1. Bioengineered miR-328-3p modulates GLUT1-mediated glucose uptake and metabolism to exert synergistic antiproliferative effects with chemotherapeutics
- Author
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Yi, Wanrong, Tu, Mei-Juan, Liu, Zhenzhen, Zhang, Chao, Batra, Neelu, Yu, Ai-Xi, and Yu, Ai-Ming
- Subjects
Pharmacology and Pharmaceutical Sciences ,Biomedical and Clinical Sciences ,Orphan Drug ,Genetics ,Rare Diseases ,Biotechnology ,Cancer ,1.1 Normal biological development and functioning ,Bioengineered RNA ,MiR-328 ,LATI ,GLUT1 ,Chemosensitivity ,2-NBDG ,2-[N-(7-nitrobenz-2-oxa-1 ,3-diazol-4-yl) amino]-2-deoxyglucose ,ABCG2 ,ATP-binding cassette subfamily G member 2 ,ACN ,acetonitrile ,Au/Uv ,absorbance unit of ultraviolet-visible spectroscopy ,BCRP ,breast cancer resistant protein ,BERA ,bioengineered miRNA agent ,CI ,combination index ,CPT ,cisplatin ,DOX ,doxorubicin ,E. coli ,Escherichia coli ,ESI ,electrospray ionization ,FPLC ,fast protein liquid chromatography ,Fa ,fraction affected ,GLUT1 ,glucose transporter protein type 1 ,HCC ,hepatocellular carcinoma ,HPLC ,high-performance liquid chromatography ,IS ,internal standard ,KRB ,Krebs–Ringer bicarbonate ,LAT1 ,LAT1 ,large neutral amino acid transporter 1 ,LC–MS/MS ,liquid chromatography–tandem mass spectroscopy ,MCT4 ,monocarboxylate transporter 4 ,MRE ,miRNA response elements ,MRM ,multiple reaction monitoring ,OS ,osteosarcoma ,PAGE ,polyacrylamide gel electrophoresis ,PTEN ,phosphatase and tensin homolog ,PVDF ,Polyvinylidene fluoride ,RAGE ,receptor for advanced glycosylation end products ,RT-qPCR ,reverse transcription quantitative real-time polymerase chain reaction ,SLC2A1 ,7A5 ,16A3 ,solute carrier family 2 member 1 ,family 7 member 5 ,family 16 member 3 ,WT ,wild type ,hBERA ,humanized bioengineered miRNA agent ,hsa ,Homo sapiens ,htRNASer ,human seryl-tRNA ,mTOR ,mammalian target of rapamycin ,miR or miRNA ,microRNA ,ncRNA ,noncoding RNAs ,nt ,nucleotide ,Pharmacology and pharmaceutical sciences - Abstract
MicroRNAs (miRNAs or miRs) are small noncoding RNAs derived from genome to control target gene expression. Recently we have developed a novel platform permitting high-yield production of bioengineered miRNA agents (BERA). This study is to produce and utilize novel fully-humanized BERA/miR-328-3p molecule (hBERA/miR-328) to delineate the role of miR-328-3p in controlling nutrient uptake essential for cell metabolism. We first demonstrated successful high-level expression of hBERA/miR-328 in bacteria and purification to high degree of homogeneity (>98%). Biologic miR-328-3p prodrug was selectively processed to miR-328-3p to suppress the growth of highly-proliferative human osteosarcoma (OS) cells. Besides glucose transporter protein type 1, gene symbol solute carrier family 2 member 1 (GLUT1/SLC2A1), we identified and verified large neutral amino acid transporter 1, gene symbol solute carrier family 7 member 5 (LAT1/SLC7A5) as a direct target for miR-328-3p. While reduction of LAT1 protein levels by miR-328-3p did not alter homeostasis of amino acids within OS cells, suppression of GLUT1 led to a significantly lower glucose uptake and decline in intracellular levels of glucose and glycolytic metabolite lactate. Moreover, combination treatment with hBERA/miR-328 and cisplatin or doxorubicin exerted a strong synergism in the inhibition of OS cell proliferation. These findings support the utility of novel bioengineered RNA molecules and establish an important role of miR-328-3p in the control of nutrient transport and homeostasis behind cancer metabolism.
- Published
- 2020