(PQC2) Niche-responsive RNA editing by ADAR1 in dormant multiple myeloma initiating cell maintenance

(PQC2) ADAR1 在休眠多发性骨髓瘤中进行生态位响应性 RNA 编辑，启动细胞维持

基本信息

批准号：
9060287
负责人：
Catriona Helen Macleod Jamieson
金额：
$ 16.09万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9060287
关键词：
1q21 Address Adenosine Aftercare Agonist Automobile Driving B-Lymphocytes Biological Assay Bone Marrow CD19 gene Cell Cycle Cell Cycle Kinetics Cell Line Cell Maintenance Cell Survival Cell physiology Cells Chromosomes Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Complex Coupled Cues Development Disease Double-Stranded RNA Drug resistance Employee Strikes Event Flow Cytometry Generations Genes Genome Stability Goals Health Hematologic Neoplasms Hematopoietic Hematopoietic Neoplasms Human In Vitro Inflammation Inflammatory Inosine Interleukin-6 Investigation Lead Life Life Expectancy Maintenance Malignant - descriptor Malignant Neoplasms Measures Mediating Modeling Molecular Molecular Evolution Multiple Myeloma Patients Pharmaceutical Preparations Plasma Plasma Cells Play Population Production Proteasome Inhibitor Proteins RNA Editing RNA Probes RNA Processing Rare Diseases Recurrence Recurrent disease Regulation Relapse Resistance Role Sampling Signal Transduction Solid Neoplasm Stem cells Stromal Cells Thalidomide Therapeutic Agents Transcript Treatment Failure Treatment Protocols Ubiquitination acquired drug resistance adenosine deaminase base bone cancer stem cell cytokine high risk in vivo knock-down knockout gene lenalidomide leukemia leukemic stem cell malignant breast neoplasm mouse model new therapeutic target novel novel diagnostics novel strategies outcome forecast prevent response self-renewal small hairpin RNA standard of care stem stem cell population targeted treatment therapy resistant transcriptome tumor

项目摘要

DESCRIPTION (provided by applicant): Despite new available therapies, including immunomodulatory drugs such as thalidomide and lenalidomide in multiple myeloma (MM), about 20-25% of patients are still considered at high risk for treatment failure. The emergence of therapeutically recalcitrant cases and disease relapse even under intensive treatment regimens suggests the existence of a dormant myeloma-initiating population within the bone marrow (BM) that is capable of drug escape. These myeloma-initiating cells are defined by the lack of the plasma cell marker CD138, while they express several hematopoietic stem (CD38) and B cell markers such as CD27 and CD19; however, the molecular and cellular mechanisms that regulate myeloma-initiating cell generation and maintenance are so far poorly understood. Recently, we and other groups showed that both NOTCH signaling and inflammation- responsive ADAR1 activation are crucial events regulating malignant stem cell maintenance in the bone marrow microenvironment, characterized by enhanced survival and self-renewal and cell cycle alterations of dormant progenitor cells. In this context, the central hypothesis of this proposal is that MM niche-derived pro- inflammatory signals induce aberrant human-specific RNA editing driven by adenosine deaminase acting on dsRNA-1 (ADAR1) in dormant myeloma-initiating cells that is accentuated by lenalidomide resistance. This project will: 1) determine whether ADAR1 activity is enhanced in myeloma-initiating cells and investigate the effects of lenalidomide treatment on ADAR1-dependent RNA editing in therapeutic resistance and relapse; 2) identify the NOTCH-regulated pro-inflammatory cytokines that activate ADAR1-dependent RNA editing in myeloma-initiating cells; and 3) determine whether direct inhibition of ADAR1 activity in MM initiating cells, or blocking microenvironmental signals that activate ADAR1, sensitizes myeloma-initiating cells to lenalidomide and prevents myeloma-initiating cell maintenance. These aims will address PQC2: What molecular or cellular events establish tumor dormancy after treatment and what leads to recurrence? We will utilize both in vitro and in vivo measures of myeloma-initiating cell function using multicolor flow cytometry and fluorescent RNA probe-based strategies to purify and profile primary MM cellular constituents, coupled with a novel diagnostic qPCR-based assay to detect endogenous RNA editing, and gene knockout (CRISPR) or lentiviral shRNA- knockdown strategies to modulate NOTCH-dependent ADAR1 activation. A fluorescent ubiquitination cell cycle indicator (FUCCI) bi-cistronic lentiviral reporer will facilitate investigations of dormant live cells, in robust bone marrow stromal co-culture models and bioluminescent humanized MM mouse models. The ultimate goal is to investigate niche-dependent ADAR1 activation as a novel mechanism driving transcriptome recoding and molecular evolution of dormant myeloma-initiating cells, laying the groundwork for targeted therapeutics with potential applications in an array of other therapeutically recalcitrant malignancies.

描述（由适用提供）：尽管有新的可用疗法，包括多发性骨髓瘤（MM）中的免疫调节药物，例如沙利度胺和多纳他胺，但仍有约20-25％的患者仍被视为治疗衰竭的高风险。即使在强化治疗方案下，生物学上顽固的病例和疾病的出现也表明，骨髓（BM）内存在休眠的骨髓瘤人群（BM），能够逃脱药物。这些骨髓瘤引发细胞的定义是缺乏浆细胞标记CD138，而它们表达了几个造血茎（CD38）和B细胞标记物（例如CD27和CD19）。然而，迄今为止对调节骨髓瘤的细胞产生和维持的分子和细胞机制迄今为止鲜为人知。最近，我们和其他小组表明，Notch信号传导和炎症反应性ADAR1激活是调节骨髓微环境中恶性干细胞维持的关键事件，其特征是增强的存活率，自我恢复和细胞周期的变化，以及细胞周期的变化。在这种情况下，这一点的中心假设提议是，MM利基衍生的促炎信号通过作用于DSRNA-1（ADAR1）的腺苷死亡素酶在休眠的骨髓瘤降低细胞中引起异常的人类特异性RNA编辑驱动，而该细胞突显了多纳米胺抗性。该项目将：1）确定在骨髓瘤发射细胞中的ADAR1活性是否增强，并研究Lenalidomide治疗对ADAR1依赖性RNA编辑对治疗耐药性和继电器的影响； 2）确定在骨髓瘤发射细胞中激活ADAR1依赖性RNA编辑的缺口调节的促炎细胞因子； 3）确定是否直接抑制MM启动细胞中ADAR1活性，或阻断激活ADAR1的微环境信号，会使骨髓瘤启动细胞对列纳奈替胺，并阻止骨髓瘤降低细胞的维持。这些目标将解决PQC2：哪些分子或细胞事件在治疗后会出现肿瘤休眠状态，什么导致复发？ We will utilize both in vitro and in vivo measurements of myeloma-initiating cell function using multicolor flow cytometry and fluorescent RNA probe-based strategies to purify and profile primary MM cellular constitutions, coupled with a novel diagnostic qPCR-based assay to detect endogenous RNA editing, and gene knockout (CRISPR) or lentiviral shRNA- knockdown strategies to modulate Notch依赖性ADAR1激活。荧光泛素化细胞周期指标（FUCCI）双月生态生产者将促进休眠活细胞的投资，在强大的骨髓基质共培养模型和生物发光的人性化MM小鼠模型中。最终的目标是研究依赖小裂依赖性的ADAR1激活，作为一种新的机制，驱动转录组重新编码和休眠骨髓瘤发射细胞的分子进化，为在其他治疗疗法的其他治疗型重新耐药的恶性肿瘤中靶向靶向治疗奠定了基础。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

RNA rewriting, recoding, and rewiring in human disease.

DOI：
10.1016/j.molmed.2015.07.001
发表时间：
2015-09
期刊：
Trends in molecular medicine
影响因子：
13.6
作者：
M. Zipeto;Qingfei Jiang;E. Melese;C. Jamieson
通讯作者：
M. Zipeto;Qingfei Jiang;E. Melese;C. Jamieson

Multiple myeloma-derived Jagged ligands increases autocrine and paracrine interleukin-6 expression in bone marrow niche.

DOI：
10.18632/oncotarget.10820
发表时间：
2016-08-30
期刊：
Oncotarget
影响因子：
0
作者：
Colombo M;Galletti S;Bulfamante G;Falleni M;Tosi D;Todoerti K;Lazzari E;Crews LA;Jamieson CH;Ravaioli S;Baccianti F;Garavelli S;Platonova N;Neri A;Chiaramonte R
通讯作者：
Chiaramonte R

Cancer Cells Exploit Notch Signaling to Redefine a Supportive Cytokine Milieu.

DOI：
10.3389/fimmu.2018.01823
发表时间：
2018
期刊：
Frontiers in immunology
影响因子：
7.3
作者：
Colombo M;Mirandola L;Chiriva-Internati M;Basile A;Locati M;Lesma E;Chiaramonte R;Platonova N
通讯作者：
Platonova N

Identification of small molecules uncoupling the Notch::Jagged interaction through an integrated high-throughput screening.

DOI：
10.1371/journal.pone.0182640
发表时间：
2017
期刊：
PloS one
影响因子：
3.7
作者：
Platonova N;Parravicini C;Sensi C;Paoli A;Colombo M;Neri A;Eberini I;Chiaramonte R
通讯作者：
Chiaramonte R