Mechanisms and targeting of aberrant Gas activation in myeloid neoplasms

骨髓肿瘤中异常气体​​激活的机制和靶向

• 批准号: 10659809
• 负责人: Eirini Papapetrou
• 金额: $ 67.09万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659809
• 关键词: Acute Myelocytic Leukemia; Address; Apoptosis; Biochemical; Biochemical Genetics; Biological Assay; Bone Marrow; CRISPR/Cas technology; Cell physiology; Cells; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Cyclic Peptides; Data; Dependence; Development; Disease; Dysmyelopoietic Syndromes; FLT3 gene; Frequencies; Future; G-Protein-Coupled Receptors; GNAS gene; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Gases; Gene Mutation; Genes; Genetic; Genetic Transcription; Goals; Hematopoietic; Hematopoietic stem cells; Human; In Vitro; Inflammatory; Knock-out; Malignant Neoplasms; Mediating; Mutate; Mutation; Myeloproliferative disease; Oncogenic; Patients; Pharmaceutical Preparations; Phase; Phenotype; Population Genetics; Process; Production; Prognosis; Protein Array; Protein Isoforms; RNA Binding; RNA Splicing; RNA-Binding Proteins; Role; SRSF2 gene; Sampling; Secondary acute myeloid leukemia; Secondary to; Signal Pathway; Signal Transduction; Signaling Molecule; Somatic Mutation; Spliced Genes; System; Testing; Therapeutic; Therapeutic Intervention; Tissue Banks; Transcript; Work; Xenograft procedure; acute myeloid leukemia cell; drug development; effective therapy; genetic approach; induced pluripotent stem cell; inhibitor; leukemic transformation; mRNA Precursor; mutant; new therapeutic target; novel; novel therapeutics; pharmacologic; programs; research clinical testing; stem cell model; therapeutic target; transcriptome sequencing; transcriptomics; Acute Myelocytic Leukemia; Address; Apoptosis; Biochemical; Biochemical Genetics; Biological Assay; Bone Marrow; CRISPR/Cas technology; Cell physiology; Cells; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Cyclic Peptides; Data; Dependence; Development; Disease; Dysmyelopoietic Syndromes; FLT3 gene; Frequencies; Future; G-Protein-Coupled Receptors; GNAS gene; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Gases; Gene Mutation; Genes; Genetic; Genetic Transcription; Goals; Hematopoietic; Hematopoietic stem cells; Human; In Vitro; Inflammatory; Knock-out; Malignant Neoplasms; Mediating; Mutate; Mutation; Myeloproliferative disease; Oncogenic; Patients; Pharmaceutical Preparations; Phase; Phenotype; Population Genetics; Process; Production; Prognosis; Protein Array; Protein Isoforms; RNA Binding; RNA Splicing; RNA-Binding Proteins; Role; SRSF2 gene; Sampling; Secondary acute myeloid leukemia; Secondary to; Signal Pathway; Signal Transduction; Signaling Molecule; Somatic Mutation; Spliced Genes; System; Testing; Therapeutic; Therapeutic Intervention; Tissue Banks; Transcript; Work; Xenograft procedure; acute myeloid leukemia cell; drug development; effective therapy; genetic approach; induced pluripotent stem cell; inhibitor; leukemic transformation; mRNA Precursor; mutant; new therapeutic target; novel; novel therapeutics; pharmacologic; programs; research clinical testing; stem cell model; therapeutic target; transcriptome sequencing; transcriptomics

PROJECT SUMMARY/ABSTRACT Myelodysplastic syndromes (MDS) are myeloid neoplasms with dismal prognosis, frequent progression to acute myeloid leukemia (AML) and no effective treatment. A decade ago, a development with transformative potential for this disease was the discovery that more than half of MDS patients have somatic mutations in genes encoding splicing factors (SFs, i.e. RNA binding proteins that regulate pre-mRNA splicing). While the high frequency and early occurrence during the disease course of SF mutations rendered them promising targets, efforts to therapeutically leverage this through splicing modulator drugs did not show promise in clinical testing and the drug development pipeline for these targets, and for MDS in general, is currently nearly empty. We developed genetically faithful human induced pluripotent stem cell (iPSC) models of SF-mutated MDS using CRISPR gene editing and performed integrative analyses of splicing (RNA-Seq) and RNA binding (eCLIP) to search for mis-spliced transcripts that are direct common targets of two of the main SF mutations (SRSF2 P95L and the U2AF1 S34F). We found that both mutant SRSF2 and mutant U2AF1 cause altered splicing of the gene GNAS, promoting the production of a longer isoform (GNAS-L), which in turn produces a longer form of the alpha subunit of the stimulatory G protein, G⍺s (G⍺s-L). G proteins are key signaling molecules involved in many important signaling pathways and cell functions, including oncogenic processes. Our preliminary data using functional, biochemical and population genetics approaches support a critical role for the long form of G⍺s (G⍺s-L) as an MDS driver and reveal a new mechanism by which SF mutations drive MDS that opens a completely novel and unexplored therapeutic avenue for MDS, AML and other cancers with SF gene mutations. The goal of this proposal is to investigate G⍺s as a therapeutic target for MDS and identify opportunities for therapeutic interventions that inhibit signaling by G⍺s-L. Specifically, we propose to: (1) Evaluate the effects of direct degradation (through a dTAG) or inhibition (by novel cyclic peptides) of G⍺s in splicing factor (SF)-mutated MDS and AML hematopoietic cells (primary and iPSC-derived) using recently developed iPSC-based models of MDS-to-sAML progression and longitudinal bone marrow samples from MDS patients who progressed to AML; (2) Characterize and target cell signaling downstream of Gs-L through candidate and unbiased approaches (Reverse Phase Protein Array, CyTOF, transcriptomics); and (3) Identify the G-protein coupled receptors (GPCRs) involved in Gs-L activation in SF-mutated MDS and AML through focused CRISPR KO screens. This study can establish a novel therapeutic target that may transform the treatment of MDS, AML and possibly other cancers.

项目摘要/摘要 骨髓增生性综合征（MDS）是髓样肿瘤，进展惨淡，经常进展 急性髓样白血病（AML），没有有效的治疗方法。十年前，具有变革性的发展 这种疾病的潜力是发现，超过一半的MDS患者在 编码剪接因子的基因（SFS，即调节前MRNA剪接的RNA结合蛋白）。而 SF突变疾病过程中的高频和早期发生使他们承诺 目标，通过剪接调节剂药物在治疗上利用这一点的努力并未在临床上显示出希望 目前，对于这些目标以及MDS的测试和药物开发方案，目前几乎是空的。 我们开发了一般忠实的人类诱导的多能干细胞（IPSC）模型的SF-Mutated模型 MDS使用CRISPR基因编辑并进行了剪接（RNA-SEQ）和RNA结合的集成分析 （Eclip）搜索s缩的成绩单，这是两个主要SF突变的直接共同目标 （SRSF2 P95L和U2AF1 S34F）。我们发现突变体SRSF2和突变体U2AF1都会改变 基因GNA的剪接，促进了更长的同工型（GNAS-L）的产生，从而产生A 刺激G蛋白的α亚基的较长形式，G⍺S（G⍺S-L）。 G蛋白是关键信号 参与许多重要的信号通路和细胞功能的分子，包括致癌过程。 我们使用功能，生化和种群遗传学方法的初步数据支持关键作用 对于长形式的G⍺S（G⍺S-L）作为MDS驱动器，并揭示了一种新机制，SF突变驱动 MD为MD，AML和其他癌症打开了完全新颖且出乎意料的治疗途径 SF基因突变。 该提案的目的是调查G⍺S作为MD的治疗目标并确定 抑制G⍺S-L信号传导的治疗干预措施的机会。具体来说，我们建议：（1） 评估直接降解（通过DTAG）或抑制（通过新型环状肽）的效果 最近使用剪接因子（SF）突变的MDS和AML造血细胞（主要和IPSC衍生）使用 开发了基于IPSC的MDS-to-SAML进展模型和MDS的纵向骨髓样品 发展为AML的患者； （2）通过GS-L的下游表征和靶细胞信号通过 候选和无偏的方法（反相蛋白阵列，cytof，转录组学）； （3）识别 SF突变的MDS中涉及GS-L激活的G蛋白偶联受体（GPCR）和AML通过 专注的CRISPR KO屏幕。 这项研究可以建立一个新的治疗靶标，该靶标可能会改变MD，AML和 其他癌症可能。