Identifying and understanding drivers of chemoresistance in small cell lung cancer

识别和了解小细胞肺癌化疗耐药的驱动因素

基本信息

批准号：
10753857
负责人：
David MacPherson
金额：
$ 54.57万
依托单位：
FRED HUTCHINSON CANCER CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10753857
关键词：
Bromodeoxyuridine Cancer Patient Cancer cell line Cell Death Chemoresistance Chromatin Cisplatin Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Complex DNA Damage Data Etoposide Family member Gene Targeting Genes Genetic Genetic Transcription Genomics Histone H3 Human In Situ Nick-End Labeling In Vitro Kinetics Knowledge Libraries MYCL1 gene MYCN gene Malignant neoplasm of lung Modeling Molecular Mus Mutation Neurosecretory Systems Parents Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Process Proliferating Proteins Proteomics RNA analysis Refractory Relapse Resistance Role SAGA System Testing Tissues cancer type chemotherapy epigenetic silencing established cell line genetic manipulation histone modification in vivo insight lentivirally transduced lung cancer cell member mutant novel overexpression patient derived xenograft model patient response prevent response screening small cell lung carcinoma tissue culture transcriptome sequencing treatment response tumor growth

Bromodeoxyuridine Cancer Patient Cancer cell line Cell Death Chemoresistance Chromatin Cisplatin Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Complex DNA Damage Data Etoposide Family member Gene Targeting Genes Genetic Genetic Transcription Genomics Histone H3 Human In Situ Nick-End Labeling In Vitro Kinetics Knowledge Libraries MYCL1 gene MYCN gene Malignant neoplasm of lung Modeling Molecular Mus Mutation Neurosecretory Systems Parents Pathway interactions Patients Pharmaceutical Preparations Phosphorylation Process Proliferating Proteins Proteomics RNA analysis Refractory Relapse Resistance Role SAGA System Testing Tissues cancer type chemotherapy epigenetic silencing established cell line genetic manipulation histone modification in vivo insight lentivirally transduced lung cancer cell member mutant novel overexpression patient derived xenograft model patient response prevent response screening small cell lung carcinoma tissue culture transcriptome sequencing treatment response tumor growth

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项目摘要

SUMMARY Small cell lung cancer (SCLC) is an aggressive and lethal neuroendocrine lung cancer type. Most patients initially respond to chemotherapy but relapse occurs within months and genetic alterations that drive chemoresistance are poorly understood. Beyond amplification of MYC family members and epigenetic silencing of SLFN11, the field has an extremely poor understanding of genes that promote SCLC chemoresistance. We developed a novel system in which we genetically alter highly chemosensitive patient derived xenograft (PDX) models of SCLC to identify perturbations that confer resistance to cisplatin/etoposide (CIS-ETO) in vivo. Lentiviral overexpression of either MYCN or MYCL caused complete switch to chemoresistance (Grunblatt et al, 2020). To systematically identify SCLC chemoresistance drivers, we expanded use of this PDX lentiviral transduction system to perform in vivo CRISPR inactivation screens. We identified sgRNAs targeting multiple components of the SAGA (Spt- Ada-Gcn5 acetyltransferase) chromatin modifying complex as screen hits and confirmed that deleting the SAGA member USP22, a deubiquitylase, indeed confers chemoresistance in two SCLC PDX models, while return of USP22 to a USP22-null SCLC PDX model re-sensitizes to chemotherapy. Our overarching hypothesis is that suppressing the expression of USP22 and SAGA complex members drives chemoresistance in SCLC, and that transcriptional changes caused by SAGA suppression are critical. Aim 1, we will interrogate how genetically perturbing multiple SAGA complex members, including USP22 and TADA1, in PDX models alters the in vivo response to chemotherapy. Aim 2 employs genomic and proteomic approaches to develop a deep molecular understanding of the USP22-regulated genes and pathways that contribute to chemotherapy response in SCLC and uses human patient data to prioritize key SAGA targets for functional study. Decades of studying chemotherapy response in SCLC cell lines grown in vitro have provided little insight into how chemoresistance emerges, suggesting that key aspects of this process are not recapitulated under tissue culture conditions. Our novel system prioritizes the study of chemoresistance using in vivo approaches with potential to provide foundational knowledge to help prevent chemoresistance or re-sensitize chemoresistant SCLC to chemotherapy.

概括小细胞肺癌（SCLC）是一种侵略性和致命的神经内分泌肺癌类型。大多数患者最初对化学疗法的反应，但复发发生在几个月内，遗传改变会驱动化学疗法知之甚少。除了扩大MYC家庭成员和SLFN11的表观遗传沉默之外，场对促进SCLC化学抗性的基因的理解非常差。我们开发了一本小说我们在遗传上改变了高度化学敏感的患者衍生异种移植（PDX）模型的系统鉴定在体内赋予顺铂/依托泊苷（顺泊替型）的扰动。慢病毒过表达 MYCN或MYCL的完整切换到化学抗性（Grunblatt等，2020）。系统地识别SCLC化学抗性驱动因素，我们扩展了该PDX慢病毒转导系统的使用体内CRISPR灭活屏幕。我们确定了针对传奇多个组件的SGRNA（spt- ADA-GCN5乙酰转移酶）染色质修饰复合物作为屏幕命中并确认删除传奇成员USP22，一种去泛素层，确实赋予了两个SCLC PDX模型中的化学耐药性，而返回 USP22至USP22-NULL SCLC PDX模型重新敏感到化学疗法。我们的总体假设是抑制USP22和SAGA复合构件的表达驱动SCLC的化学耐药性，并且由传奇抑制引起的转录变化至关重要。目标1，我们将询问遗传学在PDX模型中扰动多个SAGA复杂成员，包括USP22和TADA1，改变了体内对化学疗法的反应。 AIM 2采用基因组和蛋白质组学方法来开发深层分子了解有助于SCLC化学疗法反应的USP22调节的基因和途径并使用人类患者数据来确定关键传奇目标进行功能研究。数十年的学习体外生长的SCLC细胞系中的化学疗法反应几乎没有深入了解化学抗性出现，表明该过程的关键方面在组织培养条件下没有概括。我们的新型系统优先使用具有潜力的体内方法来研究化学抗性有助于防止化学耐药或重新敏感化学耐药性SCLC的基础知识。