Development of matrix metalloproteinase-targeted probes for lung inflammation detection with positron emission tomography

Kondo, Naoya; Temma, Takashi; Aita, Kazuki; Shimochi, Saeka; Koshino, Kazuhiro; Senda, Michio; Iida, Hidehiro

doi:10.1038/s41598-018-19890-1

Cited by 15 publications

(13 citation statements)

References 24 publications

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“…The membrane was blocked with 5% milk diluted in TBST (10 minutes, rocking) and incubated with primary antibody (Rabbit Anti GDF11/GDF8 Abcam Cat# EPR4567 38 ) at a 1:1000 dilution in 1.5% milk in TBST (2 hours, rocking). The membrane was washed 5 times with TBST (5 minutes per wash) and incubated with secondary antibody (Anti Rabbit R&D antibody Cat#HAF008 82 ) at a 1:3000 dilution in 1.5% milk in TBST (1 hour, rocking). The membrane was washed 5 times with TBST (5 minutes per wash), incubated using Pierce ECL Plus Western Blotting Substrate, and developed.…”

Section: Methodsmentioning

confidence: 99%

Variation in zygotic CRISPR/Cas9 gene editing outcomes generates novel reporter and deletion alleles at the Gdf11 locus

Goldstein

Valido

Lewandowski

et al. 2019

Sci Rep

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Recent advances in CRISPR/Cas gene editing technology have significantly expanded the possibilities and accelerated the pace of creating genetically engineered animal models. However, CRISPR/Cas-based strategies designed to precisely edit the genome can often yield unintended outcomes. Here, we report the use of zygotic CRISPR/Cas9 injections to generate a knock-in GFP reporter mouse at the Gdf11 locus. Phenotypic and genomic characterization of founder animals from these injections revealed a subset that contained the correct targeting event and exhibited GFP expression that, within the hematopoietic system, was restricted predominantly to lymphoid cells. Yet, in another subset of founder mice, we detected aberrant integration events at the target site that dramatically and inaccurately shifted hematopoietic GFP expression from the lymphoid to the myeloid lineage. Additionally, we recovered multiple Gdf11 deletion alleles that modified the C-terminus of the GDF11 protein. When bred to homozygosity, most of these alleles recapitulated skeletal phenotypes reported previously for Gdf11 knockout mice, suggesting that these represent null alleles. However, we also recovered one Gdf11 deletion allele that encodes a novel GDF11 variant protein (“GDF11-WE”) predicted to contain two additional amino acids (tryptophan (W) and glutamic acid (E)) at the C-terminus of the mature ligand. Unlike the other Gdf11 deletion alleles recovered in this study, homozygosity for the Gdf11WE allele did not phenocopy Gdf11 knockout skeletal phenotypes. Further investigation using in vivo and in vitro approaches demonstrated that GDF11-WE retains substantial physiological function, indicating that GDF11 can tolerate at least some modifications of its C-terminus and providing unexpected insights into its biochemical activities. Altogether, our study confirms that one-step zygotic injections of CRISPR/Cas gene editing complexes provide a quick and powerful tool to generate gene-modified mouse models. Moreover, our findings underscore the critical importance of thorough characterization and validation of any modified alleles generated by CRISPR, as unintended on-target effects that fail to be detected by simple PCR screening can produce substantially altered phenotypic readouts.

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Section: Methodsmentioning

confidence: 99%

Variation in zygotic CRISPR/Cas9 gene editing outcomes generates novel reporter and deletion alleles at the Gdf11 locus

Goldstein

Valido

Lewandowski

et al. 2019

Sci Rep

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“…Macrophages will secrete matrix metalloproteases (MMPs) and many other cytokines, which are all attractive options for the molecular imaging of COPD. Using a mouse model of COPD, a radiofluorinated probe, 18 F-IPFP, was developed and tested to target MMP-9 and MMP-12; the accumulation of 18 F-IPFP was 4× higher in the lungs of COPD mice than in normal mice [135]. 99m Tc-labeled RP805 is another MMP targeted radiotracer that saw significantly greater accumulation in IL-13 transgenic mice than control mice using SPECT/CT ( Figure 3) [136].…”

Section: Chronic Obstructive Pulmonary Disease (Copd)mentioning

confidence: 99%

Molecular Imaging of Inflammatory Disease

et al. 2021

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Inflammatory diseases include a wide variety of highly prevalent conditions with high mortality rates in severe cases ranging from cardiovascular disease, to rheumatoid arthritis, to chronic obstructive pulmonary disease, to graft vs. host disease, to a number of gastrointestinal disorders. Many diseases that are not considered inflammatory per se are associated with varying levels of inflammation. Imaging of the immune system and inflammatory response is of interest as it can give insight into disease progression and severity. Clinical imaging technologies such as computed tomography (CT) and magnetic resonance imaging (MRI) are traditionally limited to the visualization of anatomical information; then, the presence or absence of an inflammatory state must be inferred from the structural abnormalities. Improvement in available contrast agents has made it possible to obtain functional information as well as anatomical. In vivo imaging of inflammation ultimately facilitates an improved accuracy of diagnostics and monitoring of patients to allow for better patient care. Highly specific molecular imaging of inflammatory biomarkers allows for earlier diagnosis to prevent irreversible damage. Advancements in imaging instruments, targeted tracers, and contrast agents represent a rapidly growing area of preclinical research with the hopes of quick translation to the clinic.

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“…Temma et al developed [ 18 F] 67 (Scheme 16) for MMPs-targeted PET [108]. Non-radiolabeled 67 exhibited IC 50 in the nanomolar range (MMP-2 = 4.4, MMP-9 = 8.1, MMP-12 = 1.5, MMP-13 = 4.1 nM).…”

Section: Mmpis Labelled Imaging Agents For Chronic Obstructive Pulmentioning

confidence: 99%

“…( C ) Maximum intensity projection images of removed lungs (left) and heart (right) from the Air (upper) and CS (lower) groups. Reprinted with permission from [108] copyright © 2018, Springer Nature.…”

Section: Figures Schemes and Tablesmentioning

confidence: 99%

Molecular Imaging Probes Based on Matrix Metalloproteinase Inhibitors (MMPIs)

et al. 2019

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Matrix metalloproteinases (MMPs) are a family of zinc- and calcium-dependent endopeptidases which are secreted or anchored in the cell membrane and are capable of degrading the multiple components of the extracellular matrix (ECM). MMPs are frequently overexpressed or highly activated in numerous human diseases. Owing to the important role of MMPs in human diseases, many MMP inhibitors (MMPIs) have been developed as novel therapeutics, and some of them have entered clinical trials. However, so far, only one MMPI (doxycycline) has been approved by the FDA. Therefore, the evaluation of the activity of a specific subset of MMPs in human diseases using clinically relevant imaging techniques would be a powerful tool for the early diagnosis and assessment of the efficacy of therapy. In recent years, numerous MMPIs labeled imaging agents have emerged. This article begins by providing an overview of the MMP subfamily and its structure and function. The latest advances in the design of subtype selective MMPIs and their biological evaluation are then summarized. Subsequently, the potential use of MMPI-labeled diagnostic agents in clinical imaging techniques are discussed, including positron emission tomography (PET), single-photon emission computed tomography (SPECT) and optical imaging (OI). Finally, this article concludes with future perspectives and clinical utility.

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Development of matrix metalloproteinase-targeted probes for lung inflammation detection with positron emission tomography

Cited by 15 publications

References 24 publications

Variation in zygotic CRISPR/Cas9 gene editing outcomes generates novel reporter and deletion alleles at the Gdf11 locus

Variation in zygotic CRISPR/Cas9 gene editing outcomes generates novel reporter and deletion alleles at the Gdf11 locus

Molecular Imaging of Inflammatory Disease

Molecular Imaging Probes Based on Matrix Metalloproteinase Inhibitors (MMPIs)

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