Targeting protein–protein interactions in hematologic malignancies: still a challenge or a great opportunity for future therapies?

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/110054/1/imr12244.pd

Antioxidant-loaded mesoporous silica : an evaluation of the physicochemical properties

The dangerous effects of oxidative stress can be alleviated by antioxidants—substances with the ability to prevent damage caused by reactive oxygen species. The adsorption of antioxidants onto nanocarriers is a well-known method that might protect them against rough environ-mental conditions. The aim of this study was to investigate the adsorption and desorption of gallic acid (GA), protocatechuic acid (PCA), chlorogenic acid (CGA), and 4-hydroxybenzoic acid (4-HBA) using commercially available mesoporous silica materials (MSMs), both parent (i.e., SBA-15 and MCM-41) and surface functionalized (i.e., SBA-NH2 and SBA-SH). The MSMs loaded with active compounds were characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDX), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), thermoporometry (TPM), and powder X-ray diffraction (XRD). High-performance liquid chromatography (HPLC-CAD) was used to evaluate the performance of the adsorption and desorption processes. The antioxidant potential was investigated using the Folin–Ciocalteu (FC) spectrophotometric method. Among the studied MSMs, the highest adsorption of GA was observed for amine-modified SBA-15 mesoporous silica. The adsorption capacity of SBA-NH2 increased in the order of PCA, 4-HBA < GA < CGA. Different desorption effectiveness levels of the adsorbed compounds were observed with the antioxidant capacity preserved for all investigated compounds

Intestinal fructose and glucose metabolism in health and disease

The worldwide epidemics of obesity and diabetes have been linked to increased sugar consumption in humans. Here, we review fructose and glucose metabolism, as well as potential molecular mechanisms by which excessive sugar consumption is associated to metabolic diseases and insulin resistance in humans. To this end, we focus on understanding molecular and cellular mechanisms of fructose and glucose transport and sensing in the intestine, the intracellular signaling effects of dietary sugar metabolism, and its impact on glucose homeostasis in health and disease. Finally, the peripheral and central effects of dietary sugars on the gut–brain axis will be reviewed.Spanish MINISTERIO DE ECONOMÍA, INDUSTRIA Y COMPETITIVIDAD, grant numbers SAF2016-77871-C2-1-R and SAF2016-77871-C2-2-R to I.C-C. and G.P. respectively; the EFSD European Research Programme on New Targets for Type 2 Diabetes supported by an educational research grant from MSD to I.C-C. and G.P.; the FUNDACIÓN LA-CAIXA Y FUNDACIÓN CAJA DE BURGOS, grant number CAIXA-UBU001 to G.P

Gastrin Induces Nuclear Export and Proteasomal Degradation of Menin in Enteric Glial Cells

Background & aims: The multiple endocrine neoplasia, type 1 (MEN1) locus encodes the nuclear protein and tumor suppressor menin. MEN1 mutations frequently cause neuroendocrine tumors such as gastrinomas, characterized by their predominant duodenal location and local metastasis at time of diagnosis. Diffuse gastrin cell hyperplasia precedes the appearance of MEN1 gastrinomas, which develop within submucosal Brunner\u27s glands. We investigated how menin regulates expression of the gastrin gene and induces generation of submucosal gastrin-expressing cell hyperplasia. Methods: Primary enteric glial cultures were generated from the VillinCre:Men1FL/FL:Sst-/- mice or C57BL/6 mice (controls), with or without inhibition of gastric acid by omeprazole. Primary enteric glial cells from C57BL/6 mice were incubated with gastrin and separated into nuclear and cytoplasmic fractions. Cells were incubated with forskolin and H89 to activate or inhibit protein kinase A (a family of enzymes whose activity depends on cellular levels of cyclic AMP). Gastrin was measured in blood, tissue, and cell cultures using an ELISA. Immunoprecipitation with menin or ubiquitin was used to demonstrate post-translational modification of menin. Primary glial cells were incubated with leptomycin b and MG132 to block nuclear export and proteasome activity, respectively. We obtained human duodenal, lymph node, and pancreatic gastrinoma samples, collected from patients who underwent surgery from 1996 through 2007 in the United States or the United Kingdom. Results: Enteric glial cells that stained positive for glial fibrillary acidic protein (GFAP+) expressed gastrin de novo through a mechanism that required PKA. Gastrin-induced nuclear export of menin via cholecystokinin B receptor (CCKBR)-mediated activation of PKA. Once exported from the nucleus, menin was ubiquitinated and degraded by the proteasome. GFAP and other markers of enteric glial cells (eg, p75 and S100B), colocalized with gastrin in human duodenal gastrinomas. Conclusions: MEN1-associated gastrinomas, which develop in the submucosa, might arise from enteric glial cells through hormone-dependent PKA signaling. This pathway disrupts nuclear menin function, leading to hypergastrinemia and associated sequelae

BMI1 regulates PRC1 architecture and activity through homo- and hetero-oligomerization.

BMI1 is a core component of the polycomb repressive complex 1 (PRC1) and emerging data support a role of BMI1 in cancer. The central domain of BMI1 is involved in protein-protein interactions and is essential for its oncogenic activity. Here, we present the structure of BMI1 bound to the polyhomeotic protein PHC2 illustrating that the central domain of BMI1 adopts an ubiquitin-like (UBL) fold and binds PHC2 in a beta-hairpin conformation. Unexpectedly, we find that the UBL domain is involved in homo-oligomerization of BMI1. We demonstrate that both the interaction of BMI1 with polyhomeotic proteins and homo-oligomerization via UBL domain are necessary for H2A ubiquitination activity of PRC1 and for clonogenic potential of U2OS cells. Here, we also emphasize need for joint application of NMR spectroscopy and X-ray crystallography to determine the overall structure of the BMI1-PHC2 complex

Epigenetic regulation of IL‐12‐dependent T cell proliferation

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141412/1/jlb0601-sup-0001.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/141412/2/jlb0601.pd

Detection of disordered regions in globular proteins using 13 C‐detected NMR

Characterization of disordered regions in globular proteins constitutes a significant challenge. Here, we report an approach based on 13 C‐detected nuclear magnetic resonance experiments for the identification and assignment of disordered regions in large proteins. Using this method, we demonstrate that disordered fragments can be accurately identified in two homologs of menin, a globular protein with a molecular weight over 50 kDa. Our work provides an efficient way to characterize disordered fragments in globular proteins for structural biology applications.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/94460/1/2174_ftp.pd

A small-molecule inhibitor of the aberrant transcription factor CBFβ-SMMHC delays leukemia in mice

This is the author’s version of the work. It is posted here by permission of the AAAS for personal use, not for redistribution. The definitive version was published in Science on 2015 February 13; 347(6223): 779–784, DOI: 10.1126/science.aaa0314.Acute myeloid leukemia (AML) is the most common form of adult leukemia. The transcription factor fusion CBFβ-SMMHC (core binding factor β and the smooth-muscle myosin heavy chain), expressed in AML with the chromosome inversion inv(16)(p13q22), outcompetes wild-type CBFβ for binding to the transcription factor RUNX1, deregulates RUNX1 activity in hematopoiesis, and induces AML. Current inv(16) AML treatment with nonselective cytotoxic chemotherapy results in a good initial response but limited long-term survival. Here, we report the development of a protein-protein interaction inhibitor, AI-10-49, that selectively binds to CBFβ-SMMHC and disrupts its binding to RUNX1. AI-10-49 restores RUNX1 transcriptional activity, displays favorable pharmacokinetics, and delays leukemia progression in mice. Treatment of primary inv(16) AML patient blasts with AI-10-49 triggers selective cell death. These data suggest that direct inhibition of the oncogenic CBFβ-SMMHC fusion protein may be an effective therapeutic approach for inv(16) AML, and they provide support for transcription factor targeted therapy in other cancers

Foreword

The postsynaptic density protein-95/disks large/zonula occludens-1 (PDZ) protein domain family is one of the most common proteinprotein interaction modules in mammalian cells, with paralogs present in several hundred human proteins. PDZ domains are found in most cell types, but neuronal proteins, for example, are particularly rich in these domains. The general function of PDZ domains is to bring proteins together within the appropriate cellular compartment, thereby facilitating scaffolding, signaling, and trafficking events. The many functions of PDZ domains under normal physiological as well as pathological conditions have been reviewed recently. In this review, we focus on the molecular details of how PDZ domains bind their protein ligands and their potential as drug targets in this context