Risk for in vivo mutagenesis of the P53 gene by nucleoside analog antiviral drug

核苷类似物抗病毒药物体内 P53 基因突变的风险

基本信息

批准号：
8150962
负责人：
VERNON E WALKER
金额：
$ 7.4万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-28 至 2012-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8150962
关键词：
2&apos -Deoxythymidine Alleles Animals Antiviral Agents Antiviral Therapy Biological Assay Cancer Gene Mutation Carcinogens Cells Child Chronic Disease Control Groups Cytogenetics DNA DNA Damage DNA Fingerprinting DNA Sequence Defect Detection Development Disease Dose Drug usage Environment Exons Exposure to Ficusin Frequencies Future Gene Mutation Genes Goals HIV HIV Infections HIV-1 Health Hepatitis Viruses Herpesviridae Heterochromatin Human Infant Infection Life Link Lung Neoplasms Lymphocyte Malignant Neoplasms Methods Modality Mothers Mus Mutagenesis Mutation Nature Newborn Infant Outcome Patients Perinatal Exposure Pharmaceutical Preparations Pilot Projects Population Pregnancy Pregnant Women Prophylactic treatment Proto-Oncogenes Psoralens Regimen Relative (related person)Reporter Genes Risk Rodent Role Safety Source Specimen TP53 gene Testing Therapeutic Tissues Tumor Suppressor Genes Tumor Suppressor Proteins Umbilical Cord Blood Vertical Disease Transmission Virus Virus Diseases Work Zidovudine base body system cancer risk denaturing gradient gel electrophoresis expectation exposed human population falls fetal in utero in vivo mutant nucleoside analog prevent public health relevance response success treatment effect tumor progression

项目摘要

DESCRIPTION (provided by applicant): The last 40 years have seen the development of several classes of antiviral drugs with therapeutic value in treating life-threatening or debilitating diseases such as those caused by HIV-1, herpesviruses, and hepatitis viruses, and some of the most effecacious antiviral agents include nucleoside analogs that also carry the potential to damage DNA and impose long-term cancer risks. For example, combination antiviral therapies including nucleoside analogs have transformed HIV-1 infection from a fatal disease to a chronic illness, and dramatically reduced vertical transmission of the virus during pregnancy. However, the great benefits of regimens based upon the nucleoside analog, zidovudine (AZT) are accompanied by host cell AZT-DNA incorporation, cytogenetic effects, and mutagenic responses in reporter genes like glyophorin A and HPRT of HIV-infected mothers and their newborns. The main goal of this pilot study is to determine if in utero HIV-1 prophylaxis using AZT causes increased occurrence of P53 tumor suppressor gene mutations that may be linked in the future to cancer-related health outcomes in exposed children. We will also assess the potential of maternal HIV-infection in the absence of antiviral treatment as a potential source of mutations during fetal life. PCR-based denaturing gradient gel electrophoresis, using psoralen-clamped primers, will be used to define the frequency and nature of mutations in p53 Exons 5-9 of cord blood lymphocytes from infants exposed in utero to AZT compared with those born to healthy uninfected mothers or HIV-infected mothers receiving no antiviral treatment. We will also develop sensitive quantitative PCR-based allele-specific competitive blocker assays for frequently occurring P53 mutations to detect the mutation with DNA sequencing and to determine the proportion of mutant to wild-type DNA in specimens. The expectation is that AZT-based prophylaxis, but not fetal "exposure" to maternal HIV-1, will induce P53 gene mutations. Affirmation of this hypothesis will lay the groundwork for future research to define the magnitude of cancer risk, to assess the relative safety of nucleoside-analog sparing strategies or nucleoside analogs thought to be less toxic, and to determine the role of drug-induced P53 gene mutations in HIV-1-infected patients. PUBLIC HEALTH RELEVANCE: PROJECT NARRATIVE Treatment of HIV-infected pregnant women is needed to prevent the virus from infecting the baby; however, the current treatments that use nucleoside analogs like zidovudine (AZT) may damage DNA and impose a risk for cancer later in life. The main goal of this work is to determine exposure to AZT during pregnancy causes mutations in the P53 tumor suppressor that can be involved in the progression of cancer.

描述（由申请人提供）：过去 40 年中，已经开发出几类具有治疗价值的抗病毒药物，可用于治疗危及生命或使人衰弱的疾病，例如由 HIV-1、疱疹病毒和肝炎病毒以及某些病毒引起的疾病。最有效的抗病毒药物包括核苷类似物，它们也有可能损害 DNA 并带来长期癌症风险。例如，包括核苷类似物在内的联合抗病毒疗法已将 HIV-1 感染从致命性疾病转变为慢性疾病，并显着减少了妊娠期间病毒的垂直传播。然而，基于核苷类似物齐多夫定 (AZT) 的治疗方案的巨大益处伴随着宿主细胞 AZT-DNA 掺入、细胞遗传学效应以及感染 HIV 的母亲及其新生儿的血型蛋白 A 和 HPRT 等报告基因的诱变反应。这项试点研究的主要目标是确定使用 AZT 进行子宫内 HIV-1 预防是否会增加 P53 肿瘤抑制基因突变的发生率，这些突变可能在未来与暴露儿童的癌症相关健康结果相关。我们还将评估在没有抗病毒治疗的情况下母体艾滋病毒感染作为胎儿生命中潜在突变来源的可能性。基于 PCR 的变性梯度凝胶电泳，使用补骨脂素钳位引物，将用于确定与健康未感染母亲所生婴儿相比，子宫内暴露于 AZT 的婴儿的脐带血淋巴细胞 p53 外显子 5-9 的突变频率和性质或未接受抗病毒治疗的艾滋病毒感染母亲。我们还将针对频繁发生的 P53 突变开发灵敏的基于定量 PCR 的等位基因特异性竞争性阻断剂测定法，以通过 DNA 测序检测突变，并确定样本中突变体与野生型 DNA 的比例。预期基于 AZT 的预防，而不是胎儿“暴露”于母体 HIV-1，将诱发 P53 基因突变。这一假设的确认将为未来的研究奠定基础，以确定癌症风险的大小，评估核苷类似物保留策略或被认为毒性较小的核苷类似物的相对安全性，并确定药物诱导的 P53 基因的作用HIV-1感染者的突变。公共卫生相关性：项目叙述需要对感染艾滋病毒的孕妇进行治疗，以防止病毒感染婴儿；然而，目前使用齐多夫定 (AZT) 等核苷类似物的治疗可能会损伤 DNA，并在以后的生活中增加罹患癌症的风险。这项工作的主要目标是确定怀孕期间接触 AZT 会导致 P53 肿瘤抑制因子发生突变，从而参与癌症的进展。