RESUMEN
Several types of psoriasiform dermatitis are associated with increased IL-36 cytokine activity in the skin. A rare, but severe, psoriasis-like disorder, generalized pustular psoriasis (GPP), is linked to loss-of-function mutations in the gene encoding IL-36RA, an important negative regulator of IL-36 signaling. To understand the effects of IL-36 dysregulation in a mouse model, we studied skin inflammation induced by intradermal injections of preactivated IL-36α. We found the immune cells infiltrating IL-36α-injected mouse skin to be of dramatically different composition than those infiltrating imiquimod-treated skin. The IL-36α-induced leukocyte population comprised nearly equal numbers of CD4+ αß T cells, neutrophils, and inflammatory dendritic cells, whereas the imiquimod-induced population comprised γδ T cells and neutrophils. Ligands for chemokine receptors CCR6 and CXCR2 are increased in both GPP and IL-36α-treated skin, which led us to test an optimized small-molecule antagonist (CCX624) targeting CCR6 and CXCR2 in the IL-36α model. CCX624 significantly reduced the T cell, neutrophil, and inflammatory dendritic cell infiltrates and was more effective than saturating levels of an anti-IL-17RA mAb at reducing inflammatory symptoms. These findings put CCR6 and CXCR2 forward as novel targets for a mechanistically distinct therapeutic approach for inflammatory skin diseases involving dysregulated IL-36 signaling, such as GPP.
Asunto(s)
Antiinflamatorios/farmacología , Interleucina-1/toxicidad , Psoriasis/inmunología , Receptores CCR6/antagonistas & inhibidores , Receptores de Interleucina-8B/antagonistas & inhibidores , Animales , Interleucina-1/inmunología , Ratones , Ratones Endogámicos BALB C , Psoriasis/inducido químicamente , Piel/efectos de los fármacos , Piel/inmunologíaRESUMEN
mAbs that neutralize IL-17 or its receptor have proven efficacious in treating moderate-to-severe psoriasis, confirming IL-17 as an important driver of this disease. In mice, a rare population of T cells, γδT17 cells, appears to be a dominant source of IL-17 in experimental psoriasis. These cells traffic between lymph nodes and the skin, and are identified by their coexpression of the TCR variable regions γ4 and δ4. These cells are homologous to the Vγ9Vδ2 T cell population identified in human psoriatic plaques. In this study we report that a potent and specific small molecule antagonist of the CCR6 chemokine receptor, CCX2553, was efficacious in reducing multiple aspects of psoriasis in two different murine models of the disease. Administration of CCX2553 ameliorated skin inflammation in both the IL-23-induced ear swelling model and the topical imiquimod model, and significantly reduced the number of γδT17 cells in inflamed skin. γδT17 cells were greatly reduced in imiquimod-treated skin of CCR6-/- mice, but adoptively transferred wild-type (CCR6+/+) γδT17 cells homed normally to the skin of imiquimod-treated CCR6-/- mice. Our data suggest that γδT17 cells are completely dependent on CCR6 for homing to psoriasiform skin. Thus, CCR6 may constitute a novel target for a mechanistically distinct therapeutic approach to treating psoriasis.
Asunto(s)
Movimiento Celular/inmunología , Interleucina-17/inmunología , Psoriasis/inmunología , Receptores de Antígenos de Linfocitos T gamma-delta/inmunología , Receptores CCR6/inmunología , Piel/inmunología , Linfocitos T/inmunología , Animales , Movimiento Celular/efectos de los fármacos , Movimiento Celular/genética , Interleucina-17/genética , Subunidad p19 de la Interleucina-23/genética , Subunidad p19 de la Interleucina-23/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Psoriasis/genética , Psoriasis/patología , Receptores de Antígenos de Linfocitos T gamma-delta/genética , Receptores CCR6/genética , Piel/patología , Linfocitos T/patologíaRESUMEN
Nonalcoholic fatty liver disease (NAFLD) is a common disease, closely associated with obesity and insulin resistance. We investigated the presence of a subset of myeloid cells associated with metabolic disturbance in the liver of patients with NAFLD and a murine model of obesity-induced liver disease. Gene and protein expression in liver and serum was investigated with RT-PCR or ELISA and correlated to clinical disease. Liver-infiltrating immune cells were isolated from normal or diseased human liver for flow cytometric analysis. In animal experiments, mice were fed a high-fat diet (60% of calories from fat) for 16 wk, or high-fat diet with 30% fructose for 32 wk to induce steatohepatitis and fibrosis. A small molecule inhibitor of CC chemokine receptor 2 (CCR2), CCX872, was administered to some mice. A subset of CD11c+CD206+ immune cells was enriched in human liver tissue, and greater infiltration was observed in NAFLD. The presence of CD11c+CD206+ myeloid cells correlated with systemic insulin resistance. CD11c+CD206+ cells expressed high levels of CCR2, and liver CC chemokine ligand 2 (CCL2) expression was increased in nonalcoholic steatohepatitis and correlated with disease activity. In mice, CCR2 inhibition reduced infiltration of liver CD11b+CD11c+F4/80+ monocytes, which are functional homologs of human CD11c+CD206+ cells, and improved liver injury and glycemic control. A role for CCR2/CCL2 in human NAFLD has long been postulated. These data confirm a role for this chemokine/receptor axis, through mediating adipose and hepatic infiltration of myeloid cells. Inhibition of CCR2 improved hepatic inflammation and fibrosis in murine models of NAFLD. These data confirm the rationale for targeting CCR2 to treat NAFLD. NEW & NOTEWORTHY These data show for the first time that CD11c+CD206+ myeloid cells, previously associated with human adipose tissue inflammation, infiltrate into liver tissue in nonalcoholic fatty liver disease. These cells express CCR2. Inhibition of CCR2 in mice inhibits hepatic inflammation caused by a murine homolog of these myeloid cells and improves experimental liver disease.
Asunto(s)
Quimiotaxis , Resistencia a la Insulina , Hígado/metabolismo , Monocitos/metabolismo , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Receptores CCR2/metabolismo , Animales , Antiinflamatorios/farmacología , Glucemia/metabolismo , Antígeno CD11b/metabolismo , Antígeno CD11c/metabolismo , Quimiocina CCL2/metabolismo , Quimiotaxis/efectos de los fármacos , Modelos Animales de Enfermedad , Femenino , Hemoglobina Glucada/metabolismo , Humanos , Lectinas Tipo C/metabolismo , Receptores de Lipopolisacáridos/metabolismo , Hígado/efectos de los fármacos , Hígado/inmunología , Hígado/patología , Masculino , Receptor de Manosa , Lectinas de Unión a Manosa/metabolismo , Ratones Endogámicos C57BL , Persona de Mediana Edad , Monocitos/efectos de los fármacos , Monocitos/inmunología , Enfermedad del Hígado Graso no Alcohólico/inmunología , Enfermedad del Hígado Graso no Alcohólico/patología , Enfermedad del Hígado Graso no Alcohólico/prevención & control , Receptores CCR2/antagonistas & inhibidores , Receptores CCR2/genética , Receptores de Superficie Celular/metabolismo , Transducción de SeñalRESUMEN
While it has long been established that the chemokine receptor CCR9 and its ligand CCL25 are essential for the movement of leukocytes into the small intestine and the development of small-intestinal inflammation, the role of this chemokine-receptor pair in colonic inflammation is not clear. Toward this end, we compared colonic CCL25 protein levels in healthy individuals to those in patients with ulcerative colitis. In addition, we determined the effect of CCR9 pharmacological inhibition in the mdr1a(-/-) mouse model of ulcerative colitis. Colon samples from patients with ulcerative colitis had significantly higher levels of CCL25 protein compared to healthy controls, a finding mirrored in the mdr1a(-/-) mice. In the mdr1a(-/-) mice, CCR9 antagonists significantly decreased the extent of wasting and colonic remodeling and reduced the levels of inflammatory cytokines in the colon. These findings indicate that the CCR9:CCL25 pair plays a causative role in ulcerative colitis and suggest that CCR9 antagonists will provide a therapeutic benefit in patients with colonic inflammation.
Asunto(s)
Colitis Ulcerosa/tratamiento farmacológico , Colitis Ulcerosa/metabolismo , Receptores CCR/antagonistas & inhibidores , Receptores CCR/metabolismo , Subfamilia B de Transportador de Casetes de Unión a ATP/genética , Subfamilia B de Transportador de Casetes de Unión a ATP/metabolismo , Animales , Quimiocinas CC/genética , Quimiocinas CC/metabolismo , Colitis Ulcerosa/genética , Femenino , Humanos , Técnicas In Vitro , Ratones , Ratones Noqueados , Sulfonamidas/uso terapéuticoRESUMEN
The concentration of CXCL12/SDF-1 in the bloodstream is tightly regulated, given its central role in leucocyte and stem/progenitor cell egress from bone marrow and recruitment to sites of inflammation or injury. The mechanism responsible for this regulation is unknown. Here we show that both genetic deletion and pharmacological inhibition of CXCR7, a high-affinity CXCL12 receptor, caused pronounced increases in plasma CXCL12 levels. The rise in plasma CXCL12 levels was associated with an impairment in the ability of leucocytes to migrate to a local source of CXCL12. Using a set of complementary and highly sensitive techniques, we found that CXCR7 protein is expressed at low levels in multiple organs in both humans and mice. In humans, CXCR7 was detected primarily on venule endothelium and arteriole smooth muscle cells. CXCR7 expression on venule endothelium was also documented in immunodeficient mice and CXCR7(+/lacZ) mice. The vascular expression of CXCR7 therefore gives it immediate access to circulating CXCL12. These studies suggest that endothelial CXCR7 regulates circulating CXCL12 levels and that CXCR7 inhibitors might be used to block CXCL12-mediated cell migration for therapeutic purposes.
Asunto(s)
Quimiocina CXCL12/inmunología , Endotelio Vascular/inmunología , Regulación de la Expresión Génica/inmunología , Células Endoteliales de la Vena Umbilical Humana/inmunología , Receptores CXCR/inmunología , Animales , Movimiento Celular/inmunología , Quimiocina CXCL12/sangre , Endotelio Vascular/citología , Endotelio Vascular/metabolismo , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Leucocitos/citología , Leucocitos/inmunología , Leucocitos/metabolismo , Ratones , Especificidad de Órganos/inmunología , Receptores CXCR/biosíntesisRESUMEN
The interaction of PD-L1 with PD-1 is a major immune checkpoint that limits effector T cell function against cancer cells; monoclonal antibodies that block this pathway have been approved in multiple tumor indications. As a next generation therapy, small molecule inhibitors of PD-L1 have inherent drug properties that may be advantageous for certain patient populations compared to antibody therapies. In this report we present the pharmacology of the orally-available, small molecule PD-L1 inhibitor CCX559 for cancer immunotherapy. CCX559 potently and selectively inhibited PD-L1 binding to PD-1 and CD80 in vitro, and increased activation of primary human T cells in a T cell receptor-dependent fashion. Oral administration of CCX559 demonstrated anti-tumor activity similar to an anti-human PD-L1 antibody in two murine tumor models. Treatment of cells with CCX559 induced PD-L1 dimer formation and internalization, which prevented interaction with PD-1. Cell surface PD-L1 expression recovered in MC38 tumors upon CCX559 clearance post dosing. In a cynomolgus monkey pharmacodynamic study, CCX559 increased plasma levels of soluble PD-L1. These results support the clinical development of CCX559 for solid tumors; CCX559 is currently in a Phase 1, first in patient, multicenter, open-label, dose-escalation study (ACTRN12621001342808).
Asunto(s)
Antígeno B7-H1 , Neoplasias , Humanos , Ratones , Animales , Antígeno B7-H1/metabolismo , Inhibidores de Puntos de Control Inmunológico , Receptor de Muerte Celular Programada 1 , Macaca fascicularis , Anticuerpos Monoclonales , Neoplasias/tratamiento farmacológico , Inmunoterapia/métodosRESUMEN
A general way of improving the potency of CXCR3 antagonists with fused hetero-bicyclic cores was identified. Optimization efforts led to the discovery of a series of imidazo-pyrazine derivatives with improved pharmacokinetic properties in addition to increased potency. The efficacy of the lead compound 21 is evaluated in a mouse lung inflammation model.
Asunto(s)
Antiinflamatorios/química , Bencenoacetamidas/química , Óxidos S-Cíclicos/química , Imidazoles/química , Pirazinas/química , Receptores CXCR3/antagonistas & inhibidores , Animales , Antiinflamatorios/síntesis química , Antiinflamatorios/farmacocinética , Bencenoacetamidas/síntesis química , Bencenoacetamidas/farmacocinética , Óxidos S-Cíclicos/síntesis química , Óxidos S-Cíclicos/farmacocinética , Humanos , Imidazoles/síntesis química , Imidazoles/farmacocinética , Ratones , Conformación Molecular , Pirazinas/síntesis química , Pirazinas/farmacocinética , Ratas , Receptores CXCR3/metabolismoRESUMEN
A series of imidazole derivatives have been designed and optimized for CXCR3 antagonism, pharmacokinetic properties, and reduced formation of glutathione conjugates. Our efforts led to the discovery of potent CXCR3 antagonists with good pharmacokinetic properties. These compounds are useful tools for in vivo studies of CXCR3 function.
Asunto(s)
Imidazoles/química , Imidazoles/farmacología , Receptores CXCR3/antagonistas & inhibidores , Animales , Cromatografía Líquida de Alta Presión , Diseño de Fármacos , Humanos , Imidazoles/farmacocinética , Ratones , Ratones Noqueados , Ratas , Receptores CXCR3/genética , Receptores CXCR3/fisiología , Relación Estructura-ActividadRESUMEN
Recent literature indicates that mice deficient in the chemokine receptor CCR9 (CCR9(-/-) mice) are unable to generate oral tolerance. The present report describes how such inability can be overcome by increasing the dose of oral antigen. Pharmacological inhibition of CCR9 did not affect the generation of oral tolerance, regardless of antigen dose. These results highlight the inadequacy of genetic deletion of CCR9 when predicting the effects of pharmacological CCR9 inhibition on intestinal biology.
Asunto(s)
Antígenos/inmunología , Tolerancia Inmunológica/inmunología , Receptores CCR/antagonistas & inhibidores , Administración Oral , Animales , Antígenos/administración & dosificación , Ratones , Ratones Noqueados , Ovalbúmina/inmunología , Receptores CCR/genéticaRESUMEN
A series of quinazolinone-derived inhibitors of the CXCR3 receptor have been synthesized and their affinity for the receptor evaluated. Compounds were evaluated in a (125)I-IP10 displacement assay and in in vitro cell migration assays to IP10, ITAC, and MIG using human peripheral blood mononuclear cells.