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Principal Investigator: Laimonis A. LAIMINS
Organization: NORTHWESTERN UNIVERSITY AT CHICAGO
Fiscal Year: 2020
Award: $355,500
Funding agency: National Cancer Institute
PROJECT SUMMARY
Human papillomaviruses (HPV) are the causative agents of cervical, anal and many oral cancers. While
prophylactic vaccines to prevent HPV infections have been developed, there is no effective therapeutic
treatment for existing HPV lesions. It is therefore of critical importance to understand how the productive life
cycle of high-risk HPVs is regulated to identify potential new therapeutic targets. HPVs infect stratified
squamous epithelia and link their productive life cycles to the differentiation of the infected cell. My laboratory
recently demonstrated that the amplification of HPV genomes in differentiating cells is dependent on activation
of the ataxia-telangiectasia mutated (ATM) kinase pathway. In contrast, the ATM pathway has minimal effects
on stable maintenance replication in undifferentiated cells. In HPV positive cells, members of the ATM
pathway, such as γ−H2AX, CHK2, and NBS1, are bound to viral genomes in distinct nuclear replication foci.
Our studies have shown that E7 and E1 can activate the DNA damage response in the absence of viral
genome replication and that they act through the innate immune regulator, STAT-5 and Tip60 to induce the
phosphorylation of ATM. Additional studies using complete viral genomes have indicate that additional viral
proteins, such as E5, can also activate the DNA damage pathways. Our experiments specifically implicate
homologous recombination factors in mediating HPV amplification. In addition to the ATM pathway, we recently
found that both the ataxia telangiectasia and Rad3-related (ATR) pathway, which is important for repair of
single strand breaks, along with the p38/MK2 factors are also critical for amplification.. Additional recent
studies demonstrate that the cohesin, SMC1, is activated in HPV positive cells as part of the DNA damage
response and that it forms complexes with the transcriptional insulator, CTCF, at specific sites on HPV
genomes. Knockdown of either SMC1 or CTCF blocks differentiation-dependent HPV genome amplification
and mutation of SMC1/CTCF binding sites in HPV 31 interferes with stable maintenance of viral episomes.
While we have identified members of the DNA damage pathways that are important for differentiation
dependent amplification, the mechanism by which they act is still unclear and investigating this is a major focus
of this renewal application. The overall goal of our studies is to understand how members of the DNA damage
pathways regulate the differentiation-dependent HPV life cycle. We will address the following questions: 1).
How do ATM and ATR regulate HPV genome amplification? Are double strand breaks introduced into HPV
genomes? 2). How does the p38/MK2 pathway contribute to amplification? 3). What is the mechanism by
which HPV proteins initiate the DNA damage response?
Terms: <2-dimensional><ATM Protein><ATM Serine/Threonine Protein Kinase><ATM Signaling Pathway><ATM activation><ATM kinase><ATM pathway><ATM protein kinase><ATM signaling><ATR gene><ATR protein><ATR protein kinase><ATR serine/threonine kinase><Address><Anal><Anus><Ataxia Telangiectasia Mutated><Ataxia Telangiectasia Protein><Ataxia Telangiectasia and Rad3 Related Protein><Ataxia Telangiectasia and Rad3-Related><Ataxia-Telangiectasia and Rad3-Related Gene><Ataxia-Telangiectasia-Mutated protein kinase><Basal Cell><Basal Layer><Binding Sites><Biological Response Modifiers><Biomodulators><CDS1><CHEK2><CHEK2 gene><CHK2><CSAID-Binding Protein 1><CSAID-Binding Protein 2><CSBP2><Cancers><Causality><Cds1 kinase><Cell Body><Cell Cycle><Cell Differentiation><Cell Differentiation process><Cell Division Cycle><Cell Nucleus><Cells><Cervical><Cervical Cancer><Cervix Cancer><Chk2 protein kinase><Combining Site><Complex><Cytokine-Suppressive Antiinflammatory Drug-Binding Protein 1><Cytokine-Suppressive Antiinflammatory Drug-Binding protein 2><Cytoplasm><DNA Damage><DNA Injury><DNA Replication><DNA Synthesis><DNA biosynthesis><Deacetylase><E1 protein><Electrophoresis><Electrophoretic Fractionation><Epithelial><Epithelial Cells><Epithelium><Epithelium Part><Etiology><FRAP-Related Protein-1><FRP1><G2 Phase><G2 period><Gap Phase 2><Gene Expression><Gene Transcription><Genetic Alteration><Genetic Change><Genetic Transcription><Genetic defect><Genome><Goals><HPV><HPV 31><HPV E7><HPV infection><HPV-16 E1 protein><HPV16 E1 protein><HPV31><High Risk HPV31><Homologous Recombinational Repair><Human Papilloma Virus><Human Papillomavirus><Human papilloma virus 16 E1 protein><Human papilloma virus 31><Human papilloma virus infection><Human papillomavirus 16 E1 protein><Human papillomavirus 31><Human papillomavirus infection><Immune Mediators><Immune Mediators/Modulators><Immune Regulators><Immunomodulation><Infectious Human Wart Virus><Kinases><Laboratories><Lesion><Life Cycle><Life Cycle Stages><Link><MAPK14><MAPK14 Mitogen-Activated Protein Kinase><MAPK14 gene><MEC1><Maintenance><Malignant Cervical Neoplasm><Malignant Cervical Tumor><Malignant Neoplasm of the Cervix><Malignant Neoplasms><Malignant Oral Cavity Neoplasm><Malignant Oral Cavity Tumor><Malignant Oral Neoplasm><Malignant Tumor><Malignant Tumor of the Cervix><Malignant Tumor of the Cervix Uteri><Malignant Uterine Cervix Neoplasm><Malignant Uterine Cervix Tumor><Malignant neoplasm of cervix uteri><Mediating><Messenger RNA><Methods><Mitogen-Activated Protein Kinase 14><Mouth Cancer><Mutation><Mxi2><NBN gene><NBN protein><NBS1><NBS1 gene><NHEJ><Nibrin><Non-Homologous End Joining><Non-homologous DNA End Joining><Nonhomologous DNA End Joining><Nonhomologous End Joining><Nuclear><Nucleus><Oral Cancer><P53><PP1425><Papilloma Viruses><Papillomaviridae><Papillomavirus><Pathway interactions><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Play><Polymerase><Preventative vaccine><Preventive vaccine><Process><Prophylactic vaccine><Protein Phosphorylation><Proteins><RAD53><RNA Binding><RNA Expression><RNA bound><Rad3 Related Protein><Reactive Site><Recombination Repair><Resolution><Rete Malpighii><Role><SAPK2A><SCKL><SCKL1><SIRT1><SIRT1 gene><Second Gap Phase><Single Strand Break Repair><Sirtuin 1><Site><Stratified Squamous Epithelium><Stratum Basale><Stratum Germinativum><Stress-Activated Protein Kinase 2A><TP53><TP53 gene><TRP53><Transcript><Transcription><Transphosphorylases><Treatment Efficacy><Tumor Protein p53 Gene><Undifferentiated><Uterine Cervix Cancer><Viral><Viral Gene Products><Viral Gene Proteins><Viral Genes><Viral Genome><Viral Proteins><Virion><Virus Particle><Virus Replication><Woman><arm><ataxia telangiectasia and Rad3 related><ataxia telangiectasia mutated activation><ataxia telangiectasia mutated pathway><ataxia telangiectasia mutated protein><ataxia telangiectasia mutated signaling><ataxia telangiectasia mutated signaling pathway><causation><check point kinase 2><checkpoint kinase 2><cohesin><disease causation><experiment><experimental research><experimental study><genome mutation><high risk><homologous recombination><human papilloma virus E7><human papilloma virus type 31><human papillomavirus E7><human papillomavirus type 31><immune modulation><immune regulation><immune regulator><immunologic reactivity control><immunomodulatory><immunomodulatory biologics><immunoregulation><immunoregulatory><inhibitor><inhibitor/antagonist><insight><intervention efficacy><knock-down><knockdown><life course><mRNA><mRNA Stability><malignancy><malignant mouth neoplasm><malignant mouth tumor><member><neoplasm/cancer><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><oral cavity cancer><p38><p38 MAP Kinase><p38 MAPK Gene><p38 Mitogen Activated Protein Kinase><p38 Protein Kinase><p38 SAPK><p38-Alpha><p38Alpha><p53 Genes><p95 Protein of the MRE11/RAD50 Complex><pathway><prevent><preventing><recombinational repair><recruit><repair><repaired><response><serine-threonine-protein kinase Chk2><social role><therapeutic efficacy><therapeutically effective><therapy efficacy><two-dimensional><viral DNA><viral episome maintenance><viral multiplication><viral replication><virus DNA><virus episome maintenance><virus genome><virus multiplication><virus protein><wart virus>