1. Bioengineered miR-328-3p modulates GLUT1-mediated glucose uptake and metabolism to exert synergistic antiproliferative effects with chemotherapeutics
- Author
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Aiming Yu, Wanrong Yi, Zhenzhen Liu, Ai Xi Yu, Neelu Batra, Chao Zhang, and Mei Juan Tu
- Subjects
WT, wild type ,phosphatase and tensin homolog ,internal standard ,7A5 ,absorbance unit of ultraviolet-visible spectroscopy ,receptor for advanced glycosylation end products ,miRNA response elements ,bioengineered miRNA agent ,FPLC, fast protein liquid chromatography ,0302 clinical medicine ,Au/Uv, absorbance unit of ultraviolet-visible spectroscopy ,HCC ,General Pharmacology, Toxicology and Pharmaceutics ,noncoding RNAs ,Cancer ,Fa ,0303 health sciences ,WT ,microRNA ,E. coli, Escherichia coli ,OS ,hsa ,hBERA, humanized bioengineered miRNA agent ,solute carrier family 2 member 1 ,fast protein liquid chromatography ,nucleotide ,BERA, bioengineered miRNA agent ,RAGE, receptor for advanced glycosylation end products ,RAGE ,3. Good health ,Cell biology ,family 7 member 5 ,030220 oncology & carcinogenesis ,KRB ,BCRP ,large neutral amino acid transporter 1 ,Intracellular ,Original article ,1.1 Normal biological development and functioning ,SLC2A1 ,MCT4 ,mTOR, mammalian target of rapamycin ,doxorubicin ,03 medical and health sciences ,breast cancer resistant protein ,ABCG2, ATP-binding cassette subfamily G member 2 ,osteosarcoma ,PVDF, Polyvinylidene fluoride ,Genetics ,htRNASer, human seryl-tRNA ,CPT, cisplatin ,high-performance liquid chromatography ,Chemosensitivity ,PAGE, polyacrylamide gel electrophoresis ,mammalian target of rapamycin ,Homo sapiens ,lcsh:RM1-950 ,E. coli ,CI, combination index ,RNA ,ncRNA ,miR or miRNA, microRNA ,LAT1 ,PAGE ,Polyvinylidene fluoride ,lcsh:Therapeutics. Pharmacology ,MCT4, monocarboxylate transporter 4 ,combination index ,HPLC, high-performance liquid chromatography ,ATP-binding cassette subfamily G member 2 ,IS ,family 16 member 3 ,hBERA ,GLUT1 ,HPLC ,MRM, multiple reaction monitoring ,fraction affected ,PTEN ,Glucose uptake ,LATI ,LAT1, large neutral amino acid transporter 1 ,cisplatin ,BERA ,Au/Uv ,MiR-328 ,miR or miRNA ,ncRNA, noncoding RNAs ,IS, internal standard ,DOX ,ESI, electrospray ionization ,biology ,Chemistry ,glucose transporter protein type 1 ,3-diazol-4-yl) amino]-2-deoxyglucose ,LC–MS/MS, liquid chromatography–tandem mass spectroscopy ,CI ,hepatocellular carcinoma ,monocarboxylate transporter 4 ,MRE ,human seryl-tRNA ,mTOR ,FPLC ,wild type ,MRM ,Biotechnology ,nt, nucleotide ,multiple reaction monitoring ,KRB, Krebs–Ringer bicarbonate ,humanized bioengineered miRNA agent ,MRE, miRNA response elements ,ABCG2 ,electrospray ionization ,Krebs–Ringer bicarbonate ,liquid chromatography–tandem mass spectroscopy ,nt ,Fa, fraction affected ,hsa, Homo sapiens ,2-NBDG, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxyglucose ,LC–MS/MS ,2-[N-(7-nitrobenz-2-oxa-1 ,ESI ,Escherichia coli ,OS, osteosarcoma ,SLC2A1, 7A5, 16A3, solute carrier family 2 member 1, family 7 member 5, family 16 member 3 ,BCRP, breast cancer resistant protein ,Nutrition ,030304 developmental biology ,Cell growth ,htRNASer ,PVDF ,RT-qPCR ,ACN ,Bioengineered RNA ,16A3 ,Metabolism ,ACN, acetonitrile ,DOX, doxorubicin ,GLUT1, glucose transporter protein type 1 ,PTEN, phosphatase and tensin homolog ,Solute carrier family ,acetonitrile ,reverse transcription quantitative real-time polymerase chain reaction ,2-NBDG ,biology.protein ,CPT ,HCC, hepatocellular carcinoma ,RT-qPCR, reverse transcription quantitative real-time polymerase chain reaction ,polyacrylamide gel electrophoresis - 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., Graphical abstract Fully-humanized, bioengineered miR-328-3p agent reduces glycolysis through the suppression of protein levels of targeted transporters GLUT1 and LAT1 in human osteosarcoma cells, leading to synergistic antiproliferative effects when combined with chemotherapeutic drugs.Image 1
- Published
- 2020