Nuclear IRS-1-DNA repair and mutagenesis in medulloblastoma

髓母细胞瘤中的核 IRS-1-DNA 修复和诱变

• 批准号: 8256598
• 负责人: Krzysztof Reiss
• 金额: $ 25.25万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-06 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8256598
• 关键词: Address; Affect; Attenuated; Benign; Binding; Binding Sites; Biological Assay; Biopsy; Cell Fractionation; Cell Nucleus; Cells; Central Nervous System Neoplasms; Cerebellar Neoplasms; Childhood; Childhood Cerebellar Neoplasm; Clinical; Collaborations; Collection; Complex; Computer software; Cytoplasmic Granules; DNA Damage; DNA Repair; DNA Repair Inhibition; DNA repair protein; Defect; Development; Epigenetic Process; Estradiol; Estrogen Receptor beta; Estrogen Receptors; Evaluation; Event; Frozen Sections; Future; Gene Expression; Genetic; Genomic Instability; Green Fluorescent Proteins; Growth; Health; Human; Hyperplasia; ICI 182780; In Vitro; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Insulin-Like Growth Factor Receptor; JC Virus; Laboratories; Large T Antigen; Letters; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of cerebellum; Mediating; Methodology; Microscopy; Molecular; Molecular Target; Mutagenesis; Mutation; Neuraxis; Nuclear; Nuclear Translocation; Oncogenic; Paraffin Embedding; Pathway interactions; Patients; Polyomavirus; Proteins; Receptor Signaling; Recombinants; Recurrence; Reporter; Research; Research Personnel; Research Proposals; Sampling; Series; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Source; System; Testing; Therapeutic; Thick; Tissue Microarray; Transgenic Mice; Tumor Tissue; Universities; Viral Tumor Antigens; base; cancer cell; clinically relevant; homologous recombination; inhibitor/antagonist; insulin receptor substrate 1 protein; medulloblastoma; medulloblastoma cell line; mutant; neoplastic cell; novel therapeutics; outcome forecast; overexpression; prevent; protein distribution; protein protein interaction; research study; response; tumor; tumor progression

DESCRIPTION (provided by applicant): This is a competing continuation of a RO1 project "IRS-1 JCV T-antigen interaction in Cerebellar Tumors". The results from previous founding period demonstrate that Insulin Receptor Substrate 1 (IRS-1), which is the major cytosolic component of the IGF-I receptor signaling system, can translocate to the nucleus. Importantly, we have detected nuclear IRS-1 in medulloblastoma cell lines and in medulloblastoma clinical samples, which were positive for polyomavirus JC large T-antigen. Our further studies demonstrate that nuclear IRS-1 can inhibit faithful component of DNA repair (homologous recombination DNA repair, HRR) via a direct interaction between IRS-1 and Rad51 detected at the sites of damaged DNA. We also demonstrated that the interference with DNA repair by nuclear IRS-1, and subsequent accumulation of mutations may happen in the absence of JCV T-antigen. In this respect, estrogen receptor beta (ER¿), which is highly expressed in medulloblastomas, can translocate IRS-1 to the nucleus in JCV T-antigen negative cells. These findings indicate that the interaction between IRS-1 and ER¿ could be engaged in cellular responses to DNA damage. It also suggests that inhibition of the faithful DNA repair by nuclear IRS-1 is a common event, not restricted to the medulloblastoma cases which are JCV positive. Therefore, we have developed a research plan which will analyze fundamental mechanisms leading to the development of genomic instability in medulloblastomas. The proposal is founded on the hypothesis that highly active ER¿ found in medulloblastoma mediates translocation of IRS-1 to the nucleus causing inhibition of HRR, unfaithful DNA repair, and the development of genomic instability in actively growing medulloblastoma cells. To test this hypothesis, we have developed a series of experiments, which are outlined in three Specific Aims. In Aim #1, we will evaluate ER¿ effects on IGF-I dependent and IGF-I -independent components of HRR, and will characterize binding and subcellular localizations among ER¿, IRS-1 and Rad51 in human medulloblastoma cell lines. In Aim#2, we will evaluate how pharmacological and molecular manipulations with the IGF-IR-IRS-1-E¿ signaling axis affect DNA repair fidelity in vitro and tumor progression in RCAS/tv-a transgenic mice. Finally, in Aim #3, we will analyze protein levels, subcellular localization and co-localization among IRS-1, ER¿, and Rad51 in clinical samples obtained from patients with medulloblastoma. Since DNA damaging agents are frequently used to eliminate CNS tumors including medulloblastoma, defects in DNA repair fidelity may have strong mutagenic consequences. This in turn may result in tumor progression or tumor recurrences after genotoxic treatment. Better understanding of the signaling cross-talk between IRS-1, ER¿ and DNA repair proteins is expected to provide new molecular targets for future therapeutic strategies against genomic instability, which is common in medulloblastoma patients. PUBLIC HEALTH RELEVANCE Medulloblastomas are cerebellar tumors of the childhood, in which the spread of tumor cells within central nervous system (CNS) is associated with poor prognosis. Since DNA damaging agents are frequently used to eliminate medulloblastoma, defects in DNA repair mechanisms are strongly suspected to cause accumulation of mutations contributing to tumor progression or tumor recurrences. The proposed studies of DNA repair mechanisms in association with cell signaling pathways are critically important for the development of new therapeutic strategies against malignant dissemination of these cerebellar tumors in CNS.

描述（由适用提供）：这是RO1项目“小脑肿瘤中的IRS-1 JCV T-抗原相互作用”的竞争延续。先前成立期的结果表明，胰岛素受体底物1（IRS-1）是IGF-I受体信号系统的主要胞质成分，可以转化为核。重要的是，我们已经在髓母细胞瘤细胞系和髓母细胞瘤临床样品中检测到核IRS-1，这些样品对多抗原大型T-抗原呈阳性。我们的进一步研究表明，核IRS-1可以通过在受损的DNA部位检测到的IRS-1和RAD51之间的直接相互作用来抑制DNA修复的失败成分（同源重组DNA修复，HRR，HRR）。我们还证明了核IRS-1对DNA修复的干扰，随后在没有JCV T抗原的情况下可能会发生突变。在这方面，在髓母细胞瘤中高度表达的雌激素受体β（ER？）可以将IRS-1转移到JCV T抗原阴性细胞中的核。这些发现表明，IRS-1和ER¿之间的相互作用可能参与细胞对DNA损伤的反应。这也表明，核IRS-1对DNA修复失败的抑制是一个常见事件，不仅限于JCV阳性的髓母细胞瘤病例。因此，我们制定了一项研究计划，该计划将分析导致髓母细胞瘤基因组不稳定性发展的基本机制。该提案建立在以下假设的基础上：在髓母细胞瘤中发现的高度活性ER介导了IRS-1的易位，从而导致HRR抑制，不忠DNA修复和基因组不稳定性在主动生长的髓母细胞瘤细胞中的发展。为了检验这一假设，我们开发了一系列实验，这些实验以三个特定目的概述。在AIM＃1中，我们将评估HRR的IGF-I依赖性和IGF-I非依赖性成分的影响，并将表征人类髓母细胞瘤细胞系中ER？，IRS-1和RAD51之间的结合和亚细胞局部化。在AIM＃2中，我们将评估使用IGF-IR-IRS-1-E em轴的药物和分子操作如何在体外影响DNA修复保真度和RCAS/TV-A转基因小鼠的肿瘤进展。最后，在AIM＃3中，我们将分析来自髓母细胞瘤患者获得的临床样本中IRS-1，ER？和RAD51之间蛋白质水平，亚细胞定位和共定位。由于DNA损伤剂经常用于消除包括髓母细胞瘤在内的中枢神经系统肿瘤，因此DNA修复保真度缺陷可能具有强大的诱变后果。反过来，这可能会导致肿瘤治疗后肿瘤进展或肿瘤恢复。对IRS-1，ER？和DNA修复蛋白之间的信号传导串扰的更好理解有望为针对基因组不稳定性的未来治疗策略提供新的分子靶标，这在髓母细胞瘤患者中很常见。公共卫生相关性髓母细胞瘤是童年的小脑肿瘤，其中中枢神经系统（CNS）中肿瘤细胞的扩散与预后不良有关。由于DNA损伤剂经常用于消除髓母细胞瘤，因此强烈怀疑DNA修复机制中的缺陷会导致突变加速，从而导致肿瘤进展或肿瘤恢复。对DNA修复机制与细胞信号传导途径相关的拟议研究对于开发新的治疗策略至关重要，反对CNS中这些小脑肿瘤的恶性传播。

项目成果

Differential Effects of MicroRNAs on Glioblastoma Growth and Migration.

• DOI: 10.3390/genes4010046
• 发表时间: 2013-03-04
• 期刊: Genes
• 影响因子: 3.5
• 作者: Jeansonne D;Pacifici M;Lassak A;Reiss K;Russo G;Zabaleta J;Peruzzi F
• 通讯作者: Peruzzi F

Insulin-like growth factor-I-forkhead box O transcription factor 3a counteracts high glucose/tumor necrosis factor-α-mediated neuronal damage: implications for human immunodeficiency virus encephalitis.

• DOI: 10.1002/jnr.22542
• 发表时间: 2011-02
• 期刊: JOURNAL OF NEUROSCIENCE RESEARCH
• 影响因子: 4.2
• 作者: Wilk, Anna;Urbanska, Katarzyna;Yang, Shuo;Wang, Jin Ying;Amini, Shohreh;Del Valle, Luis;Peruzzi, Francesca;Meggs, Leonard;Reiss, Krzysztof
• 通讯作者: Reiss, Krzysztof

Fenofibrate subcellular distribution as a rationale for the intracranial delivery through biodegradable carrier.

非诺贝特亚细胞分布作为通过可生物降解载体进行颅内递送的基本原理。

• 发表时间: 2015
• 期刊: Journal of physiology and pharmacology : an official journal of the Polish Physiological Society
• 作者: Grabacka,M;Waligorski,P;Zapata,A;Blake,DA;Wyczechowska,D;Wilk,A;Rutkowska,M;Vashistha,H;Ayyala,R;Ponnusamy,T;John,VT;Culicchia,F;Wisniewska-Becker,A;Reiss,K
• 通讯作者: Reiss,K

Peroxisome proliferator activated receptor α ligands as anticancer drugs targeting mitochondrial metabolism.

• DOI: 10.2174/1389201011314030009
• 发表时间: 2013
• 期刊: Current pharmaceutical biotechnology
• 影响因子: 2.8
• 作者: Grabacka M;Pierzchalska M;Reiss K
• 通讯作者: Reiss K

Polycyclic aromatic hydrocarbons-induced ROS accumulation enhances mutagenic potential of T-antigen from human polyomavirus JC.

• DOI: 10.1002/jcp.24375
• 发表时间: 2013-11
• 期刊: JOURNAL OF CELLULAR PHYSIOLOGY
• 影响因子: 5.6
• 作者: Wilk, Anna;Waligorski, Piotr;Lassak, Adam;Vashistha, Himanshu;Lirette, David;Tate, David;Zea, Arnold H.;Koochekpour, Shahriar;Rodriguez, Paulo;Meggs, Leonard G.;Estrada, John J.;Ochoa, Augusto;Reiss, Krzysztof
• 通讯作者: Reiss, Krzysztof