Molecular tools for acute spatiotemporal manipulation of signal transduction

Ross, Brian; Mehta, Sohum; Zhang, Jin

doi:10.1016/j.cbpa.2016.08.012

Cited by 18 publications

(19 citation statements)

References 59 publications

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“…We expected there may be some free SpyCatcher-IRDye800CW, which was one of the reasons we performed the SEC purification of the ∆N-SpyCatcher to completely remove any impurities, yet we still saw an increase in kidney uptake ( Figure 4 ). Given that fluorescence and DFO-conjugated antibodies are different, we now expect that this may be an issue with the maleimide linker, which has previously been shown to be unstable in vivo [ 35 , 36 ]. This apparent instability affected the tumor uptake of 89 Zr-labeled antibody when compared with the randomly labeled probe 89 Zr-nimotuzumab [ 27 ].…”

Section: Discussionmentioning

confidence: 99%

Nimotuzumab Site-Specifically Labeled with 89Zr and 225Ac Using SpyTag/SpyCatcher for PET Imaging and Alpha Particle Radioimmunotherapy of Epidermal Growth Factor Receptor Positive Cancers

Solomon

Barreto

Bernhard

et al. 2020

Cancers

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To develop imaging and therapeutic agents, antibodies are often conjugated randomly to a chelator/radioisotope or drug using a primary amine (NH2) of lysine or sulfhydryl (SH) of cysteine. Random conjugation to NH2 or SH groups can require extreme conditions and may affect target recognition/binding and must therefore be tested. In the present study, nimotuzumab was site-specifically labeled using ∆N-SpyCatcher/SpyTag with different chelators and radiometals. Nimotuzumab is a well-tolerated anti-EGFR antibody with low skin toxicities. First, ΔN-SpyCatcher was reduced using tris(2-carboxyethyl)phosphine (TCEP), which was followed by desferoxamine-maleimide (DFO-mal) conjugation to yield a reactive ΔN-SpyCatcher-DFO. The ΔN-SpyCatcher-DFO was reacted with nimotuzumab-SpyTag to obtain stable nimotuzumab-SpyTag-∆N-SpyCatcher-DFO. Radiolabeling was performed with 89Zr, and the conjugate was used for the in vivo microPET imaging of EGFR-positive MDA-MB-468 xenografts. Similarly, ∆N-SpyCatcher was conjugated to an eighteen-membered macrocyclic chelator macropa-maleimide and used to radiolabel nimotuzumab-SpyTag with actinium-225 (225Ac) for in vivo radiotherapy studies. All constructs were characterized using biolayer interferometry, flow cytometry, radioligand binding assays, HPLC, and bioanalyzer. MicroPET/CT imaging showed a good tumor uptake of 89Zr-nimotuzumab-SpyTag-∆N-SpyCatcher with 6.0 ± 0.6%IA/cc (n = 3) at 48 h post injection. The EC50 of 225Ac-nimotuzumab-SpyTag-∆N-SpyCatcher and 225Ac-control-IgG-SpyTag-∆N-SpyCatcher against an EGFR-positive cell-line (MDA-MB-468) was 3.7 ± 3.3 Bq/mL (0.04 ± 0.03 nM) and 18.5 ± 4.4 Bq/mL (0.2 ± 0.04 nM), respectively. In mice bearing MDA-MB-468 EGFR-positive xenografts, 225Ac-nimotuzumab-SpyTag-∆N-SpyCatcher significantly (p = 0.0017) prolonged the survival of mice (64 days) compared to 225Ac-control IgG (28.5 days), nimotuzumab (28.5 days), or PBS-treated mice (30 days). The results showed that the conjugation and labeling using SpyTag/∆N-SpyCatcher to nimotuzumab did not significantly (p > 0.05) alter the receptor binding of nimotuzumab compared with a non-specific conjugation approach. 225Ac-nimotuzumab-SpyTag-∆N-SpyCatcher was effective in vitro and in an EGFR-positive triple negative breast cancer xenograft model.

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Section: Discussionmentioning

confidence: 99%

Nimotuzumab Site-Specifically Labeled with 89Zr and 225Ac Using SpyTag/SpyCatcher for PET Imaging and Alpha Particle Radioimmunotherapy of Epidermal Growth Factor Receptor Positive Cancers

Solomon

Barreto

Bernhard

et al. 2020

Cancers

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“…To overcome these challenges, many novel tools have been developed in the last decade to help visualize and manipulate these signaling molecules with greater precision. Here we provide a brief overview of these novel tools and refer readers to several excellent reviews for further information (Ross, Mehta, & Zhang, 2016;Rost, Schneider-Warme, Schmitz, & Hegemann, 2017).…”

Section: Tools For Studying Spatiotemporal Signalingmentioning

confidence: 99%

New insights into the molecular mechanisms of axon guidance receptor regulation and signaling

Zang

Chaudhari

Bashaw

2021

Current Topics in Developmental Biology

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As the nervous system develops, newly differentiated neurons need to extend their axons toward their synaptic targets to form functional neural circuits. During this highly dynamic process of axon pathfinding, guidance receptors expressed at the tips of motile axons interact with soluble guidance cues or membrane tethered molecules present in the environment to be either attracted toward or repelled away from the source of these cues. As competing cues are often present at the same location and during the same developmental period, guidance receptors need to be both spatially and temporally regulated in order for the navigating axons to make appropriate guidance decisions. This regulation is exerted by a diverse array of molecular mechanisms that have come into focus over the past several decades and these mechanisms ensure that the correct complement of surface receptors is present on the growth cone, a fan-shaped expansion at the tip of the axon. This dynamic, highly motile structure is defined by a lamellipodial network lining the periphery of the growth cone interspersed with finger-like filopodial projections that serve to explore the surrounding environment. Once axon guidance receptors are deployed at the right place and time at the growth cone surface, they respond to their respective ligands by initiating a complex set of signaling events that serve to rearrange the growth cone membrane and the actin and microtubule cytoskeleton to affect axon growth and guidance. In this review, we highlight recent advances that shed light on the rich complexity of mechanisms that regulate axon guidance receptor distribution, activation and downstream signaling.

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“…The authors also suggest the rod‐like magnetoreceptor protein, cryptochrome complex, possesses an intrinsic magnetic moment, and has the attributes of both inclination and polarity magnetoreception [Qin et al, ]. The scientific community has taken the ideas suggested by Qin et al [] into consideration [Lohmann, ; Pedersen et al, ; Ross et al, ] and has provided some critical points regarding the magnetosensitivity of the proposed rod‐like complex. For instance, Hore and Mouritsen [] point out that the presence of paramagnetic ions (e.g., iron, copper, and manganese) in the vicinity of the radicals could have the counterproductive effect of inducing spin relaxation, thus destroying spin coherence.…”

Section: Molecules That Could Be Involved In the Detection Of Geomagnmentioning

confidence: 99%

Biological effects related to geomagnetic activity and possible mechanisms

Крылов

2017

Bioelectromagnetics

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This review presents contemporary data on the biological effects of geomagnetic activity. Correlations between geomagnetic indices and biological parameters and experimental studies that used simulated geomagnetic storms to detect possible responses of organisms to these events in nature are discussed. Possible mechanisms by which geomagnetic activity influences organisms are also considered. Special attention is paid to the idea that geomagnetic activity is perceived by organisms as a disruption of diurnal geomagnetic variation. This variation, in turn, is viewed by way of a secondary zeitgeber for biological circadian rhythms. Additionally, we discuss the utility of cryptochrome as a biological detector of geomagnetic storms. The possible involvement of melatonin and protein coding by the CG8198 gene in the biological effects of geomagnetic activity are discussed. Perspectives for studying mechanisms by which geomagnetic storms affect organisms are suggested. Bioelectromagnetics. 38:497-510, 2017. © 2017 Wiley Periodicals, Inc.

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Molecular tools for acute spatiotemporal manipulation of signal transduction

Cited by 18 publications

References 59 publications

Nimotuzumab Site-Specifically Labeled with 89Zr and 225Ac Using SpyTag/SpyCatcher for PET Imaging and Alpha Particle Radioimmunotherapy of Epidermal Growth Factor Receptor Positive Cancers

Nimotuzumab Site-Specifically Labeled with 89Zr and 225Ac Using SpyTag/SpyCatcher for PET Imaging and Alpha Particle Radioimmunotherapy of Epidermal Growth Factor Receptor Positive Cancers

New insights into the molecular mechanisms of axon guidance receptor regulation and signaling

Biological effects related to geomagnetic activity and possible mechanisms

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