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Bioengineered Noncoding RNAs Selectively Change Cellular miRNome Profiles for Cancer Therapy
- Source :
- The Journal of pharmacology and experimental therapeutics, vol 365, iss 3
- Publication Year :
- 2018
- Publisher :
- American Society for Pharmacology & Experimental Therapeutics (ASPET), 2018.
-
Abstract
- Noncoding RNAs (ncRNAs) produced in live cells may better reflect intracellular ncRNAs for research and therapy. Attempts were made to produce biologic ncRNAs, but at low yield or success rate. Here we first report a new ncRNA bioengineering technology using more stable ncRNA carrier (nCAR) containing a pre-miR-34a derivative identified by rational design and experimental validation. This approach offered a remarkable higher level expression (40%–80% of total RNAs) of recombinant ncRNAs in bacteria and gave an 80% success rate (33 of 42 ncRNAs). New FPLC and spin-column based methods were also developed for large- and small-scale purification of milligrams and micrograms of recombinant ncRNAs from half liter and milliliters of bacterial culture, respectively. We then used two bioengineered nCAR/miRNAs to demonstrate the selective release of target miRNAs into human cells, which were revealed to be Dicer dependent (miR-34a-5p) or independent (miR-124a-3p), and subsequent changes of miRNome and transcriptome profiles. miRNA enrichment analyses of altered transcriptome confirmed the specificity of nCAR/miRNAs in target gene regulation. Furthermore, nCAR assembled miR-34a-5p and miR-124-3p were active in suppressing human lung carcinoma cell proliferation through modulation of target gene expression (e.g., cMET and CDK6 for miR-34a-5p; STAT3 and ABCC4 for miR-124-3p). In addition, bioengineered miRNA molecules were effective in controlling metastatic lung xenograft progression, as demonstrated by live animal and ex vivo lung tissue bioluminescent imaging as well as histopathological examination. This novel ncRNA bioengineering platform can be easily adapted to produce various ncRNA molecules, and biologic ncRNAs hold the promise as new cancer therapeutics.
- Subjects :
- 0301 basic medicine
Lung Neoplasms
Bioengineering
ABCC4
Computational biology
Cell Transformation
Cell Line
Transcriptome
Drug Discovery and Translational Medicine
Mice
03 medical and health sciences
Cell Line, Tumor
microRNA
Genetics
Animals
Pharmacology & Pharmacy
Lung
Cell Proliferation
Cancer
Pharmacology
Neoplastic
Tumor
Base Sequence
biology
Gene Expression Profiling
Pharmacology and Pharmaceutical Sciences
Non-coding RNA
Gene expression profiling
MicroRNAs
Cell Transformation, Neoplastic
030104 developmental biology
Cell culture
biology.protein
Molecular Medicine
Cyclin-dependent kinase 6
Genetic Engineering
Biotechnology
Dicer
Subjects
Details
- ISSN :
- 15210103 and 00223565
- Volume :
- 365
- Database :
- OpenAIRE
- Journal :
- Journal of Pharmacology and Experimental Therapeutics
- Accession number :
- edsair.doi.dedup.....584cbae1bac8bbe01e23d85ca3d6715a