ASPH

Abstract: phoric. 2 and 3: THF (8.5 mL) was added to a mixture of 1 (0.58 g, 0.94 mmol) and dry InCI, (0.87 g, 4.10 mmol). The color changed immediately from green to blue. The solution was stirred for 5 min, methanol (1.5 mL) was added, and the mixture was separated on a chromatography column (MPLC column 46 x 2.6 cm. silica gel 40-63 F, eluent THFjmethanol l0jl). Ayellow and a blue fraction that have not yet been able to be characterized were eluted first in low yields. The third violet fraction was treated with AsPh,CI monohydrate (100 mg. 0.229 mmol). thesolvent removed at 10-' mbar, and the residue taken upina mixtureofn-hexane.ethylacetate.andacetone(5:20:15 mL).Theviolet mother liquor was filtered off through a G3 Schlenk filter; at f4 'C 2-(AsPh,), . (CH,),CO precipitates as red-brown crystals. They are separated from the mother liquor by decantation. washed three times with diethyl ether, and dried in vacuum. Yield: 90 mg (X%O, calculated on initial amount of I). correct elemental analyses. The fourth. brown fraction was treated with AsPh,Cl monohydrate (50 mg, 0.115 mmol). the solvent removed at lo-' mbar. and the residue taken up in ethyl acetate. Excess AsPh,CI and KCI remained undissolved. The brown inother liquor was filtered through a G-3 Schlenk filter. At +4T, 3-(AsPh,), precipitated in the form of dark brown crystals. which were separated by decantation from the mother liquor, washed three times with diethyl ether, and dried at lo-' mbar. Yield: 15mg (2%, calculated on initial ainonnt of 1). correct elemental analyses. A light brown residue which cannot be eluted remains on the column.

Abstract: Aspartate β-hydroxylase (ASPH) is silent in normal adult tissues only to re-emerge during oncogenesis where its function is required for generation and maintenance of malignant phenotypes. Exosomes enable prooncogenic secretome delivering and trafficking for long-distance cell-to-cell communication. This study aims to explore molecular mechanisms underlying how ASPH network regulates designated exosomes to program development and progression of breast cancer. Stable cell lines overexpressing or knocking-out of ASPH were established using lentivirus transfection or CRISPR-CAS9 systems. Western blot, MTT, immunofluorescence, luciferase reporter, co-immunoprecipitation, 2D/3-D invasion, tube formation, mammosphere formation, immunohistochemistry and newly developed in vitro metastasis were applied. Through physical interactions with Notch receptors, ligands (JAGs) and regulators (ADAM10/17), ASPH activates Notch cascade to provide raw materials (especially MMPs/ADAMs) for synthesis/release of pro-metastatic exosomes. Exosomes orchestrate EMT, 2-D/3-D invasion, stemness, angiogenesis, and premetastatic niche formation. Small molecule inhibitors (SMIs) of ASPH’s β-hydroxylase specifically/efficiently abrogated in vitro metastasis, which mimics basement membrane invasion at primary site, intravasation/extravasation (transendothelial migration), and colonization/outgrowth at distant sites. Multiple organ-metastases in orthotopic and tail vein injection murine models were substantially blocked by a specific SMI. ASPH is silenced in normal adult breast, upregulated from in situ malignancies to highly expressed in invasive/advanced ductal carcinoma. Moderate-high expression of ASPH confers more aggressive molecular subtypes (TNBC or Her2 amplified), early recurrence/progression and devastating outcome (reduced overall/disease-free survival) of breast cancer. Expression profiling of Notch signaling components positively correlates with ASPH expression in breast cancer patients, confirming that ASPH-Notch axis acts functionally in breast tumorigenesis. ASPH-Notch axis guides particularly selective exosomes to potentiate multifaceted metastasis. ASPH’s pro-oncogenic/pro-metastatic properties are essential for breast cancer development/progression, revealing a potential target for therapy.

Abstract: A series of novel yttrium- and lanthanide-containing heteropolyoxopalladates have been prepared and isolated as hydrated sodium salts, Na5[XIIIPdII12(AsPh)8O32]⋅y H2O (X=Y (1), Pr (2), Nd (3), Sm (4), Eu (5), Gd (6), Tb (7), Dy (8), Ho (9), Er (10), Tm (11), Yb (12), Lu (13); y=15–27). The polyanions [XIIIPdII12(AsPh)8O32]5− consist of a cuboid framework of twelve PdII ions with eight phenylarsonate heterogroups located at the vertices and a central guest ion X. The compounds 1–13 have been prepared in a simple one-pot self-assembly reaction of Pd(CH3COO)2, phenylarsonic acid and the respective salt of the element X in 0.5 M aqueous sodium acetate solution (pH 6.9), and characterized in the solid state by single-crystal X-ray diffraction, elemental and thermogravimetric (TGA) analyses, and IR spectroscopy. It was demonstrated that small, medium, and also large lanthanide ions can be incorporated in the center of the novel heteropolypalladate [XIIIPdII12(AsPh)8O32]5−. The LnO bond lengths follow the expected trend decreasing from left to right in the lanthanide series. This indicates that the {PdII12O32} shell can adjust to the coordination requirements of the encapsulated guest cation. Compounds 3 and 5 were selected for electrochemical studies. Their cyclic voltammetry in a lithium acetate buffer at pH 5.9 showed a Pd0 deposition process on the glassy carbon electrode surface. Coulometry indicated that all PdII centers were reduced to Pd0. The film was stable and could be taken out of the deposition medium and characterized in pure pH 5.9 buffer. Magnetic susceptibility and EPR measurements were carried out on 5 and 6. The former was confirmed to be diamagnetic and the latter strongly paramagnetic with a S=7/2 ground state. DFT calculations for some of the polyoxometalates have been also performed.