Therapeutic targeting of IRAK4 in MDS

MDS 中 IRAK4 的治疗靶向

• 批准号: 10537837
• 负责人: Daniel Starczynowski
• 金额: $ 54.5万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-02 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10537837
• 关键词: AML/MDS; Acute Myelocytic Leukemia; Alternative Splicing; Biological Assay; C-terminal; CDK2 gene; Cell Line; Cell physiology; Cells; Cellular biology; Chronic; Clinical; Clinical Trials; Complex; Correlative Study; Data; Death Domain; Disease; Dysmyelopoietic Syndromes; Exhibits; Exons; Functional disorder; Gene Expression; Genes; Genetic; Growth; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic stem cells; IRAK4 gene; Immune; Immune signaling; In Vitro; Induced Mutation; Inflammatory; Lead; Leukemic Cell; Link; Lysine; MAP Kinase Gene; Mediating; Mutation; N-terminal; Nature; Oncogenic; Outcome; Pathway interactions; Patients; Pharmacology; Phosphotransferases; Prognosis; Protein Isoforms; Protein-Serine-Threonine Kinases; Publishing; RNA Splicing; Recurrent disease; Regulation; Role; Sampling; Signal Pathway; Signal Transduction; Spliceosomes; TRAF6 gene; Therapeutic; Ubiquitination; Work; Xenograft procedure; base; biomarker-driven; clinical predictors; clinically relevant; efficacy testing; in vivo; inhibitor; kinase inhibitor; knock-down; leukemia; mutant; novel; overexpression; patient derived xenograft model; phosphoproteomics; potential biomarker; predicting response; response; small hairpin RNA; therapeutic target; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Spliceosome mutations and alterations in RNA splicing are common in myelodysplastic syndrome (MDS), yet the specific oncogenic changes due to these alterations have not been fully identified. We demonstrated that overexpression of an active isoform of the serine/threonine kinase, IRAK4, is seen in MDS. The active long IRAK4 isoform (IRAK4-L) consists of an N-terminal death domain (DD) and C-terminal kinase domain (KD), which stochastically assembles with the Myddosome complex and results in maximal activation of innate immune signaling pathways. Inclusion of exon 4 in the long isoform is mediated by mutant splicing factor (SF), U2AF1, which results in oncogenic signaling and poor prognosis in MDS/AML (Smith et al., Nature Cell Biology, 2019). Importantly, inhibition of IRAK4-L abrogates leukemic growth in vitro and in vivo. These findings established that mutations in U2AF1 induce expression of therapeutically targetable "active" IRAK4 isoforms and provide a genetic link to activation of chronic innate immune signaling in MDS. Our preliminary RNA-sequencing studies in primary MDS samples reveal that (i) splicing factor SF3B1 mutations also lead to the active IRAK4-L isoforms via retention of exon 6, and that (ii) a sizable proportion of splicing factor (SF) wild-type MDS patients also exhibit expression of IRAK4-L. We hypothesize that oncogenic IRAK4-L isoforms are critical for the function of SF-mutant (i.e. U2AF1 and SF3B1) and a subset of SF-wildtype MDS disease propagating cells. As such, this proposal will comprehensively evaluate the mechanistic basis of IRAK4-L regulation in SF3B1 and SF-wild type MDS (Aim 1) by using isogenic cell lines expressing the SF3B1 K700E mutants and by altering the expression of splicing modulator PRMT5. Aim 2 will determine downstream regulators of oncogenic IRAK4 isoform activation in MDS. In addition to examination of NF-κB/MAPK signaling, we will perform an unbiased phospho-proteomic screen to identify direct substrates of IRAK4-L and examine their roles in hematopoietic dysfunction. Aim 3 will determine the efficacy of IRAK4-L inhibition in MDS by a clinical-grade IRAK4 kinase inhibitor. In vitro sensitivity and differentiation assays in U2AF1- and SF3B1-mutant, and SF wildtype MDS cells will be conducted. Patient derived xenografts with U2AF1/SF3B1-mutant, and SF wildtype MDS will be treated with the IRAK4 inhibitors to examine for suppression of the disease-propagating cells. We will also determine whether overexpression of IRAK4-long isoforms is predictive of response in an ongoing clinical trial of IRAK4 inhibitor. Taken together, these studies will determine the role of splicing mutation induced IRAK4-L isoform in MDS pathobiology. Importantly, these studies will lead to potential targeted clinical trials with clinically relevant inhibitors of IRAK4 in MDS.

项目总结/文摘