3 research outputs found
<sup>64</sup>Cu-Labeled Gp2 Domain for PET Imaging of Epidermal Growth Factor Receptor
This
purpose of this study is to determine the efficacy of a 45-amino
acid Gp2 domain, engineered to bind to epidermal growth factor receptor
(EGFR), as a positron emission tomography (PET) probe of EGFR in a
xenograft mouse model. The EGFR-targeted Gp2 (Gp2-EGFR) and a nonbinding
control were site-specifically labeled with 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic
acid (DOTA) chelator. Binding affinity was tested toward human EGFR
and mouse EGFR. Biological activity on downstream EGFR signaling was
examined in cell culture. DOTA-Gp2 molecules were labeled with <sup>64</sup>Cu and intravenously injected (0.6–2.3 MBq) into mice
bearing EGFR<sup>high</sup> (<i>n</i> = 7) and EGFR<sup>low</sup> (<i>n</i> = 4) xenografted tumors. PET/computed
tomography (CT) images were acquired at 45 min, 2 h, and 24 h. Dynamic
PET (25 min) was also acquired. Tomography results were verified with
gamma counting of resected tissues. Two-tailed <i>t</i> tests
with unequal variances provided statistical comparison. DOTA-Gp2-EGFR
bound strongly to human (<i>K</i><sub>D</sub> = 7 ±
5 nM) and murine (<i>K</i><sub>D</sub> = 29 ± 6 nM)
EGFR, and nontargeted Gp2 had no detectable binding. Gp2-EGFR did
not agonize EGFR nor antagonize EGF-EGFR. <sup>64</sup>Cu-Gp2-EGFR
tracer effectively localized to EGFR<sup>high</sup> tumors at 45 min
(3.2 ± 0.5%ID/g). High specificity was observed with significantly
lower uptake in EGFR<sup>low</sup> tumors (0.9 ± 0.3%ID/g, <i>p</i> < 0.001), high tumor-to-background ratios (11 ±
6 tumor/muscle, <i>p</i> < 0.001). Nontargeted Gp2 tracer
had low uptake in EGFR<sup>high</sup> tumors (0.5 ± 0.3%ID/g, <i>p</i> < 0.001). Similar data was observed at 2 h, and tumor
signal was retained at 24 h (2.9 ± 0.3%ID/g). An engineered Gp2
PET imaging probe exhibited low background and target-specific EGFR<sup>high</sup> tumor uptake at 45 min, with tumor signal retained at
24 h postinjection, and compared favorably with published EGFR PET
probes for alternative protein scaffolds. These beneficial <i>in vivo</i> characteristics, combined with thermal stability,
efficient evolution, and small size of the Gp2 domain validate its
use as a future class of molecular imaging agents
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SARS-CoV-2 RNAemia predicts clinical deterioration and extrapulmonary complications from COVID-19
BackgroundThe determinants of coronavirus disease 2019 (COVID-19) disease severity and extrapulmonary complications (EPCs) are poorly understood. We characterized relationships between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNAemia and disease severity, clinical deterioration, and specific EPCs.MethodsWe used quantitative and digital polymerase chain reaction (qPCR and dPCR) to quantify SARS-CoV-2 RNA from plasma in 191 patients presenting to the emergency department with COVID-19. We recorded patient symptoms, laboratory markers, and clinical outcomes, with a focus on oxygen requirements over time. We collected longitudinal plasma samples from a subset of patients. We characterized the role of RNAemia in predicting clinical severity and EPCs using elastic net regression.ResultsOf SARS-CoV-2-positive patients, 23.0% (44 of 191) had viral RNA detected in plasma by dPCR, compared with 1.4% (2 of 147) by qPCR. Most patients with serial measurements had undetectable RNAemia within 10 days of symptom onset, reached maximum clinical severity within 16 days, and symptom resolution within 33 days. Initially RNAemic patients were more likely to manifest severe disease (odds ratio, 6.72 [95% confidence interval, 2.45-19.79]), worsening of disease severity (2.43 [1.07-5.38]), and EPCs (2.81 [1.26-6.36]). RNA loads were correlated with maximum severity (r = 0.47 [95% confidence interval, .20-.67]).ConclusionsdPCR is more sensitive than qPCR for the detection of SARS-CoV-2 RNAemia, which is a robust predictor of eventual COVID-19 severity and oxygen requirements, as well as EPCs. Because many COVID-19 therapies are initiated on the basis of oxygen requirements, RNAemia on presentation might serve to direct early initiation of appropriate therapies for the patients most likely to deteriorate
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Using a 29-mRNA Host Response Classifier To Detect Bacterial Coinfections and Predict Outcomes in COVID-19 Patients Presenting to the Emergency Department
Clinicians in the emergency department (ED) face challenges in concurrently assessing patients with suspected COVID-19 infection, detecting bacterial coinfection, and determining illness severity since current practices require separate workflows. Here, we explore the accuracy of the IMX-BVN-3/IMX-SEV-3 29 mRNA host response classifiers in simultaneously detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and bacterial coinfections and predicting clinical severity of COVID-19. A total of 161 patients with PCR-confirmed COVID-19 (52.2% female; median age, 50.0 years; 51% hospitalized; 5.6% deaths) were enrolled at the Stanford Hospital ED. RNA was extracted (2.5 mL whole blood in PAXgene blood RNA), and 29 host mRNAs in response to the infection were quantified using Nanostring nCounter. The IMX-BVN-3 classifier identified SARS-CoV-2 infection in 151 patients with a sensitivity of 93.8%. Six of 10 patients undetected by the classifier had positive COVID tests more than 9 days prior to enrollment, and the remaining patients oscillated between positive and negative results in subsequent tests. The classifier also predicted that 6 (3.7%) patients had a bacterial coinfection. Clinical adjudication confirmed that 5/6 (83.3%) of the patients had bacterial infections, i.e., Clostridioides difficile colitis (n = 1), urinary tract infection (n = 1), and clinically diagnosed bacterial infections (n = 3), for a specificity of 99.4%. Two of 101 (2.8%) patients in the IMX-SEV-3 "Low" severity classification and 7/60 (11.7%) in the "Moderate" severity classification died within 30 days of enrollment. IMX-BVN-3/IMX-SEV-3 classifiers accurately identified patients with COVID-19 and bacterial coinfections and predicted patients' risk of death. A point-of-care version of these classifiers, under development, could improve ED patient management, including more accurate treatment decisions and optimized resource utilization. IMPORTANCE We assay the utility of the single-test IMX-BVN-3/IMX-SEV-3 classifiers that require just 2.5 mL of patient blood in concurrently detecting viral and bacterial infections as well as predicting the severity and 30-day outcome from the infection. A point-of-care device, in development, will circumvent the need for blood culturing and drastically reduce the time needed to detect an infection. This will negate the need for empirical use of broad-spectrum antibiotics and allow for antibiotic use stewardship. Additionally, accurate classification of the severity of infection and the prediction of 30-day severe outcomes will allow for appropriate allocation of hospital resources