Therapeutic targeting of RNA splicing catalysis through inhibition of Protein Arginine Methylation

通过抑制蛋白质精氨酸甲基化来治疗 RNA 剪接催化

基本信息

批准号：
10703206
负责人：
Ernesto Guccione
金额：
$ 37.59万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-15 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10703206
关键词：
Acute Myelocytic Leukemia Alleles Antisense Oligonucleotides Arginine Catalysis Cells Chronic Lymphocytic Leukemia Data Disease Drug Targeting Dysmyelopoietic Syndromes Evaluation Event Gene Expression Genes Genetic Health Heterozygote In Vitro Individual Leukemic Cell Malignant Neoplasms Mediating Methylation Modeling Molecular Mutation Myeloid Leukemia Patients Pharmaceutical Preparations Phase Phase I Clinical Trials Phenotype Point Mutation Process Protein Inhibition Protein Methylation Protein Splicing Protein-Arginine N-Methyltransferase Proteins Proteome RNA Splicing RNA methylation RNA-Binding Proteins RNA-targeting therapy Refractory Safety Small Nuclear Ribonucleoproteins Solid Neoplasm Spliceosome Assembly Pathway Spliceosomes Therapeutic Therapeutic Effect Uveal Melanoma Work cancer cell cancer type cell killing clinical development curative treatments early phase clinical trial efficacy evaluation in vivo inhibitor leukemia mRNA Precursor mutant myeloid leukemia cell novel therapeutic intervention novel therapeutics pharmacologic preclinical development small molecule snRNP Structural Core Protein synthetic lethal interaction

项目摘要

SUMMARY Mutations in splicing factors (SF) are highly enriched in a variety of cancer types, particularly myelodysplastic syndromes (MDS), acute myeloid leukemia (AML), and chronic lymphocytic leukemia (CLL), in addition to solid tumors such as uveal melanoma. Our group has identified that cells bearing SF mutations cannot tolerate further perturbations to splicing catalysis and, consistent with this, we have identified that spliceosomal mutant cancer cells are preferentially sensitive to small molecules that disrupt pre-mRNA splicing. While the above effort has resulted in an ongoing phase I clinical trial of a spliceosome modulatory compound for patients with refractory myeloid leukemias, we do not currently know the safety or efficacy of pharmacologic modulation of core spliceosome function. To this end, our group has also recently identified that inhibiting spliceosomal assembly through inhibition of arginine methylation of Sm proteins provides an alternate means of therapeutic splicing inhibition. We have identified that inhibiting either symmetric arginine methylation (mediated by the protein arginine methyltransferase 5 (PRMT5)) or asymmetric dimethyl arginine methylation (mediated by type I PRMTs (PRMT1, 4, and 6)) reduces splicing fidelity resulting in strong preferential killing of SF-mutant leukemias over their wildtype counterparts. Here we aim to determine if in leukemia, SF-mutations portend greater vulnerability to a “second hit” targeting splicing through inhibition of type I (PRMT1/4/6) and/or type II (PRMT5) PRMTs. In Aim 1 we will define the therapeutic potential of inhibiting PRMT5, type I PRMTs, and core spliceosome function, alone or together in leukemia models with or without a SF mutation. In addition, we will understand the consequences of combined PRMT inhibition on RNA splicing and gene expression relative to inhibiting PRMT5 or Type I PRMTs alone. In parallel to the above studies, in Aim 2, we will define the molecular basis for the cooperation between PRMT inhibition and SF mutations, by first determining the methylation substrates of PRMT5 or Type I PRMTs, and secondly by determining if individual spliceosomal changes mediated by inhibiting PRMTs or core spliceosome function can be mimicked by anti-sense oligonucleotides, thereby providing an orthogonal novel therapeutic approach to eliminate SF-mutant cancer cells. The significance of these studies is that inhibitors of PRMTs are now entering phase I clinical trials in patients with a variety of cancer types and defining the mechanistic effects and therapeutic utility of PRMT inhibitors for specific genetic subsets of cancers may have incredible therapeutic importance. The health relatedness is that our studies may identify new therapeutic opportunities for a variety of cancer types that have no curative therapies for the majority of patients with these diseases currently.

概括剪接因子（SF）突变在多种癌症类型中高度丰富，尤其是骨髓增生异常综合征（MDS）、急性髓性白血病（AML）和慢性淋巴细胞白血病（CLL），以及实体瘤我们的研究小组已经发现，携带 SF 突变的细胞无法耐受葡萄膜黑色素瘤等肿瘤。对剪接催化的进一步干扰，与此一致，我们已经鉴定出剪接体突变体癌细胞优先对破坏前 mRNA 剪接的小分子敏感。虽然上述努力已导致正在进行的剪接体调节化合物的 I 期临床试验对于难治性粒细胞白血病患者，我们目前不知道药物治疗的安全性或有效性为此，我们的小组最近还发现了抑制核心剪接体功能的机制。通过抑制 Sm 蛋白精氨酸甲基化进行剪接体组装提供了另一种方法我们已经确定抑制对称精氨酸甲基化。（由蛋白质精氨酸甲基转移酶 5 (PRMT5) 介导）或不对称二甲基精氨酸甲基化（由 I 型 PRMT（PRMT1、4 和 6）介导）降低剪接保真度，导致强烈优先杀死 SF 突变型白血病优于其野生型供体。在这里，我们的目标是确定在白血病中，SF 突变是否预示着更容易遭受“第二次打击”目标通过抑制 I 型 (PRMT1/4/6) 和/或 II 型 (PRMT5) PRMT 进行剪接在目标 1 中，我们将定义单独或一起抑制 PRMT5、I 型 PRMT 和核心剪接体功能的治疗潜力具有或不具有 SF 突变的白血病模型此外，我们将了解组合的后果。相对于单独抑制 PRMT5 或 I 型 PRMT，PRMT 对 RNA 剪接和基因表达的抑制。与上述研究并行，在目标 2 中，我们将定义 PRMT 之间合作的分子基础抑制和 SF 突变，首先确定 PRMT5 或 I 型 PRMT 的甲基化底物，以及其次确定个体剪接体变化是否是通过抑制 PRMT 或核心剪接体介导的功能可以通过反义寡核苷酸来模拟，从而提供正交的新型治疗方法消除 SF 突变癌细胞的方法。这些研究的意义在于 PRMT 抑制剂现已进入患者的 I 期临床试验与多种癌症类型相关，并确定 PRMT 抑制剂的机制效应和治疗效用癌症的特定遗传亚型可能具有令人难以置信的治疗重要性。我们的研究可能会为多种尚无治疗方法的癌症类型找到新的治疗机会目前大多数患有这些疾病的患者都可以接受治疗。