Single chain anti-c-Met antibody conjugated nanoparticles for in vivo tumor-targeted imaging and drug delivery.

Aberrantly expressed c-Met, the receptor for hepatocyte growth factor (HGF), has been implicated in human lung cancer as well as malignancy, metastasis and drug-resistance in other human cancers. Thus, this molecule could be a potential target for antibody-based cancer therapy. Targeting delivery of compound to tumor represented benefit for cancer detection and therapy. In this study, we utilized phage display to identify human single chain variable fragment (scFv) antibodies that specifically bound to c-Met protein. The anti-c-Met scFvs selectively bound to and internalized in several lung cancer cell lines expressing c-Met. Conjugation of anti-c-Met scFv with PEGylated liposomes enabled the efficient delivery of doxorubicin into cancer cells where it exerted cytotoxic activity by inducing apoptosis pathway. In addition, in vivo fluorescent imaging by scFv-conjugated quantum dots showed higher tumor uptake and increased tumor-normal tissue ratios. In a tumor xenograft model, anti-c-Met immunoliposome was found to selectively increase tumor accumulation of a chemotherapeutic drug and enhance its antitumor activity. Taken together, our results suggest that anti-c-Met scFv-mediated drug delivery systems show great promise in tumor-targeted therapy and imaging.

[1]  I. Kasman,et al.  MetMAb, the one-armed 5D5 anti-c-Met antibody, inhibits orthotopic pancreatic tumor growth and improves survival. , 2007, Cancer research.

[2]  J. Rubins,et al.  Follow-up and surveillance of the lung cancer patient following curative intent therapy: ACCP evidence-based clinical practice guideline (2nd edition). , 2007, Chest.

[3]  L. Naldini,et al.  Targeting the tumor and its microenvironment by a dual-function decoy Met receptor. , 2004, Cancer cell.

[4]  Han-Chung Wu,et al.  A Novel Peptide Specifically Binding to Nasopharyngeal Carcinoma For Targeted Drug Delivery , 2004, Cancer Research.

[5]  M. Hendrix,et al.  Selection of invasive and metastatic subpopulations from a human lung adenocarcinoma cell line. , 1997, American journal of respiratory cell and molecular biology.

[6]  M. Karamouzis,et al.  Targeting MET as a strategy to overcome crosstalk-related resistance to EGFR inhibitors. , 2009, The Lancet. Oncology.

[7]  D. Ribatti,et al.  Enhanced Antitumor Efficacy of Clinical-Grade Vasculature-Targeted Liposomal Doxorubicin , 2008, Clinical Cancer Research.

[8]  A. Marchetti,et al.  Increased MET gene copy number negatively affects survival of surgically resected non-small-cell lung cancer patients. , 2009, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[9]  M. Berger,et al.  Identification of Internalizing Human Single-Chain Antibodies Targeting Brain Tumor Sphere Cells , 2010, Molecular Cancer Therapeutics.

[10]  P. Scherle,et al.  Developing c-MET pathway inhibitors for cancer therapy: progress and challenges. , 2010, Trends in molecular medicine.

[11]  P. Comoglio,et al.  Overexpression and activation of hepatocyte growth factor/scatter factor in human non-small-cell lung carcinomas. , 1996, British Journal of Cancer.

[12]  Kristian Pietras,et al.  High interstitial fluid pressure — an obstacle in cancer therapy , 2004, Nature Reviews Cancer.

[13]  A. Jemal,et al.  Cancer Statistics, 2009 , 2009, CA: a cancer journal for clinicians.

[14]  L. Tanoue.  Cancer Statistics, 2009 , 2010 .

[15]  Erkki Ruoslahti,et al.  Tissue-penetrating delivery of compounds and nanoparticles into tumors. , 2009, Cancer cell.

[16]  T. Blundell,et al.  Functional map and domain structure of MET, the product of the c-met protooncogene and receptor for hepatocyte growth factor/scatter factor , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[17]  H R Hoogenboom,et al.  By-passing immunization. Human antibodies from V-gene libraries displayed on phage. , 1991, Journal of molecular biology.

[18]  C. Benz,et al.  Anti-CD166 single chain antibody-mediated intracellular delivery of liposomal drugs to prostate cancer cells , 2007, Molecular Cancer Therapeutics.

[19]  Chin-Tarng Lin,et al.  Peptide-mediated targeting to tumor blood vessels of lung cancer for drug delivery. , 2007, Cancer research.

[20]  Chien-Hsun Wu,et al.  A Novel Peptide Enhances Therapeutic Efficacy of Liposomal Anti-Cancer Drugs in Mice Models of Human Lung Cancer , 2009, PloS one.

[21]  H. Kroemer,et al.  Elucidation of the mechanism enabling tumor selective prodrug monotherapy. , 1998, Cancer research.

[22]  U. Nielsen,et al.  Tumor targeting using anti-her2 immunoliposomes. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[23]  Luca Toschi,et al.  Single-Agent and Combination Therapeutic Strategies to Inhibit Hepatocyte Growth Factor/MET Signaling in Cancer , 2008, Clinical Cancer Research.

[24]  J. Christensen,et al.  The MET receptor tyrosine kinase is a potential novel therapeutic target for head and neck squamous cell carcinoma. , 2009, Cancer research.

[25]  D. Christ,et al.  Selection of human antibody fragments by phage display , 2007, Nature Protocols.

[26]  J. Rubins,et al.  Follow-up and surveillance of the lung cancer patient following curative-intent therapy. , 2003, Chest.

[27]  Z. Han,et al.  HGF receptor up-regulation contributes to the angiogenic phenotype of human endothelial cells and promotes angiogenesis in vitro. , 2003, Blood.

[28]  R. Cardiff,et al.  Met induces mammary tumors with diverse histologies and is associated with poor outcome and human basal breast cancer , 2009, Proceedings of the National Academy of Sciences.

[29]  A. Ido,et al.  Translational research to identify clinical applications of hepatocyte growth factor , 2009, Hepatology research : the official journal of the Japan Society of Hepatology.

[30]  B. Liu,et al.  Targeted drug delivery to mesothelioma cells using functionally selected internalizing human single-chain antibodies , 2008, Molecular Cancer Therapeutics.

[31]  Weibo Cai,et al.  Preparation of peptide-conjugated quantum dots for tumor vasculature-targeted imaging , 2008, Nature Protocols.

[32]  Theresa M Allen,et al.  Internalizing antibodies are necessary for improved therapeutic efficacy of antibody-targeted liposomal drugs. , 2002, Cancer research.

[33]  Matthew R Cooperberg,et al.  Mapping Tumor Epitope Space by Direct Selection of Single-Chain Fv Antibody Libraries on Prostate Cancer Cells , 2004, Cancer Research.

[34]  Han-Chung Wu,et al.  Antiangiogenic Targeting Liposomes Increase Therapeutic Efficacy for Solid Tumors* , 2009, Journal of Biological Chemistry.

[35]  F. Dosio,et al.  Stealth liposomes: review of the basic science, rationale, and clinical applications, existing and potential , 2006, International journal of nanomedicine.

[36]  A. Gabizon,et al.  Sterically stabilized liposomes: improvements in pharmacokinetics and antitumor therapeutic efficacy. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Chin-Tarng Lin,et al.  Hepatocellular carcinoma cell-specific peptide ligand for targeted drug delivery , 2008, Molecular Cancer Therapeutics.

[38]  J. Marks,et al.  Toward selection of internalizing antibodies from phage libraries. , 1999, Biochemical and biophysical research communications.

[39]  N. Fausto.  Hepatocyte growth factor receptor and the c-met oncogene Bottaro DP, Rubin JS, Faletto DL, Chan AM-L, Kmiecik TE, Vande Woude GF, Aaronson SA. Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product. Science 1991;251:802?804 , 1991 .

[40]  W. Birchmeier,et al.  Met, metastasis, motility and more , 2003, Nature Reviews Molecular Cell Biology.

[41]  V. Torchilin.  Recent advances with liposomes as pharmaceutical carriers , 2005, Nature Reviews Drug Discovery.

[42]  N. Hanna,et al.  MET Amplification Leads to Gefitinib Resistance in Lung Cancer by Activating ERBB3 Signaling , 2008 .

[43]  A. Coxon,et al.  Safety, Pharmacokinetics, and Pharmacodynamics of AMG 102, a Fully Human Hepatocyte Growth Factor–Neutralizing Monoclonal Antibody, in a First-in-Human Study of Patients with Advanced Solid Tumors , 2010, Clinical Cancer Research.

[44]  J. Rubin,et al.  Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product. , 1991, Science.