Phage Display-Derived Peptides for Osteosarcoma Imaging

National Institute of Biomedical Imaging and Bioengineering, NIH, Bethesda, MD, USA.
Clinical Cancer Research (Impact Factor: 8.72). 08/2010; 16(16):4268-77. DOI: 10.1158/1078-0432.CCR-10-0968
Source: PubMed


Osteosarcoma represents the most common malignant primary bone tumor in childhood; however, the survival rate has remained unchanged for the past 20 years. To improve existing diagnosis and treatment methods and broaden the spectrum of imaging agents that can be used for early detection and assessment of tumor response to therapy, we performed a phage display-based screening for peptide sequences that bind specifically to osteosarcoma cells.
From the Ph.D.-12 phage display peptide library composed of 2.7 x 10(9) different displayed peptides, one peptide was enriched after four rounds of in vitro selection in 143B osteosarcoma tumor cells with 293T human embryonic kidney cells as a control. Both the peptide and the phage clone displaying the peptide were conjugated with fluorescent dyes for in vitro cell and ex vivo tumor tissue stainings. The peptide was further labeled with (18)F for positron emission tomography imaging studies. Cell uptake and efflux and ex vivo biodistribution were also done with (18)F-labeled osteosarcoma specific peptide.
ASGALSPSRLDT was the dominant sequence isolated from biopanning and named as OSP-1. OSP-1 shares a significant homology with heparinase II/III family protein, which binds and reacts with heparan sulfate proteoglycans. The fluorescence staining showed that FITC-OSP-1-phage or Cy5.5-OSP-1 had high binding with a panel of osteosarcoma cell lines, much less binding with UM-SCC1 human head and neck squamous cell carcinoma cells, and almost no binding with 293T cells, whereas the scrambled peptide OSP-S had virtually no binding to all the cell lines. (18)F-OSP-1 had significantly higher accumulation in 143B tumor cells both in vitro and in vivo than (18)F-OSP-S. (18)F-OSP-1 also had higher uptake in 143B tumors than in UM-SCC-1 tumors.
Our data suggest that OSP-1 peptide is osteosarcoma specific, and the binding site of OSP-1 might be related to heparan sulfate proteoglycans. Appropriately labeled OSP-1 peptide has the potential to serve as a novel probe for osteosarcoma imaging.

Download full-text


Available from: Nicholas Chan, Sep 26, 2014
  • Source
    • "The technique used along with the method used for pathologic evaluation of the tissue specimen determines how efficiently the disease is identified. Additional methods for enhancing diagnostic efficiency could include biomarkers or novel cell displays.18,36–56 We have evaluated the CytoViva® illumination nanotechnology which, apart from being a novel method of display, contains software that is able to learn and store new patterns. "
    [Show abstract] [Hide abstract]
    ABSTRACT: There are currently many techniques and devices available for the diagnosis of lung cancer. However, rapid on-site diagnosis is essential for early-stage lung cancer, and in the current work we investigated a new diagnostic illumination nanotechnology. Tissue samples were obtained from lymph nodes, cancerous tissue, and abnormal intrapulmonary lesions at our interventional pulmonary suites. The following diagnostic techniques were used to obtain the samples: endobronchial ultrasound bronchoscopy; flexible bronchoscopy; and rigid bronchoscopy. Flexible and rigid forceps were used because several of the patients were intubated using a rigid bronchoscope. In total, 30 tissue specimens from 30 patients were prepared. CytoViva® illumination nanotechnology was subsequently applied to each of the biopsy tissue slides. A spectral library was created for adenocarcinoma, epidermal growth factor receptor mutation-positive adenocarcinoma, squamous cell carcinoma, usual interstitial pneumonitis, non-specific interstitial pneumonitis, typical carcinoid tumor, sarcoidosis, idiopathic pulmonary fibrosis, small cell neuroendocrine carcinoma, thymoma, epithelioid and sarcomatoid mesothelioma, cryptogenic organizing pneumonia, malt cell lymphoma, and Wegener's granulomatosis. The CytoViva software, once it had created a specific spectral library for each entity, was able to identify the same disease again in subsequent paired sets of slides of the same disease. Further evaluation of this technique could make this illumination nanotechnology an efficient rapid on-site diagnostic tool.
    International Journal of Nanomedicine 11/2013; 8:4533-4542. DOI:10.2147/IJN.S54418 · 4.38 Impact Factor
  • Source
    • "Although Raman spectroscopy has been applied to distinguish two types of microcalcifications occurring in benign and malignant lesions, it is unclear whether the Raman spectroscopic method is suitable for distinguishing microcalcifications in vivo [39] [40]. Currently, a variety of imaging modalities and spectroscopic techniques are being explored to improve osteotropic cancer diagnosis and treatment [41] [42] [43] [44]. However , there is no reliable diagnostic imaging marker or medical modality to distinguish accurately between CO and HA calcifications with high affinity and specificity in biological tissues non-invasively. "
    [Show abstract] [Hide abstract]
    ABSTRACT: BACKGROUND: Although microcalcifications of hydroxyapatite can be found in both benign and malignant osteotropic tumors, they are mostly seen in proliferative lesions, including carcinoma. The aim of present study is to develop a molecular imaging contrast agent for selective identification of hydroxyapatite calcification in human osteotropic tumor tissues ex vivo and in human osteosarcoma cells in vitro. METHODS: A bioinspired biomarker, hydroxyapatite binding peptide (HABP), was designed to mimic natural protein osteocalcin property in vivo. A fluorescein isothiocyanate dye conjugated HABP (HABP-19) was utilized to characterize hydroxyapatite on human osteotropic tumor tissue sections ex vivo and to selectively image hydroxyapatite calcifications in human osteosarcoma cells in vitro. RESULTS: Using a HABP-19 molecular imaging probe, we have shown that it is possible to selectively image hydroxyapatite calcifications in osteotropic cancers ex vivo and in human SaOS-2 osteosarcoma cells in vitro. CONCLUSION: Hydroxyapatite calcifications were selectively detected in osteotropic tissues ex vivo and in the early stage of the calcification process of SaOS-2 human osteosarcoma in vitro using our HABP-19 molecular imaging probe. This new target-selective molecular imaging probe makes it possible to study the earliest events associated with hydroxyapatite deposition in various osteotropic cancers at the cellular and molecular levels. GENERAL SIGNIFICANCE: It potentially could be used to diagnosis and treat osteotropic cancer or to anchor therapeutic agents directing the local distribution of desired therapy at calcified sites.
    Biochimica et Biophysica Acta 05/2013; 1830(10). DOI:10.1016/j.bbagen.2013.05.015 · 4.66 Impact Factor
  • Source
    • "Molecular imaging technologies are considered promising methods because they obtain the information through monitoring the key molecular behaviours and host responses related to early events in disease development and progress at the cellular and molecular levels [1,3]. Compared with traditional imaging techniques which are mainly based on anatomical structures of organs, molecular imaging usually utilizes specific molecular probes targeting unique receptors (molecules) of tumor tissues or other diseased tissues to form the localized pictures of image contrast[4]. Thus, it becomes the key point to identify and generate the tumor-specific molecular ligands with high binding affinity. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Tumor vasculature expresses a distinct set of molecule signatures on the endothelial cell surface different from the resting blood vessels of other organs and tissues in the body. This makes them an attractive target for cancer therapy and molecular imaging. The current technology using the in vivo phage display biopanning allows us to quickly isolate and identify peptides potentially homing to various tumor blood vessels. Tumor-homing peptides in conjugation with chemotherapeutic drugs or imaging contrast have been extensively tested in various preclinical and clinical studies. These tumor-homing peptides have valuable potential as targeting probes for tumor molecular imaging and drug delivery. In this review, we summarize the recent advances about the applications of tumor-homing peptides selected by in vivo phage display library screening against tumor vasculature. We also introduce the characteristics of the latest discovered tumor-penetrating peptides in their potential clinical applications.
    Journal of Translational Medicine 09/2012; 10 Suppl 1(Suppl 1):S1. DOI:10.1186/1479-5876-10-S1-S1 · 3.93 Impact Factor
Show more