Ultrasound-targeted microbubble destruction to deliver siRNA cancer therapy.

Microbubble contrast agents can specifically deliver nucleic acids to target tissues when exposed to ultrasound treatment parameters that mediate microbubble destruction. In this study, we evaluated whether microbubbles and ultrasound-targeted microbubble destruction (UTMD) could be used to enhance delivery of EGF receptor (EGFR)-directed siRNA to murine squamous cell carcinomas. Custom-designed microbubbles efficiently bound siRNA and mediated RNAse protection. UTMD-mediated delivery of microbubbles loaded with EGFR-directed siRNA to murine squamous carcinoma cells in vitro reduced EGFR expression and EGF-dependent growth, relative to delivery of control siRNA. Similarly, serial UTMD-mediated delivery of EGFR siRNA to squamous cell carcinoma in vivo decreased EGFR expression and increased tumor doubling time, relative to controls receiving EGFR siRNA-loaded microbubbles but not ultrasound or control siRNA-loaded microbubbles and UTMD. Taken together, our results offer a preclinical proof-of-concept for customized microbubbles and UTMD to deliver gene-targeted siRNA for cancer therapy.

[1]  W. Saltzman,et al.  DNA diffusion in mucus: effect of size, topology of DNAs, and transfection reagents. , 2006, Biophysical journal.

[2]  D E H Tee,et al.  Book Review: Culture of Animal Cells: A Manual of Basic Technique , 1984 .

[3]  Mordecai Schwartz.  A biomathematical approach to clinical tumor growth , 1961, Cancer.

[4]  S. Fazel,et al.  Ultrasound-targeted gene delivery induces angiogenesis after a myocardial infarction in mice. , 2009, JACC. Cardiovascular imaging.

[5]  S. Ben‐Sasson,et al.  Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation , 1992, The Journal of cell biology.

[6]  F. Khuri,et al.  Tumor Growth Inhibition by Simultaneously Blocking Epidermal Growth Factor Receptor and Cyclooxygenase-2 in a Xenograft Model , 2005, Clinical Cancer Research.

[7]  G. Tortora,et al.  EGFR antagonists in cancer treatment. , 2008, The New England journal of medicine.

[8]  Zhiyi Chen,et al.  Novel ultrasound-targeted microbubble destruction mediated short hairpin RNA plasmid transfection targeting survivin inhibits gene expression and induces apoptosis of HeLa cells , 2009, Molecular Biology Reports.

[9]  R V Shohet,et al.  Echocardiographic destruction of albumin microbubbles directs gene delivery to the myocardium. , 2000, Circulation.

[10]  William R Wagner,et al.  Ultrasonic imaging of tumor angiogenesis using contrast microbubbles targeted via the tumor-binding peptide arginine-arginine-leucine. , 2005, Cancer research.

[11]  Sanjiv Kaul,et al.  Targeted tissue transfection with ultrasound destruction of plasmid-bearing cationic microbubbles. , 2003, Ultrasound in medicine & biology.

[12]  Deanna Cross,et al.  Gene therapy for cancer treatment: past, present and future. , 2006, Clinical medicine & research.

[13]  Linda Lavery,et al.  Gene therapy of carcinoma using ultrasound-targeted microbubble destruction. , 2011, Ultrasound in medicine & biology.

[14]  F. Villanueva.  Ultrasound mediated destruction of DNA-loaded microbubbles for enhancement of cell-based therapies: new promise amidst a confluence of uncertainties? , 2009, JACC. Cardiovascular imaging.

[15]  K. Fu,et al.  Modification of the effects of continuous low dose rate irradiation by concurrent chemotherapy infusion. , 1984, International journal of radiation oncology, biology, physics.

[16]  Zhi-Yi Chen,et al.  Induced apoptosis with ultrasound-mediated microbubble destruction and shRNA targeting survivin in transplanted tumors , 2009, Advances in therapy.

[17]  J. Grandis,et al.  Inhibition of human squamous cell carcinoma growth in vivo by epidermal growth factor receptor antisense RNA transcribed from the U6 promoter. , 1998, Journal of the National Cancer Institute.

[18]  Christopher U. Jones,et al.  Radiotherapy plus cetuximab for squamous-cell carcinoma of the head and neck. , 2006, The New England journal of medicine.

[19]  D. Wood,et al.  Reversal of streptozotocin-induced diabetes in rats by gene therapy with betacellulin and pancreatic duodenal homeobox-1 , 2007, Gene Therapy.

[20]  P. Grayburn,et al.  Regeneration of Pancreatic Islets in Vivo by Ultrasound-Targeted Gene Therapy , 2010, Gene Therapy.

[21]  Hui-Xiong Xu,et al.  Anti-angiogenic gene therapy for hepatocellular carcinoma mediated by microbubble-enhanced ultrasound exposure: an in vivo experimental study. , 2008, Journal of drug targeting.

[22]  Eric Tom,et al.  Myocardial Ischemic Memory Imaging With Molecular Echocardiography , 2007, Circulation.

[23]  A. McHale,et al.  Optimising ultrasound-mediated gene transfer (sonoporation) in vitro and prolonged expression of a transgene in vivo: potential applications for gene therapy of cancer. , 2009, Cancer letters.

[24]  Q. Lu,et al.  Gene transfer with microbubble ultrasound and plasmid DNA into skeletal muscle of mice: comparison between commercially available microbubble contrast agents. , 2005, Radiology.

[25]  Karen Margolis,et al.  Disclosure of Potential Conflicts of Interest , 2014 .

[26]  T. Tuschl,et al.  Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells , 2001, Nature.

[27]  Robert Langer,et al.  Knocking down barriers: advances in siRNA delivery , 2009, Nature Reviews Drug Discovery.

[28]  Raffi Bekeredjian,et al.  Ultrasound-Targeted Microbubble Destruction Can Repeatedly Direct Highly Specific Plasmid Expression to the Heart , 2003, Circulation.

[29]  Yun Chen,et al.  Tumor-targeted delivery of siRNA by self-assembled nanoparticles. , 2008, Molecular therapy : the journal of the American Society of Gene Therapy.

[30]  W. Pardridge,et al.  Intravenous RNA Interference Gene Therapy Targeting the Human Epidermal Growth Factor Receptor Prolongs Survival in Intracranial Brain Cancer , 2004, Clinical Cancer Research.

[31]  T. Jovin,et al.  Small interfering RNAs suppress the expression of endogenous and GFP-fused epidermal growth factor receptor (erbB1) and induce apoptosis in erbB1-overexpressing cells. , 2003, Experimental cell research.

[32]  A. Lee,et al.  Design of noninflammatory synthetic siRNA mediating potent gene silencing in vivo. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[33]  G. Mills,et al.  Intratumoral epidermal growth factor receptor antisense DNA therapy in head and neck cancer: first human application and potential antitumor mechanisms. , 2009, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[34]  Thomas D. Schmittgen,et al.  Analyzing real-time PCR data by the comparative CT method , 2008, Nature Protocols.

[35]  For April. , 1903 .

[36]  M. Lam,et al.  Ultrasound-contrast agent mediated naked gene delivery in the peritoneal cavity of adult rat , 2007, Gene Therapy.

[37]  Shiro Mori,et al.  Herpes simplex virus thymidine kinase-mediated suicide gene therapy using nano/microbubbles and ultrasound. , 2008, Ultrasound in medicine & biology.

[38]  V. P. Collins,et al.  Observations on growth rates of human tumors. , 1956, The American journal of roentgenology, radium therapy, and nuclear medicine.

[39]  S. Kawakami,et al.  Strategies for In Vivo Delivery of siRNAs , 2010, BioDrugs.

[40]  Raffi Bekeredjian,et al.  Optimization of ultrasound parameters for cardiac gene delivery of adenoviral or plasmid deoxyribonucleic acid by ultrasound-targeted microbubble destruction. , 2003, Journal of the American College of Cardiology.

[41]  Raffi Bekeredjian,et al.  Use of ultrasound contrast agents for gene or drug delivery in cardiovascular medicine. , 2005, Journal of the American College of Cardiology.

[42]  R. Shohet,et al.  DNA-loaded albumin microbubbles enhance ultrasound-mediated transfection in vitro. , 2002, Ultrasound in medicine & biology.

[43]  William R. Wagner,et al.  Modulating Targeted Adhesion of an Ultrasound Contrast Agent to Dysfunctional Endothelium , 2002, Annals of Biomedical Engineering.

[44]  J. Au,et al.  Delivery of siRNA Therapeutics: Barriers and Carriers , 2010, The AAPS Journal.

[45]  M. Sioud.  Induction of inflammatory cytokines and interferon responses by double-stranded and single-stranded siRNAs is sequence-dependent and requires endosomal localization. , 2005, Journal of molecular biology.