Novel DNA damage response therapeutics targeting replication protein A

针对复制蛋白 A 的新型 DNA 损伤反应疗法

基本信息

批准号：
10432115
负责人：
JOHN J. TURCHI
金额：
$ 49.69万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10432115
关键词：
1-Phosphatidylinositol 3-Kinase Address Automobile Driving Biochemical CHEK1 gene CHEK2 gene CRISPR screen Cell Death Cells Characteristics Chemicals Chromosomal Rearrangement Chromosomes Clinical Clinical Trials Combined Modality Therapy Coupled DNA Binding DNA Damage DNA Repair Pathway DNA-dependent protein kinase Data Developmental Therapeutics Program Drug Targeting Epithelial Cells Epithelial ovarian cancer Genetic Genetic Determinism Genetic Models Genomic DNA Health Human Individual Malignant Neoplasms Malignant neoplasm of lung Malignant neoplasm of ovary Measures Methodology Modeling Molecular Mutation Non-Small-Cell Lung Carcinoma Oncogenic Pathway interactions Patients Pharmacology Phosphotransferases Play Publishing Research Role SS DNA BP Series Serous Signal Transduction Single-Stranded DNA Therapeutic Therapeutic Agents Toxic effect Tumor-Derived anticancer activity ataxia telangiectasia mutated protein cancer cell cancer type chemical genetics clinically relevant driver mutation effective therapy exhaustion genome sequencing homologous recombination in vivo inhibitor inhibitor therapy novel novel therapeutics patient derived xenograft model patient response personalized medicine repaired replication factor A replication stress response small molecule success targeted agent targeted cancer therapy targeted treatment therapeutic target treatment response treatment strategy whole genome

项目摘要

Novel DNA damage response therapeutics targeting replication protein A Abstract The DNA damage response (DDR) is now considered a tractable pathway to target for cancer therapy. The DDR and DNA repair pathways are also amenable to personalized therapies by exploiting synthetic lethal interactions as evidenced by the clinical success of PARP inhibitors in homologous recombination (HR) deficient cancers. The DDR is initiated by engagement of the PI3 kinase-related kinases ATM, ATR, and DNA- PK. These kinases and the downstream checkpoint kinases CHK1 and CHK2, are clinically validated targets being actively investigated in multiple clinical trials. A novel target in the DDR pathway is the human single stranded DNA binding protein, replication protein A (RPA) which plays a critical role in the DDR to detect replication stress (RS) and signal to the ATR kinase. RS is a common feature in cancer cells and provides the therapeutic window for the anticancer activity of DDR targeted drugs. RS coupled with a DDR blockade results in replication catastrophe (RC) and eventually cell death. RPA is a critical protector from RC and depletion of RPA or “RPA exhaustion” can elicit RC and cell death when there is insufficient single-strand DNA binding capacity to protect the genomic DNA. We have identified a potent and selective small molecule RPA inhibitor (RPAi), NERx 329, which possesses biochemical RPA inhibition and cellular engagement of RPA. NERx 329 also displays no overt toxicity in vivo and possesses anticancer activity alone and in combination with DNA damaging chemotherapeutics and certain DDR targeted agents. While the anticancer activity could be the result of inhibiting RPA’s role in individual repair or replication pathways, our published and preliminary data point to a more global mechanism of action. Our overarching hypothesis is that chemical inhibition of RPA can mimic RPA exhaustion to inhibit the DDR and provide selective anticancer activity via novel genetic interactions common in lung and ovarian cancer. To address this hypothesis, we will elucidate the mechanisms and determinants of the cellular anticancer activity of our novel RPAi. We will focus on two cancers types, high grade serous epithelial ovarian cancer (EOC) and non-small cell lung cancer (NSCLC), as our analysis of clinical survival data reveals an important role for RPA in these cancers. In Aim 1 we will interrogate how chemical exhaustion of RPA impacts RS and the DDR in lung and ovarian cancer. Aim 2 will focus on elucidating the genetic determinants of sensitivity to RPAi’s. We will identify novel chemical-genetic interactions in a DDR focused CRISPR screen assessing RPAi activity. Clinically relevant genetic interactions will be identified in a unique series of PD-EOC spheroid lines we have developed. Whole genome sequencing will identify genetic alterations and chromosomal rearrangements which may correlate with response to RPAi therapy. From these studies, specific genetic alterations that impact RPAi sensitivity will be validated with in vivo PDX models of lung and ovarian cancer and DDR targeted developmental therapeutic agents. Completion of these aims will define RPAi mechanism of action and the genetic interactions that dictate RPAi sensitivity. These data are crucial towards developing novel DDR targeted cancer therapeutics and identifying the relevant genetics characteristics to maximize patient response and efficacy.

新型DNA损伤反应疗法靶向复制蛋白A 抽象的现在，DNA损伤反应（DDR）被认为是癌症治疗靶向靶标的途径。这 DDR和DNA修复途径也可以通过利用合成致死性来适应个性化疗法 PARP抑制剂在同源重组（HR）中的临床成功证明了相互作用不足的癌症。 DDR是通过PI3激酶相关激酶ATM，ATR和DNA-的参与而引发的 PK。这些激酶和下游检查点激酶CHK1和CHK2是临床验证的目标在多个临床试验中积极研究。 DDR途径中的一个新目标是人类的单身滞留的DNA结合蛋白，复制蛋白A（RPA）在DDR中起关键作用以检测复制应力（RS）并向ATR激酶发出信号。 RS是癌细胞中的常见特征，并提供 DDR靶向药物的抗癌活性的治疗窗口。 RS与DDR封锁结果结合在复制灾难（RC）中，最终是细胞死亡。 RPA是RC的关键保护者，耗尽当单链DNA结合不足时保护基因组DNA的能力。我们已经确定了潜力和选择性的小分子RPA抑制剂（RPAI），NERX 329，具有RPA的生化RPA抑制和细胞参与。 Nerx 329 还没有显示体内明显的毒性，并且仅具有抗癌活性并与DNA结合破坏化学治疗剂和某些DDR靶向剂。而抗癌活性可能是抑制RPA在个体维修或复制途径中的作用的结果，我们已发布的和初步数据指出更全球的作用机制。我们的总体假设是化学抑制 RPA可以模仿RPA耗尽以抑制DDR并通过新颖肺癌和卵巢癌常见的遗传相互作用。为了解决这一假设，我们将阐明机制和确定我们新型RPAI的细胞抗癌活性。我们将专注于两个癌症类型，高级浆液上皮卵巢癌（EOC）和非小细胞肺癌（NSCLC）我们对临床生存数据的分析揭示了RPA在这些癌症中的重要作用。在目标1中，我们将询问RPA的化学耗尽如何影响肺癌和卵巢癌的DDR。 AIM 2意志专注于阐明对RPAI敏感性的遗传决定因素。我们将确定新颖的化学基因以DDR为中心的CRISPR屏幕评估RPAI活动中的相互作用。临床相关的遗传相互作用将在我们开发的一系列独特的PD-EOC球形线系列中识别。整个基因组测序将确定可能与RPAI的反应相关的遗传改变和染色体重排治疗。从这些研究中，将通过IN验证影响RPAI敏感性的特定遗传改变肺癌和卵巢癌和DDR靶向发育疗法的体内PDX模型。完成这些目标将定义RPAI作用机理和决定RPAI敏感性的遗传相互作用。这些数据对于开发新型DDR靶向癌症治疗至关重要，并确定相关的遗传学特征可最大程度地提高患者的反应和效率。