ABSTRACT
Ulcerative colitis is a chronic condition in which a dysregulated immune response contributes to the acute intestinal inflammation of the colon. Current clinical therapies often exhibit limited efficacy and undesirable side effects. Here, programmable nanomicelles were designed for colitis treatment and loaded with RU.521, an inhibitor of the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway. STING-inhibiting micelles (SIMs) comprise hyaluronic acid-stearic acid conjugates and include a reactive oxygen species (ROS)-responsive thioketal linker. SIMs were designed to selectively accumulate at the site of inflammation and trigger drug release in the presence of ROS. Our in vitro studies in macrophages and in vivo studies in a murine model of colitis demonstrated that SIMs leverage HA-CD44 binding to target sites of inflammation. Oral delivery of SIMs to mice in both preventive and delayed therapeutic models ameliorated colitis's severity by reducing STING expression, suppressing the secretion of proinflammatory cytokines, enabling bodyweight recovery, protecting mice from colon shortening, and restoring colonic epithelium. In vivo end points combined with metabolomics identified key metabolites with a therapeutic role in reducing intestinal and mucosal inflammation. Our findings highlight the significance of programmable delivery platforms that downregulate inflammatory pathways at the intestinal mucosa for managing inflammatory bowel diseases.
Subject(s)
Colitis, Ulcerative , Membrane Proteins , Micelles , Nucleotidyltransferases , Animals , Colitis, Ulcerative/drug therapy , Colitis, Ulcerative/pathology , Colitis, Ulcerative/metabolism , Colitis, Ulcerative/chemically induced , Nucleotidyltransferases/metabolism , Nucleotidyltransferases/antagonists & inhibitors , Membrane Proteins/metabolism , Membrane Proteins/antagonists & inhibitors , Mice , Humans , Mice, Inbred C57BL , RAW 264.7 Cells , Reactive Oxygen Species/metabolismABSTRACT
Proteins carry out cellular functions. Identifying proteins within tissues, which are characteristically comprised of various cell types, is critical to understanding how the tissue functions. Being able to assess protein expression in tissues is also essential to gaining insight into how tissues change under different physiological conditions, in pathological states, in response to treatments, etc. Immunohistochemistry exploits antibody-antigen associations to identify specific proteins within tissues. This is a very powerful technique as it allows for studying tissues intact, preserving cellular relationships, and tissue structure. Here, we discuss the process of using an antibody specific for PPARγ to identify ovarian cells that express this transcription factor, and how its expression changes during the ovarian cycle.
Subject(s)
Immunohistochemistry/methods , PPAR gamma/metabolism , Paraffin Embedding/methods , Animals , Antibody Specificity , Female , Mice , PPAR gamma/immunology , Pregnancy , Protein Transport , RatsABSTRACT
Microsatellite or simple sequence repeat (SSR) markers have wide applicability for genetic analysis in crop plant improvement strategies. The objectives of this project were to isolate, characterize, and map a comprehensive set of SSR markers for maize (Zea mays L.). We developed 1051 novel SSR markers for maize from microsatellite-enriched libraries and by identification of microsatellite-containing sequences in public and private databases. Three mapping populations were used to derive map positions for 978 of these markers. The main mapping population was the intermated B73 x Mo17 (IBM) population. In mapping this intermated recombinant inbred line population, we have contributed to development of a new high-resolution map resource for maize. The primer sequences, original sequence sources, data on polymorphisms across 11 inbred lines, and map positions have been integrated with information on other public SSR markers and released through MaizeDB at URL:www.agron.missouri.edu. The maize research community now has the most detailed and comprehensive SSR marker set of any plant species.
