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Novel relationships of splicing factors in temozolomide-resistant glioblastoma

替莫唑胺耐药胶质母细胞瘤中剪接因子的新关系

基本信息

批准号：
10557860
负责人：
Deanna Marie Tiek
金额：
$ 9.85万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
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：
2020-02-01 至 2023-09-18
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10557860
关键词：
Agonist Alkylating Agents Alternative Splicing Apoptotic Arginine Biology Blood - brain barrier anatomy Brain Brain Neoplasms Cell model Cell physiology Cells DNA DNA Damage DNA Repair Diagnosis Doctor of Philosophy Drug resistance Equilibrium Event FDA approved Genes Genetic Transcription Glioblastoma Goals In Vitro Knowledge Lesion Malignant Neoplasms Mentorship Methods Methylation Methyltransferase MicroRNAs Modeling Molecular Nuclear Orphan Receptor Operative Surgical Procedures Organ Paper Pathway interactions Patients Phase Phosphorylation Phosphotransferases Porifera Postdoctoral Fellow Process Production Protein Isoforms Proteins RNA RNA Splicing RNA-Binding Protein EWS RNA-Binding Proteins Radiation therapy Regulation Research Research Personnel Research Project Grants Research Technics Resistance Role Serine Signal Transduction Testing Therapeutic Training Transcript Translating Tubulin Untranslated RNA Work Xenograft procedure anticancer research base cell growth cell motility chemotherapy circular RNA combat estrogen-related receptor experience gene product improved in silico in vivo inhibitor metabolic phenotype neoplastic cell new therapeutic target novel novel strategies novel therapeutic intervention post-doctoral training pre-doctoral programs receptor function repaired research study response success temozolomide therapeutic target transcriptome

项目摘要

Glioblastoma (GBM) is a devastating cancer, due to both our narrow understanding of its molecular drivers and limited therapeutic strategies. One potential mechanistic driver is alternative splicing. The brain contains the most alternatively spliced transcripts of any organ, and many splicing factors are upregulated between normal brain and GBM. While chemotherapeutic options are limited by the physical blood brain barrier (BBB), the DNA- damaging agent temozolomide (TMZ) is able to cross into the brain. However, most patients rapidly become resistant to TMZ and TMZ-resistant GBM is uniformly fatal. An initial goal of my PhD research was to establish novel TMZ-resistant cellular models in order to identify pathways that could be targeted for GBM treatment. My comprehensive characterization of the cell growth, motility, and metabolic phenotypes of my two new TMZ- resistant GBM models forms the basis for my initial first-author paper. During my dissertation research (Aim 1), I have conducted two complementary studies that identify novel approaches to targeting alternative splicing events in GBM. The first (Aim 1.1) is to target the alternatively spliced estrogen-related receptor beta (ERRβ). I have started to define with in silico and in vitro methods how the pro-apoptotic isoform of this gene, ERRβ2, is processed. I found that the serine/arginine (SR) rich splicing factor SRSF6 plays a role in ERRβ2 production and that inhibition of Cdc-like kinases (CLKs, which phosphorylate SR proteins) with TG-003 in combination with the ERRβ synthetic agonist DY-131 potently inhibits TMZ-resistant GBM cells in vitro and in intracranial xenografts. The second (Aim 1.2) is a broader study of splicing inhibition and regulation in TMZ-resistant GBM. I found that TMZ decreases the phosphorylation (p) of SR proteins in TMZ-sensitive, but not TMZ-resistant models. This is accompanied by mis-localization of pSR proteins, and increased baseline levels of DNA damage. In TMZ- resistant GBM cells, the RNA binding protein EWS also mis-localizes and forms aggregates that are stabilized by tubulin. My working hypothesis is that because of the increased DNA damage in TMZ-resistant GBM, the DNA damage response becomes reprogrammed which causes splicing factors (like EWS and pSR proteins) to be displaced from their normal cellular compartments and poised for aberrant aggregation. Also, that this new splicing factor/DNA damage repair axis can be therapeutically targeted with novel splicing inhibitors. During the postdoctoral training period (Aim 2), I will address a key gap in our understanding of the GBM transcriptome: the role of non-coding RNAs, specifically the noncanonical back-spliced or circular RNAs (circRNAs). I propose to define the circRNA landscape of GBM, to determine the regulatory functions and to propose potential therapeutic applications of these abundant and dynamic regulators of splicing and transcription. Together, my pre- and postdoctoral research experiences will have prepared me to balance both big picture ideas and focused studies of mechanism when I establish my own research program as an independent cancer researcher.

胶质母细胞瘤（GBM）是一种毁灭性的癌症，因为我们对其分子驱动因素的理解很狭隘