title: Nuclear Localization Signal-Enhanced Polyurethane-Short Branch Polyethylenimine-Mediated Delivery of Let-7a Inhibited Cancer Stem-Like Properties by Targeting the 3'-UTR of HMGA2 in Anaplastic Astrocytoma.
abstract: Anaplastic astrocytoma (AA) is a grade III glioma that often occurs in middle-aged patients and presents a uniformly poor prognosis. A small subpopulation of cancer stem cells (CSCs) possessing a self-renewing capacity is reported to be responsible for tumor recurrence and therapeutic resistance. An accumulating amount of microRNAs (miRNA) were found aberrantly expressed in human cancers and regulate CSCs. Efforts have been made to couple miRNAs with nonviral gene delivery approaches to target specific genes in cancer cells. However, the efficiency of delivery of miRNAs to AA-derived CSCs is still an applicability hurdle. The present study aimed to investigate the effectiveness and applicability of nonviral vector-mediated delivery of Let-7a with regard to eradication of AA and AA-derived CSC cells. Herein, our miRNA/mRNA microarray and RT-PCR analysis showed that the expression of Let-7a, a tumor-suppressive miRNA, is inversely correlated with the levels of HMGA2 and Sox2 in the AA side population (SP(+)) cells. Luciferase reporter assay showed that Let-7a directly targets the 3'-UTRs of HMGA2 in AA-SP(+) cells. Knockdown of HMGA2 significantly suppressed the protein expression of Sox2 in AA-SP(+) cells, whereas overexpression of HMGA2 upregulated Sox2 expression in AA-SP(-). Nuclear localization signal (NLS) peptides can facilitate nuclear targeting of DNA and are used to improve gene delivery. Using polyurethane-short branch polyethylenimine (PU-PEI) as a therapeutic delivery vehicle, we conjugated NLS with Let-7 and successfully delivered it to AA-SP(+) cells, resulting in significantly suppressed expression of HMGA2 and Sox2, tumorigenicity, and CSC-like abilities. This treatment facilitated the differentiation of AA-SP(+) cells into non-SP CSCs. Furthermore, PU-PEI-mediated delivery of NLS-conjugated Let-7a in AA-SP(+) cells suppressed the expression of drug-resistant and antiapoptotic genes, and increased cell sensitivity to radiation. Finally, the in vivo delivery of PU-PEI-NLS-Let-7a significantly suppressed the tumorigenesis of AA-SP(+) cells and synergistically improved the survival rate of orthotopically AA-SP(+)-transplanted immunocompromised mice when combined with radiotherapy. Therefore, PU-PEI-NLS-Let-7a is a potential novel therapeutic approach for AA.
authors.affiliation: Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan.
authors.forename: Meng-Yin
authors.initials: MY
authors.lastname: Yang
authors.forename: Ming-Teh
authors.initials: MT
authors.lastname: Chen
authors.forename: Pin-I
authors.initials: PI
authors.lastname: Huang
authors.forename: Chien-Ying
authors.initials: CY
authors.lastname: Wang
authors.forename: Yun-Chin
authors.initials: YC
authors.lastname: Chang
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Yang
authors.forename: Wen-Liang
authors.initials: WL
authors.lastname: Lo
authors.forename: Wen-Hsing
authors.initials: WH
authors.lastname: Sung
authors.forename: Yi-Wen
authors.initials: YW
authors.lastname: Liao
authors.forename: Yi-Yen
authors.initials: YY
authors.lastname: Lee
authors.forename: Yuh-Lih
authors.initials: YL
authors.lastname: Chang
authors.forename: Ling-Ming
authors.initials: LM
authors.lastname: Tseng
authors.forename: Yi-Wei
authors.initials: YW
authors.lastname: Chen
authors.forename: Hsin-I
authors.initials: HI
authors.lastname: Ma
mesh_terms: D000328:Adult; D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use / Q000633:toxicity; D001254:Astrocytoma / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D001483:Base Sequence; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D002945:Cisplatin / Q000627:therapeutic use / Q000633:toxicity; D005260:Female; D025743:HMGA2 Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000737:chemistry / Q000378:metabolism*; D008875:Middle Aged; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D019913:Nuclear Localization Signals / Q000737:chemistry*; D011094:Polyethyleneimine / Q000737:chemistry*; D011140:Polyurethanes / Q000737:chemistry*; D011839:Radiation, Ionizing; D016415:Sequence Alignment; D058985:Side-Population Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D025743:HMGA2 Protein; D035683:MicroRNAs; D019913:Nuclear Localization Signals; D011140:Polyurethanes; D011094:Polyethyleneimine; D002945:Cisplatin