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Targeting defective spliceosomal pathway in myeloid malignancies

靶向骨髓恶性肿瘤中的缺陷剪接体途径

基本信息

批准号：
10434248
负责人：
VALERIA VISCONTE
金额：
$ 23.17万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10434248
关键词：
3&apos Splice Site AML/MDS Acute Myelocytic Leukemia Adult Animal Model Autophagocytosis Binding Biochemical Biological Assay Cancer Model Cell Line Cell model Cells Chemicals Clinical Data Clinical Research Clonal Evolution Code Computer-Aided Design Crystallization Data Development Disease model Drug Targeting Dysmyelopoietic Syndromes Erythropoiesis Essential Genes Exhibits Future Gene Deletion Genes Glioma Goals Growth Hematopoietic Human IRAK4 gene Impairment In Vitro Intervention K-562 K562 Cells Lead Libraries Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of pancreas Malignant neoplasm of urinary bladder Messenger RNA Missense Mutation Modeling Mus Mutate Mutation Myeloid Cells Myeloproliferative disease NF-kappa B Oncogenic Outcome Pathway interactions Patients Pattern Pharmacology Phase I Clinical Trials Prognosis Protein Isoforms Proteins Quantitative Reverse Transcriptase PCR RNA Splicing Regulation Risk SF1 SRSF2 gene Sampling Signal Transduction Solid Neoplasm Spliceosome Assembly Pathway Spliceosomes Structure Structure-Activity Relationship Therapeutic Therapeutic Effect Therapeutic Studies Toxic effect Transgenic Mice Validation Xenograft procedure acute myeloid leukemia cell base cancer cell cancer survival chemical synthesis disease phenotype efficacy evaluation exome improved in vivo inhibitor innovation leukemia mRNA Precursor malignant breast neoplasm melanoma mouse model mutant novel novel therapeutics preclinical development preference prognostic prognostic value recruit screening small molecule therapeutics targeted agent targeted sequencing therapeutic candidate therapeutic development therapeutic target tool transcriptome transcriptomics

项目摘要

Abstract Whole-exome and targeted sequencing of MDS patients’ samples have led to discover a set of genes (SF3B1, U2AF1, SRSF2, ZRSR2 and LUC7L2) encoding mutant RNA splicing factors that alter expression patterns of protein isoforms in clonal evolution of hematopoietic cells during MDS progression. Subsequent studies found occurrences of these splicing factor mutations in AML and solid tumors. Clinical data also showed heterozygous missense mutations or hemizygous deletions of these genes are mostly mutually exclusive implying that mutations in multiple proteins in splicing regulation may be lethal in MDS cells. To date, only one compound, H3B-8800, to target SF3B1 is in phase I clinical trial to treat MDS and other myeloid malignancies. No agents targeting other mutated splicing factors are in pre-clinical development and if other mutant factors can be potential drug targets is unknown. Among them, U2AF1 mutations occurred in early MDS clones and are poor prognostic features for progression to leukemia. Studies showed U2AF1S34F hematopoietic cells and transgenic mouse models are sensitive to sudemycin that targets SF3B1. Recently, U2AF1wt was suggested to be a haplo-essential gene for the survival of cancer cells containing U2AF1 mutation. Our central hypothesis is that U2AF1wt inhibition may induce synthetic lethality to U2AF1S34F clones that depends on U2AF1wt for survival. Our rationale is that blockade of U2AF1/U2AF2 association may impair U2AF1wt function. Using fragment-based library screening, we have identified a hit compound that inhibited the U2AF1/U2AF2 binding and exhibited selective growth inhibition in K562-U2AF1S34F and human primary cells carrying U2AF1 mutations but not their wild-type counterparts. Our objective of this proposal is to develop a new class of U2AF1 inhibitors based on the hit and assess the therapeutic concept of synthetic lethality in MDS/AML disease models. Our long-term goal is to develop small-molecule therapeutics to target cancer cells defective in the spliceosome pathway. To achieve our goal, we have devised two specific aims: 1) Optimization of U2AF1 inhibitors by integrating computer-aided design with chemical syntheses; 2) Study the therapeutic effects of U2AF1 inhibitors using in vitro and in vivo U2AF1mut MDS/AML models. In Aim 1, we will improve the activity and selectivity of U2AF1 inhibitors based on the preliminary structure activity relationship and selectivity data of SF1-8 by integrating chemical syntheses, computer-aided design, biochemical assays and the co- crystal structure determination. In Aim 2, we will assess the activities of our U2AF1 inhibitors in MDS/AML cellular models, analyze transcriptome and splicing pattern changes in cell lines treated with U2AF1 inhibitors and determine the effects of our inhibitors on erythropoiesis in xenograft mouse models. Our application is innovative and significant, because it builds on our discovery of the hit to U2AF1, an emerging novel target associated with the transformation of myeloid malignancies, and has the potential to establish a new therapeutic platform to treat MDS/AML by pharmacologically targeting mutant U2AF1 cells.

抽象的 MDS 患者样本的全外显子组和靶向测序发现了一组基因（SF3B1、 U2AF1、SRSF2、ZRSR2 和 LUC7L2）编码突变 RNA 剪接因子，改变 MDS 进展期间造血细胞克隆进化中的蛋白质亚型。随后的研究发现这些剪接因子突变在 AML 和实体瘤中的发生。临床数据还显示这些基因的杂合错义突变或半合缺失大多是相互排斥的这意味着剪接调节中多种蛋白质的突变可能对MDS细胞是致命的。迄今为止，只有一个靶向 SF3B1 的化合物 H3B-8800 正处于治疗 MDS 和其他骨髓恶性肿瘤的 I 期临床试验中。没有针对其他突变剪接因子的药物处于临床前开发阶段，并且如果其他突变剪接因子能否成为潜在的药物靶点尚不清楚。其中，U2AF1突变发生在早期MDS克隆中，是进展为白血病的不良预后特征。研究表明 U2AF1S34F 造血细胞和转基因小鼠模型对针对 SF3B1 的苏德霉素敏感。最近，U2AF1wt被建议是含有 U2AF1 突变的癌细胞生存的单倍体必需基因。我们的中心假设是 U2AF1wt 抑制可能会诱导依赖于 U2AF1wt 的 U2AF1S34F 克隆的合成致死性生存。我们的理由是，阻断 U2AF1/U2AF2 关联可能会损害 U2AF1wt 功能。使用基于片段的文库筛选，我们鉴定出了抑制 U2AF1/U2AF2 结合的命中化合物并在 K562-U2AF1S34F 和携带 U2AF1 的人原代细胞中表现出选择性生长抑制突变，但不是其野生型对应物。我们此提案的目标是开发一类新的 U2AF1抑制剂基于命中并评估MDS/AML综合致死治疗概念疾病模型。我们的长期目标是开发针对有缺陷的癌细胞的小分子疗法在剪接体途径中。为了实现我们的目标，我们制定了两个具体目标：1）优化将计算机辅助设计与化学合成相结合的U2AF1抑制剂； 2）研究治疗方法使用体外和体内 U2AF1mut MDS/AML 模型研究 U2AF1 抑制剂的作用。在目标 1 中，我们将改进基于初步结构活性关系和选择性的 U2AF1 抑制剂的活性和选择性通过整合化学合成、计算机辅助设计、生化分析和协同分析，获得 SF1-8 的数据晶体结构测定。在目标 2 中，我们将评估 U2AF1 抑制剂在 MDS/AML 中的活性细胞模型，分析 U2AF1 抑制剂处理的细胞系中的转录组和剪接模式变化并确定我们的抑制剂对异种移植小鼠模型中红细胞生成的影响。我们的应用程序是创新且意义重大，因为它建立在我们发现 U2AF1（一个新兴的新靶标）的基础上与骨髓恶性肿瘤的转化相关，并有可能建立一个新的通过药理学靶向突变 U2AF1 细胞治疗 MDS/AML 的治疗平台。