Characterizing the pathogenesis and targeted therapeutics of Wilms' Tumor 1 (WT1) mutations in AML

描述 AML 中 Wilms 肿瘤 1 (WT1) 突变的发病机制和靶向治疗

• 批准号: 10459634
• 负责人: Haijiao Zhang
• 金额: $ 24.9万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10459634
• 关键词: Acute; Advisory Committees; Apoptosis; BCL2 gene; Bioinformatics; Biological Assay; Bone Marrow; Bone Marrow Cells; Bone Marrow Stem Cell; C-terminal; CEBPA gene; CRISPR screen; CSF3R gene; Candidate Disease Gene; Caring; Cell Line; Cell Lineage; Cell Proliferation; Cells; Chemoresistance; Clinic; Clustered Regularly Interspaced Short Palindromic Repeats; Colony-Forming Units Assay; Combined Modality Therapy; Cytarabine; Cytogenetics; Data; Development; Dimerization; Disease; Disease Progression; Dose; Drug resistance; Drug usage; Engineering; Exons; FLT3 gene; FLT3 inhibitor; Faculty; Frameshift Mutation; Genes; Goals; Growth; Hematologic Neoplasms; Hematopoietic stem cells; Human; Inbred BALB C Mice; Leukemic Cell; Manuscripts; Mediating; Medical; Medical Students; Mentors; Mentorship; Modality; Modeling; Molecular; Monitor; Mus; Mutation; Myeloid Leukemia; N-terminal; Oncogenes; Oncogenic; Outcome; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phenotype; Positioning Attribute; Preparation; Prognosis; Publishing; RNA; Recurrent disease; Refractory; Regulatory Element; Research; Research Personnel; Resistance; Sampling; Series; Sum; Therapeutic; Training; Transcriptional Regulation; Translations; Transplantation; Tyrosine Kinase Domain; WT1 gene; Xenograft procedure; Zinc Fingers; career; chemotherapy; clinical predictors; clinically relevant; combinatorial; deep sequencing; disease phenotype; drug candidate; drug sensitivity; functional genomics; gene therapy; genome editing; genome-wide; in vivo; inhibitor; leukemia; leukemia treatment; leukemic stem cell; loss of function; mouse model; mutant; myeloid leukemia cell; novel therapeutics; overexpression; progenitor; receptor; resistance mechanism; resistance mutation; screening; standard care; stem cell biology; targeted cancer therapy; targeted treatment; therapeutic target; transcription factor; transcriptome sequencing; translational medicine; variant of unknown significance

PROJECT SUMMARY/ABSTRACT Dr. Haijiao Zhang has been training as a medical student for 7 years with a focus on hematological malignancies and as a researcher for 6 years in the field of characterizing oncogene and drug resistance mechanisms. Her research has led to 8 published manuscripts and 4 other manuscripts in submission /preparation. The theme of Dr. Zhang's research involves systematic characterization of leukemia oncogenes and understanding leukemia drug resistance mechanisms. She has characterized 4 distinct mechanisms of CSF3R mutations and diverse mechanisms associated with drug resistance to the FLT3 inhibitor crenolanib. The WT1 (Wilms' tumor 1) gene encodes a transcription factor with C-terminal zinc-finger domains, whereas the N-terminal part contains domains that mediate receptor dimerization, RNA recognition, and transcriptional regulation. WT1 mutations are present in approximately 10–15% of AML and are associated with chemotherapy resistance and disease relapse. WT1 mutations are frameshift mutations or substitutions across all structural domains, predominantly in exons 7 and 9. The pathogenesis mediated by loss-of-function of WT1 zinc finger domain mutations has been studied; however very little is known about the oncogenic potential, drug sensitivity, and the structural basis of mutations residing in other domains of WT1. In the current proposal, Dr. Zhang will evaluate the functional consequences of these mutations and characterize the mechanisms associated with the growth/survival advantage and a higher propensity for chemotherapy resistance of WT1 mutations. She also aims to identify therapeutics with enhanced efficacy in the WT1 mutant setting. Using ex vivo inhibitor screening on primary leukemia patient samples, she has determined that inhibitors of BCL2 and RAF/ERK pathways demonstrated higher efficacy against WT1 mutant samples compared with controls. She will validate these findings using NRG mouse model. Finally, she will investigate the phenotypic, mechanistic, and therapeutics targeting WT1 combinatorial mutation. Dr. Zhang's long-term career goals include the establishment of an independent research lab focusing on systematic characterization of oncogenic mutations and chemotherapy resistance to better predict the clinical relevance of mutations and identify novel therapeutics to circumvent chemotherapy resistance. During the mentored phase, she will continue to receive excellent mentorship from Dr. Jeffrey Tyner, an expert on functional genomics and targeted therapy. She will receive co-mentorship from Dr. Ravi Majeti, an expert in leukemia stem cell biology, CRISPR editing, and high-fidelity xenotransplantation mouse models, which she will use in the current project. The proposed research will also be enhanced by guidance from Dr. Brian Druker, a pioneer in translational medicine and targeted therapies for cancer; and Dr. Shannon McWeeney who will provide guidance in bioinformatics analyses. Additionally, Dr. Tyner, Dr. Majeti, and the advisory team will assist her in navigating the transition to an independent faculty position.

项目概要/摘要 张海蛟医生已经接受了7年的医学生培训，专注于血液学 恶性肿瘤研究人员，在癌基因和耐药性特征研究领域工作了 6 年 她的研究已发表 8 篇手稿，并提交了 4 篇其他手稿。 /准备。张博士的研究主题涉及白血病癌基因的系统表征。 并了解白血病耐药机制。她描述了 4 种不同的机制。 CSF3R 突变和与 FLT3 抑制剂 crenolanib 耐药相关的多种机制。 WT1（肾母细胞瘤 1）基因编码具有 C 末端锌指结构域的转录因子，而 N 端部分包含介导受体二聚化、RNA 识别和转录的结构域 WT1 突变存在于大约 10-15% 的 AML 中，并且与 AML 相关。 化疗耐药和疾病复发是跨移码突变或替代。 所有结构域，主要位于外显子 7 和 9。WT1 功能丧失介导的发病机制 锌指结构域突变已被研究；然而，人们对其致癌潜力知之甚少， 药物敏感性，以及 WT1 其他域中突变的结构基础。 张博士将评估这些突变的功能后果并描述其机制 与 WT1 的生长/生存优势和较高的化疗耐药倾向相关 她还致力于使用 ex 来确定在 WT1 突变环境中具有增强功效的治疗方法。 她对原发性白血病患者样本进行了体内抑制剂筛选，确定了 BCL2 和 与对照相比，RAF/ERK 途径对 WT1 突变样品表现出更高的功效。 最后，她将使用 NRG 小鼠模型验证这些发现。 以及针对 WT1 组合突变的治疗方法。 张博士的长期职业目标包括建立一个独立的研究实验室，专注于 致癌突变和化疗耐药性的系统表征，以更好地预测临床 突变的相关性并确定新的治疗方法来规避化疗耐药性。 在指导阶段，她将继续得到专家Jeffrey Tyner博士的出色指导。 她将接受功能基因组学和靶向治疗专家 Ravi Majeti 博士的共同指导。 白血病干细胞生物学、CRISPR 编辑和高保真异种移植小鼠模型，她 将在当前项目中使用，布莱恩德鲁克博士的指导也将加强拟议的研究， 转化医学和癌症靶向治疗领域的先驱；以及 Shannon McWeeney 博士，他将 此外，Tyner 博士、Majeti 博士和顾问团队将提供生物信息学分析方面的指导。 协助她过渡到独立教职职位。