Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 2 de 2
Filter
Add more filters










Database
Language
Publication year range
1.
Anal Biochem ; 569: 53-58, 2019 03 15.
Article in English | MEDLINE | ID: mdl-30721669

ABSTRACT

Docking on the p53-binding site of murine double minute 2 (MDM2) by small molecules restores p53's tumor-suppressor function. We previously assessed 3244 FDA-approved drugs via "computational conformer selection" for inhibiting MDM2 and p53 interaction. Here, we developed a surface plasmon resonance method to experimentally confirm the inhibitory effects of the known MDM2 inhibitor, nutlin-3a, and two drug candidates predicted by our computational method. This p53/MDM2 interaction displayed a dosage-dependent weakening when MDM2 is pre-mixed with drug candidates. The inhibition efficiency order is nutlin-3a (IC50 = 97 nM) > bepridil (206 nM) > azelastine (307 nM). Furthermore, we verified their anti-proliferation effects on SJSA-1 (wild-type p53 and overexpressed MDM2), SW480 (mutated p53), and SaOs-2 (deleted p53) cancer cell lines. The inhibitory order towards SJSA-1 cell line is nutlin-3a (IC50 = 0.8 µM) > bepridil (23 µM) > azelastine (25 µM). Our experimental results are in line with the computational prediction, and the higher IC50 values from the cell-based assays are due to the requirement of higher drug concentrations to penetrate cell membranes. The anti-proliferation effects of bepridil and azelastine on the cell lines with mutated and deleted p53 implied some p53-independent anti-proliferation effects.


Subject(s)
Proto-Oncogene Proteins c-mdm2/metabolism , Small Molecule Libraries/chemistry , Surface Plasmon Resonance , Tumor Suppressor Protein p53/metabolism , Bepridil/chemistry , Bepridil/metabolism , Cell Line, Tumor , Humans , Imidazoles/chemistry , Imidazoles/metabolism , Piperazines/chemistry , Piperazines/metabolism , Protein Binding , Proto-Oncogene Proteins c-mdm2/antagonists & inhibitors , Proto-Oncogene Proteins c-mdm2/genetics , Small Molecule Libraries/metabolism , Tumor Suppressor Protein p53/antagonists & inhibitors , Tumor Suppressor Protein p53/genetics
2.
Cell Rep ; 19(7): 1365-1377, 2017 05 16.
Article in English | MEDLINE | ID: mdl-28514657

ABSTRACT

Brain microvascular endothelial cells (BMECs) are an essential component of the blood-brain barrier (BBB) that shields the brain against toxins and immune cells. While BBB dysfunction exists in neurological disorders, including Huntington's disease (HD), it is not known if BMECs themselves are functionally compromised to promote BBB dysfunction. Further, the underlying mechanisms of BBB dysfunction remain elusive given limitations with mouse models and post-mortem tissue to identify primary deficits. We undertook a transcriptome and functional analysis of human induced pluripotent stem cell (iPSC)-derived BMECs (iBMEC) from HD patients or unaffected controls. We demonstrate that HD iBMECs have intrinsic abnormalities in angiogenesis and barrier properties, as well as in signaling pathways governing these processes. Thus, our findings provide an iPSC-derived BBB model for a neurodegenerative disease and demonstrate autonomous neurovascular deficits that may underlie HD pathology with implications for therapeutics and drug delivery.


Subject(s)
Blood-Brain Barrier/pathology , Endothelial Cells/pathology , Huntington Disease/pathology , Induced Pluripotent Stem Cells/pathology , Microvessels/pathology , Neovascularization, Physiologic , Wnt Signaling Pathway , Gene Regulatory Networks , Humans , Huntington Disease/genetics , Induced Pluripotent Stem Cells/metabolism , Transcriptome/genetics , Transcytosis , beta Catenin/metabolism
SELECTION OF CITATIONS
SEARCH DETAIL
...