Pathogenesis of ETV6-Related Acute Lymphoblastic Leukemia

ETV6相关急性淋巴细胞白血病的发病机制

• 批准号: 10837399
• 负责人: KIM Erika NICHOLS
• 金额: $ 42.04万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10837399
• 关键词: ATAC-seq; Acute Lymphocytic Leukemia; Affect; B cell differentiation; B-Cell Acute Lymphoblastic Leukemia; B-Cell Development; B-Lymphocytes; Binding; Biological Assay; Biological Models; Biology; Blood Cells; Cancer Biology; Cancer Etiology; Cancer-Predisposing Gene; Cell Count; Cell Differentiation process; Cell Line; Cells; ChIP-seq; Child; Childhood; Childhood Acute Lymphocytic Leukemia; Chromatin; Classification; Clinical; Collaborations; Cytoplasm; DNA Binding; Data; Databases; Defect; Development; Dysplastic Megakaryocyte; ETV6 gene; Epigenetic Process; Event; Exhibits; Family; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic Variation; Germ-Line Mutation; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Heterozygote; Human; Impairment; In Vitro; Individual; Inherited; Lesion; Literature; Malignant - descriptor; Malignant Childhood Neoplasm; Malignant Neoplasms; Modeling; Mus; Mutant Strains Mice; Mutate; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Phenotype; Physicians; Physiology; Population; Positioning Attribute; Predisposition; Proteins; Public Health; Recurrence; Reporting; Saint Jude Children' s Research Hospital; Sampling; Scientist; Somatic Mutation; Syndrome; Testing; Thrombocytopenia; Transcription Repressor; Variant; autosome; cancer predisposition; cell growth; cellular development; childhood cancer mortality; cohort; derepression; disease classification; disease prognosis; genome sequencing; improved; in vitro Assay; in vivo; induced pluripotent stem cell; insight; leukemia; leukemogenesis; mouse model; novel; personalized management; prevent; progenitor; programs; stem cells; transcription factor; transcriptome; transcriptome sequencing; transmission process; treatment optimization; tumorigenesis; whole genome

PROJECT SUMMARY As the initiating genetic events in tumorigenesis, germline variants in cancer predisposing genes perturb cell growth and differentiation and set the stage for malignant transformation. Accordingly, the study of cancer predisposing genes and their associated hereditary syndromes provides critical insights into normal physiology and cancer biology. By investigating families with autosomal dominant transmission of thrombocytopenia and Bacute lymphoblastic leukemia (B-ALL), we and others identified pathogenic germline variants affecting ETV6, the gene encoding the ETS variant 6 transcriptional repressor. Subsequently, we sequenced germline samples from 4,405 children with B-ALL and detected similar variants in 0.5% of patients. Further association studies revealed a significant 22.94-fold enrichment of pathogenic ETV6 variants in this ALL cohort compared to 134,187 non-ALL controls in gnomAD (P= 2.2 × 10-16). These data firmly support our overall premise that pathogenic germline ETV6 variants predispose to childhood ALL. To better understand how germline ETV6 variants promote leukemogenesis, we used in vitro assays to interrogate their effects on the functions of the encoded ETV6 protein. Notably, each of the pathogenic ETV6 variants examined significantly reduced ETV6 transcription repressor activity, impaired ETV6 DNA binding capacity and mis-localized ETV6 to the cytoplasm. In parallel, we generated a novel mouse model harboring a recurrent B-ALL-associated Etv6 variant (Etv6R355X). Preliminary studies of Etv6R355X/+ mice reveal significant perturbations in early B cell development, as well as hematopoietic stem and progenitor cell (HSPC) number and function. Finally, we generated isogenic induced pluripotent stem cell (iPSC) lines harboring pathogenic ETV6 variants, which generate dysplastic megakaryocytes, similar to ETV6 variant positive patients. Based on these results, we hypothesize that ETV6 variants predispose to ALL by perturbing key transcriptional programs that impair hematopoietic development. In this proposal, we will make use of our unique model systems to rigorously test this hypothesis. In Aim 1, we will characterize the hematopoietic compartments of humans and mice that do or do not harbor pathogenic germline ETV6 variants. We will examine whether WT and ETV6 variant-positive iPSCs or mouse hematopoietic progenitors properly differentiate along various lineages. To establish how germline ETV6 variants influence gene expression in developing hematopoietic progenitors, in Aim 2 we will use RNA-sequencing and ATAC-sequencing to explore transcriptional landscapes and identify putative ETV6 target genes in mouse and iPSC-derived B progenitors and HSPC harboring WT or variant ETV6. Finally, in Aim 3 we will perform comprehensive whole genome and RNA-sequencing of B-ALL samples to elucidate the somatic genetic lesions that function in concert with germline ETV6 variants to drive B-leukemogenesis. We will use in vitro and in vivo approaches to assess the leukemiapromoting effects of these second hits. This project will provide new insights into the influence of germline genetic variation on hematopoiesis and development of B-ALL, the most common childhood cancer.

项目摘要 作为肿瘤发生中的遗传事件，癌症易感基因的种系变异 生长和分化，为恶性转化奠定了基础。彼此之间的研究 诱发基因及其相关的雌性综合症为正常生理提供了关键的见解 和癌症生物学。通过调查血小板减少症和B急性淋巴细胞白血病（B-All）的常染色体显性传播家族，我们和其他人鉴定 编码ETS变体6转录表示的基因。随后，我们对种系样品进行了测序 来自4,405名B-all儿童，在0.5％的患者中发现了类似的变体。进一步的关联研究 在这一切队列中，病原ETV6变体的富集显着22.94倍，而134,187 GNOMAD中的非控制器（p = 2.2×10-16）。这些数据首先支持我们的整体前提 种系ETV6变体易于童年。更好地了解种系ETV6变体 促进白血病发生，我们使用体外测定法来询问它们对编码功能的影响 ETV6蛋白。值得注意的是，每个致病性ETV6变体都显着降低了ETV6转录 抑制剂活性，ETV6 DNA结合能力受损，而将ETV6置于细胞质中。并行，我们 产生了具有复发性B al-B-All相关ETV6变体（ETV6R355X）的新型小鼠模型。初步的 ETV6R355X/+小鼠的研究显示早期B细胞发育以及造血性扰动明显 茎和祖细胞（HSPC）编号和功能。最后，我们产生了同基因诱导的多能茎 具有致病性ETV6变体的细胞（IPSC）线，产生异型质巨核细胞，类似于 ETV6变异阳性患者。基于这些结果，我们假设ETV6变体易于 所有这些都是干扰损害造血发展的关键转录程序。在此提案中， 我们将利用我们独特的模型系统来严格检验这一假设。在AIM 1中，我们将描述 人类和小鼠的造血室，它们具有致病性生殖线ETV6 变体。我们将检查WT和ETV6变体阳性IPSC还是小鼠造血祖细胞 沿各种谱系正确区分。建立种系ETV6变体如何影响基因表达 在开发造血祖细胞时，在AIM 2中，我们将使用RNA测序和ATAC进行探索 转录景观并识别小鼠和IPSC衍生的B祖细胞中的假定ETV6靶基因 HSPC携带WT或变体ETV6。最后，在AIM 3中，我们将执行全面的整个基因组和 B-ALL样品的RNA测序以阐明与种系一起起作用的体细胞遗传病变 ETV6变体驱动B-美元素发生。我们将使用体外和体内方法来评估这些第二次命中的白血病。该项目将提供有关种系影响的新见解 造血症的遗传变异和B-All的发展，B-All是最常见的儿童癌症。

项目成果

Pathogenic Variants in Adult-Onset Cancer Predisposition Genes in Pediatric Cancer: Prevalence and Impact on Tumor Molecular Features and Clinical Management.

儿童癌症中成人发病的癌症易感基因的致病变异：患病率及其对肿瘤分子特征和临床治疗的影响。

• DOI: 10.1158/1078-0432.ccr-22-2482
• 发表时间: 2023
• 期刊: Clinical cancer research : an official journal of the American Association for Cancer Research
• 作者: McGee,RoseB;Oak,Ninad;Harrison,Lynn;Xu,Ke;Nuccio,Regina;Blake,AliseK;Mostafavi,Roya;Lewis,Sara;Taylor,LeslieM;Kubal,Manish;Ouma,Annastasia;Hines-Dowell,StacyJ;Cheng,Cheng;Furtado,LarissaV;Nichols,KimE
• 通讯作者: Nichols,KimE

Immune stress suppresses innate immune signaling in preleukemic precursor B-cells to provoke leukemia in predisposed mice.

• DOI: 10.1038/s41467-023-40961-z
• 发表时间: 2023-08-24
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Isidro-Hernandez, Marta;Casado-Garcia, Ana;Oak, Ninad;Aleman-Arteaga, Silvia;Ruiz-Corzo, Belen;Martinez-Cano, Jorge;Mayado, Andrea;Sanchez, Elena G.;Blanco, Oscar;Gaspar, Ma Luisa;Orfao, Alberto;Alonso-Lopez, Diego;De Las Rivas, Javier;Riesco, Susana;Prieto-Matos, Pablo;Gonzalez-Murillo, Africa;Criado, Francisco Javier Garcia;Cenador, Maria Begona Garcia;Ramirez-Orellana, Manuel;de Andres, Belen;Vicente-Duenas, Carolina;Cobaleda, Cesar;Nichols, Kim E.;Sanchez-Garcia, Isidro
• 通讯作者: Sanchez-Garcia, Isidro