TGR5 activation induces cytoprotective changes in the heart and improves myocardial adaptability to physiologic,inotropic, and pressure‐induced stress in mice

Introduction

Administration of cholic acid, or its synthetic derivative, 6‐alpha‐ethyl‐23(S)‐methylcholic acid (INT‐777), activates the membrane GPCR, TGR5, influences whole body metabolism, reduces atherosclerosis, and benefits the cardiovascular physiology in mice. Direct effects of TGR5 agonists, and the role for TGR5, on myocardial cell biology and stress response are unknown.

Methods

Mice were fed chow supplemented with 0.5% cholic acid (CA) or 0.025% INT‐777, a specific TGR5 agonist, or regular chow for 3 weeks. Anthropometric, biochemical, physiologic (electrocardiography and echocardiography), and molecular analysis was performed at baseline. CA and INT‐777 fed mice were challenged with acute exercise‐induced stress, acute catecholamine‐induced stress, and hemodynamic stress induced by transverse aortic constriction (TAC) for a period of 8 weeks. In separate experiments, mice born with constitutive deletion of TGR5 in cardiomyocytes (CM‐TGR5^del) were exposed to exercise, inotropic, and TAC‐induced stress.

Results

Administration of CA and INT‐777 supplemented diets upregulated TGR5 expression and activated Akt, PKA, and ERK1/2 in the heart. CA and INT‐777 fed mice showed improved exercise tolerance, improved sensitivity to catecholamine and attenuation in pathologic remodeling of the heart under hemodynamic stress. In contrast, CM‐TGR5^del showed poor response to exercise and catecholamine challenge as well as higher mortality and signs of accelerated cardiomyopathy under hemodynamic stress.

Conclusions

Bile acids, specifically TGR5 agonists, induce cytoprotective changes in the heart and improve myocardial response to physiologic, inotropic, and hemodynamic stress in mice. TGR5 plays a critical role in myocardial adaptability, and TGR5 activation may represent a potentially attractive treatment option in heart failure.

     

Comparison between VDR analogs and current immunosuppressive drugs in relation to CXCL10 secretion by human renal tubular cells

During kidney allograft rejection, CXC chemokine ligand 10 (CXCL10)–CXC chemokine receptor 3 (CXCR3) trafficking between peripheral blood and tissues initiates alloresponse and perpetuates a self‐inflammatory loop; thus, CXCL10–CXCR3 axis could represent a pharmacologic target. In this perspective, immunosuppressors targeting graft‐resident cells, beside immune cells, could be very advantageous. Vitamin D receptor (VDR) agonists exhibit considerable immunomodulatory properties. This study aimed to investigate whether elocalcitol and BXL‐01‐0029 could decrease the expression of CXCL10 in activated renal tubular cells in vitro and thus be useful in kidney allograft rejection treatment. Experiments were performed in human tubular renal cells stimulated with interferon‐γ + tumor necrosis factor‐α with and without VDR agonists, tacrolimus, sirolimus, hydrocortisone, methylprednisolone, cyclosporin A and mycophenolate mofetil. CXCL10 protein secretion and gene expression were measured by ELISA and by quantitative PCR. Specific inhibitors were used to investigate intracellular pathways involved in tubular cells activation. For IC₅₀ determination and comparison, dose‐response curves with VDR agonists, tacrolimus and mycophenolic acid were performed. Elocalcitol and BXL‐01‐0029 inhibited CXCL10 secretion by renal cells, without affecting cell viability, while almost all the immunosuppressors were found to be ineffective, except for tacrolimus and mycophenolate mofetil. BXL‐01‐0029 was the most potent drug and, notably, it was found to be capable of allowing reduction in tacrolimus‐inhibitory doses. Our data suggest that BXL‐01‐0029 could potentially be a dose‐reducing agent for conventional immunosuppressors in kidney rejection management.     

1,25-Dihydroxyvitamin D3 analogs as potential therapies in transplantation

While immunosuppressive drugs now permit good control of acute allograft rejection, chronic rejection remains an important medical problem, and the induction of stable transplantation tolerance has not yet been achieved in patients. 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3), a secosteroid hormone that controls cell proliferation and differentiation and exerts immunoregulatory activities in addition to regulating calcium and bone metabolism, has the potential to contribute to the management of allograft rejection. Recent advances in understanding the immunomodulatory properties of 1,25(OH)2D3 and its analogs suggest the clinical application of these hormones in transplantation, with the aim of facilitating tolerance induction and preventing chronic graft rejection.     

A 1alpha,25-dihydroxyvitamin D(3) analog enhances regulatory T-cells and arrests autoimmune diabetes in NOD mice

Type 1 diabetes is a chronic progressive autoimmune disease characterized by mononuclear cell infiltration, dominated by interleukin-12 (IL-12)-dependent Th1 cells, of the pancreatic islets, with subsequent destruction of insulin-producing beta-cells. Here, we demonstrate that treatment of adult nonobese diabetic (NOD) mice with an analog of 1alpha,25-dihydroxyvitamin D(3), an immunomodulatory agent preventing dendritic cell maturation, decreases lipopolysaccharide-induced IL-12 and gamma-interferon production, arrests Th1 cell infiltration and progression of insulitis, and inhibits diabetes development at nonhypercalcemic doses. Arrest of disease progression is accompanied by an enhanced frequency in the pancreatic lymph nodes of CD4(+)CD25(+) regulatory T-cells that are able to inhibit the T-cell response to the pancreatic autoantigen insulinoma-associated protein 2 and to significantly delay disease transfer by pathogenic CD4(+)CD25(-) cells. Thus, a short treatment of adult NOD mice with an analog of 1,25-dihydroxyvitamin D(3) inhibits IL-12 production, blocks pancreatic infiltration of Th1 cells, enhances CD4(+)CD25(+) regulatory cells, and arrests the progression of type 1 diabetes, suggesting its possible application in the treatment of human autoimmune diabetes.     

Glia-T cell dialogue

Interactions of CD4(+) T helper (Th) cells with microglia and astrocytes are likely to play an important role in regulating immune responses as well as tissue damage and repair during infectious and autoimmune central nervous system (CNS) diseases. T cells secreting Th1-type cytokines provide inducing signals for microglia to mature into functional antigen presenting cells (APC). The ability of microglia to act as efficient APC for the restimulation of Th1 cells suggests a role for these cells in the local amplification of pro-inflammatory immune responses. Conversely, the Th2-inducing capacity of microglia and astrocytes together with their ability to produce anti-inflammatory mediators could play a role in providing counter-regulatory signals limiting CNS inflammation. In this article, we review recent studies addressing the functional significance of T cell-CNS glia interactions and present new data on the expression of cyclooxygenase-2, the inducible enzyme involved in prostanoid biosynthesis, in microglia and astrocytes during the course of experimental allergic encephalomyelitis.     

Calcitriol derivatives with two different side chains at C-20. II. Diastereoselective syntheses of the metabolically produced 24(R)-hydroxygemini

Vitamin D derivatives containing two side chains emanating at C-20 are known as gemini. We have recently synthesized two gemini which are related to calcitriol and 19-norcalcitriol containing two identical side chains. The metabolism of these species involves 24(R)-hydroxylation on one of the side chains. To determine the outcome of this diastereospecific transformation, we synthesized both C-20 epimeric pairs containing the 24(R)-hydroxy group in the gemini and 19-norgemini series. On the basis of the availability of these reference compounds, it was shown that the metabolic hydroxylation occurred at the pro-R side chain in both gemini compounds. In comparison to the parent compounds, the 24-hydroxygemini required higher doses to increase blood calcium levels in mice and to suppress INF-gamma release in MLR.     

Selective immunointervention in autoimmune diseases: lessons from multiple sclerosis.

Activation of peripheral T cells by foreign and self antigens is under stringent control by different mechanisms, both thymic and peripheral. Control of T cell reactivity is accomplished by three major types of mechanisms: 1) deletion, the physical elimination of T cells specific for a given antigen, 2) anergy, the functional incapacity of T cells to respond to antigen, 3) suppression, the inhibition of T cell function by a regulatory (suppressor) cell. Their failure may lead to autoimmune diseases. The progress in understanding T cell activation, inactivation and modulation is being translated into strategies able to induce selective immunosuppression to treat different pathological situations, notably autoimmune diseases, allergies, and allograft rejection. The medical need for selective immunosuppression is very high, as the available immunosuppressive drugs are substantially inadequate because of limited efficacy, modest selectivity, and considerable toxicity. Key attack points for selective immunointervention have been identified: modulation of antigen recognition, co-stimulation blockade, induction of regulatory cells, deviation to non-pathogenic or protective responses, neutralization of proinflammatory cytokines, induction or administration of anti-inflammatory cytokines, and modulation of leukocyte trafficking. All these forms of immunointervention have been successfully used to prevent and sometimes treat experimental autoimmune diseases. Based on these results, expectations have been raised for exploiting the same strategies to inhibit the activation of human autoreactive T cells. In this overview, we will examine recent advances towards immunointervention in multiple sclerosis (MS) as a paradigm for successes and failures of current immunotherapeutic approaches in human autoimmune diseases.     

Basic strategies of the immune system in the regulation of antibody response

Summary Three major regulatory mechanisms operating in the control of antibody response have been examined: 1. antibody feedback; 2. T cell regulation (I. regulatory interactions among T cell subsets, II. H-2 linked Ir gene control of T cell function, III. regulatory role of antigenic epitopes in T cell subsets induction); 3. idiotypic network. Analysis of the results obtained in the lysozyme system together with available data in the literature have permitted the delineation of a model of antigen-triggered events involved in the regulation of antibody response. The basic feature of the proposed model is the integration of two major specific communication systems among lymphocytes engaged in the antibody response: antigen bridge and idiotypic complementarity.