Epigenetic control of skin homeostasis by the ULK3 nuclear kinase
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Principal Investigator: GIAN-PAOLO DOTTO Organization: MASSACHUSETTS GENERAL HOSPITAL Fiscal Year: 2024 Award: $462,800 Funding agency: National Institute of Arthritis and Musculoskeletal and Skin Diseases SUMMARY Field cancerization is a major clinical condition resulting from combined keratinocyte and dermal fibroblast alterations. Identification of regulatory molecules to target alterations of both cell types is of substantial interest. We propose that the nuclear kinase ULK3 is one such a molecule, which we previously studied for its role in fibroblasts. Here we will pursue the following aims: (1) We will test the hypothesis that ULK3 impinges on key transcription regulatory networks determining keratinocyte self-renewal and oncogenic potential. ULK3 is up- regulated in keratinocyte-derived skin and head/neck Squamous Cell Carcinomas (SCCs) and silencing of the gene suppresses proliferative and tumorigenic potential of both human keratinocytes (HKCs) and SCC cells. We will explore the interplay between ULK3 and the p63 and GLI transcription networks in growth/differentiation of these cells and their oncogenicity, in orthotopic models of skin cancer expansion. 2) We will test the hypothesis that ULK3 functions at the intersection between the arginine methylome and chromatin organization. Arginine methylation is a little investigated mode of chromatin regulation in SCCs development. We have found that ULK3 associates with the arginine methylases PRMT1 and PRMT5 (PRMTs) and that ULK3 loss in SCC cells results in down-modulation of di-methylated forms of Histone 4 (H4R3), well established products of PRMTs activity. We will test whether ULK3 controls PRMT1 and 5 activity and/or links these enzymes to key substrates such as p63, GLI2 and histones (specifically H4). We will further determine whether ULK3 control of gene transcription in HKCs and SCC involves a broader impact on chromatin organization through convergent PRMT1- and 5- dependent mechanisms. 3) We will test the hypothesis that ULK3 is a target of translational relevance for skin field cancerization. We will undertake a dual genetic and pharmacological approach. Genetically, we have developed mice with Ulk3flox alleles and K14- CreER transgene for keratinocyte-specific Ulk3 deletion. We will examine the impact of Ulk3 deletion on normal skin homeostasis and, after transfer into an albino hairless background, assess the consequences for UVB-induced skin field cancerization. Pharmacologically, we found that ULK3 inhibitory compounds suppress proliferative potential of SCC cells as well as cancer/stromal cell expansion. We will extend our studies with ULK3 and PRMT1/5 inhibitors on cultured cells and further test the most promising compounds in an in vivo orthotopic model of skin cancer/stromal cells expansion. Terms: <Acetylation><Affect><Alleles><Allelomorphs><Arginine><Arginine Methylase><Assay><Autoregulation><Binding><Bioassay><Biochemical><Biological Assay><Body Tissues><CRISPR><CRISPR/Cas system><Cancer Induction><Cancers><Carcinoma Cell><Cell Body><Cell Communication and Signaling><Cell Differentiation><Cell Differentiation process><Cell Signaling><Cells><Cervical Squamous Cell Carcinoma><Cervix Squamous Cell Carcinoma><Cervix Uteri Squamous Cell Carcinoma><ChIP Sequencing><ChIP-seq><ChIPseq><Chromatin><Chronic><Clinical><Closure by Ligation><Clustered Regularly Interspaced Short Palindromic Repeats><Co-Immunoprecipitations><Co-culture><Cocultivation><Coculture><Coculture Techniques><Cultured Cells><Cutaneous Squamous Cell Carcinoma><DNA-Protein Interaction><Dermal><Development><Differentiation and Growth><Disease Management><Disorder Management><Drugs><Ensure><Enzyme Gene><Enzymes><Epidermis><Epidermoid Carcinoma><Epidermoid Skin Carcinoma><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Erinaceidae><Fibroblasts><Future><GLI Family Gene><GLI Family Protein><GLI Protein><GLI gene><GLI-Kruppel Family Member 2><GLI1><GLI1 Gene><GLI1 Protein><GLI2><GLI2 gene><Gene Deletion><Gene Expression><Gene Inactivation><Gene Silencing><Gene Transcription><Gene set enrichment analysis><Genes><Genetic><Genetic Transcription><Gli2 protein><Glioma Associated Oncogene Homolog 1 Protein><Glioma Associated Oncogene Homolog Protein><Glioma-Associated Oncogene Homolog><HNSCC><Head and Neck><Head and Neck Squamous Cell Carcinoma><Head and neck structure><Hedgehogs><Histone (Arginine) Methyltransferase><Histone H3><Histones><Homeostasis><Human><Immune><Immunes><In Vitro><Individual><Intermediary Metabolism><Intracellular Communication and Signaling><Kinases><L-Arginine><Lesion><Ligation><Link><Lung><Lung Respiratory System><Malignant Epithelial Cell><Malignant Neoplasms><Malignant Skin Neoplasm><Malignant Tumor><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediating><Medication><Metabolic Processes><Metabolism><Methylation><Mice><Mice Mammals><Modeling><Modern Man><Molecular Interaction><Morbidity><Morbidity - disease rate><Murine><Mus><Myelin Basic Protein (Arginine) Methyltransferase><Nuclear><Oncogenic><Patients><Pharmaceutical Preparations><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Physiological Homeostasis><Planocellular Carcinoma><Play><Prevent skin cancer><Prevention><Process><Promoter Regions><Promotor Regions><Protein Arginine Methyltransferase><Protein Methylase I><Protein Methyltransferase I><Protein Phosphorylation><Protein-Arginine N-Methyltransferase><Proteins><RNA Expression><Recurrent Neoplasm><Recurrent tumor><Regulation><Repression><Role><SCCHN><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Site><Skin><Skin Aging><Skin Cancer><Squamous Carcinoma><Squamous Cell Epithelioma><Squamous cell carcinoma><Stromal Cells><Testing><Tissues><Toxic effect><Toxicities><Transcription><Transcription Regulation><Transcriptional Control><Transcriptional Regulation><Transgenes><Transgenic Mice><Transphosphorylases><UVB><UVB induced><UVB radiation><Ultraviolet B><Ultraviolet B Radiation><Uterine Cervix Squamous Cell Carcinoma><Work><aged skin><biological signal transduction><cancer progression><carcinogenesis><cell type><cellular differentiation><chromatin immunoprecipitation-sequencing><clinical significance><clinically significant><cutaneous photodamage><developmental><drug/agent><epigenetically><gene deletion mutation><gene signatures><genetic promoter element><genetic promoter sequence><genetic signature><gli2 gene product><glioma associated oncogene 1><glioma associated oncogene family zinc finger 1><glioma associated protein 2><head and neck squamous carcinoma><head and neck squamous cell cancer><in vivo><inhibitor><interest><keratinocyte><malignancy><malignant skin tumor><methylome><neoplasm progression><neoplasm recurrence><neoplasm/cancer><neoplastic progression><organ transplant patient><organ transplant recipient><overexpress><overexpression><pharmacologic><prevent><preventing><proliferation potential><promoter sequence><protein kinase inhibitor><pulmonary><response><self-renew><self-renewal><skin cancer prevention><skin photoaging><skin photodamage><skin solar aging><skin squamous cell carcinoma><social role><transcription regulatory network><transcriptional silencing><transgene><tumor progression><tumorigenic><ultra violet B radiation><ultraviolet B induced><ultraviolet b radiation induced><ultraviolet light b induced>