sirolimus and Crohn-Disease

Autophagy is an intracellular process whereby cytoplasmic constituents are degraded within lysosomes. Autophagy functions to eliminate unwanted or damaged materials such as proteins and organelles as their accumulation would be harmful to the cellular system. Autophagy also acts as a defense mechanism against invading pathogens and plays an important role in innate and adaptive immunity. In physiological processes, autophagy is involved in the regulation of tissue development, differentiation and remodeling, which are essential for maintaining cellular homeostasis. Recent studies have demonstrated that autophagy is linked to various diseases and involved in pathophysiological roles, such as adaptation during starvation, anti-aging, antigen presentation, tumor suppression and cell death. The modulation of autophagy has shown greatest promise in Crohn's disease as most of autophagy drugs involved in these diseases are currently under clinical trials and some has been approved by Food and Drug Administration. This review article discusses autophagy and potential drugs that are currently available for its modulation in Crohn's disease.

Topics: Adalimumab; Anti-Inflammatory Agents; Autophagy; Certolizumab Pegol; Crohn Disease; Humans; Immunosuppressive Agents; Infliximab; Sirolimus

Mutations in genes involved in macroautophagy have been found to be associated with Morbus Crohn, also called Crohn's disease (CD), an inflammatory bowel disease. Taking this disease as an example for pathogenesis due to altered macroautophagy, we discuss here how macroautophagy supports innate and adaptive immunity. This support ranges from maintenance of components of the immune system, antigen processing for presentation to the immune system, to education of the immune system in order to distinguish self from dangerous non-self. A better understanding of these mechanisms should allow us not only to develop therapeutical strategies for CD but also to utilize macroautophagy for enhanced immunity against pathogens and tumors.

Topics: Adaptive Immunity; Animals; Antigen Presentation; Autophagy; Carrier Proteins; Crohn Disease; Cytokines; Humans; Immunity, Innate; Immunosuppressive Agents; Microtubule-Associated Proteins; Sirolimus; Small Ubiquitin-Related Modifier Proteins; T-Lymphocytes

A prospective study was undertaken to compare the efficacy of everolimus versus azathioprine or placebo in maintaining steroid-induced remission in active Crohn's disease (CD) and assess the safety and pharmacokinetics of everolimus.. This was a randomized, double-blind, placebo-controlled, proof-of-concept study in adults with moderate-to-severe active CD. The patients received oral steroids for a rapid induction of remission plus everolimus 6 mg/day, azathioprine 2.5 mg/kg/day, or placebo as maintenance treatment. The main outcome measure was the treatment success, defined as a steroid-free remission by the end of month 3 and maintained until study cutoff without the use of prohibited efficacy treatments.. Following an interim analysis, the study was terminated before enrollment was completed due to the lack of efficacy. The full intent-to-treat population comprised 138 patients. Only 96 patients who entered the study > or =7 months prior to data cutoff were included in the primary efficacy population. The treatment success was achieved in 13 of 38 everolimus patients, 22 of 36 azathioprine patients, and 8 of 22 placebo patients. Using the Kaplan-Meier estimates at month 7, the incidence of treatment success was 22.0% with everolimus group (95% confidence interval [CI] 6.7-37.3%, P= 0.610 vs placebo), 38.3% with azathioprine group (95% CI 20.6-55.9%, P= 0.500 vs placebo), and 28.8% with placebo group (95% CI 7.7-49.9%). The type and incidence of adverse events in the everolimus cohort were similar to those reported in the approved transplantation indications.. The safety and tolerability of everolimus (6 mg/day) in patients with active CD were comparable to azathioprine. At this dose, everolimus is not more efficacious in achieving a steroid-free remission in active CD than the comparators.

Topics: Adolescent; Adult; Aged; Azathioprine; Crohn Disease; Double-Blind Method; Everolimus; Female; Humans; Immunosuppressive Agents; Male; Middle Aged; Placebos; Prospective Studies; Sirolimus; Statistics, Nonparametric; Steroids; Treatment Outcome

Topics: Administration, Topical; Adolescent; Anti-Inflammatory Agents; Biopsy; Crohn Disease; Drug Therapy, Combination; Female; Genitalia; Humans; Immunohistochemistry; Immunosuppressive Agents; Injections, Intralesional; Lymphedema; Pain; Pruritus; Sirolimus; Skin; Treatment Outcome; Triamcinolone; Warts

Topics: Adult; Agammaglobulinemia; Arthralgia; Crohn Disease; Genetic Variation; Humans; Immunosuppressive Agents; Lymphocyte Count; Lymphopenia; Male; PTEN Phosphohydrolase; Sirolimus

An impairment of the intestinal barrier function is one of the major characteristics of Crohn's disease (CD). This study aimed to evaluate the impact of autophagy induction by rapamycin on the intestinal epithelial barrier function in CD model mice.. IL-10 knockout (IL-10 KO) mice were used as the human CD models in this study. All the mice were randomly assigned into four groups, (a) wild-type (WT) group; (b) IL-10 KO group; (c) IL-10 KO + rapamycin group and (d) IL-10 KO + 3-methyladenine (3-MA), containing 6 mice in each group. The disease activity index (DAI), histology, pro-inflammatory cytokines and chemotactic factors in colon tissues, intestinal and colonic permeability, distributions and expressions of tight junction (TJ) proteins, epithelial apoptosis of mice in four groups were evaluated and compared.. Autophagy induction by rapamycin treatment ameliorated DAI and histological colitis, decreased pro-inflammatory cytokines (TNF-α, IFN-γ and IL-17) and chemotactic factors (CXCL-1 and CXCL-2), decreased intestinal and colonic permeability, improved the distribution and expression of TJ proteins in IL-10 KO mice.. Autophagy induction by rapamycin significantly improved intestinal barrier function and protected IL-10 KO mice from the experimental chronic colitis.

Topics: Animals; Apoptosis; Autophagy; Crohn Disease; Disease Models, Animal; Humans; Interleukin-10; Intestinal Mucosa; Mice; Mice, Knockout; Permeability; Sirolimus

We recently observed reduced autophagy in Crohn's disease patients and an anti-inflammatory effect of autophagy stimulation in murine colitis, but both anti- and pro-fibrotic effects are associated with autophagy stimulation in different tissues, and fibrosis is a frequent complication of Crohn's disease. Thus, we analyzed the effects of pharmacological modulation of autophagy in a murine model of intestinal fibrosis and detected that autophagy inhibition aggravates, while autophagy stimulation prevents, fibrosis. These effects are associated with changes in inflammation and in collagen degradation in primary fibroblasts. Thus, pharmacological stimulation of autophagy may be useful against intestinal fibrosis.

Topics: Animals; Autophagy; Collagen; Crohn Disease; Disease Models, Animal; Fibroblasts; Fibrosis; Immunosuppressive Agents; Inflammation; Intestines; Mice; Mice, Inbred C57BL; Sirolimus

Significant studies have been carried out to understand effective management of intestinal fibrosis. However, the lack of better knowledge of fibrosis has hindered the development of a preventative drug. Primarily, finding a suitable animal model is challenging in understanding the mechanism of Crohn's-associated intestinal fibrosis pathology. Here, we adopted an effective method where TNBS chemical exposure to mice rectums produces substantially deep ulceration and chronic inflammation, and the mice then chronically develop intestinal fibrosis. Also, we describe a technique where a rapamycin injection shows inhibitory effects on TNBS-mediated fibrosis in the mouse model. To assess the underlying mechanism of fibrosis, we methodically discuss a procedure for purifying Cx3Cr1+ cells from the lamina propria of TNBS-treated and control mice. This detailed protocol will be helpful to researchers who are investigating the mechanism of fibrosis and pave the path to find a better therapeutic invention for Crohn's-associated intestinal fibrosis.

Topics: Animals; Colitis; Crohn Disease; Disease Models, Animal; Intestinal Mucosa; Intestines; Mice; Sirolimus; Trinitrobenzenesulfonic Acid

Crohn's disease (CD) is characterized by an imbalance of effector and regulatory T cells in the intestinal mucosa. The efficacy of anti-adhesion therapies led us to investigate whether impaired trafficking of T-regulatory (Treg) cells contributes to the pathogenesis of CD. We also investigated whether proper function could be restored to Treg cells by ex vivo expansion in the presence of factors that activate their regulatory activities.. We measured levels of the integrin α4β7 on Treg cells isolated from peripheral blood or lamina propria of patients with CD and healthy individuals (controls). Treg cells were expanded ex vivo and incubated with rapamycin with or without agonists of the retinoic acid receptor-α (RARA), and their gene expression profiles were analyzed. We also studied the cells in cytokine challenge, suppression, and flow chamber assays and in SCID mice with human intestinal xenografts.. We found that Treg cells from patients with CD express lower levels of the integrin α4β7 than Treg cells from control patients. The pathway that regulates the expression of integrin subunit α is induced by retinoic acid (RA). Treg cells from patients with CD incubated with rapamycin and an agonist of RARA (RAR568) expressed high levels of integrin α4β7, as well as CD62L and FOXP3, compared with cells incubated with rapamycin or rapamycin and all-trans retinoic acid. These Treg cells had increased suppressive activities in assays and migrated under conditions of shear flow; they did not produce inflammatory cytokines, and RAR568 had no effect on cell stability or lineage commitment. Fluorescently labeled Treg cells incubated with RAR568 were significantly more likely to traffic to intestinal xenografts than Treg cells expanded in control medium.. Treg cells from patients with CD express lower levels of the integrin α4β7 than Treg cells from control patients. Incubation of patients' ex vivo expanded Treg cells with rapamycin and an RARA agonist induced expression of α4β7 and had suppressive and migratory activities in culture and in intestinal xenografts in mice. These cells might be developed for treatment of CD. ClinicalTrials.gov, Number: NCT03185000.

Topics: Adult; Animals; Antineoplastic Agents; Case-Control Studies; Cell Culture Techniques; Cell Movement; Cells, Cultured; Crohn Disease; Female; Forkhead Transcription Factors; Gene Expression; Heterografts; Humans; Immunosuppressive Agents; Integrins; Intestinal Mucosa; L-Selectin; Lymphocyte Activation; Male; Mice; Mice, SCID; Middle Aged; Organic Chemicals; Retinoic Acid Receptor alpha; Sirolimus; T-Lymphocytes, Regulatory; Transcriptome; Tretinoin

Impaired Paneth cell expression of antimicrobial protein (AMP) lysozyme is found in patients with Crohn's disease with the autophagy gene ATG16L1 risk allele, in mice with mutations in autophagy genes Atg16L1, Atg5 and Atg7, and in Irgm1 knockout mice. Defective autophagy is also associated with expansion of resident Gram-negative bacteria in the intestinal lumen. These findings suggest that autophagy may control extracellular resident microbes by governing expression of lysozyme. To test the hypothesis that autophagy may have a defensive role in host response to resident extracellular microbes, we investigated the relationship between gut microbes, autophagy, and lysozyme. RAW 264.7 macrophages were treated with fecal slurry (FS), representing the resident microbial community; lipopolysaccharide (LPS); or butyrate, representing microbial products; or a representative resident Gram-negative bacterium Desulfovibrio vulgaris (DSV). FS, LPS, and DSV inhibited lysozyme expression, whereas butyrate had no effect. Induction of autophagy by rapamycin countered this inhibition, whereas silencing of the autophagy gene Irgm1 exacerbated the inhibitory effects of LPS on lysozyme expression. LPS also inhibited lysozyme activity against DSV and autophagy reversed this effect. Our results provide a novel insight into an interaction between gut bacteria, autophagy and AMP whereby autophagy may defend the host by countering the suppression of antimicrobial protein by Gram-negative bacteria.

Topics: Animals; Autophagy; Crohn Disease; Desulfovibrio vulgaris; Desulfovibrionaceae Infections; Feces; Gastrointestinal Microbiome; Gene Expression Regulation; GTP-Binding Proteins; Humans; Lipopolysaccharides; Macrophages; Mice; Mice, Knockout; Muramidase; Paneth Cells; RAW 264.7 Cells; RNA, Small Interfering; Sirolimus

Post-renal transplant de-novo inflammatory bowel disease (IBD) may develop despite the presence of mycophenolate mofetil (MMF), a drug used for treatment of IBD, in the immunosuppressive regimen. A 39-year-old man received live unrelated renal transplant, and was started postoperatively on prednisolone, MMF, and tacrolimus, which was changed to sirolimus when he developed diabetes mellitus two months post-transplant. Nine months post-transplant, the patient developed recurrent attacks of bloody diarrhea and ischio-rectal abscesses complicated by anal fistulae not responding to routine surgical treatment. Colonoscopy diagnosed IBD, a Crohn's disease-like pattern. The patient was treated with steroids and 5-aminosalicylic acid (5-ASA) in addition to a two months course of ciprofloxacin and metronidazole. He became asymptomatic and rectal lesions healed within one month of treatment. The patient continued to be asymptomatic, and he maintained normal graft function on the same immunosuppressive treatment in addition to 5-ASA. We conclude that de-novo IBD disease can develop in renal transplant recipients in spite of immunosuppressive therapy including MMF.

Topics: Adult; Aminosalicylic Acids; Ciprofloxacin; Crohn Disease; Humans; Immunosuppressive Agents; Inflammatory Bowel Diseases; Kidney Transplantation; Male; Metronidazole; Mycophenolic Acid; Sirolimus; Treatment Outcome

Scientific and medical authors tend to be biased toward submitting "statistically significant" findings for publication. Journals show a similar bias in publishing such "positive" studies. The large number of publications in medical research means that, in a field obsessed with controlling Type I error rates, we have journals with an abundance of Type I errors. Failing to publish studies that do not show a treatment or exposure effect creates a literature conspicuously absent of trials necessary for unbiased meta-analyses and systematic reviews. Furthermore, by shelving or rejecting studies with nonstatistically significant outcomes, authors and editors censor the most important contributors to medical research: our consenting volunteers.

Topics: Biomedical Research; Clinical Trials as Topic; Crohn Disease; Data Interpretation, Statistical; Everolimus; Humans; Immunosuppressive Agents; Publication Bias; Publishing; Sirolimus; Treatment Failure

We present the case of a 37-year-old woman with severe refractory colonic and perianal Crohn's disease who had lost response to second-line, steroid-sparing treatments azathioprine, methotrexate and infliximab. For many such patients extensive surgery has often been considered the only option. New insights provided by the results of genome-wide association scanning in Crohn's disease highlight autophagy, a cellular process implicated in the clearance of intracellular bacteria, as a key process in Crohn's disease pathogeneses. Sirolimus (rapamycin) is a drug used to upregulate autophagy in cell culture in the laboratory, and in clinical practice to prevent rejection following organ transplantation due to independent immunosuppressive action. Our patient was treated with sirolimus for 6 months at a dose that maintained serum trough levels of 5 ng/ml. There was marked and sustained improvement in Crohn's disease symptoms with the Harvey-Bradshaw index falling from 13 to 3, in serum markers of inflammation (C-reactive protein fell from 79 to 2) and endoscopic appearance. This is the first reported case of the use of sirolimus to treat Crohn's disease.

Topics: Adult; Biomarkers; C-Reactive Protein; Colonoscopy; Crohn Disease; Female; Humans; Immunosuppressive Agents; Severity of Illness Index; Sirolimus

Topics: Adult; Crohn Disease; Everolimus; Female; Humans; Immunosuppressive Agents; Sirolimus

A limited number of therapeutic strategies are currently available for patients with inflammatory bowel disease (IBD). In particular, the maintenance therapy after remission in Crohn's disease (CD) is not satisfactory and new approaches are needed. Interleukin-10 gene-deficient (IL-10-/-) mice, a well-characterized experimental model of CD, develop severe chronic colitis due to an aberrant Th1 immune response. Everolimus, an inhibitor of the mammalian target of rapamycin (mTOR), a new immunosuppressive reagent, has been used successfully in animal models for heart, liver, lung and kidney transplantation. In the present study, we examined the efficacy of everolimus in the treatment of chronic colitis in an IL-10-/- mouse model. Everolimus was administered orally for a period of 4 weeks to IL-10-/- mice with clinical signs of colitis. The gross and histological appearances of the colon and the numbers, phenotype and cytokine production of lymphocytes were compared with these characteristics in a control group. The 4-week administration of everolimus resulted in a significant decrease in the severity of colitis, together with a significant reduction in the number of CD4+ T cells in the colonic lamina propria as well as IFN-gamma production in colonic lymphocytes. Everolimus treatment of established colitis in IL-10-/- mice ameliorated the colitis, probably as a result of decreasing the number of CD4+ T cells in the colonic mucosa and an associated reduction in IFN-gamma production.

Topics: Animals; Body Weight; CD4-Positive T-Lymphocytes; Cells, Cultured; Crohn Disease; Cytokines; Disease Models, Animal; Everolimus; Female; Immunosuppressive Agents; Interferon-gamma; Interleukin-10; Intestinal Mucosa; Lymph Nodes; Lymphocyte Count; Male; Mice; Mice, Inbred C57BL; Sirolimus; Spleen; T-Lymphocyte Subsets; Treatment Outcome