Role of RNA methylation in chemoresistant cancer cells

RNA 甲基化在化疗耐药癌细胞中的作用

基本信息

批准号：
10083714
负责人：
Shobha Vasudevan
金额：
$ 19.26万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
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：
2020-01-15 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10083714
关键词：
3&apos Untranslated Regions ATM Signaling Pathway Acute Myelocytic Leukemia Adenosine Affect Affinity Chromatography Antibodies Biological Assay Blast Cell Blood Cancer Etiology Cancer Relapse Cell Cycle Cell Line Cell Survival Cells Chemicals Chemoresistance Clinical Co-Immunoprecipitations Complex Coupled Data Disease remission Gene Expression Genes Genetic Transcription Goals Heterogeneity Immunoprecipitation Lead Malignant Neoplasms Maps Mediating Messenger RNA Methods Methylation Methyltransferase Modification N(6)-ribosyladenine Normal Cell Outcome Patients Population Positioning Attribute Post-Transcriptional Regulation Proteome Proteomics RNA RNA methylation RNA-Binding Proteins Reader Recurrence Regulation Resistance Role Sampling Signal Transduction Site Testing Therapeutic Translating Translations acute myeloid leukemia cell base biological heterogeneity cancer cell cancer stem cell chemotherapy crosslink cytokine immune resistance in vivo in vivo Model inhibitor/antagonist leukemic stem cell mRNA Expression new therapeutic target protein degradation stem cell biomarkers targeted treatment therapeutic biomarker transcriptome tumor

项目摘要

Summary Role of RNA methylation in chemoresistant cancer cells Goal Based on our data, we propose that transiently quiescent populations in acute myeloid leukemia (AML) are maintained in part by RNA methylation, permitting synthesis of survival and tumor promoting regulators necessary for chemoresistance and subsequent AML persistence. The primary objective is to characterize the role of RNA methylation in regulating gene expression in resistant AML, which contributes to chemosurvival. Significance AML is a serious malignancy that displays clinical resistance due to heterogeneity. Resistant AML cells include quiescent (G0) cells that are transiently arrested states, and thus resist clinical therapy that targets only cycling cells; these cells re-enter proliferation to cause AML persistence. A primary issue is that resistance regulators remain to be uncovered. A second need is to identify markers to detect resistant cells. Such cells show distinct gene expression that permits chemosurvival (PNAS 2014, Mol. Cell 2016, Biorxiv/ 418715). Canonical translation is decreased; yet specific mRNAs are expressed by unknown mechanisms. Regulation of specific gene expression in resistant AML needs to be uncovered & targeted to curtail AML. Premise Profiling in resistant cells revealed expression of factors that affect cell survival. Expressed mRNAs in G0 resistant cells have extended 3? untranslated regions (UTRs) to include regulatory sites, such as for RNA methylation. Methylation at N6 position of adenosines (m6A) are unique marks on RNAs that regulate gene expression via RNA binding proteins called readers, to control distinct cell states. Role of m6A in resistant AML remains to be uncovered. We find the m6A methyltransferase increases due to therapy induced signals. Our data reveal that m6A is needed for specific gene expression & resistance. We developed inhibitors to block therapy induced signals & antisense to block unique m6A sites on extended 3’UTRs, which reduce resistance. These data suggest that m6A regulates specific gene expression to enable therapy survival. Characterization of mRNAs that are modified & expressed in resistant cells, in vivo & in patient samples by m6A, their readers, their role in resistance, & of inhibitors that block m6A, will provide markers & therapeutics to curb resistance. Method Aim I will characterize m6A readers that regulate known modified mRNAs, by in vivo crosslinking coupled RNA affinity purification in resistant AML cells & in vivo models. Chemical inhibitors to therapy induced signaling that promotes m6A and antisense to block mapped m6A sites will be tested to curb resistance. Aim II will globally identify m6A targets that contribute to resistance in cell lines & in vivo models, using m6A antibody & methyltransferase immunoprecipitation. Role of readers & m6A mRNAs in resistant cells & in vivo is tested by depletions coupled with chemosurvival assays; their expression will be verified in resistant patient samples. Outcomes mRNAs expressed by m6A, their readers, their role in resistance in AML cells & in vivo, & inhibitors of m6A to curb resistance will be identified. Their expression & efficacy in patient samples will be uncovered.

RNA甲基化在化学抗性癌细胞中的摘要作用基于我们的数据的目标，我们提出急性髓样白血病（AML）中的瞬时种群部分通过RNA甲基化来维持，允许合成生存和促进调节剂的肿瘤对于化学抗性和随后的AML持久性所必需的。主要目的是表征 RNA甲基化在调节抗性AML中基因表达中的作用，这有助于化学生存。显着性AML是一种严重的恶性肿瘤，由于异质性而显示出临床抗性。抵抗的 AML细胞包括瞬时停滞状态的静态（G0）细胞，因此抵抗临床治疗仅靶向循环细胞；这些细胞重新进入增殖以引起AML持久性。主要问题是阻力调节剂仍有待发现。第二个需求是识别标记以检测电阻细胞。这种细胞显示出允许化学生存的独特基因表达（PNAS 2014，Mol。Cell2016，Biorxiv/ 418715）。规范翻译得到改善；然而，特定的mRNA通过未知机制表达。需要发现耐药AML中特定基因表达的调节，以减少AML。抗性细胞中的前提谱图揭示了影响细胞存活的因素的表达。表示mRNA G0抗性细胞已延长3？未翻译区域（UTR）包括调节位点，例如RNA 甲基化。腺苷N6位置（M6A）位置的甲基化是调节基因的RNA的独特标记通过RNA结合蛋白的表达称为读取器，以控制不同的细胞态。 M6A在抗性AML中的作用还有要发现的。我们发现由于治疗诱导的信号，M6A甲基转移酶增加。我们的数据表明，特定基因表达和抗性需要M6a。我们开发了抑制剂以阻止治疗引起的信号和反义可阻止延长的3'UTRS上的独特M6A位点，从而降低耐药性。这些数据表明M6A调节特定的基因表达以实现治疗生存。表征在抗性细胞，体内和患者样品中通过M6A（其读者）进行修饰和表达的mRNA的mRNA 它们在阻止M6a的抗药性和抑制剂中的作用将提供标记和治疗以遏制抗性。方法目的我将通过体内交联来表征调节已知改性mRNA的M6A读取器抗性AML细胞和体内模型中的RNA亲和纯化耦合。诱导治疗的化学抑制剂促进M6A和反义以阻断映射的M6A位点的信号将进行测试以遏制电阻。目标II 使用M6A抗体和甲基转移酶免疫沉淀。测试了读者和M6A mRNA在抗性细胞和体内的作用通过耗竭加化学生存测定；它们的表达将在耐药的患者样本中得到验证。 M6A表达的结果mRNA，其读者，它们在AML细胞和体内的抗性中的作用，并抑制剂将确定M6A到遏制的抑制性。它们在患者样品中的表达和效率将被发现。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

NR4A1 regulates expression of immediate early genes, suppressing replication stress in cancer.

DOI：
10.1016/j.molcel.2021.09.016
发表时间：
2021-10-07
期刊：
Molecular cell
影响因子：
16
作者：
Guo H;Golczer G;Wittner BS;Langenbucher A;Zachariah M;Dubash TD;Hong X;Comaills V;Burr R;Ebright RY;Horwitz E;Vuille JA;Hajizadeh S;Wiley DF;Reeves BA;Zhang JM;Niederhoffer KL;Lu C;Wesley B;Ho U;Nieman LT;Toner M;Vasudevan S;Zou L;Mostoslavsky R;Maheswaran S;Lawrence MS;Haber DA
通讯作者：
Haber DA

Long-read sequencing reveals complex patterns of wraparound transcription in polyomaviruses.