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年来，在治疗威胁生命或使人衰弱的疾病（例如由HIV-1引起的疾病）中的治疗价值的几类抗病毒药物的发展开发，疱疹病毒和肝炎病毒病毒病毒和肝炎病毒病毒的疾病以及一些最有效的抗病毒体的抗病毒体可能会造成癌症癌症的危害和造成癌症的风险。例如，包括核苷类似物在内的组合抗病毒疗法已将HIV-1感染从致命疾病转化为慢性疾病，并在怀孕期间大大降低了病毒的垂直传播。然而，基于核苷类似物的方案的巨大益处，齐多夫丁（AZT）伴随着宿主细胞AZT-DNA掺入，细胞遗传学作用以及诸如HIV感染母亲和新生儿的HIV感染者A和HPRT（例如甘氨酸A和HPRT）中的基因中的诱变基因。这项试验研究的主要目的是确定在子宫内预防中，使用AZT导致p53肿瘤抑制基因突变的发生增加，这将来可能与暴露儿童中与癌症相关的健康结果联系起来。在没有抗病毒治疗的情况下，我们还将评估母体HIV感染的潜力，这是胎儿生命过程中突变的潜在来源。基于PCR使用牛皮岩钳制底漆的基于PCR的降低梯度凝胶电泳将用于定义p53外显子外显子中突变的频率和性质，与在健康未感染的母亲或接受的不受欢迎的母亲相比，在子宫内暴露于子宫内暴露于子宫内的婴儿的脐带血淋巴细胞。我们还将开发敏感的定量PCR特异性竞争阻滞剂测定法，以用于常见发生的p53突变，以检测使用DNA测序的突变，并确定样品中突变体与野生型DNA的比例。期望基于AZT的预防，而不是胎儿“暴露于母亲HIV-1”，将诱导p53基因突变。这一假设的肯定将为未来的研究奠定基础，以定义癌症风险的大小，以评估核苷 - 酰胺抗差异策略的相对安全性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基因突变在HIV-1感染患者中的作用。公共卫生相关性：需要对HIV感染的孕妇进行项目叙事治疗，以防止病毒感染婴儿；但是，当前使用齐多夫丁（AZT）等核苷类似物的治疗可能会损害DNA并在以后的生活中施加癌症的风险。这项工作的主要目的是确定怀孕期间暴露于AZT会导致p53肿瘤抑制剂的突变，该突变可能参与癌症的进展。