scholarly journals Development of an orally-administrable tumor vasculature-targeting therapeutic using annexin A1-binding D-peptides

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0241157
Author(s):  
Motohiro Nonaka ◽  
Hideaki Mabashi-Asazuma ◽  
Donald L. Jarvis ◽  
Kazuhiko Yamasaki ◽  
Tomoya O. Akama ◽  
...  
Keyword(s):  
Brain Tumor ◽  
Near Infrared ◽  
Tumor Vasculature ◽  
Mirror Image ◽  
Whole Body ◽  
Whole Body Imaging ◽  
Annexin A1 ◽  
N Terminus ◽  
Peptide Phage Display

We previously reported that IF7 peptide, which binds to the annexin A1 (ANXA1) N-terminus, functions as a tumor vasculature-targeted drug delivery vehicle after intravenous injection. To enhance IF7 stability in vivo, we undertook mirror-image peptide phage display using a synthetic D-peptide representing the ANXA1 N-terminus as target. We then identified peptide sequences, synthesized them as D-amino acids, and designated the resulting peptide dTIT7, which we showed bound to the ANXA1 N-terminus. Whole body imaging of mouse brain tumor models injected with near infrared fluorescent IRDye-conjugated dTIT7 showed fluorescent signals in brain and kidney. Furthermore, orally-administered dTIT7/geldanamycin (GA) conjugates suppressed brain tumor growth. Ours is a proof-of-concept experiment showing that ANXA1-binding D-peptide can be developed as an orally-administrable tumor vasculature-targeted therapeutic.

scholarly journals Development of an orally-administrable tumor vasculature-targeting therapeutic using annexin A1-binding D-peptides

2020 ◽  
Author(s):  
Motohiro Nonaka ◽  
Hideaki Mabashi-Asazuma ◽  
Donald L. Jarvis ◽  
Kazuhiko Yamasaki ◽  
Tomoya O. Akama ◽  
...  
Keyword(s):  
Near Infrared ◽  
Peptide Binding ◽  
Tumor Vasculature ◽  
Whole Body ◽  
Luciferase Expression ◽  
Whole Body Imaging ◽  
Annexin A1 ◽  
Binding Assays ◽  
N Terminus

ABSTRACTIF7 peptide, which binds to the annexin A1 (ANXA1) N-terminal domain, functions as a tumor vasculature-targeted drug delivery vehicle after intravenous injection. To enhance IF7 stability in vivo, we undertook mirror-image peptide phage display using a synthetic D-peptide representing the Anxa1 N-terminus as target. Peptide sequences were identified, synthesized as D-amino acids, and designated as dTIT7, which was shown to bind the ANXA1 N-terminus. Whole body imaging of mouse brain tumors modeled with near infrared fluorescent IRDye-conjugated dTIT7 showed fluorescent signals in brain and kidney. Furthermore, orally-administered geldanamycin (GA)-conjugated dTIT7 suppressed brain tumor growth. Ours is a proof-of-concept experiment showing that Anxa1-binding D-peptide could be developed as an orally-administrable, tumor vasculature-targeted therapeutic.Role of each author: MN designed and performed experiments, analyzed data, and wrote the manuscript; HMA and DLJ produced recombinant ANXA1 protein; KY conducted NMR analysis and data analysis; TOA designed, performed and analyzed LC-MS/MS data; MN, TS, IKT, YS, and TY analyzed peptide-binding assays and performed in silico structural analysis; CTU produced lentivirus for luciferase expression; CNH performed peptide binding assays, tissue culture and animal experiments; and MNF supervised the project and wrote the manuscript.

Assessment of the near-Infrared Fluorescent Protein FP635 for Dynamic In Vivo Imaging in Models of Adaptive Immune Responses.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2436-2436
Author(s):  
Simone S Riedel ◽  
Marco Herold ◽  
Markus Hirschberg ◽  
Christian Brede ◽  
Carina A Baeuerlein ◽  
...  
Keyword(s):  
T Cells ◽  
Near Infrared ◽  
Whole Body ◽  
Whole Body Imaging ◽  
Signal Attenuation ◽  
Tissue Penetration ◽  
Body Imaging ◽  
Signal Intensities

Abstract Abstract 2436 Poster Board II-413 Sensitive in vivo imaging methods have advanced the fields of stem cell transplantation, graft-versus–host disease (GVHD) and graft-versus-tumor responses (GVT). Near-infrared (NIF) fluorescent proteins (FP) appear advantageous for deeper tissue penetration due to minimized absorbance by hemoglobin, water and lipids. Therefore we tested whether a recently published NIF FP (FP635, “Katushka”) could serve as a single reporter for whole body and single cell imaging. To compare signal intensities of eGFP and FP635 we generated fluorescent MOSEC cell lines (mouse ovarian cancer), titrated them in vitro and subcutaneously (s.c.) in vivo in Balb/c nu/nu mice. MOSEC FP635 showed twice the signal intensities compared to MOSEC eGFP in vitro by spectral fluorescence imaging (FLI). In vivo the eGFP signal was attenuated >60% in contrast to only 20% for FP635 from subcutaneous sites. However, FP635 signals from deep tissue layers were quenched. To address whether reduced signal attenuation of FP635 may allow sensitive visualization of immune processes by FLI and multiphoton-laser-scanning-microscopy (MPM) we generated transgenic mice in the genetic C57Bl/6 (B6) background, expressing FP635 under the ubiquitin promoter. Transgenic founders were selected upon signal intensities of leukocyte populations measured by flow cytometry in the PerCP channel. Combination of FP635 with colors other than red were possible for multiparameter flow cytometry. Next, eGFP, DsRed and FP635 splenocytes from transgenic donors were titrated as described above. In vitro signal intensities of FP635 splenocytes were >5 times lower compared to the other two FPs. FP635 signal absorption in vivo was low (30%) which is consistent with MOSEC titration results. In vivo DsRed detection was most sensitive and signals were similarly attenuated as FP635 in contrast to eGFP (60%). Subsequently, we aimed to visualize FP635 in a model of GVHD, where alloreactive T cells undergo massive expansion. Balb/c nu/nu mice were lethally irradiated and transplanted with 5×106 B6.WT bone marrow cells plus either 2×107 B6.DsRed+Luciferase+ or 2×107 B6.FP635 splenocytes. Sensitivity for DsRed cell detection was superior over FP635 cells. FP635 signal was only weakly detectable in lymph nodes (LN) by ex vivo FLI, where DsRed signals were detectable at earlier timepoints and LNs were even visualized by in vivo FLI. DsRed+ Luciferase+ double transgenic splenocytes allowed direct comparison of bioluminescence imaging (BLI) to FLI. Timely in vivo visualization of immune cells in deep tissues was feasible only by BLI. After whole body imaging the suitability of FP635 for MPM was checked by co-injecting eGFP B cells and either DsRed or FP635 T cells intravenously into RAG-/- mice. As FP635 is a NIF FP we expected to achieve deeper tissue penetration in hemoglobin rich organs, such as the spleen, in single cell microscopy. After 6 weeks of adoptive cell transfer we imaged spleens by MPM. Tissue penetration depths of DsRed or FP635 T cells were compared to eGFP B cells. No advantage in penetration depth of FP635 over DsRed was measured. Photobleaching is an important factor for microscopy, especially if cells are to be tracked over long time. FP635 transfected 293T cells bleached faster (t1/2=108 sec) than 293T cells transfected with eGFP (t1/2>900 sec) or DsRed (t1/2=411 sec). These experiments indicate that very high expression levels of FP635 need to be achieved for imaging. The signal attenuation of FP635 is low which may increase the sensitivity but in our hands DsRed showed comparable characteristics. Yet, the fast photobleaching of FP635 compared to the broadly established FPs DsRed and eGFP may be disadvantageous for long term microscopic tracking of cells. Our data indicate that BLI is by far superior over FLI in sensitivity and tissue penetration for whole body imaging of immune cells. However, FLI of red or near-infrared clonally selectable tumor cell lines may provide a welcome color addition to study immune cell-tumor interactions in combined models of BLI and FLI. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Imaging Depths of Near-Infrared Quantum Dots in First and Second Optical Windows

2012 ◽  
Vol 11 (4) ◽  
pp. 7290.2011.00057 ◽  
Author(s):  
Nayoun Won ◽  
Sanghwa Jeong ◽  
Kangwook Kim ◽  
Jungheon Kwag ◽  
Joonhyuck Park ◽  
...  
Keyword(s):  
Near Infrared ◽  
Incidence Angle ◽  
Small Animal ◽  
Apparent Size ◽  
Whole Body ◽  
Whole Body Imaging ◽  
Optical Window ◽  
Small Animal Models ◽  
Imaging Depth

Potential advantages of quantum dot (QD) imaging in the second optical window (SOW) at 1,000 to 1,400 nm over the first optical window (FOW) at 700 to 900 nm have attracted much interest. QDs that emit at 800 nm (800QDs) and QDs that emit at 1,300 nm (1,300QDs) are used to investigate the imaging depths at the FOW and SOW. QD images in biologic tissues are processed binarized via global thresholding method, and the imaging depths are determined using the criteria of contrast to noise ratio and relative apparent size. Owing to the reduced scattering in the SOW, imaging depth in skin can be extended by approximately three times for 1,300QD/SOW over 800QD/FOW. In liver, excitation of 1,300QD/SOW can be shifted to longer wavelengths; thus, the imaging depth can be extended by 1.4 times. Effects of quantum yield (QY), concentration, incidence angle, polarization, and fluence rate F on imaging depth are comprehensively studied. Under F approved by the Food and Drug Administration, 1,300QDs with 50% QY can reach imaging depths of 29.7 mm in liver and 17.5 mm in skin. A time-gated excitation using 1,000 times higher F pulses can obtain the imaging depth of ≈ 5 cm. To validate our estimates, in vivo whole-body imaging experiments are performed using small-animal models.

scholarly journals Whole-Body Imaging to Assess Cell-Based Immunotherapy: Preclinical Studies with an Update on Clinical Translation

2021 ◽  
Author(s):  
Noriko Sato ◽  
Peter L. Choyke
Keyword(s):  
T Cell ◽  
Cell Tracking ◽  
Checkpoint Inhibitors ◽  
Cell Receptor ◽  
Clinical Translation ◽  
Whole Body ◽  
Whole Body Imaging ◽  
Body Imaging ◽  
Cell Therapies

AbstractIn the past decades, immunotherapies against cancers made impressive progress. Immunotherapy includes a broad range of interventions that can be separated into two major groups: cell-based immunotherapies, such as adoptive T cell therapies and stem cell therapies, and immunomodulatory molecular therapies such as checkpoint inhibitors and cytokine therapies. Genetic engineering techniques that transduce T cells with a cancer-antigen-specific T cell receptor or chimeric antigen receptor have expanded to other cell types, and further modulation of the cells to enhance cancer targeting properties has been explored. Because cell-based immunotherapies rely on cells migrating to target organs or tissues, there is a growing interest in imaging technologies that non-invasively monitor transferred cells in vivo. Here, we review whole-body imaging methods to assess cell-based immunotherapy using a variety of examples. Following a review of preclinically used cell tracking technologies, we consider the status of their clinical translation.

scholarly journals Radioanalytical Techniques to Quantitatively Assess the Biological Uptake and In Vivo Behavior of Hazardous Substances

Molecules ◽  
2020 ◽  
Vol 25 (17) ◽  
pp. 3985
Author(s):  
Jae Young Lee ◽  
Sajid Mushtaq ◽  
Jung Eun Park ◽  
Hee Soon Shin ◽  
So-Young Lee ◽  
...  
Keyword(s):  
Nuclear Imaging ◽  
Hazardous Substances ◽  
Environmental Research ◽  
Whole Body ◽  
Whole Body Imaging ◽  
Toxic Substances ◽  
Industrial Chemicals ◽  
Biological Uptake ◽  
Radioanalytical Methods

Concern about environmental exposure to hazardous substances has grown over the past several decades, because these substances have adverse effects on human health. Methods used to monitor the biological uptake of hazardous substances and their spatiotemporal behavior in vivo must be accurate and reliable. Recent advances in radiolabeling chemistry and radioanalytical methodologies have facilitated the quantitative analysis of toxic substances, and whole-body imaging can be achieved using nuclear imaging instruments. Herein, we review recent literature on the radioanalytical methods used to study the biological distribution, changes in the uptake and accumulation of hazardous substances, including industrial chemicals, nanomaterials, and microorganisms. We begin with an overview of the radioisotopes used to prepare radiotracers for in vivo experiments. We then summarize the results of molecular imaging studies involving radiolabeled toxins and their quantitative assessment. We conclude the review with perspectives on the use of radioanalytical methods for future environmental research.

scholarly journals Properties of Fluorescent Far-Red Anti-TNF Nanobodies

Antibodies ◽  
2018 ◽  
Vol 7 (4) ◽  
pp. 43 ◽  
Author(s):  
Ekaterina Gorshkova ◽  
Grigory Efimov ◽  
Ksenia Ermakova ◽  
Ekaterina Vasilenko ◽  
Diana Yuzhakova ◽  
...  
Keyword(s):  
Autoimmune Diseases ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Humanized Mice ◽  
Whole Body ◽  
Whole Body Imaging ◽  
Theranostic Agent ◽  
Variable Heavy

Upregulation of the expression of tumor necrosis factor (TNF-α, TNF) has a significant role in the development of autoimmune diseases. The fluorescent antibodies binding TNF may be used for personalized therapy of TNF-dependent diseases as a tool to predict the response to anti-TNF treatment. We generated recombinant fluorescent proteins consisting of the anti-TNF module based on the variable heavy chain (VHH) of camelid antibodies fused with the far-red fluorescent protein Katushka (Kat). Two types of anti-TNF VHH were developed: one (BTN-Kat) that was bound both human or mouse TNF, but did not neutralize their activity, and a second (ITN-Kat) that was binding and neutralizing human TNF. BTN-Kat does not interfere with TNF biological functions and can be used for whole-body imaging. ITN-Kat can be evaluated in humanized mice or in cells isolated from humanized mice. It is able to block human TNF (hTNF) activities both in vitro and in vivo and may be considered as a prototype of a theranostic agent for autoimmune diseases.

scholarly journals Homogentisic acid-derived pigment as a biocompatible label for optoacoustic imaging of macrophages

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Ina Weidenfeld ◽  
Christian Zakian ◽  
Peter Duewell ◽  
Andriy Chmyrov ◽  
Uwe Klemm ◽  
...  
Keyword(s):  
Single Cells ◽  
Cell Types ◽  
Homogentisic Acid ◽  
Tissue Imaging ◽  
Whole Body ◽  
Whole Body Imaging ◽  
Optoacoustic Imaging ◽  
Pigment Formation ◽  
Functionally Diverse

Abstract Macrophages are one of the most functionally-diverse cell types with roles in innate immunity, homeostasis and disease making them attractive targets for diagnostics and therapy. Photo- or optoacoustics could provide non-invasive, deep tissue imaging with high resolution and allow to visualize the spatiotemporal distribution of macrophages in vivo. However, present macrophage labels focus on synthetic nanomaterials, frequently limiting their ability to combine both host cell viability and functionality with strong signal generation. Here, we present a homogentisic acid-derived pigment (HDP) for biocompatible intracellular labeling of macrophages with strong optoacoustic contrast efficient enough to resolve single cells against a strong blood background. We study pigment formation during macrophage differentiation and activation, and utilize this labeling method to track migration of pro-inflammatory macrophages in vivo with whole-body imaging. We expand the sparse palette of macrophage labels for in vivo optoacoustic imaging and facilitate research on macrophage functionality and behavior.

scholarly journals Novel Robust Hepatitis C Virus Mouse Efficacy Model

2006 ◽  
Vol 50 (10) ◽  
pp. 3260-3268 ◽  
Author(s):  
Qing Zhu ◽  
Yoko Oei ◽  
Dirk B. Mendel ◽  
Evelyn N. Garrett ◽  
Montesa B. Patawaran ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Protease Inhibitor ◽  
Small Animal ◽  
Rna Replication ◽  
Treatment Withdrawal ◽  
Whole Body ◽  
Hcv Rna ◽  
Whole Body Imaging

ABSTRACT The lack of a robust small-animal model for hepatitis C virus (HCV) has hindered the discovery and development of novel drug treatments for HCV infections. We developed a reproducible and easily accessible xenograft mouse efficacy model in which HCV RNA replication is accurately monitored in vivo by real-time, noninvasive whole-body imaging of gamma-irradiated SCID mice implanted with a mouse-adapted luciferase replicon-containing Huh-7 cell line (T7-11). The model was validated by demonstrating that both a small-molecule NS3/4A protease inhibitor (BILN 2061) and human alpha interferon (IFN-α) decreased HCV RNA replication and that treatment withdrawal resulted in a rebound in replication, which paralleled clinical outcomes in humans. We further showed that protease inhibitor and IFN-α combination therapy was more effective in reducing HCV RNA replication than treatment with each compound alone and supports testing in humans. This robust mouse efficacy model provides a powerful tool for rapid evaluation of potential anti-HCV compounds in vivo as part of aggressive drug discovery efforts.

scholarly journals Whole-Body Imaging of Sequestration of Plasmodium falciparum in the Rat

2005 ◽  
Vol 73 (11) ◽  
pp. 7736-7746 ◽  
Author(s):  
Fredrik Pettersson ◽  
Anna M. Vogt ◽  
Cathrine Jonsson ◽  
Bobo W. Mok ◽  
Alireza Shamaei-Tousi ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Plasmodium Falciparum ◽  
Severe Malaria ◽  
Whole Body ◽  
Surgical Removal ◽  
Whole Body Imaging ◽  
Rat Lung ◽  
Body Imaging

ABSTRACT The occlusion of vessels by packed Plasmodium falciparum-infected (iRBC) and uninfected erythrocytes is a characteristic postmortem finding in the microvasculature of patients with severe malaria. Here we have employed immunocompetent Sprague-Dawley rats to establish sequestration in vivo. Human iRBC cultivated in vitro and purified in a single step over a magnet were labeled with 99mtechnetium, injected into the tail vein of the rat, and monitored dynamically for adhesion in the microvasculature using whole-body imaging or imaging of the lungs subsequent to surgical removal. iRBC of different lines and clones sequester avidly in vivo while uninfected erythrocytes did not. Histological examination revealed that a multiadhesive parasite adhered in the larger microvasculature, inducing extensive intravascular changes while CD36- and chondroitin sulfate A-specific parasites predominantly sequester in capillaries, inducing no or minor pathology. Removal of the adhesive ligand Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1), preincubation of the iRBC with sera to PfEMP1 or preincubation with soluble PfEMP1-receptors prior to injection significantly reduced the sequestration. The specificity of iRBC binding to the heterologous murine receptors was confirmed in vitro, using primary rat lung endothelial cells and rat lung cryosections. In offering flow dynamics, nonmanipulated endothelial cells, and an intact immune system, we believe this syngeneic animal model to be an important complement to existing in vitro systems for the screening of vaccines and adjunct therapies aiming at the prevention and treatment of severe malaria.

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