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

• 批准号: 10317276
• 负责人: JOHN J. TURCHI
• 金额: $ 50.9万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 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/10317276
• 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/antagonist; 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

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.

靶向复制蛋白A的新型DNA损伤应答治疗剂 摘要 DNA损伤反应（DDR）现在被认为是一个易于处理的途径，以靶向癌症治疗。的 DDR和DNA修复途径也适用于个性化治疗， PARP抑制剂在同源重组（HR）中的临床成功证明了相互作用 缺陷型癌症DDR是由PI 3激酶相关激酶ATM、ATR和DNA-1的参与启动的。 PK.这些激酶和下游检查点激酶CHK 1和CHK 2是临床验证的靶标 正在多个临床试验中积极研究。DDR途径中的一个新靶点是人类单核苷酸受体。 链DNA结合蛋白，复制蛋白A（RPA），其在DDR中起关键作用以检测 复制应激（RS）和信号的ATR激酶。RS是癌细胞的一个共同特征， DDR靶向药物抗癌活性的治疗窗口。斯普斯卡共和国加上解除武装、复员和重返社会的封锁， 复制灾难（RC）和最终的细胞死亡。RPA是防止RC和 当单链DNA结合不足时，RPA或“RPA耗尽”可引起RC和细胞死亡 保护基因组DNA的能力我们已经确定了一种有效的选择性小分子RPA抑制剂 （RPAi），NERx 329，其具有生化RPA抑制和RPA的细胞接合。NERx 329 在体内也没有明显的毒性，并且单独和与DNA组合具有抗癌活性 损害化疗药物和某些DDR靶向剂。虽然抗癌活性可能是 由于抑制RPA在个体修复或复制途径中的作用，我们发表的和初步的数据 指向一个更加全球化的行动机制。我们的总体假设是，化学抑制 RPA可以模拟RPA耗尽以抑制DDR，并通过新的途径提供选择性抗癌活性。 在肺癌和卵巢癌中常见的遗传相互作用。为了解决这一假设，我们将阐明 我们的新型RPAi的细胞抗癌活性的机制和决定因素。我们将重点关注两个 高级别浆液性上皮性卵巢癌（EOC）和非小细胞肺癌（NSCLC）， 我们对临床存活数据的分析揭示了RPA在这些癌症中的重要作用。在目标1中， 询问RPA的化学耗竭如何影响肺癌和卵巢癌中的RS和DDR。目标2将 重点是阐明RPAi敏感性的遗传决定因素。我们将发现新的化学遗传 在评估RPAi活性的DDR聚焦的CRISPR筛选中的相互作用。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球体线中进行鉴定。全基因组测序 将鉴定可能与RPAi反应相关的遗传改变和染色体重排 疗法从这些研究中，影响RPAi敏感性的特定遗传改变将通过以下方法进行验证： 肺癌和卵巢癌的体内PDX模型和DDR靶向发育治疗剂。完成 这些目标将定义RPAi的作用机制和决定RPAi敏感性的遗传相互作用。 这些数据对于开发新的DDR靶向癌症疗法和确定相关的药物至关重要。 遗传学特征，以最大限度地提高患者的反应和疗效。