Mechanism of Folate Deficiency as a Co-Factor for HPV16-induced Carcinogenesis

叶酸缺乏作为 HPV16 诱发癌变的辅助因素的机制

基本信息

批准号：
8441816
负责人：
Asok Antony
金额：
--
依托单位：
RLR VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-01-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8441816
关键词：
Affinity Applications Grants Benign Binding Capsid Capsid Proteins Cells Clone Cells DNA DNA Fragmentation DNA Integration DNA Maintenance Dependovirus Developing Countries Diet Dose Elderly Ensure Exposure to Folate Folic Acid Deficiency Frequencies Gene Expression Generations Genes Genome Genomics HIV HPV-High Risk Health Healthcare Heterogeneous-Nuclear Ribonucleoproteins Human Papillomavirus Human papillomavirus 16 Immunodeficient Mouse Implant In Vitro Individual Infection Investigation L2 viral capsid protein Lead Leucovorin Malignant Neoplasms Measures Methotrexate Minor Modeling Molecular Mus Normal Cell Oncogenes Operon Papillomavirus Patients Physiological Positioning Attribute RNA RNA-Protein Interaction Risk Series Staging Testing Time Tissues Transcript Uracil Veterans Viral Viral Load result Vitamin B 12 Deficiency carcinogenesis ds-DNA feeding implantation in vivo keratinocyte monolayer mutant novel nutrition permissiveness prevent subcutaneous

Affinity Applications Grants Benign Binding Capsid Capsid Proteins Cells Clone Cells DNA DNA Fragmentation DNA Integration DNA Maintenance Dependovirus Developing Countries Diet Dose Elderly Ensure Exposure to Folate Folic Acid Deficiency Frequencies Gene Expression Generations Genes Genome Genomics HIV HPV-High Risk Health Healthcare Heterogeneous-Nuclear Ribonucleoproteins Human Papillomavirus Human papillomavirus 16 Immunodeficient Mouse Implant In Vitro Individual Infection Investigation L2 viral capsid protein Lead Leucovorin Malignant Neoplasms Measures Methotrexate Minor Modeling Molecular Mus Normal Cell Oncogenes Operon Papillomavirus Patients Physiological Positioning Attribute RNA RNA-Protein Interaction Risk Series Staging Testing Time Tissues Transcript Uracil Veterans Viral Viral Load result Vitamin B 12 Deficiency carcinogenesis ds-DNA feeding implantation in vivo keratinocyte monolayer mutant novel nutrition permissiveness prevent subcutaneous

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项目摘要

DESCRIPTION (provided by applicant): We have recently identified that homocysteinylation of heterogeneous nuclear ribonucleoprotein-E1 (hnRNP-E1) during physiological folate deficiency, activates a nutrition-sensitive posttranscriptional RNA operon that also includes an important interaction with two loci in Human Papillomavirus type-16 (HPV16) RNA. This HPV16 RNA-protein interaction with homocysteinylated-hnRNP-E1 led to a profound perturbation in the generation of both HPV16 major (L1) and minor (L2) viral capsid proteins in vitro; in HPV16-harboring keratinocytes that were propagated as monolayers; as well as when these HPV16-keratinocytes were developed into organotypic rafts in physiologic low-folate medium. Despite a similar HPV16 DNA viral load in HPV16-high folate- and low folate-organotypic rafts, the latter contained a high-level of integration of HPV16 DNA into genomic DNA. Subcutaneous implantation of 18-day HPV16-low folate-organotypic rafts in Beige Nude XID immunodeficient mice led to an aggressive HPV16-induced cancer within 12 weeks. Thus, we have developed a new model of HPV16-induced carcinogenesis within a time frame of less than 4 months. Because folate deficiency can induce single-strand nicks in genomic DNA and also double-strand DNA fragmentation, our overarching hypothesis is that the unstable genomic DNA in HPV16-low folate-organotypic rafts is primarily responsible for the high-level integration of HPV16 DNA into genomic DNA and transformation of this benign tissue to cancer. So we will test various aspects of this hypothesis using three specific aims: In Specific Aim #1, we will characterize the time-course relationship, extent, and consequence of integration of HPV16 DNA into the genomic DNA of HPV16-organotypic rafts in vitro and in vivo, and assess the frequency of transformation of benign HPV16-organotypic rafts into cancer in immunodeficient mice. In Specific Aim #2 we will assess the potential of amplified 'capsid-less' HPV16 DNA to integrate into unperturbed and intact [stable] genomic DNA after transduction and expression of novel genes encoding various strengths of molecular mimics of homocysteinylated-hnRNP-E1 into HPV16-harboring keratinocytes that are subsequently developed into organotypic rafts under high-folate conditions. In Specific Aim #3, we will assess the permissiveness of unperturbed and transiently perturbed genomic DNA to integrate HPV16 DNA in AAV2-transduced [high folate] HPV16-organotypic rafts that either do or do not contain an abundance of 'capsid-less' HPV16 DNA, and then evaluate the potential for such genomic HPV16 DNA integration to induce carcinogenesis within implants of rafts in Beige Nude XID mice. Such investigations will ultimately provide a better understanding of the mechanism of transformation of HPV16-infected tissues to cancer and benefit HPV-infected elderly Veterans with poor nutrition; Veterans with HPV16 and human immunodeficiency virus (HIV); and those in developing countries where the combination of poor nutrition, and co-infection with HIV and HPV16 places individuals at high risk for HPV-induced cancers

描述（由申请人提供）：我们最近确定，在生理叶酸缺乏期间，异质性核核糖核蛋白-E1（HNRNP-E1）的同型同型静态化，激活了对营养敏感的转录后RNA操纵子，该操纵子也包括与人类Papillomavirus type-16（HPV16）中的两个基因座的重要相互作用。这种HPV16 RNA蛋白质与同型凝固性的HNRNP-E1的相互作用导致HPV16 Major（L1）和Minar（L2）病毒capsid蛋白在体外产生深远的扰动；在HPV16-HARBORING角质形成细胞中，它们被传播为单层；以及当这些HPV16-甲状腺细胞发展为生理低叶酸培养基中的器官筏时。尽管HPV16 DNA病毒载量相似，但在HPV16高和低叶酸 - 叶酸 - 构型筏中，后者含有将HPV16 DNA整合到基因组DNA中的高级整合。米色裸体XID免疫缺陷小鼠中18天的HPV16-low叶酸 - 叶酸 - 构型筏的皮下植入，导致了12周内侵略性HPV16诱导的癌症。因此，我们在少于4个月的时间范围内开发了一种新的HPV16诱导的致癌作用模型。 Because folate deficiency can induce single-strand nicks in genomic DNA and also double-strand DNA fragmentation, our overarching hypothesis is that the unstable genomic DNA in HPV16-low folate-organotypic rafts is primarily responsible for the high-level integration of HPV16 DNA into genomic DNA and transformation of this benign tissue to cancer. 因此，我们将检验该假设的各个方面使用三个特定目标：在特定的目标＃1中，我们将表征时刻关系的关系，将HPV16 DNA整合到体外和体内HPV16- Oranotypic Rafts的基因组DNA中的程度和结果，并评估了免疫缺陷小鼠中良性HPV16-有机筏向癌症转化为癌症的频率。在特定目的＃2中，我们将评估放大“无冠状” HPV16 DNA的潜力，以在转导和表达各种编码分子模拟物的各种分子模拟物的分子模拟物中的分子模拟物中的分子模拟物中，将同生构基化的hnrnp-e1纳入HPV16-HARBORITITICTHARBORITITICTHARBORITITICTYERIPTICY，将基因组成的基因组合和表达分子进行了分子模拟的各种分子模拟的各种强度后，将其整合到不受干扰且完整的[稳定]基因组DNA中的潜力。高叶酸条件下的筏。在特定的目标＃3中，我们将评估不受干扰和瞬时扰动的基因组DNA的允许性，以整合AAV2转导的[高叶酸] HPV16-构造筏中的HPV16 DNA，这些割草的功能或不包含'capsid-capsid-capsid-of-capv16 dna，并不包含大量的基因hpv16 dNA，并不包含hpv16 dna的范围。诱导米色裸Xid小鼠木筏植入物中的致癌作用。此类研究最终将更好地理解HPV16感染组织转化为癌症的机理，并使感染HPV的老年退伍军人的营养不良。具有HPV16和人类免疫缺陷病毒（HIV）的退伍军人；在发展中国家营养不良以及与艾滋病毒和HPV16共同感染的发展中国家，将HPV引起的癌症的个人高风险