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Functional and Pharmacologic Investigation of the NUP98 Fusion Oncoprotein Interactome

NUP98 融合癌蛋白相互作用组的功能和药理学研究

基本信息

批准号：
10724053
负责人：
Nicole Michmerhuizen
金额：
$ 14.1万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10724053
关键词：
Acute Myelocytic Leukemia Address Binding Bioinformatics Biological Assay Bone Marrow Transplantation C-terminal CRISPR screen CRISPR/Cas technology Cell Death Cell Nucleus Cell Survival Cells Childhood Acute Myeloid Leukemia Childhood Leukemia Chromatin Chromatin Structure Chromosomal Rearrangement Chromosomal translocation Colony-forming units Complex DNA Binding Data Data Set Developmental Gene Disadvantaged Disease Epigenetic Process Fusion Oncogene Proteins Gene Expression Gene Expression Profile Gene Expression Regulation Gene Fusion Genes Goals Guide RNA HOXA9 gene Hematopoietic Hematopoietic stem cells Histone Acetylation Immunoprecipitation In Vitro Investigation Lead Libraries Liquid substance Lysine MLL gene Malignant - descriptor Malignant Neoplasms Mass Spectrum Analysis Mediating Mediator Membrane Modeling Mus N-terminal Nuclear Pore Complex Proteins Oncogene Deregulation Oncogenic Organelles Outcome Patient-Focused Outcomes Pharmacology Phase Phenotype Property Proteins Reader Recurrence Research Research Proposals Role Saint Jude Children&apos s Research Hospital Scientist Testing Therapeutic Therapeutic Intervention Time Training Transcriptional Regulation Validation career cell growth cell transformation chimeric gene chromatin remodeling design drug efficacy effective therapy epigenomics experimental study exportin 1 protein fitness functional genomics high risk histone acetyltransferase homeodomain in vivo leukemia leukemogenesis member mouse model novel novel therapeutic intervention nucleocytoplasmic transport pharmacologic post-doctoral training pre-doctoral promoter quantitative imaging self-renewal small molecule small molecule inhibitor stem-like cell structural biology targeted treatment therapeutic target therapeutically effective therapy resistant

项目摘要

PROJECT SUMMARY Chromosomal translocations involving Nucleoporin 98 (NUP98) are observed in approximately 5% of pediatric acute myeloid leukemia (AML) and are associated with resistance to therapy and poor outcome, with approximately 35% 5 year overall survival. NUP98 rearrangements lead to expression of oncogenic chimeric gene fusions involving the intrinsically disordered, N-terminal region of NUP98 and the C-terminal region of one of over 30 identified partner genes. The partner genes commonly have domains with key functional properties, including homeodomain moieties (e.g. HOXA9) and roles in transcriptional regulation (e.g. NSD1, KDM5A). In complex with other machinery needed for gene regulation, NUP98 fusion oncoproteins (FOs) bind to the promoters of many developmental genes. This leads to changes in chromatin structure, increased expression of target genes, and aberrant hematopoietic self-renewal. Recent studies, including my own, have shown that the ability of NUP98 FOs to localize within the nucleus in membrane-less organelles, or “puncta” formed through liquid-liquid phase separation (LLPS), is necessary for transformation and deregulated gene expression phenotypes. Nevertheless, which proteins interact with NUP98 FOs in puncta and the importance of puncta formation for effective therapeutic targeting of NUP98-rearranged cells is not known. This research proposal seeks to identify the proteins found in NUP98 FO-associated puncta, uncover how puncta alter gene regulation, and determine if puncta disruption correlates with effective treatment of NUP98-rearranged cells. Aim 1 will examine the role of histone acetyltransferase (HAT) complex members, which my preliminary data identified as key NUP98 FO interacting proteins, in NUP98::KDM5A FO-driven cell transformation. I will perform CRISPR/Cas9 editing of HAT complex genes in hematopoietic stem and progenitor cells (HSPCs) from our Nup98::Kdm5a mouse model and study the in vitro and in vivo consequences of these alterations. I will also examine gene expression and chromatin remodeling in Nup98::Kdm5a HSPCs with and without HAT complex disruption. Aim 2 will determine whether effective therapeutic targeting of NUP98 FOs leads to puncta disruption. I will perform co-localization experiments for FO with proteins involved in nuclear transport and gene regulation. I will then pharmacologically inhibit these interacting proteins using available small molecule inhibitors and assess changes in puncta features and cell viability over time to determine if puncta disruption correlates with drug efficacy. In Aim 3, I will identify interacting proteins that are vulnerabilities in NUP98-rearranged cells and use pharmacologic inhibition of crucial interactors to identify how they are involved in cell transformation, gene regulation, and LLPS. Together, these studies will identify critical interacting proteins in leukemias bearing NUP98 gene fusions, examine how they contribute to leukemogenesis, and uncover how they might be targeted for therapeutic benefit.

项目总结