The role of PARP10 in alleviating replication stress and promoting cellular proliferation and tumorigenesis

PARP10在缓解复制应激、促进细胞增殖和肿瘤发生中的作用

基本信息

批准号：
10552014
负责人：
Claudia M Nicolae
金额：
$ 36.09万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
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：
2021-02-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10552014
关键词：
ADP Ribose Transferases Address Adenosine Diphosphate Ribose Apoptosis Biochemical Biological Biological Assay Breast Bypass CRISPR/Cas technology Cell Cycle Progression Cell Line Cell Proliferation Cell physiology Cells Cellular biology Chromosome Fragile Sites Clinic Coupled DNA DNA Biochemistry DNA Repair DNA Structure DNA biosynthesis DNA lesion DNA replication fork DNA sequencing DNA-Directed DNA Polymerase Down-Regulation Enzymes Epithelium Exposure to Family Fiber Genes Genetic Models Genetic Transcription Genome Stability Genomic DNA Genomic Instability Genomics Growth Hela Cells Human Immunocompromised Host In Vitro Knock-out Malignant Neoplasms Measures Mediating Mitochondria Modeling Molecular Mus Mutagenesis Mutation Normal Cell Nucleic Acids Oncogenes Oncogenic Ovarian Pathway interactions Phenotype Poly(ADP-ribose) Polymerases Polymerase Post-Translational Protein Processing Process Proliferating Property Proteins Resistance Role S phase Signal Transduction Structure Testing Transgenic Mice Tumor Promotion Ubiquitination Validation brca gene cancer cell carcinogenesis cell motility design genetic manipulation genome editing genomic tools in vivo inhibitor member mouse genetics mouse model mutant next generation next generation sequencing novel overexpression oxidation personalized cancer therapy recruit replication stress targeted cancer therapy transcriptome sequencing transcriptomics tumor tumor growth tumor xenograft tumorigenesis

项目摘要

Project Summary Identification of molecular pathways that are preferentially employed and relied upon by cancer cells compared to normal cells is key to designing novel personalized cancer therapies. PARP10 is a poorly characterized member of the PARP family. We previously showed that PARP10 promotes translesion synthesis (TLS)- mediated bypass of DNA lesions during DNA replication, thereby alleviating replication stress. More recently, we also showed that PARP10 is a novel oncogene. We found that the PARP10 gene is amplified and/or overexpressed in a large number of tumors including breast and ovarian, with very few observed occurrences of downregulation or loss. We found that the PARP10 gene is amplified and/or overexpressed in a large proportion of human tumors including breast and ovarian, with almost no occurrences of downregulation or loss. Moreover, we found that PARP10 overexpression in, non-transformed human epithelial RPE1 cells results in enhanced proliferation, resistance to replication stress, and increased xenograft tumor formation in immunocompromised mice. The opposing phenotypes were found upon knockout of PARP10 in cancer HeLa cells. These findings suggest that PARP10 is a putative oncogene and its expression promotes tumor formation and growth. Mutagenic TLS has been previously proposed to promote transformation by both allowing hyper-proliferation and inducing genomic instability. Thus, we hypothesize that PARP10 expression suppresses replication stress through TLS-mediated bypass of replication arresting structures, thereby allowing hyper-proliferation of cancer cells. We propose here to directly test this, in three specific aims which address the hypothesis at three different levels: Aim 1 will investigate the mechanism employed by PARP10 to modulate PCNA-dependent TLS at the molecular level, using biochemical and cellular localization and interaction assays. Aim 2 will functionally test the impact of this mechanism of cellular processes including genomic stability and DNA replication. Aim 3 will employ a mouse genetic model to unambiguously investigate if Parp10 expression induces tumor formation or promotes tumor growth. Using state-of-the-art cellular, molecular and genomic tools (including: CRISPR/Cas9-mediated genome editing; molecular DNA fiber combing to measure fork stability; next generation sequencing approaches to measure mutagenesis and mutation burden) we will investigate here the molecular mechanisms underlying this novel oncogenic function of PARP10. This may eventually result in validation of a new target for cancer therapy.

项目概要比较癌细胞优先采用和依赖的分子途径的鉴定对正常细胞的影响是设计新型个性化癌症疗法的关键。 PARP10 的特征很差 PARP家族的成员。我们之前表明 PARP10 促进跨损伤合成 (TLS) - 在 DNA 复制过程中介导绕过 DNA 损伤，从而减轻复制压力。最近，我们还表明 PARP10 是一种新型癌基因。我们发现 PARP10 基因被扩增和/或在包括乳腺癌和卵巢在内的大量肿瘤中过度表达，但观察到的情况很少下调或损失。我们发现 PARP10 基因在大量细胞中被扩增和/或过度表达。包括乳腺癌和卵巢癌在内的人类肿瘤的比例，几乎没有发生下调或损失。此外，我们发现 PARP10 在非转化的人上皮 RPE1 细胞中过度表达导致增殖增强、对复制应激的抵抗力以及异种移植肿瘤形成的增加免疫功能低下的小鼠。在癌症 HeLa 中敲除 PARP10 后发现了相反的表型细胞。这些发现表明 PARP10 是一种假定的癌基因，其表达可促进肿瘤发生形成和生长。之前已提出诱变 TLS 来促进两者的转化允许过度增殖并诱导基因组不稳定。因此，我们假设 PARP10 表达通过 TLS 介导的复制抑制结构旁路抑制复制应激，从而允许癌细胞过度增殖。我们在这里建议直接测试这一点，以三个具体目标在三个不同层面解决该假设：目标 1 将研究 PARP10 采用的机制利用生化和细胞定位在分子水平上调节 PCNA 依赖性 TLS，相互作用测定。目标 2 将在功能上测试这种细胞过程机制的影响，包括基因组稳定性和 DNA 复制。目标 3 将采用小鼠遗传模型来明确研究如果 Parp10 表达诱导肿瘤形成或促进肿瘤生长。使用最先进的蜂窝技术，分子和基因组工具（包括：CRISPR/Cas9 介导的基因组编辑；分子 DNA 纤维梳理以测量货叉稳定性；下一代测序方法来测量诱变和突变负担）我们将在这里研究这种新的致癌功能背后的分子机制 PARP10 的。这可能最终导致癌症治疗新靶标的验证。