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.

项目摘要/摘要 克隆造血（CH）是一种常见现象，定义为在 造血干细胞（HSPC）及其在没有明显血液学的情况下膨胀 疾病。据信，CH突变的成熟，髓样细胞会产生炎症微环境 促进突变HSPC的健身优势。这些反过来将以更高的速度扩展并区分 进入更高数量的髓样细胞，从而在炎症之间建立正反馈回路 信号传导和克隆膨胀。但是，CH突变的HSPC是否也可以触发细胞自主炎症 信号为自己提供选择性克隆优势的信号仍然未知。 CH增加了 血液学恶性肿瘤，心血管疾病和实体瘤死亡率。由于这些逆境 结局和CH的高度流行率在古老的情况下，需要开发新的疗法。 针对突变的HSPC中的炎症 - 避免破坏一般免疫反应 - 可能 成为一种潜在的有效策略。我的研究前研究（AIM 1）旨在确定 CH-MUTANT HSPC并评估其作为恢复寡克隆造血的治疗靶标的潜力。到 在CH中找到新颖的细胞自主炎症途径，我设计了一个SGRNA库来靶向 炎症相关基因。使用此库进行高通量CRISPR/CAS9筛选，我已经确定了 Ch-突变鼠HSPC的一般和基因型特异性炎症依赖性。在此提案中， 特定目标1.1试图验证负面选择的命中，证明当存在时，命中基因 会议对Ch-Mutant HSPC与野生型的选择性优势，并描述特定角色 克隆扩张中的凭据热门单曲。在特定的目标1.2中，我将使用针对的小分子抑制剂 候选基因 - 在体内和小鼠中 - 以识别基因表达和细胞因子谱的变化 跨越造血细胞子集。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标抑制方面的效率。我的博士后研究（AIM 2）将重点 关于炎症在衰老和CH的联系中的作用，通过发现转录和表观遗传学 与年龄相关的炎症的机制促进了CH和潜在的恶性转化为 白血病。总体而言，这两个项目将使用人类样本来验证鼠标发现，将导致 开发针对创新的新疗法，以阻止或恢复CH并减轻其临床后遗症。我， 申请人将在纪念斯隆·克特林癌症的罗斯·莱文博士实验室中提出这一建议 中心（MSK）是世界领域的癌症治疗和研究领域的领先机构之一。 MSK丰富的环境 并在Gerstner Sloan Kettering研究生院的支持下，大量资源， 确保成功完成拟议的研究和职业发展计划。