Synthesis and functionalization of protease-activated nanoparticles with tissue plasminogen activator peptides as targeting moiety and diagnostic tool for pancreatic cancer

Sophie Dobiasch, Szilard Szanyi, Aleko Kjaev, Jens Werner, Albert Strauss, Christian Weis, Lars Grenacher, Katya Kapilov-Buchman, Liron Limor Israel, Jean Paul Lellouche, Erica Locatelli, Mauro Comes Comes Franchini, Jennifer Vandooren, Ghislain Opdenakker, Klaus Felix

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15 Scopus citations

Abstract

Background: Functionalized nanoparticles (NPs) are one promising tool for detecting specific molecular targets and combine molecular biology and nanotechnology aiming at modern imaging. We aimed at ligand-directed delivery with a suitable target-biomarker to detect early pancreatic ductal adenocarcinoma (PDAC). Promising targets are galectins (Gal), due to their strong expression in and on PDAC-cells and occurrence at early stages in cancer precursor lesions, but not in adjacent normal tissues. Results: Molecular probes (10-29 AA long peptides) derived from human tissue plasminogen activator (t-PA) were selected as binding partners to galectins. Affinity constants between the synthesized t-PA peptides and Gal were determined by microscale thermophoresis. The 29 AA-long t-PA-peptide-1 with a lactose-functionalized serine revealed the strongest binding properties to Gal-1 which was 25-fold higher in comparison with the native t-PA protein and showed additional strong binding to Gal-3 and Gal-4, both also over-expressed in PDAC. t-PA-peptide-1 was selected as vector moiety and linked covalently onto the surface of biodegradable iron oxide nanoparticles (NPs). In particular, CAN-doped maghemite NPs (CAN-Mag), promising as contrast agent for magnetic resonance imaging (MRI), were selected as magnetic core and coated with different biocompatible polymers, such as chitosan (CAN-Mag-Chitosan NPs) or polylactic co glycolic acid (PLGA) obtaining polymeric nanoparticles (CAN-Mag@PNPs), already approved for drug delivery applications. The binding efficacy of t-PA-vectorized NPs determined by exposure to different pancreatic cell lines was up to 90%, as assessed by flow cytometry. The in vivo targeting and imaging efficacy of the vectorized NPs were evaluated by applying murine pancreatic tumor models and assessed by 1.5 T magnetic resonance imaging (MRI). The t-PA-vectorized NPs as well as the protease-activated NPs with outer shell decoration (CAN-Mag@PNPs-PEG-REGAcp-PEG/tPA-pep1Lac) showed clearly detectable drop of subcutaneous and orthotopic tumor staining-intensity indicating a considerable uptake of the injected NPs. Post mortem NP deposition in tumors and organs was confirmed by Fe staining of histopathology tissue sections. Conclusions: The targeted NPs indicate a fast and enhanced deposition of NPs in the murine tumor models. The CAN-Mag@PNPs-PEG-REGAcp-PEG/tPA-pep1Lacinterlocking steps strategy of NPs delivery and deposition in pancreatic tumor is promising.

Original languageEnglish
Article number81
JournalJournal of Nanobiotechnology
Volume14
Issue number1
DOIs
StatePublished - 19 Dec 2016

Bibliographical note

Publisher Copyright:
© 2016 The Author(s).

Funding

Research funding: This project has received funding from the European Union’s Seventh Program for research, technological development and dem‑ onstration under grant agreement no 263307 (SaveMe large‑scale collabora‑ tive project). Human tissue Tissue samples were obtained from patients admitted to the Department of General, Visceral and Transplantation Surgery, University of Hei‑ delberg. The study was approved by the Ethical Committee of the University of Heidelberg (case number 301/2001) and conducted in accordance with the Helsinki Declaration; written informed consent was obtained from all patients. FFPE‑tissue samples of patients with pathologically confirmed PDAC were obtained from the Biobank of the European Pancreas Center (EPZ), Depart‑ ment of General Surgery, University Hospital Heidelberg [Pancobank team of Prof. M.W. Büchler, Dr. N.A. Giese, E. Soyka, and M. Stauch supported by BMBF grants 01GS08114 and 01ZX1305C, Heidelberger Stiftung Chirurgie and BMBH (Prof. P. Schirmacher; BMBF grant 01EY1101)].

FundersFunder number
BMBH01EY1101
European Union’s Seventh Program
Seventh Framework Programme263307
Bundesministerium für Bildung und Forschung01GS08114, 01ZX1305C
Heidelberger Stiftung Chirurgie
Universitätsklinikum Heidelberg

    Keywords

    • Galectins
    • Nanotheranostics
    • Pancreatic cancer
    • Tissue plasminogen activator

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