Inferring predominant pathways in cellular models of breast cancer using limited sample proteomic profiling

<p>Abstract</p> <p>Background</p> <p>Molecularly targeted drugs inhibit aberrant signaling within oncogenic pathways. Identifying the predominant pathways at work within a tumor is a key step towards tailoring therapies to the patient. Clinical samples pose significant challenges for proteomic profiling, an attractive approach for identifying predominant pathways. The objective of this study was to determine if information obtained from a limited sample (i.e., a single gel replicate) can provide insight into the predominant pathways in two well-characterized breast cancer models.</p> <p>Methods</p> <p>A comparative proteomic analysis of total cell lysates was obtained from two cellular models of breast cancer, BT474 (HER2+/ER+) and SKBR3 (HER2+/ER-), using two-dimensional electrophoresis and MALDI-TOF mass spectrometry. Protein interaction networks and canonical pathways were extracted from the Ingenuity Pathway Knowledgebase (IPK) based on association with the observed pattern of differentially expressed proteins.</p> <p>Results</p> <p>Of the 304 spots that were picked, 167 protein spots were identified. A threshold of 1.5-fold was used to select 62 proteins used in the analysis. IPK analysis suggested that metabolic pathways were highly associated with protein expression in SKBR3 cells while cell motility pathways were highly associated with BT474 cells. Inferred protein networks were confirmed by observing an up-regulation of IGF-1R and profilin in BT474 and up-regulation of Ras and enolase in SKBR3 using western blot.</p> <p>Conclusion</p> <p>When interpreted in the context of prior information, our results suggest that the overall patterns of differential protein expression obtained from limited samples can still aid in clinical decision making by providing an estimate of the predominant pathways that underpin cellular phenotype.</p

RA-MAP, molecular immunological landscapes in early rheumatoid arthritis and healthy vaccine recipients

Rheumatoid arthritis (RA) is a chronic inflammatory disorder with poorly defined aetiology characterised by synovial inflammation with variable disease severity and drug responsiveness. To investigate the peripheral blood immune cell landscape of early, drug naive RA, we performed comprehensive clinical and molecular profiling of 267 RA patients and 52 healthy vaccine recipients for up to 18 months to establish a high quality sample biobank including plasma, serum, peripheral blood cells, urine, genomic DNA, RNA from whole blood, lymphocyte and monocyte subsets. We have performed extensive multi-omic immune phenotyping, including genomic, metabolomic, proteomic, transcriptomic and autoantibody profiling. We anticipate that these detailed clinical and molecular data will serve as a fundamental resource offering insights into immune-mediated disease pathogenesis, progression and therapeutic response, ultimately contributing to the development and application of targeted therapies for RA.</p

Herpes Simplex Virus-1 Tracheitis and Failure to Wean From Mechanical Ventilation in an Immunocompetent Host

SESSION TITLE: Monday Medical Student/Resident Case Report Posters SESSION TYPE: Med Student/Res Case Rep Postr PRESENTED ON: 10/21/2019 02:30 PM - 03:15 PM INTRODUCTION: Herpes simplex virus-1 (HSV -1) is an ever-ubiquitous pathogen with the ability to infect multiple organ systems. In immunocompetent adult hosts, clinical infection with HSV-1 in the respiratory system is rare, though HSV-1 has been detected in respiratory secretions of mechanically ventilated, immunocompetent patients. It has also been implicated as a rare and reversible cause of tracheal stenosis in patients with failure to wean from mechanical ventilation. CASE PRESENTATION: We present a 42 year old woman with a past medical history of asthma, history of pulmonary embolism and prior history of tracheostomy, who presented to our intensive care unit (ICU) as a transfer from an outside hospital where she had been treated for non-productive cough, fever, chills and acute hypoxic respiratory failure. On admission to our facility, she had increased oxygen requirements which escalated from high flow nasal cannula to mechanical ventilation. A CT scan revealed diffuse bilateral airspace and interstitial disease as well as tracheal stenosis at the site of her prior tracheostomy. Though the patient had been treated appropriately with broad-spectrum antibiotics for an extensive period of time, she had worsening respiratory failure with persistent leukocytosis and fevers. A bronchoscopy with bronchoalveolar lavage was performed, revealing a negative microbial work up with exception of polymerase chain reaction (PCR) detection of HSV-1. She was started on acyclovir, and in under 48 hours, was subsequently weaned from the ventilator. DISCUSSION: Respiratory failure with tracheobronchial involvement secondary to HSV infection in immunocompetent hosts is likely under recognized and under-reported. Based on our case and limited literature, HSV-1 has the potential to cause tracheal stenosis with tracheitis and respiratory failure with failure to wean from ventilation. Current treatment with acyclovir is effective in resolving the infection, and may have the additional benefit of reversing tracheal stenosis. Post treatment bronchoscopy has even demonstrated resolution of the inflammation and narrowing in some reports. CONCLUSIONS: In immunocompetent hosts with respiratory failure with tracheal stenosis and failure to wean from ventilation, HSV infection of the trachea or bronchial tree should be considered. Testing of HSV via PCR is available, and further case series and reports can add to this growing body of literature

Spontaneous coronary artery dissection and its association with heritable connective tissue disorders

Objective: Spontaneous coronary artery dissection (SCAD) is an under-recognised but important cause of myocardial infarction and sudden cardiac death. We sought to determine the role of medical and molecular genetic screening for connective tissue disorders in patients with SCAD.Methods: We performed a single-centre retrospective descriptive analysis of patients with spontaneous coronary artery disease who had undergone medical genetics evaluation 1984-2014 (n=116). The presence or absence of traits suggestive of heritable connective tissue disease was extracted. Genetic testing for connective tissue disorders and/or aortopathies, if performed, is also reported.Results: Of the 116 patients (mean age 44.2 years, 94.8% women and 41.4% with non-coronary fibromuscular dysplasia (FMD)), 59 patients underwent genetic testing, of whom 3 (5.1%) received a diagnosis of connective tissue disorder: a 50-year-old man with Marfan syndrome; a 43-year-old woman with vascular Ehlers-Danlos syndrome and FMD; and a 45-year-old woman with vascular Ehlers-Danlos syndrome. An additional 12 patients (20.3%) had variants of unknown significance, none of which was thought to be a definite disease-causing mutation based on in silico analyses.Conclusions: Only a minority of patients with SCAD who undergo genetic evaluation have a likely pathogenic mutation identified on gene panel testing. Even fewer exhibit clinical features of connective tissue disorder. These findings underscore the need for further studies to elucidate the molecular mechanisms of SCAD

Sex-, race-and ethnicity-based differences in thromboembolic events among adults hospitalized with COVID-19

BACKGROUND: Patients hospitalized with COVID-19 have an increased risk of thromboembolic events. Whether sex, race or ethnicity impacts these events is unknown. We studied the association between sex, race, and ethnicity and venous and arterial thromboembolic events among adults hospitalized with COVID-19. METHODS AND RESULTS: We used the American Heart Association Cardiovascular Disease COVID-19 registry. Primary expo-sures were sex and race and ethnicity, as defined by the registry. Primary outcomes were venous thromboembolic events and arterial thromboembolic events. We used logistic regression for risk adjustment. We studied 21 528 adults hospitalized with COVID-19 across 107 centers (54.1% men; 38.1% non-Hispanic White, 25.4% Hispanic, 25.7% non-Hispanic Black, 0.5% Native American, 4.0% Asian, 0.4% Pacific Islander, and 5.9% other race and ethnicity). The rate of venous thromboembolic events was 3.7% and was more common in men (4.2%) than women (3.2%; P\u3c0.001), and in non-Hispanic Black patients (4.9%) than other races and ethnicities (range, 1.3%– 3.8%; P\u3c0.001). The rate of arterial thromboembolic events was 3.9% and was more common in men (4.3%) than women (3.5%; P=0.002), and in non-Hispanic Black patients (5.0%) than other races and ethnicities (range, 2.3%– 4.7%; P\u3c0.001). Compared with men, women were less likely to experience venous thromboem-bolic events (adjusted odds ratio [OR], 0.71; 95% CI, 0.61– 0.83) and arterial thromboembolic events (adjusted OR, 0.76; 95% CI, 0.66– 0.89). Compared with non-Hispanic White patients, non-Hispanic Black patients had the highest likelihood of venous thromboembolic events (adjusted OR, 1.27; 95% CI, 1.04–1.54) and arterial thromboembolic events (adjusted OR, 1.35; 95% CI, 1.11–1.65). CONCLUSIONS: Men and non-Hispanic Black adults hospitalized with COVID-19 are more likely to have venous and arterial thromboembolic events. These subgroups may represent at-risk patients more susceptible to thromboembolic COVID-19 complications