Bioengineered miR-328-3p modulates GLUT1-mediated glucose uptake and metabolism to exert synergistic antiproliferative effects with chemotherapeutics.
10.1016/j.apsb.2019.11.001
- Author:
Wanrong YI
1
;
Mei-Juan TU
2
;
Zhenzhen LIU
2
;
Chao ZHANG
2
;
Neelu BATRA
2
;
Ai-Xi YU
1
;
Ai-Ming YU
2
Author Information
1. Department of Orthopaedic Trauma and Microsurgery, Zhongnan Hospital of Wuhan University, Wuhan 430072, China.
2. Department of Biochemistry & Molecular Medicine, UC Davis School of Medicine, Sacramento 95817, CA, USA.
- Publication Type:Journal Article
- Keywords:
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;
Bioengineered RNA;
CI, combination index;
CPT, cisplatin;
Cancer;
Chemosensitivity;
DOX, doxorubicin;
E. coli, Escherichia coli;
ESI, electrospray ionization;
FPLC, fast protein liquid chromatography;
Fa, fraction affected;
GLUT1;
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;
MiR-328;
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
- From:
Acta Pharmaceutica Sinica B
2020;10(1):159-170
- CountryChina
- Language:English
-
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/), we identified and verified large neutral amino acid transporter 1, gene symbol solute carrier family 7 member 5 (LAT1/) 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.
