Transcriptional Determinants of L-Asparaginase Response in Leukemia

白血病 L-天冬酰胺酶反应的转录决定因素

基本信息

批准号：
10316164
负责人：
Robert Thomas Williams
金额：
$ 3.52万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
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：
2020-01-13 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10316164
关键词：
Acute Lymphocytic Leukemia Acute T Cell Leukemia Address Affect Amino Acids Asparagine Aspartate-Ammonia Ligase BTB/POZ Domain Binding Biological Assay CRISPR/Cas technology Cancer cell line Cell Line Cells ChIP-seq Childhood Chromatin DNA Binding DNA-Directed RNA Polymerase Drug Targeting Electrophoretic Mobility Shift Assay Enzymes Exhibits Face Fellowship Genes Genetic Screening Genetic Transcription Genome Guide RNA Hematology Human Immunocompromised Host Immunoprecipitation Impairment In Vitro Individual Investigation Knock-out Laboratories Leukemia Acute Lymphoblastic Chemotherapy Leukemic Cell Libraries Luciferases MLL gene Malignant Childhood Neoplasm Malignant neoplasm of lung Mass Spectrum Analysis Massive Parallel Sequencing Mediating Mus Oncology Patients Physicians Play Prognostic Marker Proteins RNA Polymerase II Recombinants Regulation Relapse Reporter Residencies Resistance Resistance development Risk Role Scientist Serum Subgroup Survival Rate T-Lymphocyte Testing Training Transcription Elongation Transcription Initiation Transcriptional Regulation Transposase Universities Up-Regulation Work Zinc Fingers activating transcription factor 4 acute lymphoblastic leukemia cell asparaginase career cell growth chromatin immunoprecipitation clinically relevant deprivation design experimental study genome-wide high risk histone modification in vivo insight leukemia malignant breast neoplasm pancreatic cancer cells programs promoter recruit resistance mechanism response therapeutic target transcription factor

项目摘要

Project Summary/Abstract L-asparaginase is a bacterial enzyme that depletes serum asparagine to inhibit leukemic cell growth in acute lymphoblastic leukemia. Some patients do not respond to L-asparaginase and the cellular mechanisms underlying non-response remain poorly understood. Under asparagine deprivation, cells activate a transcriptional program known as the amino acid deprivation response that is mediated by activating transcription factor 4 (ATF4). This program culminates in the expression of asparagine synthetase (ASNS), the enzyme responsible for the synthesis of asparagine. Other than ATF4, the transcriptional regulators involved in this response are poorly defined. To identify transcriptional genes involved in the response to L-asparaginase, I developed a CRISPR-Cas9 genetic screening approach to individually knock out each transcription-related gene in the genome. This approach allowed determination of genes that were required for the resistance of an ALL cell line to L- asparaginase. The top-scoring gene in my preliminary genetic screen was a poorly described transcription factor, ZBTB1. Knockout of ZBTB1 sensitized this ALL cell line to treatment with L-asparaginase. Preliminary work suggested that ZBTB1 enriches in the promoter of ASNS to promote transcription. I hypothesize that ZBTB1 and the regulation of ASNS expression may be a suitable drug target in asparaginase resistant leukemia. A lack of insight into the mechanistic role of ZBTB1 in mediating transcription, however, precludes investigation of this hypothesis. In this proposal, building on my preliminary work, I will test the hypothesis that ZBTB1 positively regulates the transcription of ASNS and may be a therapeutic target for L-asparaginase resistant ALLs. In Aim 1, I will investigate the mechanism by which ZBTB1 regulates the expression of ASNS. In Aim 2, I will determine whether ZBTB1 knockout sensitizes human ALL cell lines and primary human ALLs to treatment with L- asparaginase in vivo. I anticipate that these studies will determine: 1) the mechanistic role of ZBTB1 in the response to L-asparaginase in leukemic cells and 2) the potential of ZBTB1 as a therapeutic target in L- asparaginase resistant ALL. This work will be completed in the laboratory of Dr. Kivanc Birsoy with the co- advisement of Dr. Robert Roeder at the Rockefeller University. The training plan outlined in this proposal is designed to best prepare me for a career as an independent physician-scientist following residency and fellowship training in hematology and oncology.

项目概要/摘要 L-天冬酰胺酶是一种细菌酶，可消耗血清天冬酰胺以抑制急性白血病细胞的生长淋巴细胞白血病。一些患者对 L-天冬酰胺酶没有反应及其细胞机制潜在的不回应仍然知之甚少。在缺乏天冬酰胺的情况下，细胞激活称为氨基酸剥夺反应的转录程序，由激活介导转录因子 4 (ATF4)。该程序最终导致天冬酰胺合成酶 (ASNS) 的表达，负责合成天冬酰胺的酶。除 ATF4 外，转录调控因子还参与该响应的定义不明确。为了鉴定参与 L-天冬酰胺酶反应的转录基因，我开发了 CRISPR-Cas9 基因筛选方法单独敲除基因组中每个转录相关基因。这该方法可以确定 ALL 细胞系对 L- 的抗性所需的基因。天冬酰胺酶。在我的初步基因筛选中得分最高的基因是一个描述不佳的转录因素，ZBTB1。 ZBTB1 的敲除使该 ALL 细胞系对 L-天冬酰胺酶治疗敏感。初步的研究表明，ZBTB1 富集 ASNS 启动子以促进转录。我假设 ZBTB1 和 ASNS 表达的调节可能是天冬酰胺酶耐药的合适药物靶点白血病。然而，缺乏对 ZBTB1 在介导转录中的机制作用的了解，排除了对这一假设的调查。在本提案中，基于我的前期工作，我将测试 ZBTB1 积极调节的假设 ASNS 的转录，可能是 L-天冬酰胺酶耐药 ALL 的治疗靶点。在目标 1 中，我将研究ZBTB1调节ASNS表达的机制。在目标 2 中，我将确定 ZBTB1 敲除是否会使人类 ALL 细胞系和原代人类 ALL 对 L-治疗敏感体内的天冬酰胺酶。我预计这些研究将确定：1）ZBTB1 在白血病细胞对 L-天冬酰胺酶的反应以及 2) ZBTB1 作为 L-天冬酰胺酶治疗靶点的潜力天冬酰胺酶耐药性 ALL。这项工作将在 Kivanc Birsoy 博士的实验室与合作者完成洛克菲勒大学罗伯特·罗德博士的建议。本提案中概述的培训计划是旨在为我在住院医师实习后成为一名独立医师科学家的职业生涯做好最好的准备血液学和肿瘤学进修培训。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Dietary thiamine influences l-asparaginase sensitivity in a subset of leukemia cells.

膳食硫胺素影响白血病细胞亚群的 L-天冬酰胺酶敏感性。

DOI：
发表时间：
2020
期刊：
影响因子：
13.6
作者：
Guarecuco, Rohiverth;Williams, Robert T;Baudrier, Lou;La, Konnor;Passarelli, Maria C;Ekizoglu, Naz;Mestanoglu, Mert;Alwaseem, Hanan;Rostandy, Bety;Fidelin, Justine;Garcia;Molina, Henrik;Birsoy, Kıvanç
通讯作者：
Birsoy, Kıvanç

ZBTB1 Regulates Asparagine Synthesis and Leukemia Cell Response to L-Asparaginase.

DOI：
10.1016/j.cmet.2020.03.008
发表时间：
2020-04-01
期刊：
Cell metabolism
影响因子：
29
作者：
Robert T. Williams;Rohiverth Guarecuco;Leah A. Gates;D. Barrows;M. C. Passarelli;B. Carey;Lou Baudrier;Swarna K A Jeewajee;Konnor C. La;Benjamin Prizer;S. Malik;J. Garcia;Xiphias Ge Zhu;J. Cantor;H. Molina;T. Carroll;R. Roeder;O. Abdel;C. Allis;Kıvanç Birsoy
通讯作者：
Kıvanç Birsoy