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: 10673864
• 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/10673864
• 关键词: Acceleration; Acute; Advisory Committees; Apoptosis; BCL2 gene; Bioinformatics; Biological Assay; Bone Marrow; Bone Marrow Cells; Bone Marrow Stem Cell; C-terminal; CEBPA gene; CRISPR screen; 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; Granulocyte Colony-Stimulating Factor Receptors; Growth; Hematologic Neoplasms; Hematopoietic; Human; Inbred BALB C Mice; Inhibition of Cell Proliferation; 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; Proliferating; Publishing; RNA; Recurrent disease; Refractory; Regulatory Element; Research; Research Personnel; Resistance; Sampling; Series; Therapeutic; Training; Transcriptional Regulation; Transplantation; Tyrosine Kinase Domain; WT1 gene; Xenograft procedure; Zinc Fingers; candidate validation; 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; stem cell biology; targeted cancer therapy; targeted treatment; therapeutic target; therapy resistant; transcription factor; transcriptome sequencing; translational medicine; translational therapeutics; 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.

项目总结/摘要 博士张海娇作为一名医学生接受了7年的培训，重点是血液学 恶性肿瘤，并在表征癌基因和耐药性领域担任了6年的研究员 机制等她的研究已导致8个出版的手稿和4个其他手稿提交 /制备。张博士的研究主题涉及白血病癌基因的系统表征 了解白血病耐药机制。她描述了四种不同的机制， CSF 3R突变和与FLT 3抑制剂crenolanib耐药性相关的多种机制 WT 1（Wilms' tumor 1）基因编码具有C-末端锌指结构域的转录因子，而 N-末端部分含有介导受体二聚化、RNA识别和转录的结构域， 调控WT 1突变存在于大约10-15%的AML中，并且与 化疗耐药性和疾病复发。WT 1突变是跨基因组的移码突变或取代。 所有结构域，主要在外显子7和9中。WT 1功能丧失介导的发病机制 已经对锌指结构域突变进行了研究;然而对致癌潜力知之甚少， 药物敏感性，以及WT 1其他结构域突变的结构基础。在目前的提案中， 博士Zhang将评估这些突变的功能后果并描述其机制 与生长/生存优势和WT 1的化疗耐药倾向较高相关 突变。她还旨在确定在WT 1突变体环境中具有增强功效的疗法。使用离 在原发性白血病患者样品的体内抑制剂筛选中，她已经确定BCL 2和BCL 3的抑制剂可以用于治疗白血病。 与对照相比，RAF/ERK途径对WT 1突变样品表现出更高的功效。她 将使用NRG小鼠模型验证这些发现。最后，她将调查表型，机械， 和靶向WT 1组合突变的治疗剂。 博士张的长期职业目标包括建立一个独立的研究实验室， 系统表征致癌突变和化疗耐药性，以更好地预测临床 突变的相关性，并确定新的治疗方法，以规避化疗耐药性。 在指导阶段，她将继续接受专家杰弗里泰纳博士的出色指导 功能性基因组学和靶向治疗她将接受拉维·马盖蒂博士的共同指导， 白血病干细胞生物学、CRISPR编辑和高保真异种移植小鼠模型， 将用于当前项目。拟议中的研究也将得到布莱恩·德鲁克博士的指导， 转化医学和癌症靶向治疗的先驱;香农·麦克维尼博士将 为生物信息学分析提供指导。此外，泰纳博士、马盖蒂博士和顾问小组将 帮助她导航过渡到一个独立的教师职位。