Insulin Hexamer-Caged Gadolinium Ion as MRI Contrast-o-phore.
ABSTRACT: High-relaxivity protein-complexes of GdIII are being pursued as MRI contrast agents in hope that they can be used at much lower doses that would minimize toxic-side effects of GdIII release from traditional contrast agents. We construct here a new type of protein-based MRI contrast agent, a proteinaceous cage based on a stable insulin hexamer in which GdIII is captured inside a water filled cavity. The macromolecular structure and the large number of "free" GdIII coordination sites available for water binding lead to exceptionally high relaxivities per one GdIII ion. The GdIII slowly diffuses out of this cage, but this diffusion can be prevented by addition of ligands that bind to the hexamer. The ligands that trigger structural changes in the hexamer, SCN- , Cl- and phenols, modulate relaxivities through an outside-in signaling that is allosterically transduced through the protein cage. Contrast-o-phores based on protein-caged metal ions have potential to become clinical contrast agents with environmentally-sensitive properties.
Project description:A PCN theranostic platform comprises a doxorubicin (DXR)-loaded liposomal core and an acid-sensitive polymer shell that is functionalized with Herceptin and GdIII-based MRI contrast agents. In vitro testing reveals a 14-fold increase in DXR-based cytotoxicity versus a non-targeted analogue and an 120-fold improvement in cellular GdIII-uptake in comparison with clinically approved DOTA-GdIII, leading to significant T1 MRI contrast enhancement.
Project description:Magnetic resonance imaging (MRI) has emerged as a leading diagnostic technique in clinical and preclinical settings. However, the application of MRI to assess specific disease markers for diagnosis and monitoring drug effect has been severely hampered by the lack of desired contrast agents with high relaxivities, and optimized in vivo retention time. We have reported the development of protein-based MRI contrast agents (ProCA1) by rational design of Gd(3+) binding sites into a stable protein resulting in significantly increased longitudinal (r(1)) and transverse (r(2)) relaxivities compared to Gd-DTPA. Here, we report a further improvement of protein contrast agents ProCA1 for in vivo imaging by protein modification with various sizes of polyethylene glycol (PEG) chain. PEGylation results in significant increases of both r(1) and r(2) relaxivities (up to 200%), and these high relaxivities persist even at field strengths up to 9.4 T. In addition, our experimental results demonstrate that modified contrast agents have significant improvement of in vivo MR imaging and biocompatibilities including dose efficiency, protein solubility, blood retention time and decreased immunogenicity. Such improvement can be important to the animal imaging and pre-clinical research at high or ultra-high field where there is an urgent need for molecular imaging probes and optimized contrast agent.
Project description:Nanoscale metal-organic frameworks (NMOFs) based on Gd3+ centers and benzenedicarboxylate and benzenetricarboxylate bridging ligands were synthesized using reverse microemulsions and characterized using SEM, PXRD, and TGA. These NMOFs exhibit extraordinarily large R1 and R2 relaxivities because of the presence of up to tens of millions of Gd3+ centers in each nanoparticle and are thus efficient T1 and T2 contrast agents for MRI. The NMOFs can also be made highly luminescent by doping with Eu3+ or Tb3+ centers. The results from this work suggest that NMOFs can be used as potential contrast agents for multimodal imaging.
Project description:One essential requirement for more sensitive gadolinium-based MRI contrast agents is to slow the molecular tumbling of the gadolinium(III) ion, which increases the gadolinium's relaxivity (i.e., its ability to speed up the NMR relaxation of nearby water molecules). One route to this is through conjugation to high-molecular-weight polymers such as dendrimers. In this work, amine-functionalized TREN-bis(1,2-HOPO)-TAM-ethylamine and TREN-bis(1-Me-3,2-HOPO)-TAM-ethylamine ligands have been synthesized and attached to biocompatible 40 kDa esteramide (EA)- and poly-l-lysine (PLL)-based dendrimers capable of binding up to eight gadolinium complexes. These conjugates have T(1) relaxivities of up to 38.14 ± 0.02 mM(-1) s(-1) per gadolinium at 37 °C, corresponding to relaxivities of up to 228 mM(-1) s(-1) per dendrimer molecule. This relaxivity expressed on a "per Gd" basis is several times that of the small-molecule complexes and an order of magnitude higher than that of current commercial agents. Because of their high performance and low toxicity, these macromolecules may constitute an attractive complement to currently available gadolinium(III)-based contrast agents.
Project description:Particle-based magnetic resonance imaging (MRI) contrast agents have been the focus of recent studies, primarily due to the possibility of preparing multimodal particles capable of simultaneously targeting, imaging, and treating specific biological tissues in vivo. In addition, particle-based MRI contrast agents often have greater sensitivity than commercially available, soluble agents due to decreased molecular tumbling rates following surface immobilization, leading to increased relaxivities. Mesoporous silica particles are particularly attractive substrates due to their large internal surface areas. In this study, we immobilized a unique phosphonate-containing ligand onto mesoporous silica particles with a range of pore diameters, pore volumes, and surface areas, and Gd(III) ions were then chelated to the particles. Per-Gd(III) ionic relaxivities ranged from ?2 to 10 mM(-1) s(-1) (37 °C, 60 MHz), compared to 3.0-3.5 mM(-1) s(-1) for commercial agents. The large surface areas allowed many Gd(III) ions to be chelated, leading to per-particle relaxivities of 3.3 × 10(7) mM(-1) s(-1), which is the largest value measured for a biologically suitable particle.
Project description:Conventional T1- or T2-weighted single mode contrast-enhanced magnetic resonance imaging (MRI) may produce false results. Thereby, there is a need to develop dual contrast agents, T1- and T2-weighted, for more accurate MRI imaging. The dual contrast agents should possess high magnetic resonance (MR) relaxivities, targeted tumor linking, and minimum recognition by the immune system. We have developed nitrodopamine-PEG grafted single core truncated cubic iron oxide nanoparticles (ND-PEG-tNCIOs) capable of producing marked dual contrasts in MRI with enhanced longitudinal and transverse relaxivities of 32 ± 1.29 and 791 ± 38.39 mM-1 s-1, respectively. Furthermore, the ND-PEG-tNCIOs show excellent colloidal stability in physiological buffers and higher cellular internalization in cancerous cells than in phagocytic cells, indicating the immune evasive capability of the nanoparticles. These findings indicate that tNCIOs are strong candidates for dual contrast MRI imaging, which is vital for noninvasive real-time detection of nascent cancer cells in vivo and for monitoring stem cells transplants.
Project description:Eight-coordinate hydroxypyridinone/terephthalamide GdIII complexes display high relaxivities due to their two inner sphere water molecules. This relaxivity can be further increased by functionalizing the terephthalamide moiety with an amine. A significant hydrogen bonding interaction between the amine and another water molecule close to the GdIII apparently facilitates its coordination on the open site of the metal. The resulting nine-coordinate complex has three inner sphere water molecules, while maintaining high stability and fast ligand exchange rates.
Project description:Metal-free magnetic resonance imaging (MRI) agents could overcome the established toxicity associated with metal-based agents in some patient populations and enable new modes of functional MRI in vivo. Herein, we report nitroxide-functionalized brush-arm star polymer organic radical contrast agents (BASP-ORCAs) that overcome the low contrast and poor in vivo stability associated with nitroxide-based MRI contrast agents. As a consequence of their unique nanoarchitectures, BASP-ORCAs possess per-nitroxide transverse relaxivities up to ?44-fold greater than common nitroxides, exceptional stability in highly reducing environments, and low toxicity. These features combine to provide for accumulation of a sufficient concentration of BASP-ORCA in murine subcutaneous tumors up to 20 h following systemic administration such that MRI contrast on par with metal-based agents is observed. BASP-ORCAs are, to our knowledge, the first nitroxide MRI contrast agents capable of tumor imaging over long time periods using clinical high-field 1H MRI techniques.
Project description:A DTPA-Gd containing polymer was grown in the interior of a heat shock protein cage resulting in T(1) particle relaxivities of 4200 mM(-1) sec(-1) for the 12 nm particle. Relaxivity parameters were determined, and this analysis suggests that the rotational correlation time has been optimized while the water exchange lifetime is longer than optimal. This synthetic approach holds much promise for the development of next generation contrast agents and this report will aid in their design.
Project description:Considerable efforts have been focused on the exploitation of macromolecule ligands for synthesis of magnetic Fe3O4 nanoparticles as T1 magnetic resonance imaging (MRI) contrast agents, but studies that concern macromolecule ligands with different charges and coordination groups are still limited. Herein, we used poly(acrylic acid) (PAA), poly(allylamine hydrochloride) (PAH), and polyvinyl alcohol (PVA), which possess negative, positive and neutral charges with carboxylic acid, amino and hydroxyl groups respectively, as templates and stabilizers to fabricate Fe3O4 nanoparticles through coprecipitation reaction. The obtained Fe3O4-PAA, Fe3O4-PAH, and Fe3O4-PVA nanoparticles showed T1 contrast performance with r1 relaxivities of 23.4, 60.3, and 30.6 mM s-1 at 0.5 T (25 °C), and a r2/r1 ratio of 2.62, 3.82, and 7.26, respectively. The cell viability assay revealed that Fe3O4-PAA and Fe3O4-PVA exhibited good biocompatibility, while Fe3O4-PAH displayed high cytotoxicity. In vivo T1-weighted (1 T) mice showed that both Fe3O4-PAA and Fe3O4-PVA were able to display remarkably brighten the contrast enhancement for the mice tumor and kidney sites, but Fe3O4-PAA had better contrast performance. This work highlights that the macromolecule ligands play an important role in the biocompatibility and T1 contrast performance of magnetic Fe3O4 nanoparticles.