Immuno-NanoDecoder — Nanostructured molecular decoders for the quantitative, multiplexed, layer-by-layer detection of disease-associated proteins

Results in brief

Nanostructured molecular decoders for the quantitative, multiplexed, layer-by-layer detection of disease-associated proteins

Immunofluorescence and in situ hybridization imaging are established techniques for detecting tissue-specific, cumulative changes in protein or nucleic acid profiles associated with cancer development and progression. However, the current use of fluorochromes (typically one per biomarker) does not permit the co-localization of more than four different biomarkers in the same histological section, due to the paucity of fluorochromes with non- overlapping absorption/emission spectra. In addition, fluorochromes have qualitatively distinct fluorescence properties, which limits the determination of relative amounts of biomarkers to semiquantitative visual scoring. The proposed method intends to overcome the aforementioned limits of fluorescence-based imaging to achieve definitive, multiplexed biomarker detection and quantification in situ. The value of the approach undertaken in this project is based on the premise that the quantification of only one or several markers is imprecise, but that the analysis of a larger set of defined biomarkers would permit accurate prognostic classification of the disease phenotype. This project primarily aims to develop a prototypical, functional nanodevice and to demonstrate its potential as an innovative tool for detecting multiple disease-associated proteins belonging to two widespread, and etiologically different diseases: melanoma and glycogenosis type II, the latter being a rare genetic lysosomal storage disorder. In particular, we will focus our immuno-nanodecoders to (i) the analysis of Melanoma Antigen GEnes (MAGE) for an increased capability for disease classification, and (ii) the detection acid alpha-glucosidase (GAA) and selected proteins involved in the autophagic process to in vitro assess the efficacy of small molecules to revert the cellular pathologic phenotype of glycogenosis type II. Due to the complex molecular profiles involved in such disease (see below), their study will provide an ideal testing workbench for our immuno-nanodecoders.

Data: CORDIS, © European Union

Project objective

The long-range goal of this project is to develop a molecular nanodevice, based on nucleic acid-protein conjugates, for the multiplexed, quantitative imaging of biomarkers in tissue samples and cultured cells.The Immuno-nanodecoder will be used for the accurate molecular characterization of skin cancer (melanoma) and glycogenosis type II cellular models and to evaluate the in vitro response to experimental therapies.The nanodevices (nanodecoders) will consist of self-assembled DNA nanostructures that can reversibly change their fluorescence signal output in response to hybridization to nucleic acid sequences, or to a specific enzymatic reaction. Each nanodevice will be coupled to a specific molecular probe, such as an antibody, peptide, or protein that uniquely recognize disease biomarkers. The coupling will allow the nanodecoder to detect biomarker presence and distribution in cells and tissues, in a layer-by-layer fashion, using optical fluorescence microscopy. The number of biomarkers that can be detected will be limited only by the capacity to design nanodecoders with differing specificities, which is essentially unlimited. The creation of nanodecoders and optimization of their function will greatly advance biomarker imaging, which currently lacks a high-throughput, convenient method for the in situ, quantitative microscopic analysis of altered tissue regions. This project will be driven by knowledge exchange and the expertise of an interdisciplinary team comprised of both early-stage and experienced university and hospital researchers.Complementary research programs, ranging from nanotechnology to molecular medicine and pathology will support each step of the developmental and applicative parts of the project towards the achievement of its objectives. Funding of this program will enable long-term, transformative collaborations that will contribute to the integration and collaboration of research groups between European Countries and key Third Countries.

Original text from CORDIS.

Participants

UNIVERSITA DEGLI STUDI DI ROMA TOR VERGATA · RomaCoordinatorItaly
AZIENDA SANITARIA UNIVERSITARIA FRIULI CENTRALE · UdineItaly
CONSEJO NACIONAL DE INVESTIGACIONES CIENTIFICAS Y TECNICAS (CONICET) · Buenos AiresArgentina
TEMPLE UNIVERSITY OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION · PhiladelphiaUnited States
UNIVERSITY OF LINCOLN · LincolnUnited Kingdom

Immuno-NanoDecoder · Nanostructured molecular decoders for the quantitative, multiplexed, layer-by-layer detection of disease-associated proteins

Results in brief

Nanostructured molecular decoders for the quantitative, multiplexed, layer-by-layer detection of disease-associated proteins

Project objective

Participants

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