bluuebunny/crossref_metadata_2025 huggingface dataset:
Myosin light chain kinase (MYLK) coding polymorphisms modulate human lung endothelial cell barrier responses via altered tyrosine phosphorylation, spatial localization, and lamellipodial protrusions
Authors: Wang Ting , Brown Mary E. , Kelly Gabriel T. , Camp Sara M. , Mascarenhas Joseph B. , Sun Xiaoguang , Dudek Steven M. , Garcia Joe G.N.
Published: February 2018

Sphingosine 1‐phosphate (S1P) is a potent bioactive endogenous lipid that signals a rearrangement of the actin cytoskeleton via the regulation of non‐muscle myosin light chain kinase isoform (nmMLCK). S1P induces critical nmMLCK Y464 and Y471 phosphorylation resulting in translocation of nmMLCK to the periphery where spatially‐directed increases in myosin light chain (MLC) phosphorylation and tension result in lamellipodia protrusion, increased cell‐cell adhesion, and enhanced vascular barrier integrity. MYLK, the gene encoding nmMLCK, is a known candidate gene in lung inflammatory diseases, with coding genetic variants (Pro21His, Ser147Pro, Val261Ala) that confer risk for inflammatory lung injury and influence disease severity. The functional mechanisms by which these MYLK coding single nucleotide polymorphisms (SNPs) affect biologic processes to increase disease risk and severity remain elusive. In the current study, we utilized quantifiable cell immunofluorescence assays to determine the influence of MYLK coding SNPs on S1P‐mediated nmMLCK phosphorylation and translocation to the human lung endothelial cell (EC) periphery. These disease‐associated MYLK variants result in reduced levels of S1P‐induced Y464 phosphorylation, a key site for nmMLCK enzymatic regulation and activation. Reduced Y464 phosphorylation resulted in attenuated nmMLCK protein translocation to the cell periphery. We further conducted EC kymographic assays which confirmed that lamellipodial protrusion in response to S1P challenge was retarded by expression of a MYLK transgene harboring the three MYLK coding SNPs. These data suggest that ARDS/severe asthma‐associated MYLK SNPs functionally influence vascular barrier‐regulatory cytoskeletal responses via direct alterations in the levels of nmMLCK tyrosine phosphorylation, spatial localization, and lamellipodial protrusions.
Endothelial dysfunction in pulmonary arterial hypertension: loss of cilia length regulation upon cytokine stimulation
Authors: Dummer Anneloes , Rol Nina , Szulcek Robert , Kurakula Kondababu , Pan Xiaoke , Visser Benjamin I. , Bogaard Harm Jan , DeRuiter Marco C. , Goumans Marie‐José , Hierck Beerend P.
Published: February 2018

Pulmonary arterial hypertension (PAH) is a syndrome characterized by progressive lung vascular remodelling, endothelial cell (EC) dysfunction, and excessive inflammation. The primary cilium is a sensory antenna that integrates signalling and fine tunes EC responses to various stimuli. Yet, cilia function in the context of deregulated immunity in PAH remains obscure. We hypothesized that cilia function is impaired in ECs from patients with PAH due to their inflammatory status and tested whether cilia length changes in response to cytokines. Primary human pulmonary and mouse embryonic EC were exposed to pro‐ (TNFα, IL1β, and IFNγ) and/or anti‐inflammatory (IL‐10) cytokines and cilia length was quantified. Chronic treatment with all tested inflammatory cytokines led to a significant elongation of cilia in both control human and mouse EC (by ∼1 µm, P < 0.001). This structural response was PKA/PKC dependent. Intriguingly, withdrawal of the inflammatory stimulus did not reduce cilia length. IL‐10, on the other hand, blocked and reversed the pro‐inflammatory cytokine‐induced cilia elongation in healthy ECs, but did not influence basal length. Conversely, primary cilia of ECs from PAH patients were significantly longer under basal conditions compared to controls (1.86 ± 0.02 vs. 2.43 ± 0.08 µm, P = 0.002). These cilia did not elongate further upon pro‐inflammatory stimulation and anti‐inflammatory treatment did not impact cilia length. The missing length modulation was specific to cytokine stimulation, as application of fluid shear stress led to increased cilia length in the PAH endothelium. We identified loss of cilia length regulation upon cytokine stimulation as part of the endothelial dysfunction in PAH.
A direct comparison of decision rules for early discharge of suspected acute coronary syndromes in the era of high sensitivity troponin
Authors: Chew Pei Gee , Frost Fredrick , Mullen Liam , Fisher Michael , Zadeh Heidar , Grainger Ruth , Albouaini Khaled , Dodd James , Patel Bilal , Velavan Periaswamy , Kunadian Babu , Rawat Anju , Obafemi Toba , Tong Sarah , Jones Julia , Khand Aleem
Published: February 2018

Background:

We tested the hypothesis that a single high sensitivity troponin at limits of detection (LOD HSTnT) (<5 ng/l) combined with a presentation non-ischaemic electrocardiogram is superior to low-risk Global Registry of Acute Coronary Events (GRACE) (<75), Thrombolysis in Myocardial Infarction (TIMI) (≤1) and History, ECG, Age, Risk factors and Troponin (HEART) score (≤3) as an aid to early, safe discharge for suspected acute coronary syndrome.

Methods:

In a prospective cohort study, risk scores were computed in consecutive patients with suspected acute coronary syndrome presenting to the Emergency Room of a large English hospital. Adjudication of myocardial infarction, as per third universal definition, involved a two-physician, blinded, independent review of all biomarker positive chest pain re-presentations to any national hospital. The primary and secondary outcome was a composite of type 1 myocardial infarction, unplanned coronary revascularisation and all cause death (MACE) at six weeks and one year.

Results:

Of 3054 consecutive presentations with chest pain 1642 had suspected acute coronary syndrome (52% male, median age 59 years, 14% diabetic, 20% previous myocardial infarction). Median time from chest pain to presentation was 9.7 h. Re-presentations occurred in eight hospitals with 100% follow-up achieved. Two hundred and eleven (12.9%) and 279 (17%) were adjudicated to suffer MACE at six weeks and one year respectively. Only HEART ≤3 (negative predictive value MACE 99.4%, sensitivity 97.6%, %discharge 53.4) and LOD HSTnT strategy (negative predictive value MACE 99.8%, sensitivity 99.5%, %discharge 36.9) achieved pre-specified negative predictive value of >99% for MACE at six weeks. For type 1 myocardial infarction alone the negative predictive values at six weeks and one year were identical, for both HEART ≤3 and LOD HSTnT at 99.8% and 99.5% respectively.

Conclusion:

HEART ≤3 or LOD HSTnT strategy rules out short and medium term myocardial infarction with ≥99.5% certainty, and short-term MACE with >99% certainty, allowing for early discharge of 53.4% and 36.9% respectively of suspected acute coronary syndrome. Adoption of either strategy has the potential to greatly reduce Emergency Room pressures and minimise follow-up investigations. Very early presenters (<3 h), due to limited numbers, are excluded from these conclusions.
Human foreskin fibroblasts: from waste bag to important biomedical applications
Authors: Oliveira Thomaz , Costa Ilana , Marinho Victor , Carvalho Valécia , Uchôa Karla , Ayres Carla , Teixeira Silmar , Vasconcelos Daniel F P
Published: February 2018

Circumcision is one of the most performed surgical procedures worldwide, and it is estimated that one in three men worldwide is circumcised, which makes the preputial skin removed after surgery an abundant material for possible applications. In particular, it is possible efficiently to isolate the cells of the foreskin, with fibroblasts being the most abundant cells of the dermis and the most used in biomedical research. This work aimed to review the knowledge and obtain a broad view of the main applications of human foreskin fibroblast cell culture. A literature search was conducted, including clinical trials, preclinical basic research studies, reviews and experimental studies. Several medical and laboratory applications of human foreskin fibroblast cell culture have been described, especially when it comes to the use of human foreskin fibroblasts as feeder cells for the cultivation of human embryonic stem cells, in addition to co-culture with other cell types. The culture of foreskin fibroblasts has also been used to: obtain induced pluripotent stem cells; the diagnosis of Clostridium difficile; to test the toxicity and effect of substances on normal cells, especially the toxicity of possible antineoplastic drugs; in viral culture, mainly of the human cytomegalovirus, study of the pathogenesis of other microorganisms; varied studies of cellular physiology and cellular interactions. Fibroblasts are important for cell models for varied application cultures, demonstrating how the preputial material can be reused, making possible new applications.

Level of evidence: Not applicable for this multicentre audit.
An Exploration of Shift Work, Fatigue, and Gender Among Police Officers: The BCOPS Study
Authors: Violanti John M. , Owens Sherry L. , Fekedulegn Desta , Ma Claudia C. , Charles Luenda E. , Andrew Michael E.
Published: February 2018

The present study examined the association between shift work and fatigue among male ( n = 230) and female ( n = 78) police officers. A 15-year work history database was used to define dominant shifts as day, afternoon, or night. A 10-item questionnaire created from the Standard Shiftwork Index (SSI) assessed fatigue. Gender-stratified analyses of variance and covariance and Poisson regression were used to compare means and prevalence of individual items across shifts. No significant differences in total fatigue scores were observed across shifts. However, the prevalence of the fatigue item “feelings of tiredness” was 89% higher among male officers working the afternoon shift compared with officers working the day shift (prevalence ratio [PR] = 1.89, 95% confidence interval [CI] = [1.12, 3.23], p = .020), after adjustment for covariates. Women reported a lower prevalence of tiredness than men on the afternoon shift. Organizations with afternoon shift workers should consider reducing fatigue at work through education and other methods.
Aerococcus urinae Mitral Valve Endocarditis-Related Stroke: A Case Report and Literature Review
Authors: Adomavicius Darius , Bock Mark , Vahl Christian-Friedrich , Siegel Ekkehard
Published: February 2018

Background. Aerococcus urinae is a rare causative pathogen of infective endocarditis that results in a high risk of embolic events. The mortality rate for A urinae endocarditis is high. Old age and underlying urologic conditions are the best-known risk factors for infection. Case Description. We report the clinical course of the disease in a 49-year-old man who presented symptoms of a urinary tract infection. A few days later, transthoracic echocardiography showed a conspicuous mitral valve with myxomatous alterations. Following the detection of a cerebral embolism with associated stroke symptoms, as well as at the beginning of cardiac failure, the emergency indication for the surgical treatment of mitral valve endocarditis was given. On the second day following the operation, circulatory collapse rapidly developed. Following an unsuccessful attempt at cardiopulmonary resuscitation, the patient died. Review of the Literature. From 1991 to 2017, 29 cases of A urinae–induced endocarditis have been described in PubMed and Medline. One or 2 new cases are published annually. We review all reported cases of A urinae endocarditis, with an emphasis on the predisposing factors, course, and outcomes of the disease. Conclusion. A urinae endocarditis is a rare disease primarily affecting elderly men with urinary tract pathologies and comorbidities. The course of the disease is severe, and the outcome is often fatal. A 16S rDNA polymerase chain reaction investigation of bacterial genome provides proof of the presence of A urinae. Because of the high risk of embolism, rapid treatment should focus on the diseased heart valve. Based on existing data and the experience gained from handling cases, treatment with β-lactam and aminoglycosides is recommended. It is also recommended that operative therapy take place as soon as possible.
Dynamic Course of Serratia marcescens Pulmonic Valve Endocarditis Resulting in Submassive PE and Valve Replacement
Authors: Meyer Chloe Grace , Vacek Thomas Paul , Bansal Amit , Gurujal Ravi , Parikh Analkumar
Published: February 2018

This report illustrates a case of a 42-year-old male with a history of intravenous drug abuse who presented with septic shock. Diagnostic studies, including a transthoracic echocardiogram, chest computed tomography angiography, transesophageal echocardiogram, and blood cultures ultimately revealed Serratia marcescens pulmonic valve infective endocarditis that was treated with intravenous antibiotics. In addition to the rare form of endocarditis and bacterium involved, this case brings into awareness the dynamic nature of the hospital course that requires vigilance in responding to hypotensive episodes for consideration of pulmonary embolism. Surgical valve replacement was opted for due to such a complication in addition to the large size of the vegetation, 2.5 cm.
Maximum Occlusal Force and Incident Functional Disability in Older Adults: The Tsurugaya Project
Authors: Ohi T. , Komiyama T. , Miyoshi Y. , Murakami T. , Tsuboi A. , Tomata Y. , Tsuji I. , Watanabe M. , Hattori Y.
Published: February 2018

The purpose of the current study was to investigate the association between maximum occlusal force, which is an objective predictor of masticatory performance, and incident functional disability in an elderly Japanese population. A prospective cohort study was conducted targeting 815 (51.7% female) community-dwelling older adults aged ≥70 y residing in the Tsurugaya district, Sendai, Japan. The outcome measurement was incident functional disability, defined as a first certification of long-term care insurance in Japan, which is determined on the basis of a strictly established, uniform, nationwide standard. During a median follow-up of 7.9 y (interquartile range, 4.8–7.9 y), information on long-term care insurance was obtained from the Sendai Municipal Authority. Bilateral maximum occlusal forces of the participants were measured using a horseshoe-shaped pressure-indicating film, and the participants were categorized into quartiles based on occlusal force. Adjusted hazard ratios for functional disability were estimated with Cox proportional hazard models, adjusted for age, sex, body mass index, medical history, smoking status, alcohol consumption, duration of education, depressive symptoms, cognitive impairment, physical functioning, marital status, history of falls, and number of remaining teeth. The multiple-adjusted hazard ratios and 95% confidence intervals (CIs) for incident functional disability compared to the greatest occlusal force quartile were 1.53 (95% CI, 1.02–2.33), 1.64 (95% CI, 1.06–2.55), and 1.64 (95% CI, 1.01–2.68) for the third, second, and first quartiles, respectively ( P for trend = 0.011). A lower maximum occlusal force was significantly associated with an increased risk of functional disability independently of possible confounders, including the number of remaining teeth. Occlusal force may be a useful indicator of the relationship between oral function and geriatric health.

Knowledge Transfer Statement:This prospective cohort study demonstrated that lower maximum occlusal force was associated with an increased risk of functional disability in older adults, even after adjustment for possible confounding factors, including the number of remaining teeth. This strengthens the rationale regarding the association between oral function and geriatric health. Particularly in older adults, occlusal force is reduced by several factors other than tooth loss, such as the absence of a dental prostheses, sarcopenia in the masticatory muscle, poor periodontal condition, and orofacial pain. Our findings suggest that maximum occlusal force may be a useful biomarker associated with diverse parameters aside from the number of remaining teeth.
Ames mutagenicity, structural alerts of carcinogenicity, Hansch QSAR parameters (ClogP, CMR, MgVol), tumor site concordance/multiplicity, and tumorigenicity rank in NTP 2-year rodent studies
Authors: Smith Carr J , Perfetti Thomas A , Ko Gene M , Garg Rajni
Published: February 2018

Since its inception in 1976, the National Toxicology Program (NTP) has conducted 594 2-year studies on rats and mice by a number of routes of administration including inhalation, feed, drinking water, dermal, and intraperitoneal injection. Of these studies, the results on 470 chemicals were of adequate technical quality to be incorporated into final technical reports. In this study, the 470 chemicals were categorized from 1 to 48 by the level of “clear” neoplastic evidence in male and female rats, and in male and female mice, and given an ordinal rank from 1 to 135 following additional considerations regarding tumor site concordance and tumor multiplicity. The resultant tumorigenicity category score and ordinal rank score were examined for associations with results in the Ames Salmonella mutagenicity assay; presence or absence of structural alerts of carcinogenicity; and three Hansch Quantitative Structure-Activity Relationship (QSAR) parameters, namely, calculated base 10 logarithm of the octanol–water partition coefficient (ClogP), calculated molar refractivity (CMR), and McGowan molecular volume (MgVol). Smaller molecular volumes were found to be associated with higher levels of tumorigenicity. Whereas lower rather than higher levels of lipophilicity were found to be associated with higher levels of tumorigenicity. Positive Ames test results were positively correlated with overall tumorigenicity and with possession of structural alerts. Since larger organic molecules have more chemical reaction centers, it was not surprising that higher ClogP values were positively correlated with the number of structural alerts. The results from this study demonstrate the ability to devise rational rules for relative tumorigenicity that correlate, in biologically plausible ways, with known parameters of toxicity.
Reverse Shoulder Arthroplasty Using a Bone-preserving Glenoid Component: Clinical and Radiographic Outcomes
Authors: Sanchez-Sotelo Joaquin , Nguyen Ngoc Tram V , Morrey Mark E
Published: February 2018

Introduction

Primary reverse shoulder arthroplasty has been reported to provide good outcomes. The majority of designs utilize a large portion of the glenoid. Some studies have reported worrisome rates of notching and stress fractures of the acromion or scapular spine. The purpose of this study was to determine the outcome and complications of a bone-preserving reverse prosthesis with limited ingrowth glenoid surface paired with an onlay humeral component.

Materials and methods

Between 2014 and 2015, 96 reverse arthroplasties were implanted using a bone-preserving reverse with a 135° onlay humeral bearing. Six patients died. The remaining 87 patients (90 shoulders) were followed for 2 years or until revision. Mean age was 69.3 years. The underlying diagnosis was cuff tear arthropathy (13), irreparable cuff tear (39), osteoarthritis (32), and other (6). Shoulders were followed for 2 years or until revision surgery.

Results

Reverse shoulder arthroplasty resulted in significant improvements in pain, motion, and function. At most recent follow-up, 74 shoulders had no or mild pain. Elevation was 131° ± 31° and external rotation 46° ± 21°. The mean American Shoulder and Elbow Surgeons (ASES) score was 76. There were 4 reoperations for deep infection (2), periprosthetic fracture, and glenoid loosening. None of the remaining arthroplasties were loose. There was one stress fracture of the acromion and one of the spine of the scapula. No radiographic notching or dislocation was observed.

Discussion

Primary reverse arthroplasty using a bone-preserving glenoid component and an anatomic onlay humeral bearing provides satisfactory clinical outcomes with a very low rate of failure and complications.
Inhibition of the IRE-1α/XBP-1 pathway prevents chronic GVHD and preserves the GVL effect in mice
Authors: Schutt Steven D. , Wu Yongxia , Tang Chih-Hang Anthony , Bastian David , Nguyen Hung , Sofi M. Hanief , Zhang MengMeng , Liu Chen , Helke Kris , Wilson Carole , Schnapp Lynn M. , Del Valle Juan R. , Hu Chih-Chi Andrew , Yu Xue-Zhong
Published: February 2018

Key Points

Targeting XBP-1 on B cells is sufficient to prevent cGVHD. Pharmacologic inhibition of IRE-1α/XBP-1 prevents cGVHD while preserving GVL activity.
Addressing Western dietary pattern in obesity and NAFLD
Authors: Trovato Francesca M. , Martines Giuseppe Fabio , Catalano Daniela
Published: February 2018

Background

Western diet and sedentary life are imputed for non-alcoholic fatty liver disease (NAFLD). Relationship of obesity and bright liver, i.e., the ultrasound hallmarks of fatty liver, evaluated as Bright Liver Score (BLS), with Western diet pattern score (WDS) and with the level of physical exercise, assessed by the Baecke questionnaire, were investigated.

Methods

One hundred fifty (72 females, 78 males) NAFLD non-diabetic patients, 47.89 ± 13.85 years old, were referred to a diagnostic-therapeutic unit with the lone evidence of fatty liver as detected by an ultrasound. WDS, Adherence to Mediterranean Diet Score (AMDS), physical exercise, insulin resistance (HOMA-IR), and BMI were assessed also after 6 months of comprehensive lifestyle intervention.

Results

BLS, BMI, HOMA, and WDS were significantly greater, while AMDS and physical exercise were lower at the greater severity of ultrasound (US)-detected NAFLD. Positive linear correlation of WDS, BMI, and HOMA vs. BLS was present at patients’ referral. After intervention, decrease of liver dimension and of Bright Liver Score concurrent with a reduction of BMI, HOMA-IR, WDS, LDL cholesterol, and triglycerides was observed. A reciprocal increase of AMDS, physical exercise, and HDL cholesterol was also reported.

Conclusions

The assessment of some features of the Western diet is possible; its effect, along with overweight, is associated with a greater severity of fatty liver as evaluated by ultrasound in NAFLD.

Trial registration

NCT02670590. Registered on February 2, 2016.
Tumor-infiltrating lymphocytes to predict survival to anthracycline/taxane-based adjuvant chemotherapy in triple negative breast cancer.
Authors: Ou Qiyun , Yu Yunfang , Xiao Xiaoyun , Luo Baoming
Published: February 2018

1

Background: Previous clinical data suggested that the tumour-infiltrating lymphocytes (TILs) were predictive in breast cancer treated with adjuvant chemotherapy; however, clinical relevance has yet to be established. We hypothesized that TILs would be associated with overall survival (OS) and disease-free survival (DFS) after anthracycline/taxane-based adjuvant chemotherapy in breast cancer subtypes. Methods: PubMed and EMBASE were searched until March 2017 for studies that investigated the association of TILs with survival for anthracycline/taxane-based adjuvant chemotherapy in breast cancer. OS and DFS were combined using random-effects meta-analysis and calculated as combined hazard ratio (HR) with 95% credible intervals (CIs). The PROSPERO registry number is CRD42017072133. Results: Twelve studies comprising 9,023 patients were eligible for analysis. Six were prospective adjuvant trials (n = 7686) and six were retrospective studies (n = 1337). Pooled analysis indicated that high TILs have no significant predictive association in overall population (DFS, HR = 0.87, 95% CI, 0.70 – 1.08; OS, HR = 0.98, 95% CI, 0.89 – 1.07), Luminal A/B (DFS, HR = 0.99, 95% CI, 0.94 – 1.04; OS, HR = 1.01, 95% CI, 0.92 – 1.12), and HER2–positive patients (DFS, HR = 0.84, 95% CI, 0.71 – 1.00; OS, HR = 0.89, 95% CI, 0.77 – 1.02), but were related to improved DFS (HR, 0.81; 95% CI, 0.73 – 0.89) and OS (HR, 0.74; 95% CI, 0.66 – 0.84) in triple negative breast cancer (TNBC) patients. Additionally, increasing TILs were not significantly associated with reduced risk of relapse in HER2-positive patient through adjuvant trastuzumab (HR, 0.97; 95% CI, 0.93 – 1.01). Conclusions: This meta-analysis provides an evidence that TILs predicts survival to anthracycline/taxane-based adjuvant chemotherapy in TNBC patients and suggests that predictive benefit seemed to be influenced by breast cancer subtypes as well.
High durable CR rates and preliminary safety profile for JCAR017 in R/R aggressive b-NHL (TRANSCEND NHL 001 Study): A defined composition CD19-directed CAR T-cell product with potential for outpatient administration.
Authors: Abramson Jeremy S. , Siddiqi Tanya , Palomba Maria Lia , Gordon Leo I. , Lunning Matthew Alexander , Arnason Jon E. , Wang Michael , Forero-Torres Andres , Albertson Tina , Dehner Christine , Garcia Jacob , Li Daniel , Xie Benhuai , Maloney David G.
Published: February 2018

120

Background: JCAR017 is a defined composition, CD19-directed 4-1BB CAR T cell product administered at a precise dose of CD8 and CD4 CAR T cells in a seamless design Phase 1 pivotal trial of relapsed/refractory (R/R) B-cell NHL (TRANSCEND NHL 001; NCT02631044). Methods: Patients (pts) with R/R DLBCL NOS, PMBCL, FL grade 3B, or MCL and adequate organ function are eligible. The FULL dataset includes all pts in the DLBCL cohort (i.e. excludes MCL). The CORE dataset includes pts meeting inclusion for a planned pivotal DLBCL cohort (DLBCL NOS [ de novo or transformed from follicular lymphoma], ECOG 0-1, no prior allo-SCT). Results: As of July 7, 2017, 69 pts were treated in the DLBCL cohort and evaluable for safety. In the FULL dataset, 21 pts (30%) had cytokine release syndrome (CRS), with 1 serious CRS event (1%; Gr 4). Neurotoxicity (NT) at any grade developed in 14 (20%), including 10 (14%) with Gr 3-4 events. There were no Gr 5 CRS or NT events. Median time to onset of first CRS and NT was 5 days and 10 days, respectively, 13 (19%) received anti-cytokine therapy and only one required vasopressor support. In the CORE dataset (n = 49), similar rates of CRS and NT were shown. The majority of pts, 64% (44/69), had no CRS or NT, and median onset of CRS and NT were 5 and 11 days, respectively, suggesting outpatient delivery of JCAR017 may be feasible. In the DLBCL cohort, 68 pts were evaluable for efficacy; best overall response, 3-month, and 6-month response rates in the FULL dataset were 75% (51/68), 49% (27/55), and 40% (14/35), respectively; and, in the CORE dataset, were 84% (41/49), 65% (26/40), and 57% (13/23). The best overall, 3-month, and 6-month CR rates were 61% (30/49), 53% (21/40), and 52% (12/23), respectively, in the CORE dataset. Among 16 double/triple hit pts, best ORR was 81%, and 3-month CR rate was 60%. A trend toward improved response rate at 3 months was observed in pts treated at DL2 compared to DL1. Conclusions: At the symposium, we will report updated efficacy and the preliminary safety profile for JCAR017, which supports potential outpatient administration in R/R Aggressive B-NHL patients. Clinical trial information: NCT02631044.
Natural pattern-recognition-receptor agonists as adjuvants for in situ vaccination lymphoma immunotherapy.
Authors: Aleynick Mark , Peng Paul , Hammerich Linda , Upadhyay Ranjan , Marshall Netonia , Agudo Judith , Yellin Michael Jay , Keler Tibor , Davis Thomas A. , Brown Brian , Brody Joshua
Published: February 2018

123

Background: In patients with low-grade lymphoma, in situ vaccination has yielded both partial and complete remissions in clinical trials. Though clinical responses have been observed with multiple pattern recognition receptor agonists (PRRa), the optimal immune stimulant is unknown. We hypothesize that natural PRRa, such as the attenuated pathogens or subunits found in common prophylactic vaccines, could target multiple PRR in a physiologically relevant context and lead to a more robust activation of dendritic cells (DCs) versus synthetic PRRa. Methods: 20 vaccines, including BCG, Typhim Vi, MMR-II, etc. were screened in vitro, where DC phenotype and function were evaluated by flow cytometry. Flt3L-mobilized DC ability to phagocytose, process, present, and cross-present soluble protein or tumor derived antigen, were assessed using CRISPR gene-edited, β2m(-/-) GFP-lymphoma cells and a novel GFP-specific (‘JEDI’) CD8 T cell system. Vaccine mechanism of immune activation was elucidated using a library of PRR-null macrophage cell lines. Potent vaccines were also evaluated in vivo in a Flt3L-primed in situ vaccination using the A20 murine lymphoma model. Results: Several vaccines induced robust DC activation and several showed significant increases in subsequent T cell activation, proliferation, and tumor killing, suggesting increased antigen processing and cross-presentation by DCs. Some vaccines, either as single agents or in combination, were significantly more effective than synthetic PRRa in activating DCs and inducing a T cell response. In vivo, vaccine combination therapies induced tumor regression in a majority of animals, suggesting synergistic immune activation. Conclusions: This data suggests prophylactic vaccines are effective clinical-grade DC activators and can be repurposed for use in the in situ vaccination maneuver, with immediate translation into the clinic. Additionally, by extensive in vitro evaluation in parallel with in vivo studies, this work aims to identify a predictive in vitro molecular immune signature that correlates closely with adjuvant efficacy in vivo.
Correlation of β-catenin expression in non-small cell lung cancer to prognosis and CD8 positive cells infiltration.
Authors: Muto Satoshi , Ozaki Yuki , Inoue Takuya , Yamaura Takumi , Fukuhara Mitsuro , Hasegawa Takeo , Shio Yutaka , Suzuki Hiroyuki
Published: February 2018

142

Background: It was reported that β-catenin expression in tumor cells suppresses dendritic cell infiltration into tumor in mouse model of melanoma. It results in less of tumor infiltrating lymphocytes. In non-small cell lung cancer (NSCLC), it is not well understood that β-catenin is involved in cancer immunity in that context. Methods: We studied the association between β-catenin expression and CD8 positive cells infiltration in NSCLC. Ninety-one patients with NSCLC who had complete surgical resection during January 2013 to June 2015 were subjected. β-catenin expression of tumor cells and CD8 positive cells infiltration were analyzed by immunohistochemistry. Survival analysis including over-all survival (OS) and progression-free survival (PFS) were also statistically examined by log-rank test. Results: Fourteen cases (15%) were β-catenin positive. Among 14 β-catenin positive cases, only 2 cases (14%) had CD8 positive cells infiltration. Seventy-five cases (97%) had CD8 positive cells infiltration in β-catenin negative tumors (p < 0.01). Squamous cell carcinoma was relatively higher level of expression than others (44% vs. 9%) and mean primary tumor size was larger in β-catenin positive tumors (3.6 cm) than those in negative tumors (2.7 cm). Median OS was 43.8 months and median PFS was 37.0 months in β-catenin positive group. They were not reached in β-catenin negative group. Both OS and PFS were shorter in β-catenin positive cases (p < 0.01). Conclusions: In NSCLC, β-catenin expression was negatively associated with CD8 positive cells infiltration and poor prognosis. We furthermore will focus on the association between β-catenin expression in tumor and clinical efficacy or resistant mechanism of immune checkpoint blockades.
A preclinical study of tumor-infiltrating lymphocytes in NSCLC.
Authors: Federico Lorenzo , Haymaker Cara L. , Forget Marie-Andree , Ravelli Andrea , Bhatta Ankit , Karpinets Tatiana , Zhang Ran , Weissferdt Annikka , Fang Bingliang , Zhang Jianjun , Cascone Tina , Vaporciyan Ara A. , Futreal Andrew , Wistuba Ignacio Ivan , Sepesi Boris , Heymach John , Gibbons Don Lynn , Bernatchez Chantale ,   ICON Team
Published: February 2018

161

Background: Multiple clinical studies have shown that adoptive cell transfer (ACT) of autologous tumor-infiltrating lymphocytes (TIL) is remarkably effective in melanoma patients. Non-small cell lung cancer (NSCLC) shares similarities with melanoma in terms of mutational burden and sensitivity to immune checkpoint inhibitors. We therefore sought to test whether TIL ACT may represent a viable option for the treatment of NSCLC patients. We utilized tissue collected from patients enrolled on the prospective ImmunogenomiC prOfiling of Non-small cell lung cancer (ICON) study. Methods: TIL and tissue-infiltrating lymphocytes were expanded ex vivo from 97 freshly resected early-stage localized NSCLC tumors and 39 matched uninvolved lung tissues. Growth and functional characteristics of TIL were assessed via flow cytometry, TIL-tumor reactivity assays, and analysis of TCRβ sequencing data. Results: NSCLC showed an increased proportion of CD3+ lymphocytes within the tumor-infiltrating leukocyte component as compared to matched normal lung tissue. The TIL compartment included a suppressed CD8+ T cell subset expressing significantly higher levels of PD-1 and lacking cytolytic potential compared to T cells expanded from normal tissue. TIL contained a higher proportion of proliferating (Ki67+) CD8+CD103+ tissue-resident memory (TRM) cells expressing activation markers such as CTLA4, LAG3, PD1 and ICOS, and increased CD4+ Tregs. Despite a highly immunosuppressive environment, TIL expansion was achieved with a success rate of 68% (n = 97) but appeared hindered in patients undergoing neoadjuvant chemotherapy treatment prior to surgery (56.2%, n = 16 vs 72.5% success rate in therapy-naïve patients). In addition, expansion efficiency (number expanded and time of culture) of TIL and matched lung residing lymphocytes were significantly associated (r = 0.379, p = 0.017, n = 39). Importantly, expanded CD8+ TIL products were oligoclonal and showed reactivity toward autologous tumors. Conclusions: Although NSCLC TIL are functionally inhibited in vivo they can be successfully expanded ex vivo and demonstrate recognition of autologous tumor cells. These data suggest that TIL can potentially be used for adoptive T cell-based immunotherapy in NSCLC.
Potential risk factors of pneumonitis associated with consolidation pembrolizumab after chemoradiation in unresectable NSCLC patients.
Authors: Ahmed Shahid Sattar , Durm Greg Andrew , Donatelli John , Yao Huan , Liu Yongmei , DesRosiers Colleen , Liu Ziyue , Kong Feng Ming , Hanna Nasser H.
Published: February 2018

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Background: We have previously shown that consolidation pembrolizumab after chemoradiation is safe in a 93 patient-clinical trial of unresectable stage III NSCLC patients (Durm, ASCO 2016). However 15 patients (16%) developed grade ≥2 pneumonitis. There is limited data on risk factors for developing pneumonitis in this population. We conducted a retrospective review of these patients to identify potential clinical, biologic, radiographic and radiation treatment related risk factors for developing pneumonitis. Methods: We evaluated ECOG PS, age, gender, smoking status, stage, histology, chemo regimen, consolidation chemotherapy, TILs, PDL-1 status, PFTs, PET SUVs, grade of pre-existing ILD, COPD and Radiation dosimetry plans. Logistic regression and fisher’s-exact test were used for the odds ratio and p-values. Results: See Table. [1] [1] Age, histology, stage, chemo, PFTs, SUVs and PDL1 were not significant [2] n = sample size p = pneumonitis [3] Moderate/severe COPD was not seen in any patients with pneumonitis despite 94% of participants being smokers. Conclusions: This preliminary report highlights the utility of a multi-disciplinary approach to develop risk-stratification models for pneumonitis in an at-risk patient population. However larger prospective studies are needed to validate their meaningful use in the clinical setting.[Table: see text]
Therapeutic checkpoint targets: Evaluation of co-expression profiles in individual tumor and immune cells in the tumor microenvironment of FFPE tissue.
Authors: Laeremans Annelies , Li Na , Kim Jeff , Ma Xiao-Jun , Park Emily
Published: February 2018

176

Background: Interactions between tumor and immune cells in the tumor microenvironment (TME) play a key role in tumor progression and treatment response with accumulating evidence indicating a crucial role for tumor infiltrating immune cells. Although infiltrating cytotoxic T lymphocytes (CTLs) have been correlated with improved clinical outcome, they are ineffective in eradicating tumors due to their inhibition by immune checkpoint molecules. Immune checkpoint inhibitors have demonstrated therapeutic efficacy and durable response for several tumor types including non-small cell lung cancer (NSCLC). However, the majority of patients are resistant or relapse after initial response. Characterizing the TME for checkpoint expression with single-cell and spatial resolution can provide critical insight into new immunotherapeutic strategies and identify new predictive biomarkers for stratifying and identifying patients most likely to benefit from immunotherapy including PD-1/PD-L1 immune checkpoint blockade. Methods: Using RNAscope in situ hybridization, we evaluated in situ co-expression profiles of therapeutic checkpoint targets at single-cell level in the TME of 56 archived NSCLC FFPE tissues. Results: Checkpoint molecules including PD1, PD-L1, PD-L2, TIM3, LAG3, CTLA-4 and GITR were visualized in a highly specific and sensitive manner in individual cells within tissue morphological context. Multiple checkpoint molecules were detected in the same immune environment, especially in highly inflamed tumors. In addition to PD-L1, tumor cell-intrinsic expression of PD1, TIM3, LAG3, and PD-L2 was observed in a subset of samples. Furthermore, co-expression of therapeutic checkpoint targets including PD1, LAG3, and TIM3 was observed in infiltrating immune cells and tumor cells. Conclusions: Single-cell co-expression profiles of checkpoint molecules could shed light on how cancer cells evade the host immune surveillance and develop resistance against checkpoint blockades. Also, they could reveal valuable insights into combinatorial therapies for checkpoint markers co-expressed by the patient’s immune cells in the TME.
Systemic effect of radiotherapy (RT) followed by programmed death 1 (PD-1) inhibitors in non-small-cell lung cancer (NSCLC).
Authors: Bassanelli Maria , Giannarelli Diana , Gelibter Alain , Cecere Fabiana Letizia , Chiari Rita , Giacinti Silvana , Scagnoli Simone , Milella Michele , Ricciuti Biagio , Roberto Michela , Magri Valentina , Ferraresi Virginia , Brambilla Marta , Santarelli Mario , Marchetti Paolo , Cortesi Enrico , Ceribelli Anna
Published: February 2018

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Background: RT-induced abscopal effect seems increase the efficacy of immunotherapy. The aim of this study was to evaluate the outcome of patients (pts) with NSCLC previously undergone to RT before receiving PD-1 inhibitors. Methods: We conducted an observational, retrospective analysis of 63 consecutive pts with advanced NSCLC who received RT followed by PD-1 inhibitors at five Italian institutions. Tumor response to treatment was defined according to Response Evaluation Criteria in Solid Tumors version 1.1. Median overall survival (OS) and the 95% confidence interval (CI) were estimated with the Kaplan -Meier method. Results: 63 pts (median age 66 years; male:60.3%) with advanced NSCLC (adenocarcinoma [adc]:71.4%; squamous cells [sqc]:28.6%) were treated with PD-1 inhibitors after RT. RT was delivered a median of 70 days before the start of immunotherapy.47 pts(74.5%)received extracranical RT and 16 pts (25.7%) performed cranial irradiation. Median OS was 11.7 months (mo) [95% CI, 4.7-18.7] (adc: 13.0 mo [95% CI,7.3-18.7], sqc 5.1 mo [95%CI,2.9-7.3]). Median progression free survival (PFS) was 5.1 mo [95% CI,2.5-7.7] (adc: 9.0 mo[ 95% CI,2.6-15.3]; sqc: 3.5 mo [95% CI,2.0-5.1]). A better performance status (PS) according to ECOG scale was associated with an improved OS (PS 0[21 pts]: 15.4 mo [95% CI,10.8-19.9]; PS1[33 pts]: 11.7 mo [95%CI,4.0-19.4]; PS2[9 pts]: 4.1 mo [95% CI,0.9-7.3]). Median OS in 46 pts who received ≤ 1 previous systemic therapy was 13.0 mo [95% CI, 9.4-16.6] and in 17 pts who received ≥2 previous treatments was 7.4 mo [95% CI, 3.1-11.7]. Median OS in 45 pts aged < 70 years was 11.7 mo [95% CI, 4.7-18.7] and in 18 elderly (≥ 70 years) was 6.5 mo [95% CI, 0.0-17.7]. 9(14.3%) partial response, 23(36.5%) stable disease and 25(39.7%) progressive disease have been observed. Conclusions: The synergic combination of RT and PD-1 inhibitors leads to an enhancement of systemic antitumor immune responses, triggering the abscopal effect, resulting in prolonged OS.
Neoadjuvant combination immunotherapy with pembrolizumab and high dose IFN-α2b in locally/regionally advanced melanoma.
Authors: Tarhini Ahmad A. , Lin Yan , Drabick Joseph J. , Neves Rogerio Izar , Ernstoff Marc S. , Puzanov Igor , Vallabhaneni Priyanka , Pingpank James F. , Holtzman Matthew Peter , Sander Cindy , Kirkwood John M.
Published: February 2018

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Background: Neoadjuvant pembrolizumab at 200 mg in combination with high dose IFNα2b (HDI) for locally/regionally advanced or recurrent melanoma may improve the clinical outcomes of these high risk patients (pts), and provide access to blood and tumor pre/post pembro-HDI to illuminate the host effector and suppressor immune mechanisms. Methods: Pts were treated with pembro 200 mg IV every 3 weeks (wk) x 2 doses followed by definitive surgery, then every 3 wks for up to one year. HDI (20 MU/m²/d IV x 5 days (d)/wk for 4 wks then 10 MU/m²/d SC every other d TIW for 48 wks) was given concurrently. Tumor samples were obtained at baseline and at definitive surgery (wk 6-8) and serum/PBMC at baseline, 6 wks, 3, 6, 12 months (mo). Results: Twenty evaluable pts (14 male, 6 female, 14 cutaneous primary, 4 unknown, 3 mucosal), age 29-82 were treated. 5 had Stage IIIB (N1b, N2b, M2c), 11 IIIC (N3) and 4 IV (M1a, M1b) melanoma. Over 230 cycles have been delivered to date (median 14). Worst toxicities included grade (Gr) 3: fatigue (8; 40%), ↑CPK (5; 25%), hypophosphatemia (5; 25%), ↑lipase (3; 15%), lymphopenia (3; 15%), hypertension (2; 10%), diarrhea/colitis (1; 5%), arthralgia (1, 5%), syncope (1, 5%), hyponatremia (1, 5%), neutropenia (1; 5%), anemia (1, 5.00%) nausea (2, 10%), flu like symptoms (1, 5%). There were 3 Gr 4 events (CPK, hyperglycemia, lymphopenia). One suspected grade 5 event occurred 6 months after completion of therapy with autopsy evidence of pneumonia and myocarditis. Among 20 evaluable pts, 4 relapsed and 1 died. Median follow-up for pts who have not relapsed is 11 months. The radiologic preoperative response rate (WHO; unconfirmed) was 65%. The pathologic complete response rate (no viable tumor on histologic assessment) was 35%. Conclusions: Neoadjuvant pembro-HDI exhibited promising clinical activity. Longer follow up is underway in order to define the long term benefits and risks. Clinical trial information: NCT02339324.
PD-L1 and CD8 expression and association with outcomes in patients (pts) with BRAF V600E/K–mutant metastatic melanoma (MM) who received dabrafenib + trametinib (D+T) in the randomized phase 3 COMBI-v study.
Authors: Schadendorf Dirk , Long Georgina V. , Grob Jean Jacques , Nathan Paul D. , Ribas Antoni , Davies Michael A. , Flaherty Keith , Squires Matthew , Jaeger Savina , Powell William , Tan Puay , Robert Caroline
Published: February 2018

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Background: The phase 3 COMBI-v study (NCT01597908) showed that D+T significantly improved outcomes vs vemurafenib in pts with BRAF V600E/K–mutant MM. Checkpoint inhibitors also provide clinical benefit in some pts with MM. To date, characterization of markers associated with response to anti–PD-1 therapy has identified positive associations with PD-L1 expression and immune cell infiltration. Here we describe expression of PD-L1 and the T-cell marker CD8 in tumor samples from pts randomized to receive D+T in COMBI-v and associations with clinical outcomes. Methods: Biopsies from 74 of 352 D+T pts (21%) in COMBI-v were assessed for expression of PD-L1 (PD-L1 IHC 22C3 pharmDx assay; Dako) and CD8 (anti-CD8 antibody, clone C8/144B; Dako). PD-L1 positivity was determined as a percentage of stained tumor cells and MEL score (staining on both tumor and mononuclear inflammatory cells; 0 or 1 [negative]: < 1% staining; 2-5 [positive]: ≥ 1% staining; Daud et al. J Clin Oncol. 2016). Results: Of 74 pts analyzed, 54 (73%) had PD-L1–positive tumors, and the largest MEL score subgroup was 2 (45%). A significant association ( P < .0001) was observed between PD-L1 and CD8 expression. Overall response rate, tumor shrinkage, progression-free survival, overall survival (OS), and duration of response with D+T were not associated with PD-L1 (MEL score of ≥ 2) or CD8 positivity. However, a significant association ( P = .04) with improved OS was observed in tumors with high PD-L1 expression (≥ 20% of cells PD-L1 positive), and a trend ( P = .06) was seen in pts with a MEL score of ≥ 3. Among PD-L1–negative pts, improved OS was seen in those with high CD8 positivity ( P = .03), particularly in the stromal compartment. Conclusions: These data, representing PD-L1 and CD8 expression profiles for a BRAF-mutant MM population in the context of outcomes following D+T, showed that clinical benefit was maintained regardless of immune phenotype. The results also suggest that an immune component has an impact on outcomes following targeted therapy. Clinical trial information: NCT01597908.
Genomic and immune landscape of metastatic melanoma (MM) treated with pembrolizumab (PEM).
Authors: Kanjanapan Yada , Lien Scott , Yang Cindy , Clouthier Derek L. , Elston Sawako , Jiang Haiyan , Butler Marcus O. , Hogg David , Ohashi Pamela S , Pugh Trevor John , Siu Lillian L. , Spreafico Anna
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Background: Predictive biomarkers for PEM in MM is an active area of research. Methods: INSPIRE (NCT02644369) is a biomarker-driven Princess Margaret Cancer Centre initiative to evaluate genomic and immunologic changes in tumor and blood of patient (pts) treated with PEM at 200 mg IV Q3W. Baseline (BL) and on treatment (post 2-3 doses of PEM) fresh tumor biopsies were collected for DNA/RNA sequencing, immune-profiling, and tumor PD-L1 expression by immunohistochemistry (QualTek), with longitudinal blood collection for immunophenotyping. Results: As of 9/2017, 11 MM pts (9 cutaneous, 1 choroidal, 1 mucosal) were enrolled. Median treatment duration was 6 months (0.75 – 14.25); 7 pts remain on study. 7/11 (64%) pts had partial response (PR), 3/11 (27%) progressive disease (PD) and 1 was not evaluable (NE) per RECIST 1.1. Based on data for 8 pts (6 PR, 2 PD), higher percentage (%) PD-1 (mean 70 vs 12%), TIGIT (82 vs 43%) and 4-1BB (23 vs 4%) positivity on CD8 T cells in tumor vs blood was seen (all p < 0.05). In BL blood, lower % TIGIT positive CD8 T cells was detected in pts with PR vs PD (38 vs 58%, p < 0.05). T cell profile of 4 pts with paired BL and post PEM samples is shown in Table 1. Mean tumor PD-L1 score was 24% (0 - 90%), with no correlation to PEM response. Genomic analysis on 8 pts (5 PR, 2 PD, 1 NE) found mean tumor mutation burden (TMB) of 538. B2M deletion was seen in 2/2 PD and 0/6 PR pts. Among pts with PRs, 4/6 (67%) had TAP1 and TAP2 amplification post PEM therapy. HLA-A and HLA-B amplifications were each found in 3/6 (50%) PR pts, with only 1 present at BL. TMB was not associated with response to PEM. Conclusions: In this exploratory analysis of MM pts treated with PEM, there was significantly discordant T cell phenotype in tumor vs blood at BL. Our findings suggest B2M deletion to be associated with PEM resistance. The observed development of HLA-A/B and TAP1/2 amplification in 50-67% pts with PR post PEM suggests potential predictive significance, although further validation is warranted. Clinical trial information: NCT02644369. [Table: see text]