Therapeutic delivery of microRNAs discovered to target deregulated glioblastoma pathways inhibits tumor growth in mice
Shekhar Saha, Ying Zhang, Myron K. Gibert Jr., Collin Dube, Farina Hanif, Elizabeth Qian Xu Mulcahy, Sylwia Bednarek, Yunan Sun, Pawel Marcinkiewicz, Xiantao Wang, Gijung Kwak, Ahsan Polash, Haolin Li, Kadie Hudson, Manikarna Dinda, Tapas Saha, Matthew McCord, Fadila Guessous, Nichola Cruickshanks, Rossymar Rivera Colon, Lily Dell’Olio, Rajitha Anbu, Wenjie Liu, Songy Choi, Benjamin Kefas, Pankaj Kumar, Alexander L. Klibanov, David Schiff, Jung Soo Suk, Justin Hanes, Jamie Mata, Markus Hafner, Roger Abounader
Shekhar Saha, Ying Zhang, Myron K. Gibert Jr., Collin Dube, Farina Hanif, Elizabeth Qian Xu Mulcahy, Sylwia Bednarek, Yunan Sun, Pawel Marcinkiewicz, Xiantao Wang, Gijung Kwak, Ahsan Polash, Haolin Li, Kadie Hudson, Manikarna Dinda, Tapas Saha, Matthew McCord, Fadila Guessous, Nichola Cruickshanks, Rossymar Rivera Colon, Lily Dell’Olio, Rajitha Anbu, Wenjie Liu, Songy Choi, Benjamin Kefas, Pankaj Kumar, Alexander L. Klibanov, David Schiff, Jung Soo Suk, Justin Hanes, Jamie Mata, Markus Hafner, Roger Abounader
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Research Article Cell biology Oncology

Therapeutic delivery of microRNAs discovered to target deregulated glioblastoma pathways inhibits tumor growth in mice

Abstract

Glioblastoma is a fatal primary malignant brain tumor, with an average survival of 15 months despite surgical resection, chemotherapy, and radiation therapy. Due to the concurrent deregulation of numerous genes in glioblastoma, molecular monotherapies have not improved clinical outcomes. Evidence suggests that targeting multiple deregulated molecules is essential for better therapies; however, this is limited by the lack of suitable drugs and increased toxicity of combination therapies. To address this, we hypothesized that miRNAs, small gene-regulatory RNAs that suppress mRNA, could simultaneously inhibit multiple deregulated genes in glioblastoma and be used for more effective therapies. We identified regulatory miRNAs — those that target several deregulated genes in glioblastoma — using a combination of PAR-CLIP screening, TCGA data analyses, and an algorithm to rank target importance and miRNA therapeutic potential. We selected 2 tumor-suppressive miRNAs, miR-340 and miR-382, and 1 oncogenic miRNA, miR-17, and showed that they targeted critical glioblastoma pathways and altered cell growth, survival, invasion, and in vivo tumor growth. We developed and successfully applied a miRNA therapeutic delivery approach using brain-penetrating nanoparticles combined with MRI-guided focused ultrasound and microbubbles, to inhibit established tumor growth and extend animal survival. This strategy offers a promising approach for translating miRNA-based therapies into clinical trials for glioblastoma and other cancers.

Authors

Shekhar Saha, Ying Zhang, Myron K. Gibert Jr., Collin Dube, Farina Hanif, Elizabeth Qian Xu Mulcahy, Sylwia Bednarek, Yunan Sun, Pawel Marcinkiewicz, Xiantao Wang, Gijung Kwak, Ahsan Polash, Haolin Li, Kadie Hudson, Manikarna Dinda, Tapas Saha, Matthew McCord, Fadila Guessous, Nichola Cruickshanks, Rossymar Rivera Colon, Lily Dell’Olio, Rajitha Anbu, Wenjie Liu, Songy Choi, Benjamin Kefas, Pankaj Kumar, Alexander L. Klibanov, David Schiff, Jung Soo Suk, Justin Hanes, Jamie Mata, Markus Hafner, Roger Abounader

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Figure 5

Effects of miR-17 inhibition on cell proliferation, invasion, and neurosphere formation in glioblastoma.

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Effects of miR-17 inhibition on cell proliferation, invasion, and neuros...
(A–C) A172, U87, and U251 were transfected with either scrambled control miRNA or a miR-17 inhibitor. Cells were counted 48 hours posttransfection using trypan blue exclusion at various intervals to assess viability. (D–I) A172, U87, and U251 cells were transfected with control or miR-17 inhibitor. Invasion assay was carried out, the invaded cells were stained with crystal violet, and images were captured and analyzed using ImageJ software for quantification. (D, F, J, and L) Original magnification, ×10. (J–M) Glioma stem cell lines GSC-28 and GSC-34, plated on poly-ornithine–coated, 6-well plates, were transfected with either a scrambled control miRNA or an miR-17 inhibitor. Neurospheres were imaged 72 hours posttransfection in neurobasal complete growth medium. Images from 5 microscopic fields were taken, and neurosphere sizes were categorized into large, medium, and small for quantification using ImageJ software. Data are presented as mean ± SEM from 3 independent experiments. *P < 0.05 by 2-tailed Student’s t test.