Document text
Principal Investigator: Claudia M Nicolae
Organization: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
Fiscal Year: 2024
Award: $349,812
Funding agency: National Cancer Institute
Project Summary
Identification of molecular pathways that are preferentially employed and relied upon by cancer cells compared
to normal cells is key to designing novel personalized cancer therapies. PARP10 is a poorly characterized
member of the PARP family. We previously showed that PARP10 promotes translesion synthesis (TLS)-
mediated bypass of DNA lesions during DNA replication, thereby alleviating replication stress. More recently,
we also showed that PARP10 is a novel oncogene. We found that the PARP10 gene is amplified and/or
overexpressed in a large number of tumors including breast and ovarian, with very few observed occurrences
of downregulation or loss. We found that the PARP10 gene is amplified and/or overexpressed in a large
proportion of human tumors including breast and ovarian, with almost no occurrences of downregulation or
loss. Moreover, we found that PARP10 overexpression in, non-transformed human epithelial RPE1 cells
results in enhanced proliferation, resistance to replication stress, and increased xenograft tumor formation in
immunocompromised mice. The opposing phenotypes were found upon knockout of PARP10 in cancer HeLa
cells. These findings suggest that PARP10 is a putative oncogene and its expression promotes tumor
formation and growth. Mutagenic TLS has been previously proposed to promote transformation by both
allowing hyper-proliferation and inducing genomic instability. Thus, we hypothesize that PARP10 expression
suppresses replication stress through TLS-mediated bypass of replication arresting structures, thereby
allowing hyper-proliferation of cancer cells. We propose here to directly test this, in three specific aims which
address the hypothesis at three different levels: Aim 1 will investigate the mechanism employed by PARP10 to
modulate PCNA-dependent TLS at the molecular level, using biochemical and cellular localization and
interaction assays. Aim 2 will functionally test the impact of this mechanism of cellular processes including
genomic stability and DNA replication. Aim 3 will employ a mouse genetic model to unambiguously investigate
if Parp10 expression induces tumor formation or promotes tumor growth. Using state-of-the-art cellular,
molecular and genomic tools (including: CRISPR/Cas9-mediated genome editing; molecular DNA fiber
combing to measure fork stability; next generation sequencing approaches to measure mutagenesis and
mutation burden) we will investigate here the molecular mechanisms underlying this novel oncogenic function
of PARP10. This may eventually result in validation of a new target for cancer therapy.
Terms: <ADP Ribose><ADP Ribose Transferases><ADP-Ribosyltransferase><ADPRTs><ART Transferases><ARTases><Address><Adenosine 5'-(trihydrogen diphosphate), P'-5-ester with D-ribose><Adenosine 5'-Diphosphoribose><Adenosine Diphosphate Ribose><Adenosine Diphosphoribose><Apoptosis><Apoptosis Pathway><Assay><BRCA 1/2><BRCA1/2><Bioassay><Biochemical><Biological><Biological Assay><Breast><Bypass><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cancer Genes><Cancer Induction><Cancer-Promoting Gene><Cancers><Cas nuclease technology><Cell Body><Cell Communication and Signaling><Cell Cycle Progression><Cell Function><Cell Growth in Number><Cell Line><Cell Locomotion><Cell Migration><Cell Movement><Cell Multiplication><Cell Physiology><Cell Process><Cell Proliferation><Cell Signaling><CellLine><Cells><Cellular Function><Cellular Migration><Cellular Motility><Cellular Physiology><Cellular Process><Cellular Proliferation><Cellular biology><Chromosome Fragile Sites><Clinic><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Coupled><DNA Biochemistry><DNA Damage Repair><DNA Nucleic Acid Biochemistry><DNA Polymerases><DNA Repair><DNA Replication><DNA Structure><DNA Synthesis><DNA biosynthesis><DNA fiber analysis><DNA fiber approach><DNA fiber assay><DNA fiber combing><DNA fiber method><DNA fiber spreading><DNA fiber technique><DNA lesion><DNA replication fork><DNA seq><DNA sequencing><DNA-Dependent DNA Polymerases><DNA-Directed DNA Polymerase><DNAseq><Down-Regulation><Enzyme Gene><Enzymes><Epithelium><Exposure to><Family><Fragile Site><Gene Transcription><Generalized Growth><Genes><Genetic Alteration><Genetic Change><Genetic Models><Genetic Transcription><Genetic defect><Genetics-Mutagenesis><Genome Instability><Genome Stability><Genomic DNA><Genomic Instability><Genomic Stability><Genomics><Growth><HeLa><Hela Cells><Human><Immunocompromised><Immunocompromised Host><Immunocompromised Patient><Immunosuppressed Host><In Vitro><Intracellular Communication and Signaling><Knock-out><Knockout><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Measures><Mediating><Mice><Mice Mammals><Mitochondria><Modeling><Modern Man><Molecular><Murine><Mus><Mutagenesis><Mutagenesis Molecular Biology><Mutation><NGS Method><NGS system><Normal Cell><Nucleic Acid Biochemistry, DNA - Deoxyribonucleic Acid><Nucleic Acids><Oncogenes><Oncogenesis><Oncogenic><Ovarian><PARP Polymerase><PARP protein><PARS><Pathway interactions><Phenotype><Poly(ADP-ribose) Polymerases><Poly(ADPribose) Polymerase><Polymerase><Post-Translational Modification Protein/Amino Acid Biochemistry><Post-Translational Modifications><Post-Translational Protein Modification><Post-Translational Protein Processing><Posttranslational Modifications><Posttranslational Protein Processing><Process><Programmed Cell Death><Proliferating><Property><Protein Modification><Proteins><RNA Expression><RNA Seq><RNA sequencing><RNAseq><Resistance><Role><S Period><S phase><Signal Transduction><Signal Transduction Systems><Signaling><Strains Cell Lines><Structure><Subcellular Process><Synthesis Period><Synthesis Phase><Testing><Tissue Growth><Transcription><Transforming Genes><Transgenic Mice><Tumor Promotion><Ubiquitilation><Ubiquitination><Ubiquitinoylation><Unscheduled DNA Synthesis><Validation><biologic><biological signal transduction><brca gene><cancer cell><carcinogenesis><cell biology><cell motility><cultured cell line><design><designing><gDNA><gene manipulation><genetic manipulation><genetically manipulate><genetically perturb><genome editing><genome mutation><genomic editing><genomic tools><immunosuppressed patient><in vivo><individualized cancer therapy><inhibitor><malignancy><member><mitochondrial><mouse genetics><mouse model><murine model><mutant><neoplasm/cancer><next gen sequencing><next generation><next generation sequencing><nextgen sequencing><novel><ontogeny><overexpress><overexpression><oxidation><pathway><personalized cancer therapy><personalized cancer treatment><poly ADP polymerase><poly ADP ribose synthetase><recruit><replication fork><replication stress><resistant><social role><targeted cancer therapy><transcriptome sequencing><transcriptomic sequencing><transcriptomics><tumor><tumor growth><tumor xenograft><tumorigenesis><ubiquination><ubiquitin conjugation><validations>