Localized RNAi therapeutics of chemoresistant grade IV glioma using hyaluronan-grafted lipid-based nanoparticles.

Glioblastoma multiforme (GBM) is one of the most infiltrating, aggressive, and poorly treated brain tumors. Progress in genomics and proteomics has paved the way for identifying potential therapeutic targets for treating GBM, yet the vast majority of these leading drug candidates for the treatment of GBM are ineffective, mainly due to restricted passages across the blood-brain barrier. Nanoparticles have been emerged as a promising platform to treat different types of tumors due to their ability to transport drugs to target sites while minimizing adverse effects. Herein, we devised a localized strategy to deliver RNA interference (RNAi) directly to the GBM site using hyaluronan (HA)-grafted lipid-based nanoparticles (LNPs). These LNPs having an ionized lipid were previously shown to be highly effective in delivering small interfering RNAs (siRNAs) into various cell types. LNP's surface was functionalized with hyaluronan (HA), a naturally occurring glycosaminoglycan that specifically binds the CD44 receptor expressed on GBM cells. We found that HA-LNPs can successfully bind to GBM cell lines and primary neurosphers of GBM patients. HA-LNPs loaded with Polo-Like Kinase 1 (PLK1) siRNAs (siPLK1) dramatically reduced the expression of PLK1 mRNA and cumulated in cell death even under shear flow that simulate the flow of the cerebrospinal fluid compared with control groups. Next, a human GBM U87MG orthotopic xenograft model was established by intracranial injection of U87MG cells into nude mice. Convection of Cy3-siRNA entrapped in HA-LNPs was performed, and specific Cy3 uptake was observed in U87MG cells. Moreover, convection of siPLK1 entrapped in HA-LNPs reduced mRNA levels by more than 80% and significantly prolonged survival of treated mice in the orthotopic model. Taken together, our results suggest that RNAi therapeutics could effectively be delivered in a localized manner with HA-coated LNPs and ultimately may become a therapeutic modality for GBM.

[1]  D. Peer,et al.  Overcoming multidrug resistance with nanomedicines , 2015, Expert opinion on drug delivery.

[2]  F. Zanella,et al.  Fluorescence-guided surgery with 5-aminolevulinic acid for resection of malignant glioma: a randomised controlled multicentre phase III trial. , 2006, The Lancet. Oncology.

[3]  L. Ricci-Vitiani,et al.  Combined PDK1 and CHK1 inhibition is required to kill glioblastoma stem-like cells in vitro and in vivo , 2014, Cell Death and Disease.

[4]  B. MacVicar,et al.  Lipid Nanoparticle Delivery of siRNA to Silence Neuronal Gene Expression in the Brain , 2013, Molecular therapy. Nucleic acids.

[5]  Saliha Şahin,et al.  Olea europaea leaf extract improves the treatment response of GBM stem cells by modulating miRNA expression. , 2014, American journal of cancer research.

[6]  Q. Zhang,et al.  Liposome formulated with TAT-modified cholesterol for improving brain delivery and therapeutic efficacy on brain glioma in animals. , 2011, International journal of pharmaceutics.

[7]  Volker Seifert,et al.  Intraoperative MRI guidance and extent of resection in glioma surgery: a randomised, controlled trial. , 2011, The Lancet. Oncology.

[8]  F. Szoka,et al.  Anticancer therapeutics: targeting macromolecules and nanocarriers to hyaluronan or CD44, a hyaluronan receptor. , 2008, Molecular pharmaceutics.

[9]  D. Peer,et al.  Polysaccharides as building blocks for nanotherapeutics. , 2012, Chemical Society reviews.

[10]  Keith Bowman,et al.  Potent and persistent in vivo anti-HBV activity of chemically modified siRNAs , 2005, Nature Biotechnology.

[11]  B. Pineda,et al.  Application of Nanoparticles on Diagnosis and Therapy in Gliomas , 2013, BioMed research international.

[12]  Hyaluronan grafted lipid-based nanoparticles as RNAi carriers for cancer cells. , 2013, Cancer letters.

[13]  Shinn-Zong Lin,et al.  Targeting Cancer Stem Cells for Treatment of Glioblastoma Multiforme , 2013, Cell transplantation.

[14]  D. Peer,et al.  Loading mitomycin C inside long circulating hyaluronan targeted nano‐liposomes increases its antitumor activity in three mice tumor models , 2004, International journal of cancer.

[15]  Y. Mardor,et al.  Salirasib (farnesyl thiosalicylic acid) for brain tumor treatment: a convection-enhanced drug delivery study in rats , 2008, Molecular Cancer Therapeutics.

[16]  Ashley M. Jacobi,et al.  RNA Inhibition Highlights Cyclin D1 as a Potential Therapeutic Target for Mantle Cell Lymphoma , 2012, PloS one.

[17]  C. Wirtz,et al.  Combined Inhibition of HER1/EGFR and RAC1 Results in a Synergistic Antiproliferative Effect on Established and Primary Cultured Human Glioblastoma Cells , 2013, Molecular Cancer Therapeutics.

[18]  Shigeo Matsuda,et al.  Maximizing the Potency of siRNA Lipid Nanoparticles for Hepatic Gene Silencing In Vivo** , 2012, Angewandte Chemie.

[19]  Z. Ram,et al.  Adenovirus-mediated gene therapy with sitimagene ceradenovec followed by intravenous ganciclovir for patients with operable high-grade glioma (ASPECT): a randomised, open-label, phase 3 trial. , 2013, The Lancet. Oncology.

[20]  D. Peer,et al.  Paclitaxel-clusters coated with hyaluronan as selective tumor-targeted nanovectors. , 2010, Biomaterials.

[21]  D. Peer,et al.  Hyaluronan-coated nanoparticles: the influence of the molecular weight on CD44-hyaluronan interactions and on the immune response. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[22]  Dan Peer,et al.  Tumor targeting profiling of hyaluronan-coated lipid based-nanoparticles. , 2014, Nanoscale.

[23]  Qin He,et al.  Lactoferrin modified doxorubicin-loaded procationic liposomes for the treatment of gliomas. , 2011, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[24]  J. Kuratsu,et al.  Antitumor Effect of NK012, a 7-Ethyl-10-Hydroxycamptothecin–Incorporating Polymeric Micelle, on U87MG Orthotopic Glioblastoma in Mice Compared with Irinotecan Hydrochloride in Combination with Bevacizumab , 2010, Clinical Cancer Research.

[25]  Wei Wu,et al.  Sulforaphane Inhibits Invasion via Activating ERK1/2 Signaling in Human Glioblastoma U87MG and U373MG Cells , 2014, PloS one.

[26]  Dan Peer,et al.  Modulation of drug resistance in ovarian adenocarcinoma using chemotherapy entrapped in hyaluronan-grafted nanoparticle clusters. , 2014, ACS nano.

[27]  H. Harashima,et al.  Improvement of doxorubicin efficacy using liposomal anti-polo-like kinase 1 siRNA in human renal cell carcinomas. , 2014, Molecular pharmaceutics.

[28]  Xiao-wei Zhu,et al.  Clinicopathological significance of Polo-like kinase 1 (PLK1) expression in human malignant glioma. , 2012, Acta histochemica.

[29]  G. Faccani,et al.  Glioblastoma cancer stem cells: heterogeneity, microenvironment and related therapeutic strategies , 2010, Cell biochemistry and function.

[30]  K. Kawakami,et al.  Glycogen synthase kinase 3β inhibition sensitizes human glioblastoma cells to temozolomide by affecting O6-methylguanine DNA methyltransferase promoter methylation via c-Myc signaling. , 2013, Carcinogenesis.

[31]  M. Weller,et al.  Programmed death ligand 1 expression and tumor-infiltrating lymphocytes in glioblastoma. , 2015, Neuro-oncology.

[32]  Daniel G. Anderson,et al.  Therapeutic siRNA silencing in inflammatory monocytes , 2011, Nature Biotechnology.

[33]  H. Too,et al.  MicroRNA as potential modulators in chemoresistant high-grade gliomas , 2014, Journal of Clinical Neuroscience.

[34]  D. Peer,et al.  Modulating cancer multidrug resistance by sertraline in combination with a nanomedicine. , 2014, Cancer letters.

[35]  Minhyung Lee,et al.  Combined delivery of BCNU and VEGF siRNA using amphiphilic peptides for glioblastoma , 2014, Journal of drug targeting.

[36]  F. Szoka,et al.  Liposome-encapsulated doxorubicin targeted to CD44: a strategy to kill CD44-overexpressing tumor cells. , 2001, Cancer research.

[37]  R. Alon,et al.  Chapter 14. Real-time in vitro assays for studying the role of chemokines in lymphocyte transendothelial migration under physiologic flow conditions. , 2009, Methods in enzymology.

[38]  J. Phillips.  Novel Therapeutic Targets in the Brain Tumor Microenvironment , 2012, Oncotarget.

[39]  B. Davidson,et al.  Transvascular delivery of small interfering RNA to the central nervous system , 2007, Nature.

[40]  Diane D. Liu,et al.  A High Notch Pathway Activation Predicts Response to γ Secretase Inhibitors in Proneural Subtype of Glioma Tumor‐Initiating Cells , 2014, Stem cells.

[41]  AGR2 Expression is Regulated by HIF-1 and Contributes to Growth and Angiogenesis of Glioblastoma , 2013, Cell Biochemistry and Biophysics.

[42]  T. Honda,et al.  Systemic Leukocyte-Directed siRNA Delivery Revealing Cyclin D 1 as an Anti-Inflammatory Target , 2022 .

[43]  K. Dawson,et al.  Detecting Cryptic Epitopes Created by Nanoparticles , 2006, Science's STKE.

[44]  P. Hammond,et al.  Controlling in vivo stability and biodistribution in electrostatically assembled nanoparticles for systemic delivery. , 2011, Nano letters.

[45]  Y. Cheng,et al.  Durable Therapeutic Efficacy Utilizing Combinatorial Blockade against IDO, CTLA-4, and PD-L1 in Mice with Brain Tumors , 2014, Clinical Cancer Research.

[46]  Jun Wang,et al.  Targeted Delivery of PLK1-siRNA by ScFv Suppresses Her2+ Breast Cancer Growth and Metastasis , 2012, Science Translational Medicine.

[47]  D. Peer,et al.  Tumor-targeted hyaluronan nanoliposomes increase the antitumor activity of liposomal Doxorubicin in syngeneic and human xenograft mouse tumor models. , 2004, Neoplasia.

[48]  J. Karp,et al.  Nanocarriers as an Emerging Platform for Cancer Therapy , 2022 .

[49]  Xiangyang Xie,et al.  Improving efficiency of adriamycin crossing blood brain barrier by combination of thermosensitive liposomes and hyperthermia. , 2011, Biological & pharmaceutical bulletin.

[50]  D. Peer,et al.  Hyaluronan-grafted particle clusters loaded with Mitomycin C as selective nanovectors for primary head and neck cancers. , 2011, Biomaterials.

[51]  Raghu Raghavan,et al.  Poor drug distribution as a possible explanation for the results of the PRECISE trial. , 2010, Journal of neurosurgery.

[52]  D. Peer,et al.  Systemic Leukocyte-Directed siRNA Delivery Revealing Cyclin D1 as an Anti-Inflammatory Target , 2008, Science.

[53]  Z L Gokaslan,et al.  A multivariate analysis of 416 patients with glioblastoma multiforme: prognosis, extent of resection, and survival. , 2001, Journal of neurosurgery.