RESUMEN
Infectious and inflammatory diseases in the intestine remain a serious threat for patients world-wide. Reprogramming of the intestinal epithelium towards a protective effector state is important to manage inflammation and immunity and can be therapeutically targeted. The role of epigenetic regulatory enzymes within these processes is not yet defined. Here, we use a mouse model that has an intestinal-epithelial specific deletion of the histone demethylase Lsd1 (cKO mice), which maintains the epithelium in a fixed reparative state. Challenge of cKO mice with bacteria-induced colitis or a helminth infection model both resulted in increased pathogenesis. Mechanistically, we discovered that LSD1 is important for goblet cell maturation and goblet-cell effector molecules such as RELMß. We propose that this may be in part mediated by directly controlling genes that facilitate cytoskeletal organization, which is important in goblet cell biology. This study therefore identifies intestinal-epithelial epigenetic regulation by LSD1 as a critical element in host protection from infection.
Asunto(s)
Infecciones por Enterobacteriaceae/inmunología , Células Caliciformes/inmunología , Histona Demetilasas/inmunología , Mucosa Intestinal/metabolismo , Tricuriasis/inmunología , Animales , Citrobacter rodentium , Células Caliciformes/metabolismo , Histona Demetilasas/metabolismo , Mucosa Intestinal/inmunología , Ratones , Ratones Noqueados , TrichurisRESUMEN
Cadherin-based intercellular adhesions are essential players in epithelial homeostasis, but their dynamic regulation during tissue morphogenesis and remodeling remain largely undefined. Here, we characterize an unexpected role for the membrane-anchored metalloproteinase MT2-MMP in regulating epithelial cell quiescence. Following co-immunoprecipitation and mass spectrometry, the MT2-MMP cytosolic tail was found to interact with the zonula occludens protein-1 (ZO-1) at the apical junctions of polarized epithelial cells. Functionally, MT2-MMP localizes in the apical domain of epithelial cells where it cleaves E-cadherin and promotes epithelial cell accumulation, a phenotype observed in 2D polarized cells as well as 3D cysts. MT2-MMP-mediated cleavage subsequently disrupts apical E-cadherin-mediated cell quiescence resulting in relaxed apical cortical tension favoring cell extrusion and re-sorting of Src kinase activity to junctional complexes, thereby promoting proliferation. Physiologically, MT2-MMP loss of function alters E-cadherin distribution, leading to impaired 3D organoid formation by mouse colonic epithelial cells ex vivo and reduction of cell proliferation within intestinal crypts in vivo Taken together, these studies identify an MT2-MMP-E-cadherin axis that functions as a novel regulator of epithelial cell homeostasis in vivo.
Asunto(s)
Cadherinas/metabolismo , Homeostasis/fisiología , Mucosa Intestinal/metabolismo , Metaloproteinasa 15 de la Matriz/metabolismo , Uniones Adherentes/metabolismo , Cadherinas/genética , Movimiento Celular/fisiología , Proteínas del Citoesqueleto/metabolismo , Células Epiteliales/metabolismo , Humanos , Uniones Intercelulares/metabolismo , Uniones Estrechas/metabolismoRESUMEN
RATIONALE: Aortic dissection or rupture resulting from aneurysm causes 1% to 2% of deaths in developed countries. These disorders are associated with mutations in genes that affect vascular smooth muscle cell differentiation and contractility or extracellular matrix composition and assembly. However, as many as 75% of patients with a family history of aortic aneurysms do not have an identified genetic syndrome. OBJECTIVE: To determine the role of the protease MMP17/MT4-MMP in the arterial wall and its possible relevance in human aortic pathology. METHODS AND RESULTS: Screening of patients with inherited thoracic aortic aneurysms and dissections identified a missense mutation (R373H) in the MMP17 gene that prevented the expression of the protease in human transfected cells. Using a loss-of-function genetic mouse model, we demonstrated that the lack of Mmp17 resulted in the presence of dysfunctional vascular smooth muscle cells and altered extracellular matrix in the vessel wall; and it led to increased susceptibility to angiotensin-II-induced thoracic aortic aneurysm. We also showed that Mmp17-mediated osteopontin cleavage regulated vascular smooth muscle cell maturation via c-Jun N-terminal kinase signaling during aorta wall development. Some features of the arterial phenotype were prevented by re-expression of catalytically active Mmp17 or the N-terminal osteopontin fragment in Mmp17-null neonates. CONCLUSIONS: Mmp17 proteolytic activity regulates vascular smooth muscle cell phenotype in the arterial vessel wall, and its absence predisposes to thoracic aortic aneurysm in mice. The rescue of part of the vessel-wall phenotype by a lentiviral strategy opens avenues for therapeutic intervention in these life-threatening disorders.
Asunto(s)
Aneurisma de la Aorta Torácica/genética , Disección Aórtica/genética , Metaloproteinasas de la Matriz Asociadas a la Membrana/fisiología , Mutación Missense , Adulto , Sustitución de Aminoácidos , Angiotensina II , Animales , Aorta/embriología , Aorta/patología , Aneurisma de la Aorta Torácica/patología , Aneurisma de la Aorta Torácica/terapia , Rotura de la Aorta/etiología , Matriz Extracelular/patología , Proteínas de la Matriz Extracelular/metabolismo , Predisposición Genética a la Enfermedad , Terapia Genética , Vectores Genéticos/uso terapéutico , Células HEK293 , Humanos , Lentivirus/genética , Masculino , Metaloproteinasas de la Matriz Asociadas a la Membrana/química , Metaloproteinasas de la Matriz Asociadas a la Membrana/deficiencia , Metaloproteinasas de la Matriz Asociadas a la Membrana/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Músculo Liso Vascular/patología , Osteopontina/metabolismo , Conformación ProteicaRESUMEN
Postnatal development of the gastrointestinal tract involves the establishment of the commensal microbiota, the acquisition of immune tolerance via a balanced immune cell composition, and maturation of the intestinal epithelium. While studies have uncovered an interplay between the first two, less is known about the role of the maturing epithelium. Here we show that intestinal-epithelial intrinsic expression of lysine-specific demethylase 1A (LSD1) is necessary for the postnatal maturation of intestinal epithelium and maintenance of this developed state during adulthood. Using microbiota-depleted mice, we find plasma cells, innate lymphoid cells (ILCs), and a specific myeloid population to depend on LSD1-controlled epithelial maturation. We propose that LSD1 controls the expression of epithelial-derived chemokines, such as Cxcl16, and that this is a mode of action for this epithelial-immune cell interplay in local ILC2s but not ILC3s. Together, our findings suggest that the maturing epithelium plays a dominant role in regulating the local immune cell composition, thereby contributing to gut homeostasis.
Asunto(s)
Microbioma Gastrointestinal , Histona Demetilasas , Mucosa Intestinal , Intestino Delgado , Animales , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Mucosa Intestinal/metabolismo , Ratones , Histona Demetilasas/metabolismo , Histona Demetilasas/genética , Microbioma Gastrointestinal/inmunología , Intestino Delgado/inmunología , Intestino Delgado/microbiología , Ratones Endogámicos C57BL , Inmunidad Innata , Linfocitos/inmunología , Linfocitos/metabolismo , Ratones Noqueados , Femenino , Masculino , HomeostasisRESUMEN
Intestinal epithelial homeostasis is maintained by intrinsic and extrinsic signals. The extrinsic signals include those provided by mesenchymal cell populations that surround intestinal crypts and is further facilitated by the extracellular matrix (ECM), which is modulated by proteases such as matrix metalloproteinases (MMPs). Extrinsic signals ensure an appropriate balance between intestinal epithelial proliferation and differentiation. This study explores the role of MMP17, which is preferentially expressed by smooth muscle cells in the intestine, in intestinal homeostasis and during immunity to infection. Mice lacking MMP17 expressed high levels of goblet-cell associated genes and proteins, such as CLCA1 and RELM-ß, which are normally associated with immune responses to infection. Nevertheless, Mmp17 KO mice did not have altered resistance during a bacterial Citrobacter rodentium infection. However, when challenged with a low dose of the helminth Trichuris muris, Mmp17 KO mice had increased resistance, without a clear role for an altered immune response during infection. Mechanistically, we did not find changes in traditional modulators of goblet cell effectors such as the NOTCH pathway or specific cytokines. We found MMP17 expression in smooth muscle cells as well as lamina propria cells such as macrophages. Together, our data suggest that MMP17 extrinsically alters goblet cell maturation which is sufficient to alter clearance in a helminth infection model.
Asunto(s)
Metaloproteinasa 17 de la Matriz , Tricuriasis , Animales , Ratones , Colon , Células Caliciformes/metabolismo , Metaloproteinasa 17 de la Matriz/metabolismo , Infección Persistente , TrichurisRESUMEN
Smooth muscle is an essential component of the intestine, both to maintain its structure and produce peristaltic and segmentation movements. However, very little is known about other putative roles that smooth muscle cells may have. Here, we show that smooth muscle cells may be the dominant suppliers of BMP antagonists, which are niche factors essential for intestinal stem cell maintenance. Furthermore, muscle-derived factors render epithelium reparative and fetal-like, which includes heightened YAP activity. Mechanistically, we find that the membrane-bound matrix metalloproteinase MMP17, which is exclusively expressed by smooth muscle cells, is required for intestinal epithelial repair after inflammation- or irradiation-induced injury. Furthermore, we propose that MMP17 affects intestinal epithelial reprogramming after damage indirectly by cleaving diffusible factor(s) such as the matricellular protein PERIOSTIN. Together, we identify an important signaling axis that establishes a role for smooth muscle cells as modulators of intestinal epithelial regeneration and the intestinal stem cell niche.
Asunto(s)
Metaloproteinasa 17 de la Matriz/metabolismo , Músculo Liso/metabolismo , Regeneración/fisiología , Nicho de Células Madre/fisiología , Animales , Humanos , Mucosa Intestinal/metabolismo , Intestinos/citología , Intestinos/patología , Transducción de Señal/fisiología , Células Madre/metabolismoRESUMEN
How helminths influence the pathogenesis of sexually transmitted viral infections is not comprehensively understood. Here, we show that an acute helminth infection (Nippostrongylus brasiliensis [Nb]) induced a type 2 immune profile in the female genital tract (FGT). This leads to heightened epithelial ulceration and pathology in subsequent herpes simplex virus (HSV)-2 infection. This was IL-5-dependent but IL-4 receptor alpha (Il4ra) independent, associated with increased FGT eosinophils, raised vaginal IL-33, and enhanced epithelial necrosis. Vaginal eosinophil accumulation was promoted by IL-33 induction following targeted vaginal epithelium damage from a papain challenge. Inhibition of IL-33 protected against Nb-exacerbated HSV-2 pathology. Eosinophil depletion reduced IL-33 release and HSV-2 ulceration in Nb-infected mice. These findings demonstrate that Nb-initiated FGT eosinophil recruitment promotes an eosinophil, IL-33, and IL-5 inflammatory circuit that enhances vaginal epithelial necrosis and pathology following HSV-2 infection. These findings identify a mechanistic framework as to how helminth infections can exacerbate viral-induced vaginal pathology.
Asunto(s)
Eosinófilos/inmunología , Helmintiasis/inmunología , Herpes Simple/inmunología , Receptores de Superficie Celular/inmunología , Vagina/inmunología , Enfermedades Vaginales/inmunología , Animales , Eosinófilos/patología , Femenino , Helmintiasis/complicaciones , Helmintos , Herpes Simple/complicaciones , Herpes Simple/patología , Herpes Simple/virología , Herpesvirus Humano 2/inmunología , Inmunidad , Interleucina-33 , Interleucina-5 , Necrosis , Nippostrongylus , Receptores de Superficie Celular/genética , Vagina/patología , Vagina/virología , Enfermedades Vaginales/parasitología , Enfermedades Vaginales/virologíaRESUMEN
Intestinal epithelial homeostasis is maintained by adult intestinal stem cells, which, alongside Paneth cells, appear after birth in the neonatal period. We aimed to identify regulators of neonatal intestinal epithelial development by testing a small library of epigenetic modifier inhibitors in Paneth cell-skewed organoid cultures. We found that lysine-specific demethylase 1A (Kdm1a/Lsd1) is absolutely required for Paneth cell differentiation. Lsd1-deficient crypts, devoid of Paneth cells, are still able to form organoids without a requirement of exogenous or endogenous Wnt. Mechanistically, we find that LSD1 enzymatically represses genes that are normally expressed only in fetal and neonatal epithelium. This gene profile is similar to what is seen in repairing epithelium, and we find that Lsd1-deficient epithelium has superior regenerative capacities after irradiation injury. In summary, we found an important regulator of neonatal intestinal development and identified a druggable target to reprogram intestinal epithelium toward a reparative state.
Asunto(s)
Mucosa Intestinal , Células de Paneth , Diferenciación Celular/genética , Histona Demetilasas/genética , Humanos , Recién Nacido , OrganoidesRESUMEN
Matrix metalloproteinases are involved in vascular remodeling. Little is known about their immune regulatory role in atherosclerosis. Here we show that mice deficient for MT4-MMP have increased adherence of macrophages to inflamed peritonea, and larger lipid deposits and macrophage burden in atherosclerotic plaques. We also demonstrate that MT4-MMP deficiency results in higher numbers of patrolling monocytes crawling and adhered to inflamed endothelia, and the accumulation of Mafb+ apoptosis inhibitor of macrophage (AIM)+ macrophages at incipient atherosclerotic lesions in mice. Functionally, MT4-MMP-null Mafb+AIM+ peritoneal macrophages express higher AIM and scavenger receptor CD36, are more resistant to apoptosis, and bind acLDL avidly, all of which contribute to atherosclerosis. CCR5 inhibition alleviates these effects by hindering the enhanced recruitment of MT4-MMP-null patrolling monocytes to early atherosclerotic lesions, thus blocking Mafb+AIM+ macrophage accumulation and atherosclerosis acceleration. Our results suggest that MT4-MMP targeting may constitute a novel strategy to boost patrolling monocyte activity in early inflammation.
Asunto(s)
Aterosclerosis/enzimología , Aterosclerosis/patología , Metaloproteinasa 17 de la Matriz/deficiencia , Monocitos/metabolismo , Animales , Antígeno CD11b/metabolismo , Humanos , Macrófagos Peritoneales/metabolismo , Factor de Transcripción MafB/metabolismo , Masculino , Metaloproteinasa 17 de la Matriz/metabolismo , Ratones Endogámicos C57BL , Placa Aterosclerótica/metabolismo , Placa Aterosclerótica/patología , Receptores CCR5/metabolismo , Receptores Depuradores/metabolismoRESUMEN
Emerging evidence indicates that the metalloproteinase Adamts-1 plays a significant role in the pathophysiology of vessel remodeling, but little is known about the signaling pathways that control Adamts-1 expression. We show that vascular endothelial growth factor (VEGF), angiotensin-II, interleukin-1ß, and tumor necrosis factor α, stimuli implicated in pathological vascular remodeling, increase Adamts-1 expression in endothelial and vascular smooth muscle cells. Analysis of the intracellular signaling pathways implicated in this process revealed that VEGF and angiotensin-II upregulate Adamts-1 expression via activation of differential signaling pathways that ultimately promote functional binding of the NFAT or C/EBPß transcription factors, respectively, to the Adamts-1 promoter. Infusion of mice with angiotensin-II triggered phosphorylation and nuclear translocation of C/EBPß proteins in aortic cells concomitantly with an increase in the expression of Adamts-1, further underscoring the importance of C/EBPß signaling in angiotensin-II-induced upregulation of Adamts-1. Similarly, VEGF promoted NFAT activation and subsequent Adamts-1 induction in aortic wall in a calcineurin-dependent manner. Our results demonstrate that Adamts-1 upregulation by inducers of pathological vascular remodeling is mediated by specific signal transduction pathways involving NFAT or C/EBPß transcription factors. Targeting of these pathways may prove useful in the treatment of vascular disease.
Asunto(s)
Proteínas ADAM/metabolismo , Proteína beta Potenciadora de Unión a CCAAT/fisiología , Factores de Transcripción NFATC/metabolismo , Remodelación Vascular , Proteínas ADAM/genética , Proteína ADAMTS1 , Animales , Aorta/enzimología , Secuencia de Bases , Calcineurina/metabolismo , Células Cultivadas , Células Endoteliales de la Vena Umbilical Humana/fisiología , Humanos , Ratones Noqueados , Datos de Secuencia Molecular , Transducción de Señal , Activación Transcripcional , Factor A de Crecimiento Endotelial Vascular/fisiologíaRESUMEN
The response to environmental cues such as inflammatory stimuli requires coordinated cellular functions. Certain proteins have functions on both sides of the plasma membrane to allow coordination between the extracellular and intracellular milieus. The membrane-anchored matrix metalloproteinase MT1-MMP is well positioned to sense and modify the extracellular environment by processing matrix components, transmembrane proteins and soluble factors. Recent findings show, however, that MT1-MMP also plays unexpected intracellular roles in macrophages through its location at the plasma membrane, the Golgi or the nucleus, impacting cell motility, metabolism and gene transcription. MT1-MMP is thus an example of the evolutionary diversification of protein function, allowing optimal coordination between extracellular stimuli and cellular responses. It remains to be determined whether these new MT1-MMP functions are specific to macrophages, professional phagocytes involved in inflammation, or are present in other inflammation-responsive cells. In this review, we will summarize these site-specific MT1-MMP functions in macrophages and comment on the possible conservation of these functions in endothelial cells.