases; Cells; ChIP-seq; Chemicals; Chronic; Clinical; Clonal Expansion; Coculture Techniques; Complex; Credentialing; Custom; Dependence; Development; Development Plans; Doctor of Philosophy; Elderly; Endothelial Cells; Environment; Epigenetic Process; Feedback; Foundations; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Genotype; Goals; Hematologic Neoplasms; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; High Prevalence; Human; Immune response; Incidence; Inflammaging; Inflammation; Inflammation Mediators; Inflammatory; Institution; Interleukin-1; Interleukin-6; Knock-out; Knowledge; Laboratories; Libraries; Ligands; Link; Malignant - descriptor; Mediator; Memorial Sloan-Kettering Cancer Center; Mus; Mutation; Myeloid Cells; Non-Hematologic Malignancy; Pathology; Pathway interactions; Patients; Postdoctoral Fellow; Research; Resources; Risk; Role; Sampling; Signal Pathway; Signal Transduction; Solid Neoplasm; Somatic Mutation; System; TNF gene; Testing; Training; Work; acute myeloid leukemia cell; adverse outcome; anticancer research; cancer therapy; career development; cell type; cohort; cytokine; design; epigenomics; fitness; graduate school; in vivo engraftment; inflammatory milieu; inhibitor; leukemia; leukemic transformation; mortality; mouse model; multiple omics; mutant; new therapeutic target; novel; novel therapeutics; pre-doctoral; precision medicine; pressure; prevent; rational design; research and development; self-renewal; small molecule; small molecule inhibitor; therapeutic target; therapeutically effective; transcriptomics

PROJECT SUMMARY/ABSTRACT Clonal hematopoiesis (CH) is a common phenomenon defined as the presence of somatic mutations in hematopoietic stem and progenitor cells (HSPCs) and their expansion in the absence of overt hematological disease. CH-mutant mature, myeloid cells are believed to generate an inflammatory microenvironment promoting the fitness advantage of mutant HSPCs. These, in turn, would expand at higher rates and differentiate into more elevated numbers of myeloid cells, thereby establishing a positive feedback loop between inflammatory signaling and clonal expansion. Yet, whether CH-mutant HSPCs can also trigger cell-autonomous inflammatory signaling to provide a selective clonal advantage for themselves remains unknown. CH increases the risk of hematological malignancy, cardiovascular disease, and mortality from solid tumors. Due to these adverse outcomes and the high prevalence of CH in the elderly, there is an unmet need to develop novel therapies. Targeting inflammation specifically in mutant HSPCs —to avoid disruption of general immune responses— may be a potentially effective strategy. My predoctoral research (Aim 1) aims to identify inflammatory mediators of CH-mutant HSPCs and evaluate their potential as therapeutic targets to restore oligoclonal hematopoiesis. To find novel, cell-autonomous inflammatory pathways in CH, I have designed an sgRNA library to target inflammation-associated genes. Using this library for high-throughput CRISPR/Cas9 screening, I have identified both general and genotype-specific inflammatory dependencies of CH-mutant murine HSPCs. In this proposal, Specific Aim 1.1 seeks to validate the negative selection hits, demonstrate that, when present, the hit genes confer a selective advantage to CH-mutant HSPCs versus wild-type counterparts, and delineate the specific role of credential top hits in clonal expansion. In Specific Aim 1.2, I will use small molecule inhibitors targeting the candidate genes —both ex vivo and in mice— to identify gene expression and cytokine profile changes spanning the hematopoietic cell subsets. I will then assess differences between genetic and chemical approaches concerning their efficiency in achieving adequate target inhibition. My postdoctoral research (Aim 2) will focus on the role of inflammation at the nexus of aging and CH by uncovering the transcriptional and epigenetic mechanisms by which age-related inflammation promotes CH and potential malignant transformation to leukemia. Overall, these two projects, which will use human samples to validate the mouse findings, will lead to developing new therapies targeting inflammation to halt or revert CH and mitigate its clinical sequelae. I, the applicant, will conduct this proposal in the laboratory of Dr. Ross Levine at Memorial Sloan Kettering Cancer Center (MSK), one of the world's leading institutions in cancer treatment and research. MSK's rich environment and abundant resources in conjunction with the support of the Gerstner Sloan Kettering Graduate School, guarantee the successful completion of the proposed research and career development plans.

项目总结/摘要 克隆性造血（CH）是一种常见的现象，定义为在造血干细胞中存在体细胞突变。 造血干细胞和祖细胞（HSPC）及其在缺乏明显血液学特征的情况下的扩增 疾病CH-突变的成熟骨髓细胞被认为产生炎症微环境 促进突变体HSPC的适应性优势。反过来，它们将以更高的速度扩张， 转化为更多数量的髓样细胞，从而在炎症和炎症之间建立正反馈回路。 信号传导和克隆扩增。然而，CH-突变的HSPC是否也可以触发细胞自主炎症反应， 为自身提供选择性克隆优势的信号传导仍然未知。CH会增加 血液恶性肿瘤、心血管疾病和实体瘤死亡率。由于这些不利的 然而，由于CH在老年人中的高患病率和预后，开发新疗法的需求尚未得到满足。 特异性靶向突变HSPC中的炎症-以避免破坏一般免疫应答-可以 这是一个潜在的有效策略。我的博士前研究（目标1）旨在确定炎症介质， CH-突变HSPC和评估其作为治疗靶点恢复寡克隆造血的潜力。到 为了在CH中发现新的细胞自主炎症通路，我设计了一个sgRNA文库， 炎症相关基因使用这个文库进行高通量CRISPR/Cas9筛选，我已经鉴定出 CH突变型鼠HSPC的一般和基因型特异性炎症依赖性。在这项提案中， 具体目标1.1旨在验证阴性选择命中，证明当存在时，命中基因 赋予CH-突变HSPCs相对于野生型对应物的选择性优势，并描绘了其特定作用， 在克隆扩张中的最佳点击率。在具体目标1.2中，我将使用小分子抑制剂靶向 候选基因-体外和小鼠-以鉴定基因表达和细胞因子谱变化 跨越造血细胞亚群。然后，我将评估遗传和化学方法之间的差异 关于它们在实现足够的靶抑制方面的效率。我的博士后研究（目标2）将集中在 通过揭示转录和表观遗传的作用， 年龄相关性炎症促进CH和潜在恶性转化的机制， 白血病总的来说，这两个项目将使用人类样本来验证小鼠的发现， 开发针对炎症的新疗法，以停止或逆转CH并减轻其临床后遗症。一世 申请人将在纪念斯隆-凯特琳癌症中心的Ross Levine博士的实验室进行本提案 中心（MSK），世界领先的癌症治疗和研究机构之一。MSK丰富的环境 和丰富的资源，在郭士纳斯隆凯特琳研究生院的支持下， 保证顺利完成拟议的研究和职业发展计划。