RESUMEN
Influenza A Virus (IAV) is a recurring respiratory virus with limited availability of antiviral therapies. Understanding host proteins essential for IAV infection can identify targets for alternative host-directed therapies (HDTs). Using affinity purification-mass spectrometry and global phosphoproteomic and protein abundance analyses using three IAV strains (pH1N1, H3N2, H5N1) in three human cell types (A549, NHBE, THP-1), we map 332 IAV-human protein-protein interactions and identify 13 IAV-modulated kinases. Whole exome sequencing of patients who experienced severe influenza reveals several genes, including scaffold protein AHNAK, with predicted loss-of-function variants that are also identified in our proteomic analyses. Of our identified host factors, 54 significantly alter IAV infection upon siRNA knockdown, and two factors, AHNAK and coatomer subunit COPB1, are also essential for productive infection by SARS-CoV-2. Finally, 16 compounds targeting our identified host factors suppress IAV replication, with two targeting CDK2 and FLT3 showing pan-antiviral activity across influenza and coronavirus families. This study provides a comprehensive network model of IAV infection in human cells, identifying functional host targets for pan-viral HDT.
Asunto(s)
COVID-19 , Subtipo H5N1 del Virus de la Influenza A , Virus de la Influenza A , Gripe Humana , Humanos , Virus de la Influenza A/genética , Gripe Humana/genética , Subtipo H5N1 del Virus de la Influenza A/genética , Subtipo H3N2 del Virus de la Influenza A/metabolismo , Proteómica , Replicación Viral/genética , SARS-CoV-2 , Antivirales/metabolismo , Interacciones Huésped-Patógeno/genéticaRESUMEN
SARS-CoV-2 variants of concern (VOCs) emerged during the COVID-19 pandemic. Here, we used unbiased systems approaches to study the host-selective forces driving VOC evolution. We discovered that VOCs evolved convergent strategies to remodel the host by modulating viral RNA and protein levels, altering viral and host protein phosphorylation, and rewiring virus-host protein-protein interactions. Integrative computational analyses revealed that although Alpha, Beta, Gamma, and Delta ultimately converged to suppress interferon-stimulated genes (ISGs), Omicron BA.1 did not. ISG suppression correlated with the expression of viral innate immune antagonist proteins, including Orf6, N, and Orf9b, which we mapped to specific mutations. Later Omicron subvariants BA.4 and BA.5 more potently suppressed innate immunity than early subvariant BA.1, which correlated with Orf6 levels, although muted in BA.4 by a mutation that disrupts the Orf6-nuclear pore interaction. Our findings suggest that SARS-CoV-2 convergent evolution overcame human adaptive and innate immune barriers, laying the groundwork to tackle future pandemics.
Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos , COVID-19/virología , Inmunidad Innata/genética , Pandemias , SARS-CoV-2/genéticaRESUMEN
BACKGROUND: Estimating surgical case duration accurately is an important operating room efficiency metric. Current predictive techniques in spine surgery include less sophisticated approaches such as classical multivariable statistical models. Machine learning approaches have been used to predict outcomes such as length of stay and time returning to normal work, but have not been focused on case duration. OBJECTIVE: The primary objective of this 4-year, single-academic-center, retrospective study was to use an ensemble learning approach that may improve the accuracy of scheduled case duration for spine surgery. The primary outcome measure was case duration. METHODS: We compared machine learning models using surgical and patient features to our institutional method, which used historic averages and surgeon adjustments as needed. We implemented multivariable linear regression, random forest, bagging, and XGBoost (Extreme Gradient Boosting) and calculated the average R2, root-mean-square error (RMSE), explained variance, and mean absolute error (MAE) using k-fold cross-validation. We then used the SHAP (Shapley Additive Explanations) explainer model to determine feature importance. RESULTS: A total of 3189 patients who underwent spine surgery were included. The institution's current method of predicting case times has a very poor coefficient of determination with actual times (R2=0.213). On k-fold cross-validation, the linear regression model had an explained variance score of 0.345, an R2 of 0.34, an RMSE of 162.84 minutes, and an MAE of 127.22 minutes. Among all models, the XGBoost regressor performed the best with an explained variance score of 0.778, an R2 of 0.770, an RMSE of 92.95 minutes, and an MAE of 44.31 minutes. Based on SHAP analysis of the XGBoost regression, body mass index, spinal fusions, surgical procedure, and number of spine levels involved were the features with the most impact on the model. CONCLUSIONS: Using ensemble learning-based predictive models, specifically XGBoost regression, can improve the accuracy of the estimation of spine surgery times.
RESUMEN
BACKGROUND: Days before surgery, add-ons may be scheduled to fill unused surgical block time at an outpatient surgery center. At times, outpatient surgery centers have time limitations for end of block time and discharge from the postanesthesia care unit (PACU). The objective of our study was to develop machine learning models that predicted the following composite outcome: (1) surgery finished by end of operating room block time and (2) patient was discharged by end of recovery room nursing shift. We compared various machine learning models to logistic regression. By evaluating various performance metrics, including F1 scores, we hypothesized that models using ensemble learning will be superior to logistic regression. METHODS: Data were collected from patients at an ambulatory surgery center. The primary outcome measurement was determined to have a value of 1 (versus 0) if they met both criteria: (1) surgery ends by 5 pm and (2) patient is discharged from the recovery room by 7 pm. We developed models to determine if a procedure would meet both criteria if it were scheduled at 1 pm, 2 pm, 3 pm, or 4 pm. We implemented regression, random forest, balanced random forest, balanced bagging, neural network, and support vector classifier, and included the following features: surgery, surgeon, service line, American Society of Anesthesiologists score, age, sex, weight, and scheduled case duration. We evaluated model performance with Synthetic Minority Oversampling Technique (SMOTE). We compared the following performance metrics: F1 score, area under the receiver operating characteristic curve (AUC), specificity, sensitivity, precision, recall, and Matthews correlation coefficient. RESULTS: Among 13,447 surgical procedures, the median total perioperative time (actual case duration and PACU length stay) was 165 minutes. When SMOTE was not used, when predicting whether surgery will end by 5 pm and patient will be discharged by 7 pm, the average F1 scores were best with random forest, balanced bagging, and balanced random forest classifiers. When SMOTE was used, these models had improved F1 scores compared to no SMOTE. The balanced bagging classifier performed best with F1 score of 0.78, 0.80, 0.82, and 0.82 when predicting our outcome if cases were to start at 1 pm, 2 pm, 3 pm, or 4 pm, respectively. CONCLUSIONS: We demonstrated improvement in predicting the outcome at a range of start times when using ensemble learning versus regression techniques. Machine learning may be adapted by operating room management to allow for a better determination whether an add-on case at an outpatient surgery center could be appropriately booked.
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Procedimientos Quirúrgicos Ambulatorios , Sala de Recuperación , Humanos , Aprendizaje Automático , Redes Neurales de la Computación , Alta del PacienteRESUMEN
BACKGROUND: The objective of this study is to develop predictive models for persistent opioid use following lower extremity joint arthroplasty and determine if ensemble learning and an oversampling technique may improve model performance. METHODS: We compared various predictive models to identify at-risk patients for persistent postoperative opioid use using various preoperative, intraoperative, and postoperative data, including surgical procedure, patient demographics/characteristics, past surgical history, opioid use history, comorbidities, lifestyle habits, anesthesia details, and postoperative hospital course. Six classification models were evaluated: logistic regression, random forest classifier, simple-feedforward neural network, balanced random forest classifier, balanced bagging classifier, and support vector classifier. Performance with Synthetic Minority Oversampling Technique (SMOTE) was also evaluated. Repeated stratified k-fold cross-validation was implemented to calculate F1-scores and area under the receiver operating characteristics curve (AUC). RESULTS: There were 1042 patients undergoing elective knee or hip arthroplasty in which 242 (23.2%) reported persistent opioid use. Without SMOTE, the logistic regression model has an F1 score of 0.47 and an AUC of 0.79. All ensemble methods performed better, with the balanced bagging classifier having an F1 score of 0.80 and an AUC of 0.94. SMOTE improved performance of all models based on F1 score. Specifically, performance of the balanced bagging classifier improved to an F1 score of 0.84 and an AUC of 0.96. The features with the highest importance in the balanced bagging model were postoperative day 1 opioid use, body mass index, age, preoperative opioid use, prescribed opioids at discharge, and hospital length of stay. CONCLUSIONS: Ensemble learning can dramatically improve predictive models for persistent opioid use. Accurate and early identification of high-risk patients can play a role in clinical decision making and early optimization with personalized interventions.
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Artroplastia de Reemplazo de Cadera , Artroplastia de Reemplazo de Rodilla , Trastornos Relacionados con Opioides , Analgésicos Opioides/efectos adversos , Artroplastia de Reemplazo de Cadera/efectos adversos , Artroplastia de Reemplazo de Rodilla/efectos adversos , Humanos , Extremidad Inferior , Aprendizaje Automático , Trastornos Relacionados con Opioides/diagnóstico , Trastornos Relacionados con Opioides/epidemiología , Trastornos Relacionados con Opioides/prevención & controlRESUMEN
The emergence of SARS-CoV-2 variants of concern suggests viral adaptation to enhance human-to-human transmission1,2. Although much effort has focused on the characterization of changes in the spike protein in variants of concern, mutations outside of spike are likely to contribute to adaptation. Here, using unbiased abundance proteomics, phosphoproteomics, RNA sequencing and viral replication assays, we show that isolates of the Alpha (B.1.1.7) variant3 suppress innate immune responses in airway epithelial cells more effectively than first-wave isolates. We found that the Alpha variant has markedly increased subgenomic RNA and protein levels of the nucleocapsid protein (N), Orf9b and Orf6-all known innate immune antagonists. Expression of Orf9b alone suppressed the innate immune response through interaction with TOM70, a mitochondrial protein that is required for activation of the RNA-sensing adaptor MAVS. Moreover, the activity of Orf9b and its association with TOM70 was regulated by phosphorylation. We propose that more effective innate immune suppression, through enhanced expression of specific viral antagonist proteins, increases the likelihood of successful transmission of the Alpha variant, and may increase in vivo replication and duration of infection4. The importance of mutations outside the spike coding region in the adaptation of SARS-CoV-2 to humans is underscored by the observation that similar mutations exist in the N and Orf9b regulatory regions of the Delta and Omicron variants.
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COVID-19/inmunología , COVID-19/virología , Evolución Molecular , Evasión Inmune , Inmunidad Innata/inmunología , SARS-CoV-2/genética , SARS-CoV-2/inmunología , COVID-19/transmisión , Proteínas de la Nucleocápside de Coronavirus/química , Proteínas de la Nucleocápside de Coronavirus/metabolismo , Humanos , Inmunidad Innata/genética , Interferones/inmunología , Proteínas del Complejo de Importación de Proteínas Precursoras Mitocondriales/metabolismo , Fosfoproteínas/química , Fosfoproteínas/metabolismo , Fosforilación , Proteómica , ARN Viral/genética , RNA-Seq , SARS-CoV-2/clasificación , SARS-CoV-2/crecimiento & desarrolloRESUMEN
Emergence of SARS-CoV-2 variants, including the globally successful B.1.1.7 lineage, suggests viral adaptations to host selective pressures resulting in more efficient transmission. Although much effort has focused on Spike adaptation for viral entry and adaptive immune escape, B.1.1.7 mutations outside Spike likely contribute to enhance transmission. Here we used unbiased abundance proteomics, phosphoproteomics, mRNA sequencing and viral replication assays to show that B.1.1.7 isolates more effectively suppress host innate immune responses in airway epithelial cells. We found that B.1.1.7 isolates have dramatically increased subgenomic RNA and protein levels of Orf9b and Orf6, both known innate immune antagonists. Expression of Orf9b alone suppressed the innate immune response through interaction with TOM70, a mitochondrial protein required for RNA sensing adaptor MAVS activation, and Orf9b binding and activity was regulated via phosphorylation. We conclude that B.1.1.7 has evolved beyond the Spike coding region to more effectively antagonise host innate immune responses through upregulation of specific subgenomic RNA synthesis and increased protein expression of key innate immune antagonists. We propose that more effective innate immune antagonism increases the likelihood of successful B.1.1.7 transmission, and may increase in vivo replication and duration of infection.