Natural product derivative Gossypolone inhibits Musashi family of RNA-binding proteins.

BACKGROUND: The Musashi (MSI) family of RNA-binding proteins is best known for the role in post-transcriptional regulation of target mRNAs. Elevated MSI1 levels in a variety of human cancer are associated with up-regulation of Notch/Wnt signaling. MSI1 binds to and negatively regulates translation of Numb and APC (adenomatous polyposis coli), negative regulators of Notch and Wnt signaling respectively. METHODS: Previously, we have shown that the natural product (-)-gossypol as the first known small molecule inhibitor of MSI1 that down-regulates Notch/Wnt signaling and inhibits tumor xenograft growth in vivo. Using a fluorescence polarization (FP) competition assay, we identified gossypolone (Gn) with a > 20-fold increase in Ki value compared to (-)-gossypol. We validated Gn binding to MSI1 using surface plasmon resonance, nuclear magnetic resonance, and cellular thermal shift assay, and tested the effects of Gn on colon cancer cells and colon cancer DLD-1 xenografts in nude mice. RESULTS: In colon cancer cells, Gn reduced Notch/Wnt signaling and induced apoptosis. Compared to (-)-gossypol, the same concentration of Gn is less active in all the cell assays tested. To increase Gn bioavailability, we used PEGylated liposomes in our in vivo studies. Gn-lip via tail vein injection inhibited the growth of human colon cancer DLD-1 xenografts in nude mice, as compared to the untreated control (P < 0.01, n = 10). CONCLUSION: Our data suggest that PEGylation improved the bioavailability of Gn as well as achieved tumor-targeted delivery and controlled release of Gn, which enhanced its overall biocompatibility and drug efficacy in vivo. This provides proof of concept for the development of Gn-lip as a molecular therapy for colon cancer with MSI1/MSI2 overexpression.

Hybrid sequencing reveals the full-length Nephila pilipes pyriform spidroin 1 (PySp1).

Araneid spider silk glands can spin seven silk types that have task-specific properties owing to the higher order structure of spider silk proteins. This gives silks superior potential as novel biomaterials. Nephila pilipes, the giant golden orb-weaver, is one of the largest spiders and spins silk with exceptional torsional deformation, toughness, and other properties to support its mass; further investigation relies on a complete amino acid sequence. However, there are no full-length N. pilipes spidroin sequences; in fact, across species, most sequences remain fragmentary because of repetitive region assembly difficulties in short-read sequencing. Here, we develop a hybrid sequencing method that utilizes short-read sequencing to identify seven spidroin terminals in N. pilipes, and long-read sequencing to confirm the full-length pyriform spidroin 1 (PySp1) gene. PySp1 is 11,181 base pairs, with a single exon encoding a 3,726 amino acid protein, the QQ(x)4Qx motif, and lower repeat homogenization, distinct characteristics of genera Nephilinae PySp1. The full-length N. pilipes PySp1 sequences sheds light on spidroin evolution and demonstrates a helpful strategy to find full-length spidroins.