Kinetic Resolution of Aromatic β-Amino Acids by ω-Transaminase. -(BEA, H.-S.; PARK, H.-J.; LEE, S.-H.; YUN*, H.; Chem. Commun. (Cambridge) 47 (2011) 20, 5894-5896, http://dx.doi.org/10.1039/c1cc11528f ; Sch. Biotechnol., Yeungnam Univ., Gyeongsan 712-749, S. Korea; Eng.) -R. Staver 36-078
Stem cell therapy has been proposed as a potential therapeutic strategy for acute kidney injury (AKI). By exploiting genome editing and cell sheet technology, we aimed to generate mesenchymal stem cells (MSCs) secreting angiogenic factors [vascular endothelial growth factor (VEGF) and angiopoietin‐1 (ANG1)] or anti‐inflammatory factors [erythropoietin (EPO) and a‐melanocyte stimulating hormone (a‐MSH)] for therapeutic application in AKI. To integrate each gene expression cassette into the safe harbor locus, AAVS1, of the human umbilical cord‐derived MSCs (hUC‐MSCs) chromosome, AAVS1‐targeting Zinc Finger Nuclease (ZFN) or AAVS1‐targeting CRISPR/Cas9 system was exploited. Junction PCR analysis demonstrated the ZFN‐ or CRISPR/Cas9‐aided gene integration in hUC‐MSCs. Flow cytometry and osteogenic and adipogenic differentiation assay revealed that stemness was maintained despite genome engineering. Protein measurement in conditioned media by ELISA and immunoblotting confirmed the production and secretion of each integrated gene product. In vitro assay demonstrated the angiogenic functions (increased HUVEC migration and increased mRNA expression of MMP‐9 and Tie‐2 in co‐cultured HUVEC) of genome‐engineered hUC‐MSCs secreting VEGF or ANG1. For the stem cell therapy in AKI, a scaffold‐free cell sheet system was established using a temperature‐responsive polymer (poly(N‐isopropylacrylamide). Taken together, cell sheet system of hUC‐MSCs secreting angiogenic or anti‐inflammatory factors is successfully established. This is to be examined in animal models of AKI to demonstrate the therapeutic effects of stem cell‐based regenerative strategy against AKI.Support or Funding InformationThis study was supported by the National Research Foundation of Korea Grants 2014R1A5A2009242 and 2016R1A2B4009365 funded by the Ministry of Science, ICT and Future Planning, KoreaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
We recently demonstrated that vacuolar protein sorting‐associated protein 35 (Vps35), a component of the retromer complex, interacts with the carboxyl terminus of aquaporin‐2, and depletion of Vps35 was associated with decreased AQP2 trafficking and increased lysosomal degradation of AQP2. Sorting nexin 27 (SNX27), a PDZ domain‐containing protein, is known to cooperate with the retromer complex in the recycling of early‐endosome to the plasma membrane. Since carboxyl terminus of aquaporin‐2 has the class I PDZ‐interacting motif (X‐Thr/Ser‐X‐Φ), we aimed to examine whether SNX27 could also play a role in the regulation of AQP2 trafficking and protein abundance in the kidney collecting duct cells. Immunohistochemistry revealed that SNX27 was diffusely labeled throughout the cytoplasm in the inner medullary collecting duct cells of rat kidney. Co‐immunoprecipitation assay of rat kidney inner medulla demonstrated that SNX27 interacted with AQP2. The role of SNX27 in the dDAVP‐responsive regulation of AQP2 trafficking and protein abundance was examined in mouse collecting duct mpkCCDc14 cells under siRNA‐mediated knockdown of SNX27. Cell surface biotinylation assay revealed that dDAVP‐induced translocation of AQP2 to the apical plasma membrane was decreased under SNX27 knockdown. Moreover, dDAVP‐induced AQP2 up‐regulation was significantly blunted in the cells with the siRNA‐mediated SNX27 knockdown. During the withdrawal period after dDAVP stimulation, the decrease of AQP2 protein abundance in the cells with SNX27 knockdown was attenuated by lysosomal inhibition with chloroquine co‐treatment. Taken together, SNX27 is an interacting protein with AQP2 and is likely to act as a component of the AQP2 sorting machinery after endocytosis, partly by regulating the lysosomal degradation of AQP2 protein abundance.Support or Funding InformationThis study was supported by the National Research Foundation of Korea Grants 2014R1A5A2009242 and 2016R1A2B4009365 funded by the Ministry of Science, ICT and Future Planning, KoreaThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.