Targeting Cancer Cells Using LNA-Modified Aptamer-siRNA Chimeras.
Aptamers are chimerized with drug or antisense oligos or nanoparticles to generate targeted therapeutics for cancer. Aptamer chimerized siRNA rescues nonspecific delivery and, thereby, enhances the availability of siRNA to target cells. EpCAM RNA aptamer (EpApt or Ep) has potential for siRNA chimerization due to its secondary structure. Stathmin and survivin proteins are reported to aid oncogenicity in retinoblastoma (RB), breast cancer and other cancers. Thus, chimerization of EpCAM Apt with siRNA against survivin and stathmin, respectively, was performed by incorporating Locked Nucleic Acid (LNA) modification. The LNA-modified chimeric aptamers were stable until 96 h and got internalized into RB, WERI-Rb1 and breast cancer, MDAMB453 cell lines. The constructs were studied using the recombinant dicer enzyme for the siRNA generation. Quantitative polymerase chain reaction and immunofluorescence by microscopic analysis of chimeras in vitro exhibited silencing of stathmin and survivin in the RB and breast cancer model. The chimeric constructs showed significant inhibition of cell proliferation of breast cancer cells. Thus, LNA-modified aptamer-based siRNA delivery aids in cell targeting and necessitates further studies in animal models.
1 Department of Nanobiotechnology, Kamalnayan Bajaj Institute for Research in Vision and Ophthalmology , Chennai, Tamil Nadu, India .|2 Nanomedicine Laboratory of Immunology and Molecular Biomedical Research (NLIMBR), School of Medicine (SoM), Centre of Molecular and Medical Research (C-MMR), Faculty of Health, Deakin University , Geelong, Victoria, Australia .
Nithya
N
Subramanian
2 Nanomedicine Laboratory of Immunology and Molecular Biomedical Research (NLIMBR), School of Medicine (SoM), Centre of Molecular and Medical Research (C-MMR), Faculty of Health, Deakin University , Geelong, Victoria, Australia .
Jagat R
JR
Kanwar
2 Nanomedicine Laboratory of Immunology and Molecular Biomedical Research (NLIMBR), School of Medicine (SoM), Centre of Molecular and Medical Research (C-MMR), Faculty of Health, Deakin University , Geelong, Victoria, Australia .
Rupinder K
RK
Kanwar
1 Department of Nanobiotechnology, Kamalnayan Bajaj Institute for Research in Vision and Ophthalmology , Chennai, Tamil Nadu, India .|3 Department of L&T Ocular Pathology, Vision Research Foundation, Kamalnayan Bajaj Institute for Research in Vision and Ophthalmology , Chennai, Tamil Nadu, India .
Subramanian
S
Krishnakumar
D000951:Antigens, Neoplasm / Q000235:genetics; D052157:Aptamers, Nucleotide / Q000235:genetics / Q000627:therapeutic use*; D015815:Cell Adhesion Molecules / Q000235:genetics; D045744:Cell Line, Tumor; D000071858:Epithelial Cell Adhesion Molecule; D020868:Gene Silencing; D006801:Humans; D009369:Neoplasms / Q000473:pathology*; D009841:Oligonucleotides / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*
D000951:Antigens, Neoplasm; D052157:Aptamers, Nucleotide; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D009841:Oligonucleotides; D034741:RNA, Small Interfering; C477371:locked nucleic acid