RESUMO
B-cell lymphoma 6 (BCL6) is a zinc finger transcriptional repressor possessing a BTB-POZ (BR-C, ttk, and bab for BTB; pox virus and zinc finger for POZ) domain, which is required for homodimerization and association with corepressors. BCL6 has multiple roles in normal immunity, autoimmunity, and some types of lymphoma. Mice bearing disrupted BCL6 loci demonstrate suppressed high-affinity antibody responses to T-dependent antigens. The corepressor binding groove in the BTB-POZ domain is a potential target for small compound-mediated therapy. Several inhibitors targeting this binding groove have been described, but these compounds have limited or absent in vivo activity. Biophysical studies of a novel compound, GSK137, showed an in vitro pIC50 of 8 and a cellular pIC50 of 7.3 for blocking binding of a peptide derived from the corepressor silencing mediator for retinoid or thyroid hormone receptors to the BCL6 BTB-POZ domain. The compound has good solubility (128 µg/ml) and permeability (86 nM/s). GSK137 caused little change in cell viability or proliferation in four BCL6-expressing B-cell lymphoma lines, although there was modest dose-dependent accumulation of G1 phase cells. Pharmacokinetic studies in mice showed a profile compatible with achieving good levels of target engagement. GSK137, administered orally, suppressed immunoglobulin G responses and reduced numbers of germinal centers and germinal center B cells following immunization of mice with the hapten trinitrophenol. Overall, we report a novel small-molecule BCL6 inhibitor with in vivo activity that inhibits the T-dependent antigen immune response.
Assuntos
Proteínas Proto-Oncogênicas c-bcl-6 , Animais , Linfócitos B/metabolismo , Humanos , Camundongos , Transcrição Gênica , Dedos de ZincoRESUMO
We demonstrate the usefulness of synthetic lethal screening of a conditionally BCL6-deficient Burkitt lymphoma cell line, DG75-AB7, with a library of small molecules to determine survival pathways suppressed by BCL6 and suggest mechanism-based treatments for lymphoma. Lestaurtinib, a JAK2 inhibitor and one of the hits from the screen, repressed survival of BCL6-deficient cells in vitro and reduced growth and proliferation of xenografts in vivo BCL6 deficiency in DG75-AB7 induced JAK2 mRNA and protein expression and STAT3 phosphorylation. Surface IL10RA was elevated by BCL6 deficiency, and blockade of IL10RA repressed STAT3 phosphorylation. Therefore, we define an IL10RA/JAK2/STAT3 pathway each component of which is repressed by BCL6. We also show for the first time that JAK2 is a direct BCL6 target gene; BCL6 bound to the JAK2 promoter in vitro and was enriched by ChIP-seq. The place of JAK2 inhibitors in the treatment of diffuse large B-cell lymphoma has not been defined; we suggest that JAK2 inhibitors might be most effective in poor prognosis ABC-DLBCL, which shows higher levels of IL10RA, JAK2, and STAT3 but lower levels of BCL6 than GC-DLBCL and might be usefully combined with novel approaches such as inhibition of IL10RA.
Assuntos
Linfoma de Burkitt/tratamento farmacológico , Carbazóis/farmacologia , Subunidade alfa de Receptor de Interleucina-10/metabolismo , Janus Quinase 2/antagonistas & inibidores , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Proteínas Proto-Oncogênicas c-bcl-6/biossíntese , Fator de Transcrição STAT3/metabolismo , Animais , Linfoma de Burkitt/genética , Linfoma de Burkitt/metabolismo , Linhagem Celular Tumoral , Furanos , Humanos , Subunidade alfa de Receptor de Interleucina-10/genética , Janus Quinase 2/genética , Janus Quinase 2/metabolismo , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/metabolismo , Camundongos , Camundongos SCID , Proteínas Proto-Oncogênicas c-bcl-6/genética , Fator de Transcrição STAT3/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BCL6 is a transcriptional repressor that is over-expressed due to chromosomal translocations, or other abnormalities, in â¼40% of diffuse large B-cell lymphoma. BCL6 interacts with co-repressor, SMRT, and this is essential for its role in lymphomas. Peptide or small molecule inhibitors, which prevent the association of SMRT with BCL6, inhibit transcriptional repression and cause apoptosis of lymphoma cells in vitro and in vivo. In order to discover compounds, which have the potential to be developed into BCL6 inhibitors, we screened a natural product library. The ansamycin antibiotic, rifamycin SV, inhibited BCL6 transcriptional repression and NMR spectroscopy confirmed a direct interaction between rifamycin SV and BCL6. To further determine the characteristics of compounds binding to BCL6-POZ we analyzed four other members of this family and showed that rifabutin, bound most strongly. An X-ray crystal structure of the rifabutin-BCL6 complex revealed that rifabutin occupies a partly non-polar pocket making interactions with tyrosine58, asparagine21 and arginine24 of the BCL6-POZ domain. Importantly these residues are also important for the interaction of BLC6 with SMRT. This work demonstrates a unique approach to developing a structure activity relationship for a compound that will form the basis of a therapeutically useful BCL6 inhibitor.
Assuntos
Antibacterianos/farmacologia , Proteínas de Ligação a DNA/antagonistas & inibidores , Correpressor 2 de Receptor Nuclear/metabolismo , Proteínas Repressoras/antagonistas & inibidores , Rifabutina/farmacologia , Rifamicinas/farmacologia , Antibacterianos/química , Cristalografia por Raios X , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Células HEK293 , Humanos , Modelos Moleculares , Correpressor 2 de Receptor Nuclear/química , Mapas de Interação de Proteínas/efeitos dos fármacos , Estrutura Terciária de Proteína , Proteínas Proto-Oncogênicas c-bcl-6 , Proteínas Repressoras/química , Proteínas Repressoras/metabolismo , Rifabutina/química , Rifamicinas/química , Transcrição Gênica/efeitos dos fármacosRESUMO
INTRODUCTION: Obesity and asthma have become increasingly prevalent conditions in recent years; they often coexist and place a significant burden on the National Health Service. Asthma in the obese is more difficult to treat than in those with a normal body mass index (BMI) and is associated with resistance to traditional asthma therapies and increased use of healthcare resources. Weight loss can improve asthma control in such patients. The degree of weight loss achieved through dietary strategies, however, is often only modestly successful in this group. Bariatric surgery is increasingly used to achieve sustained significant weight loss in morbid obesity. It may offer under-recognized benefit in the difficult asthma-obesity phenotype. CASE STUDY: We describe the case of a 32-year-old female with difficult asthma who had a BMI of 45 kg/m(2) at the time of referral to our clinic. Her asthma was uncontrolled despite maximal inhaled therapy, oral therapy with Zafirlukast, and daily high-dose (25 mg) oral prednisolone. Additional therapies (subcutaneous Terbutaline and the steroid-sparing agent Methotrexate) had little impact on asthma control and she remained morbidly obese. She underwent gastric bypass surgery and, over the following 18 months, her BMI dropped to 27.7 kg/m(2), her corticosteroid dose was reduced to 7.5 mg (adrenal insufficiency proven), and maintenance inhaled therapy and oral medications were stopped as she maintained good asthma control. CONCLUSION: This case demonstrates the dramatic improvement that bariatric surgery can have on asthma symptoms and medication use in morbidly obese patients with very difficult to control asthma.
Assuntos
Asma/fisiopatologia , Cirurgia Bariátrica/métodos , Obesidade Mórbida/fisiopatologia , Obesidade Mórbida/cirurgia , Adulto , Asma/cirurgia , Cirurgia Bariátrica/normas , Índice de Massa Corporal , Feminino , Humanos , Redução de PesoRESUMO
Skin wound healing is a complex process requiring the coordinated, temporal orchestration of numerous cell types and biological processes to regenerate damaged tissue. Previous work has demonstrated that a functional ß-adrenergic receptor autocrine/paracrine network exists in skin, but the role of ß2-adrenergic receptor (ß2AR) in wound healing is unknown. A range of in vitro (single-cell migration, immunoblotting, ELISA, enzyme immunoassay), ex vivo (rat aortic ring assay), and in vivo (chick chorioallantoic membrane assay, zebrafish, murine wild-type, and ß2AR knockout excisional skin wound models) models were used to demonstrate that blockade or loss of ß2AR gene deletion promoted wound repair, a finding that is, to our knowledge, previously unreported. Compared with vehicle-only controls, ß2AR antagonism increased angiogenesis, dermal fibroblast function, and re-epithelialization, but had no effect on wound inflammation in vivo. Skin wounds in ß2AR knockout mice contracted and re-epithelialized faster in the first few days of wound repair in vivo. ß2AR antagonism enhanced cell motility through distinct intracellular signalling mechanisms and increased vascular endothelial growth factor secretion from keratinocytes. ß2AR antagonism promoted wound repair processes in the early stages of wound repair, revealing a possible new avenue for therapeutic intervention.