Dual-Receptor Targeted Nanoparticles for Photodynamic Therapy of Brain Cancer

Summary

Principal Investigator: Ann Marie Broome
Abstract: DESCRIPTION (provided by applicant): Brain cancer is a life threatening disease characterized by low survival rates. The development of selectively targeted nanoparticles conjugated with drugs is critical for improving the treatment and monitoring of this aggressive type of cancer. Photodynamic therapy (PDT) is a localized treatment modality that is promising for brain tumor treatment. Although improvements in survival were reported, the widespread use of PDT in brain tumor therapy has been partially hampered by non-targeted phototoxicity towards healthy tissue. Improving the selectivity of tumor targeting and sustained delivery of PDT drugs will dramatically enhance the success of brain cancer therapy. Pc 4 is a highly promising PDT drug, approved for clinical trials, characterized by virtual non-toxicity in the dark, high phototoxicity, and well defined chemical structure and purity. This drug operates in the near infrared (NIR) spectral range, which penetrates brain tissue most efficiently for both diagnostic optical imaging and phototherapy. The objective of the proposed research is to develop Pc 4 loaded gold nanoparticle (Au NP-Pc 4) conjugates with a thiolated PEG coating for targeted imaging-guided therapy of glioma brain cancers. We will adapt a novel cross-disciplinary approach to synthesize PEG-coated gold nanoparticles conjugated to Pc 4 and tethered peptide ligands for targeting epidermal growth factor (EGF) and transferrin (Tf) cell surface receptors, which are overexpressed in brain cancer cells. By applying genetic cell engineering, we will develop model cell lines and animal systems with human EGF and Tf receptors, expressed separately or jointly as viable cancer biomarkers on rat 9L glioma cells. We will perform state of the art in vitro and in vivo fluorescence imaging to characterize the delivery and targeting of the conjugates, as well as determining their therapeutic (PDT) efficacy. The central hypothesis is that using a dual-targeting ligand concept will dramatically improve PDT nanoparticle selectivity to brain cancers. To test this hypothesis, we will develop and test both in vitro and in vivo nanoparticle PDT efficacy to fulfill the following aims: Specific Aim 1: Development and characterization of PEGylated Au NP-Pc 4 conjugates containing EGFR and TfR binding peptide ligands. Targeted NPs loaded with the PDT drug Pc 4 will be synthesized, fully characterized (in terms of structure, ligand density, and drug loading), and optimized for selective targeting and drug release. To support the optimization process, we will perform Au NP: receptor interaction studies of each NP conjugate design, including not only equilibrium data, but also kinetic parameters of the interactions using surface plasmon resonance (SPR) Biacore technology. Specific Aim 2: Targeted nanoparticle conjugate delivery and PDT efficacy testing in vitro in Tf and EGF receptor-bearing 9L glioma cell lines. The ability to target overexpressed human receptors will be studied using engineered 9L glioma cell lines overexpressing human EGFR and TFR, separately and in combination. We will examine the uptake and localization of targeted NPs using various experimental techniques, including silver enhancement immunohistochemistry, real time confocal laser scanning microscopy, and transmission electron microscopy. The cells will then be subjected to PDT, and cellular viability will be assessed using the MTT assay. Since intracellular localization of the PDT drug is a precursor to downstream cellular events, such as apoptosis, we will also assess the mechanism of Pc 4-mediated programmed cell death using a mitochondrial membrane potential assay, TUNEL and DNA fragmentation assays, and cell permeability assays, including trypan blue staining and Annexin V/ propidium iodide flow cytometry. Specific Aim 3: In vivo translation of PDT therapy and post-therapy monitoring in glioma tumor bearing mice. We will investigate the NP targeting and the PDT efficacy of the NP conjugates in vivo in receptor overexpressing 9L tumor bearing mice using 3-dimensional fluorescence molecular tomography (FMT). We will determine circulation, biodistribution, and clearance of the targeted Au NPs and the drug Pc 4 using ICP/AAS elemental analysis and silver enhancement immunohistochemistry, and fluorescence imaging to evaluate relative concentrations of Pc 4 and the Au NPs. We will also examine the pathology of the tumors after PDT using dynamic fluorescent imaging over a seven day period. The ultimate goal of this project is a drastic improvement of combined brain cancer treatment and monitoring using a highly targeted, virtually non-toxic PDT sensitizer that can be locally activated and interrogated in real time with high spatio-temporal resolution. PUBLIC HEALTH RELEVANCE: Malignant gliomas are the most common primary brain tumors and among the most lethal cancers in man. Cell surface receptor-targeted gold nanoparticles when conjugated with Pc 4, a photodynamic therapy drug, can be molecular imaging agents used to improve the specificity of detection of these brain cancers. Our multidisciplinary research plan involves chemical synthesis and characterization of targeted nanoparticles, in vitro cell culture studies, and in vivo studies of mouse models of human glioma carcinomas. By improving the selectivity of tumor targeting, we can potentially sustain local delivery of PDT drugs, thus dramatically enhancing the success of brain cancer therapy.
Funding Period: 2010-09-30 - 2014-07-31
more information: NIH RePORT

Top Publications

  1. pmc Deep penetration of a PDT drug into tumors by noncovalent drug-gold nanoparticle conjugates
    Yu Cheng
    Department of Chemistry, Case Western Reserve University, Cleveland, Ohio 44106, USA
    J Am Chem Soc 133:2583-91. 2011
  2. pmc Addressing brain tumors with targeted gold nanoparticles: a new gold standard for hydrophobic drug delivery?
    Yu Cheng
    Department of Chemistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA, Fax 1 216 368 3006
    Small 7:2301-6. 2011
  3. pmc Nanoparticle mediated non-covalent drug delivery
    Tennyson Doane
    Department of Chemistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA
    Adv Drug Deliv Rev 65:607-21. 2013
  4. pmc An optical probe for noninvasive molecular imaging of orthotopic brain tumors overexpressing epidermal growth factor receptor
    Richard S Agnes
    Departments of Radiology, Biomedical Engineering, and Pathology, NFCR Center for Molecular Imaging at Case, Case Western Reserve University, Wearn Building, Room B 42, 11100 Euclid Avenue, Cleveland, OH 44106, USA
    Mol Cancer Ther 11:2202-11. 2012
  5. pmc Nanoparticles for imaging and treating brain cancer
    Joseph D Meyers
    Departments of Biomedical Engineering and Radiology, Case Western Reserve University, Cleveland, OH 44106, USA
    Nanomedicine (Lond) 8:123-43. 2013

Research Grants

  1. Mayo Clinic Breast Cancer SPORE
    JAMES NEWELL INGLE; Fiscal Year: 2013
  2. Mechanisms and Markers of Prostate Cancer Metastases
    ROBERT LOUIS VESSELLA; Fiscal Year: 2013
  3. WINCART: Weaving an Islander Network for Cancer Awareness, Research and Training
    Sora P Tanjasiri; Fiscal Year: 2013
  4. In Vivo Cellular and Molecular Imaging Center@Stanford
    Sanjiv S Gambhir; Fiscal Year: 2013
  5. Biology and Therapy of High Risk Neuroblastoma
    ROBERT CHARLES SEEGER; Fiscal Year: 2013
  6. CENTER FOR BIOMEDICAL RESEARCH
    Timothy Turner; Fiscal Year: 2013

Detail Information

Publications5

  1. pmc Deep penetration of a PDT drug into tumors by noncovalent drug-gold nanoparticle conjugates
    Yu Cheng
    Department of Chemistry, Case Western Reserve University, Cleveland, Ohio 44106, USA
    J Am Chem Soc 133:2583-91. 2011
    ..This study suggests that noncovalent delivery via Au NPs provides an attractive approach for cancer drugs to penetrate deep into the center of tumors...
  2. pmc Addressing brain tumors with targeted gold nanoparticles: a new gold standard for hydrophobic drug delivery?
    Yu Cheng
    Department of Chemistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA, Fax 1 216 368 3006
    Small 7:2301-6. 2011
    ..This delivery system holds promise for future delivery of a wider range of hydrophobic therapeutic drugs for the treatment of hard-to-reach cancers. ..
  3. pmc Nanoparticle mediated non-covalent drug delivery
    Tennyson Doane
    Department of Chemistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA
    Adv Drug Deliv Rev 65:607-21. 2013
    ..Through a critical evaluation of advances in drug delivery within the last decade, an outlook for biomedical applications of nanoscale transport vectors will be presented...
  4. pmc An optical probe for noninvasive molecular imaging of orthotopic brain tumors overexpressing epidermal growth factor receptor
    Richard S Agnes
    Departments of Radiology, Biomedical Engineering, and Pathology, NFCR Center for Molecular Imaging at Case, Case Western Reserve University, Wearn Building, Room B 42, 11100 Euclid Avenue, Cleveland, OH 44106, USA
    Mol Cancer Ther 11:2202-11. 2012
    ..The data presented here represent the first demonstration of differential quantitation of tumors expressing EGFR in live animals by a targeted NIR fluorescence probe using a molecular imaging device...
  5. pmc Nanoparticles for imaging and treating brain cancer
    Joseph D Meyers
    Departments of Biomedical Engineering and Radiology, Case Western Reserve University, Cleveland, OH 44106, USA
    Nanomedicine (Lond) 8:123-43. 2013
    ..This review serves to present the reader with an overview of NPs for treating brain cancer and to provide an outlook on what may come in the future. For NPs, just like the blood-brain-tumor barrier, the future is wide open...

Research Grants30

  1. Mayo Clinic Breast Cancer SPORE
    JAMES NEWELL INGLE; Fiscal Year: 2013
    ..abstract_text> ..
  2. Mechanisms and Markers of Prostate Cancer Metastases
    ROBERT LOUIS VESSELLA; Fiscal Year: 2013
    ..It also conducts pre-clinical xenograft studies and provides statistical support to the overall P01 program. Core B is the Administrative Core which provides overall administrative support to the investigators. ..
  3. WINCART: Weaving an Islander Network for Cancer Awareness, Research and Training
    Sora P Tanjasiri; Fiscal Year: 2013
    ....
  4. In Vivo Cellular and Molecular Imaging Center@Stanford
    Sanjiv S Gambhir; Fiscal Year: 2013
    ..The ICMIC@Stanford team is committed to cancer research and clinical translation of state-of-the-art molecular imaging strategies for improving clinical cancer management. ..
  5. Biology and Therapy of High Risk Neuroblastoma
    ROBERT CHARLES SEEGER; Fiscal Year: 2013
    ..nant.org), which includes 15 pediatric oncology institutions across the US and in Canada. ..
  6. CENTER FOR BIOMEDICAL RESEARCH
    Timothy Turner; Fiscal Year: 2013
    ..abstract_text> ..