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Principal Investigator: GIORGIO TRINCHIERI
Organization: DIVISION OF BASIC SCIENCES - NCI
Fiscal Year: 2022
Award: $741,872
Funding agency: National Cancer Institute
Progress has been made in several of the aims of the project. Tristetraprolin expression by keratinocytes protects against skin carcinogenesis: Cancer is caused primarily by genomic alterations resulting in deregulation of gene regulatory circuits in key growth, apoptosis, or DNA repair pathways. Multiple genes associated with the initiation and development of tumors are also regulated at the level of mRNA decay, through the recruitment of RNA-binding proteins to AU-rich elements (AREs) located in their 3'-untranslated regions. One of these ARE-binding proteins, tristetraprolin (TTP; encoded by Zfp36), is consistently dysregulated in many human malignancies. Herein, using regulated overexpression or conditional ablation in the context of cutaneous chemical carcinogenesis, we show that TTP represents a critical regulator of skin tumorigenesis. We provide evidence that TTP controlled both tumor-associated inflammation and key oncogenic pathways in neoplastic epidermal cells. We identify Areg as a direct target of TTP in keratinocytes and show that EGFR signaling potentially contributed to exacerbated tumor formation. Finally, single-cell RNA-Seq analysis indicated that ZFP36 was downregulated in human malignant keratinocytes. We conclude that TTP expression by epidermal cells played a major role in the control of skin tumorigenesis. Distinct contributions of cathelin-related antimicrobial peptide (CRAMP) derived from epithelial cells and macrophages to colon mucosal homeostasis: The cathelin-related antimicrobial peptide CRAMP protects the mouse colon from inflammation, inflammation-associated carcinogenesis, and disrupted microbiome balance, as shown in systemic Cnlp(-/-) mice (also known as Camp(-/-) mice). However, the mechanistic basis for the role and the cellular source of CRAMP in colon pathophysiology are ill defined. This study, using either epithelial or myeloid conditional Cnlp(-/-) mice, demonstrated that epithelial cell-derived CRAMP played a major role in supporting normal development of colon crypts, mucus production, and repair of injured mucosa. On the other hand, myeloid cell-derived CRAMP potently supported colon epithelial resistance to bacterial invasion during acute inflammation with exacerbated mucosal damage and higher rate of mouse mortality. Therefore, a well concerted cooperation of epithelial- and myeloid-derived CRAMP is essential for colon mucosal homeostasis. We also reported that normal (wild-type, WT) mouse macrophages increased their expression of cathelin-related antimicrobial peptide (CRAMP, encoded by Camp) after infection by viable E. coli or stimulation with inactivated E. coli and its product lipopolysaccharide (LPS), a process involving activation of NF-kB followed by protease-dependent conversion of CRAMP from an inactive precursor to an active form. The active CRAMP was required by WT macrophages for elimination of phagocytosed E. coli, with participation of autophagy-related proteins ATG5, LC3-II and LAMP-1, as well as for aggregation of the bacteria with p62 (also known as SQSTM1). This process was impaired in CRAMP(-/-) macrophages, resulting in retention of intracellular bacteria and fragmentation of macrophages. These results indicate that CRAMP is a critical component in autophagy-mediated clearance of intracellular E. coli by mouse macrophages. Myd88 in Cd11b+ cells protects from colitis-associated colon cancer through intestinal microbial homeostasis: Myeloid cells arise from the haematopoetic stem cells in the bone marrow and differentiate into natural killer (NK) cells, among others. These cells are an important component of the innate immune system which maintains homeostasis in the gut by maintaining a balance between pro- and anti-inflammatory responses and by promoting the integrity of the barrier function of the epithelium. Armed with an arsenal of pattern-recognition receptors called Toll-like receptors, these cells can respond to the multitudes of microbial antigens on the mucosal surface. Myeloid differentiation factor 88 (Myd88) is an important signaling molecule which contributes to signal transduction triggered by microbial products via recognition mediated by members of the Toll-like receptor and IL-1 receptor families. Using the azoxymethane (AOM)/DSS model of colitis-associated colorectal cancer, we previously showed that Myd88-/- mice are unable to heal colonic ulcers thereby increasing perpetuating the inflammatory microenvironment and fostering the dramatic increase in adenoma and adenocarcinoma formation in the colon. In this study, we decipher the compartment responsible for driving the anti-tumorigenic effects of MyD88. We hypothesized that MyD88 signaling from Cd11b+ myeloid cells mediates anti-tumor responses in the colon by reducing myeloid cell infiltration into the lamina propria and by preventing the expansion of GUS-type beta-glucuronidase- and invasin-containing microbes. Indeed, deletion of Myd88 in Cd11b+ cells (Myd88CD11b-/- mice) increased the number of adenomas formed relative to their corresponding floxed littermate controls. Remarkably, wild type mice co-housed with Myd88CD11b-/- animals or recipients of microbial transfer from these animals developed greater polyps in quantity and size. The contribution of CD11b+ cells to microbial homeostasis includes the negative regulation of E. coli strain O7:H7, which contributes to carcinogenesis through beta-glucuronidase and invasin activity. Taken together, MyD88 signaling in myeloid cells favors the development of a protective anti-tumoral response from the host and the gut microbiota. We also showed that enterochromaffin (EC) cells, which function as chemosensors on the gut epithelium, are known to translate environmental cues into serotonin (5-HT) production, contributing to intestinal physiology. However, how immune signals participate in gut sensation and neuroendocrine response remains unclear. Interleukin-33 (IL-33) acts as an alarmin cytokine by alerting the system of potential environmental stresses. We here demonstrate that IL-33 induced instantaneous peristaltic movement and facilitated Trichuris muris expulsion. We found that IL-33 could be sensed by EC cells, inducing release of 5-HT. IL-33-mediated 5-HT release activated enteric neurons, subsequently promoting gut motility. Mechanistically, IL-33 triggered calcium influx via a non-canonical signaling pathway specifically in EC cells to induce 5-HT secretion. Our data establish an immune-neuroendocrine axis in calibrating rapid 5-HT release for intestinal homeostasis.
Terms: <(TNF)-α><3' Untranslated Regions><3'UTR><5-HT><5-Hydroxytryptamine><5HT><Ablation><Acute><Adenocarcinoma><Animals><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-inflammatory><Antiinflammatories><Antiinflammatory Agents><Antitumor Response><Apoptosis><Apoptosis Pathway><Automobile Driving><Autophagocytosis><Autoregulation><Azoxymethane><Bacteria><Bacteria resistance><Bacteria resistant><Bacterial resistant><Beta-glucuronidase><Binding Proteins><Body Tissues><Bone Marrow><Bone Marrow Reticuloendothelial System><CAP-18><CAP18><CAP18 lipopolysaccharide-binding protein><CD107a antigen><CD11b><CR3A><CRAMP protein><CSIF><CSIF-10><CTLA-8><CTLA8><Cachectin><Calcium><Cancer Biology><Cancer Induction><Cancer Treatment><Cancers><Cell Body><Cell Communication and Signaling><Cell Growth in Number><Cell Locomotion><Cell Migration><Cell Movement><Cell Multiplication><Cell Proliferation><Cell Signaling><Cells><Cellular Migration><Cellular Motility><Cellular Proliferation><Chemical Models><Cnlp><Colitis associated colon cancer><Colitis associated colorectal cancer><Colitis induced colorectal cancer><Colon><Colonic inflammation><Communicable Diseases><Cues><Cutaneous><Cytokine Synthesis Inhibitory Factor><Cytotoxic T-Lymphocyte-Associated Antigen 8><Cytotoxic T-Lymphocyte-Associated Serine Esterase 8><Cytotoxic cell><DNA Alteration><DNA Repair Pathway><DNA Sequence Alteration><DNA mutation><DSS colitis><DSS model><DSS mouse model><DSS-induced acute colitis><DSS-induced colitis><Data><Dendritic Cells><Development><Diazene, dimethyl-, 1-oxide><Dysfunction><E coli><E. coli><EC Cell><EGF Receptor><EGFR><ERBB Protein><Edodekin Alfa><Effector Cell><Elements><Enteral><Enteramine><Enteric><Enterochromaffin><Enterochromaffin Cells><Environment><Epidermal Growth Factor Receptor><Epidermal Growth Factor Receptor Kinase><Epidermal Growth Factor Receptor Protein-Tyrosine Kinase><Epidermal Growth Factor-Urogastrone Receptors><Epithelial><Epithelial Cells><Equilibrium><Escherichia coli><Esteroproteases><Esthesia><Family><Fostering><Functional disorder><G0-G1 switch regulatory protein 24><GI microbiota><GOS24 protein><Gastrointestinal microbiota><Gene Deregulation><Generalized Growth><Genes><Genetic Models><Genetic mutation><Glucuronidase><Growth><Gut Epithelium><HER1><Hepatocarcinogenesis><Hippophaine><Homeostasis><Human><IFN><IL-1 Receptors><IL-10><IL-12><IL-17><IL-17A><IL-22><IL-23><IL1 Receptors><IL10><IL10A><IL12><IL17 Protein><IL17A><ITGAM><ITGAM gene><Immune><Immune response><Immune signaling><Immunes><Immunity><Immunoglobulin Enhancer-Binding Protein><Immunological response><Impairment><Infection><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Infiltration><Inflammation><Inflammation Mediators><Inflammatory><Inflammatory Response><Innate Immune System><Interferon Type I><Interferons><Interleukin 10 Precursor><Interleukin 17 (Cytotoxic T-Lymphocyte-Associated Serine Esterase 8)><Interleukin 17 Precursor><Interleukin-1 Receptors><Interleukin-10><Interleukin-12><Interleukin-17><Interleukins><Intestinal><Intestines><Intracellular Communication and Signaling><K lymphocyte><LAMP-1><LL37><Lamina Propria><Ligand Binding Protein><Ligand Binding Protein Gene><Light><Lipopolysaccharides><Liver Carcinogenesis><LoxP-flanked allele><MAC1A><MO1A><Macrophage Activation><Macrophage-Derived TNF><Malignant><Malignant - descriptor><Malignant Adenoma><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Mediating><Mice><Mice Mammals><Microbe><Modern Man><Molecular><Monocyte-Derived TNF><Morphogenesis><Motility><Movement><Mucosa><Mucosal Tissue><Mucous Membrane><Mucous body substance><Mucus><Murine><Mus><Myelogenous><Myeloid><Myeloid Cells><Mφ><NF-kB><NF-kappa B><NF-kappaB><NFKB><NK Cells><NKSF><Natural Killer Cell Stimulatory Factor><Natural Killer Cells><Natural Resistance><Nerve Cells><Nerve Unit><Neural Cell><Neurocyte><Neuroendocrine><Neuroendocrine System><Neurons><Neurosecretory Systems><NuP475 protein><Nuclear Factor kappa B><Nuclear Transcription Factor NF-kB><Oncogene Deregulation><Oncogenesis><Oncogenic><Organism><Palsy><Paralysed><Pathway interactions><Pattern recognition receptor><Peptidases><Peptide Hydrolases><Photoradiation><Physiologic><Physiological><Physiological Homeostasis><Physiology><Physiopathology><Play><Plegia><Polyps><Preventative vaccine><Preventive vaccine><Process><Production><Progenitor Cells><Programmed Cell Death><Prophylactic treatment><Prophylactic vaccine><Prophylaxis><Protease Gene><Proteases><Protein Binding><Proteinases><Proteins><Proteolytic Enzymes><RNA-Binding Proteins><Receptor Cell><Receptor Protein><Regulation><Regulator Genes><Reporting><Resistance><Resistance to infection><Role><STAT3><STAT3 gene><Sensation><Sequence Alteration><Serotonin><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Signaling Molecule><Skin><Skin Carcinogenesis><Source><Surface><System><T-Cells><T-Lymphocyte><TGF-alpha Receptor><TIS11 protein><TLR protein><TNF><TNF A><TNF Alpha><TNF gene><TNF-α><TNFA><TNFα><TTP protein><Time><Tissue Growth><Tissues><Toll-Like Receptor Family Gene><Toll-like receptors><Transcription Factor NF-kB><Transcriptional Regulatory Elements><Transforming Growth Factor alpha Receptor><Translating><Trichocephalus><Trichuris><Tumor Necrosis Factor><Tumor Necrosis Factor-alpha><Ulcer><Ulceration><Urogastrone Receptor><Veiled Cells><Wild Type Mouse><ZFP36 protein><acquired immunity><adaptive immunity><adenoma><anti-cancer therapy><anti-tumor response><anticancer therapy><antiinflammatory><autophagy><bacterial resistance><balance><balance function><beta-D-Glucuronoside glucuronosohydrolase><biological signal transduction><body movement><bound protein><bowel><c-erbB-1><c-erbB-1 Protein><cancer progression><cancer therapy><cancer-directed therapy><carcinogenesis><carcinogenesis in the liver><cathelicidin antimicrobial peptide><cathelin-like protein><cathelin-related antimicrobial peptide><cell motility><cell type><chemical carcinogenesis><colitis induced colon cancer><colitis-induced dysbiosis><colo-rectal carcinogenesis><colon carcinogenesis><colonic crypt><colorectal carcinogenesis><cytokine><developmental><dextran sulfate sodium colitis><dextran sulfate sodium induced colitis><dextran sulfate sodium model><dextran sulfate sodium mouse model><differentiation factors><driving><enteric microbial community><enteric microbiota><environmental stresses><environmental stressor><erbB-1><erbB-1 Proto-Oncogene Protein><erbBl><floxed><floxed allele><gastrointestinal epithelium><gastrointestinal homeostasis><gastrointestinal microbial flora><gene product><genomic alteration><gut commensal><gut community><gut flora><gut microbe community><gut microbial community><gut microbial composition><gut microbial consortia><gut microbiota><gut microbiotic><gut microflora><healing><hepatic carcinogenesis><hepatocellular carcinogenesis><host response><immune system response><immunoresponse><infection resistance><inflamed colon><inflammatory mediator><inflammatory modulation><injury and repair><interleukin-22><interleukin-23><intestinal flora><intestinal homeostasis><intestinal microbes><intestinal microbiota><intestinal microflora><intestinal tract microflora><kappa B Enhancer Binding Protein><keratinocyte><liver cancer pathogenesis><liver tumorigenesis><living system><mRNA Decay><macrophage><malignancy><member><microbial><microbial antigen><microbiome><microorganism antigen><morphogenetic process><morphogenic factors><morphogens><mortality><mouse model><mucous><murine model><naturally resistant><neoplasm progression><neoplasm/cancer><neoplastic><neoplastic progression><neuronal><nuclear factor kappa beta><ontogeny><overexpress><overexpression><paralysis><paralytic><pathophysiology><pathway><pleiotropic effect><pleiotropism><pleiotropy><prevent><preventing><programs><proto-oncogene protein c-erbB-1><receptor><recruit><regulatory gene><resistance to Bacteria><resistance to Bacterial><resistant><resistant to Bacteria><resistant to Bacterial><response><scRNA-seq><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><social role><stem cells><thymus derived lymphocyte><trans acting element><tristetraprolin><tumor><tumor progression><tumorigenesis><tumorigenic><wildtype mouse><β-D-Glucuronoside glucuronosohydrolase><β-glucuronidase>