bluuebunny/crossref_metadata_2025 huggingface dataset:
Epidemiology and Management of invasive infections among people who Use drugs (EMU): protocol for a prospective, multicentre cohort study
Authors: Attwood Lucy O , Bryant Mellissa , Lee Sue J , Vujovic Olga , Higgs Peter , Doyle Joseph S , Stewardson Andrew J
Published: April 2023

Introduction

People who inject drugs (PWID) are at risk of invasive infections such as bloodstream infections, endocarditis, osteomyelitis and septic arthritis. Such infections require prolonged antibiotic therapy, but there is limited evidence about the optimal care model to deliver to this population. The Epidemiology and Management of invasive infections among people who Use drugs (EMU) study aims to (1) describe the current burden, clinical spectrum, management and outcomes of invasive infections in PWID; (2) determine the impact of currently available models of care on completion of planned antimicrobials for PWID admitted to hospital with invasive infections and (3) determine postdischarge outcomes of PWID admitted with invasive infections at 30 and 90 days.

Methods and analysis

EMU is a prospective multicentre cohort study of Australian public hospitals who provide care to PWIDs with invasive infections. All patients who have injected drugs in the previous six months and are admitted to a participating site for management of an invasive infection are eligible. EMU has two components: (1) EMU-Audit will collect information from medical records, including demographics, clinical presentation, management and outcomes; (2) EMU-Cohort will augment this with interviews at baseline, 30 and 90 days post-discharge, and data linkage examining readmission rates and mortality. The primary exposure is antimicrobial treatment modality, categorised as inpatient intravenous antimicrobials, outpatient antimicrobial therapy, early oral antibiotics or lipoglycopeptide. The primary outcome is confirmed completion of planned antimicrobials. We aim to recruit 146 participants over a 2-year period.

Ethics and dissemination

EMU has been approved by the Alfred Hospital Human Research Ethics Committee (Project number 78815.) EMU-Audit will collect non-identifiable data with a waiver of consent. EMU-Cohort will collect identifiable data with informed consent. Findings will be presented at scientific conferences and disseminated by peer-review publications.

Trial registration number

ACTRN12622001173785; Pre-results.
Diabetic retinopathy progression in patients under monitoring for treatment or vision loss: external validation and update of a multivariable prediction model
Authors: Haider Sajjad , Adderley Nicola , Tallouzi Mohammad O , Sadiq Salman Naveed , Steel David H , Chavan Randhir , Sheikh Ijaz , Nirantharakumar Krishnarajah , Snell Kym I E
Published: April 2023

Introduction

The number of people with diabetes mellitus is increasing globally and consequently so too is diabetic retinopathy (DR). Most patients with diabetes are monitored through the diabetic eye screening programme (DESP) until they have signs of retinopathy and these changes progress, requiring referral into hospital eye services (HES). Here, they continue to be monitored until they require treatment. Due to current pressures on HES, delays can occur, leading to harm. There is a need to triage patients based on their individual risk. At present, patients are stratified according to retinopathy stage alone, yet other risk factors like glycated haemoglobin (HbA1c) may be useful. Therefore, a prediction model that combines multiple prognostic factors to predict progression will be useful for triage in this setting to improve care.

We previously developed a Diabetic Retinopathy Progression model to Treatment or Vision Loss (DRPTVL-UK) using a large primary care database. The aim of the present study is to externally validate the DRPTVL-UK model in a secondary care setting, specifically in a population under care by HES. This study will also provide an opportunity to update the model by considering additional predictors not previously available.

Methods and analysis

We will use a retrospective cohort of 2400 patients with diabetes aged 12 years and over, referred from DESP to the NHS hospital trusts with referable DR between 2013 and 2016, with follow-up information recorded until December 2021.

We will evaluate the external validity of the DRPTVL-UK model using measures of discrimination, calibration and net benefit. In addition, consensus meetings will be held to agree on acceptable risk thresholds for triage within the HES system.

Ethics and dissemination

This study was approved by REC (ref 22/SC/0425, 05/12/2022, Hampshire A Research Ethics Committee). The results of the study will be published in a peer-reviewed journal, presented at clinical conferences.

Trial Registration number

ISRCTN 10956293.
Degree of loss in the tissue thickness, microvascular density, specific perimetry and standard perimetry in early glaucoma
Authors: Hirasawa Kazunori , Yamaguchi Jun , Nagano Koichi , Kanno Junji , Kasahara Masayuki , Shoji Nobuyuki
Published: April 2023

Objective

To identify the degree of loss of the circumpapillary retinal nerve fibre layer (cpRNFL), the layer from the macular RNFL to the inner plexiform layer (mGCL++), circumpapillary (cpVD) and macular vascular density (mVD), Pulsar perimetry and standard perimetry in early glaucoma.

Methods

In this cross-sectional study, one eye from each of 96 healthy controls and 90 eyes with open-angle glaucoma were measured with cpRNFL, mGCL++, cpVD, mVD, Pulsar perimetry with Octopus P32 test (Pulsar) and standard perimetry with Humphrey field analyser 24-2 test (HFA). For direct comparison, all parameters were converted to relative change values adjusted in both their dynamic range and age-corrected normal value.

Results

The degree of loss in mGCL++ (−24.7%) and cpRNFL (−25.8%) was greater than that in mVD (−17.3%), cpVD (−14.9%), Pulsar (−10.1%) and HFA (−5.9%) (each p<0.01); the degree of loss in mVD and cpVD was greater than that in Pulsar and HFA (each p<0.01); and the degree of loss in Pulsar was greater than that in HFA (p<0.01). The discrimination ability between glaucomatous and healthy eyes (area under the curve) was higher for mGCL++ (0.90) and cpRNFL (0.93) than for mVD (0.78), cpVD (0.78), Pulsar (0.78) and HFA (0.79).

Conclusion

The degree of loss of cpRNFL and mGCL++ thickness preceded by approximately 7%–10% and 15%–20% compared with the micro-VD and visual fields in early glaucoma, respectively.

Trial registration number

UMIN Clinical Trials Registry (http://www.umin.ac.jp/; R000046076 UMIN000040372).
Improving type 2 diabetes mellitus management in Ministry of Defense Hospitals in the Kingdom of Saudi Arabia 2018–2021
Authors: Alotaibi Yasser K , Al-Nowaiser Noura , Al Harbi Turki J , Tourkmani Ayla M , Moharram Maha
Published: April 2023

Diabetes mellitus is a metabolic disease characterised by elevated levels of blood glucose and is a leading cause of disability and mortality. Uncontrolled type 2 diabetes leads to complications such as retinopathy, nephropathy and neuropathy. Improved treatment of hyperglycaemia is likely to delay the onset and progression of microvascular and neuropathic complications.

This article describes the efforts of 18 governmental hospitals in the Kingdom of Saudi Arabia that enrolled in a collaborative improvement project to improve the poor glycaemic control (HbA1c >9% to be less than 15%) of patients with diabetes by the end of 2021 among all the chronic illness clinics in the enrolled military hospitals. Enrolled hospitals were required to implement an evidence-based change package that included the implementation of diabetes clinical practice guidelines with standardised assessment and care planning tools. Furthermore, care delivery was standardised using a standard clinic scope of service that focused on multidisciplinary care teams. Finally, hospitals were required to implement diabetes registries that were used by case managers for poorly controlled patients.

The project timetable was from October 2018 to December 2021. Diabetes poor control (HbA1c >9%) showed improved mean difference of 12.7% (34.9% baseline, 22.2% after) with a p value of 0.01. Diabetes optimal testing significantly improved from 41% at the start of the project in the fourth quarter of 2018, reaching 78% by the end of the fourth quarter of 2021. Variation between hospitals showed a significant reduction in the first quarter of 2021.

The collaborative multilevel approach of standardising the care based on the best available evidence through policies, guidelines and protocols, patient-focused care and integrated care plan by a multidisciplinary team was associated with noticeable improvement in all key performance indicators of the project.
Integrated model of primary and mental healthcare for the refugee population served by an academic medical centre
Authors: Daniel Katharine E , Blackstone Sarah R , Tan Joseph S , Merkel Richard L , Hauck Fern R , Allen Claudia W
Published: April 2023

Refugees are at increased risk for developing mental health concerns due to high rates of trauma exposure and postmigration stressors. Moreover, barriers to accessing mental health services result in ongoing suffering within this population. Integrated care—which combines primary healthcare and mental healthcare into one cohesive, collaborative setting—may improve refugees’ access to comprehensive physical and mental health services to ultimately better support this uniquely vulnerable population. Although integrated care models can increase access to care by colocating multidisciplinary services, establishing an effective integrated care model brings unique logistic (eg, managing office space, delineating roles between multiple providers, establishing open communication practices between specialty roles) and financial (eg, coordinating across department-specific billing procedures) challenges. We therefore describe the model of integrated primary and mental healthcare used in the International Family Medicine Clinic at the University of Virginia, which includes family medicine providers, behavioural health specialists and psychiatrists. Further, based on our 20-year history of providing these integrated services to refugees within an academic medical centre, we offer potential solutions for addressing common challenges (eg, granting specialty providers necessary privileges to access visit notes entered by other specialty providers, creating a culture where communication between providers is the norm, establishing a standard that all providers ought to be CC’ed on most visit notes). We hope that our model and the lessons we have learned along the way can help other institutions that are interested in developing similar integrated care systems to support refugees’ mental and physical health.
Targeted multiomics in childhood-onset SLE reveal distinct biological phenotypes associated with disease activity: results from an explorative study
Authors: Wahadat Mohamed Javad , van Tilburg Sander J , Mueller Yvonne M , de Wit Harm , Van Helden-Meeuwsen Cornelia G , Langerak Anton W , Gruijters Marike J , Mubarak Amani , Verkaaik Marleen , Katsikis Peter D , Versnel Marjan A , Kamphuis Sylvia
Published: April 2023

Objective

To combine targeted transcriptomic and proteomic data in an unsupervised hierarchical clustering method to stratify patients with childhood-onset SLE (cSLE) into similar biological phenotypes, and study the immunological cellular landscape that characterises the clusters.

Methods

Targeted whole blood gene expression and serum cytokines were determined in patients with cSLE, preselected on disease activity state (at diagnosis, Low Lupus Disease Activity State (LLDAS), flare). Unsupervised hierarchical clustering, agnostic to disease characteristics, was used to identify clusters with distinct biological phenotypes. Disease activity was scored by clinical SELENA-SLEDAI (Safety of Estrogens in Systemic Lupus Erythematosus National Assessment-Systemic Lupus Erythematosus Disease Activity Index). High-dimensional 40-colour flow cytometry was used to identify immune cell subsets.

Results

Three unique clusters were identified, each characterised by a set of differentially expressed genes and cytokines, and by disease activity state: cluster 1 contained primarily patients in LLDAS, cluster 2 contained mainly treatment-naïve patients at diagnosis and cluster 3 contained a mixed group of patients, namely in LLDAS, at diagnosis and disease flare. The biological phenotypes did not reflect previous organ system involvement and over time, patients could move from one cluster to another. Healthy controls clustered together in cluster 1. Specific immune cell subsets, including CD11c+ B cells, conventional dendritic cells, plasmablasts and early effector CD4+ T cells, differed between the clusters.

Conclusion

Using a targeted multiomic approach, we clustered patients into distinct biological phenotypes that are related to disease activity state but not to organ system involvement. This supports a new concept where choice of treatment and tapering strategies are not solely based on clinical phenotype but includes measuring novel biological parameters.
Methodology to develop a capital improvement plan using optimization technique: case study
Authors: Saha Promothes
Published: April 2023

Capital Improvement Plan (CIP) plays a critical role in maintaining and upgrading a road system by determining which road segments should receive treatment for a year. The funding provided by the local government is considered as the available budget, and by synchronizing the available budget, the CIP ensures that the overall pavement condition is optimized. The primary factors included in the methodology are the following: budget, Pavement Surface Evaluation and Rating (PASER), deterioration model, cost matrix, and improvement associated with selecting the treatment. A combined priority index was determined for each roadway segment based on the functional classification of roadways and the maintenance treatment type. The linear integer optimization technique was applied to identify the best projects for improvement. The overall methodology was applied to the City of Fort Wayne’s Road system to verify the method’s applicability as a case study.
Tradeoff in Design of Communication Structures in Wireless Sensor Network
Authors: Shamai Liron Cohen , Segal Michael
Published: April 2023

In this paper, we investigate the topic of designing a sensor network by simultaneously optimizing its data energy efficiency and latency. We propose to design a network structure based on the number of nearest neighbors of wireless nodes and provide an analytic bound for the network measurements such as energy consumption and hop-diameter. We also show how to obtain the tradeoff between both criteria. To the best of our knowledge, no non-trivial sublogarithmic hop-diameter bound has been known before. Moreover, our structure is highly fault resilient. We also present simulation results to validate our claims.

1
Study on Joint Damage of Double Prefabricated Fragments Penetrating Finite Thickness Concrete
Authors: Wang Zhenning , Yang Junhui , Yin Jianping , Li Xudong , Shi Rui , Yi Jianya
Published: April 2023

In order to study the joint damage mechanism of multiple prefabricated fragments to finite thickness concrete targets, experiments on the damaging effect of a single fragment on a 300 × 300 × 100 mm concrete target were carried out, and the reliability of the simulation calculation model of single fragment damage to concrete was verified. On this basis, according to the trajectory of two fragments penetrating concrete, the double fragment penetration is divided into two penetration situations, that is, coplanar and heterogeneous. The orthogonal optimization method is used to carry out the joint damage simulation calculation of the double fragments to the concrete target by changing the fragment velocity, penetration angle, fragment spacing, and other factors. The simulation results show that the relationship between joint damage and fragment spacing is the largest when the fragment trajectories are coplanar, and the partial least squares regression coefficients affecting the joint damage time and surface joint length are 0.70 and 0.68 respectively. When the trajectory is different, the joint damage mode is relatively complex. Based on this, the joint damage degree analysis method between fragments is established, and each variable can explain 73.8% of the joint damage degree. It is found that the joint damage of the front pit area is the largest when the fragment is in different planes, and the PLS regression coefficient is −0.44. The hypothesis that joint damage is easy to occur in the area of the intersection line on the back of the target is analyzed and verified.
A Robust Residual Shrinkage Balanced Network for Image Recognition from Japanese Historical Documents
Authors: Qiu Hong , Dong Jing
Published: April 2023

Kuzushiji, as characters written in a cursive style, had been continuously used for both publishing and handwriting in Japan before the end of the 19th century. However, well into the twentieth century, most modern Japanese lost the ability to read Kuzushiji due to changes in writing systems. Therefore, how to develop advanced machine learning algorithms to identify Japanese historical characters has great significance in preserving cultural knowledge and promoting sustainable development in urban living. Unlike traditional digit image recognition, Kuzushiji image recognition is a more challenging task due to the complex script for cursive handwriting, multiple variations between characters, and imbalanced data. To address such issues, a novel deep learning‐based character recognition method called residual shrinkage balanced network (RSBN) is developed for classifying Japanese historical characters. A novel residual shrinkage structure with a soft threshold function is constructed to reduce redundant features using specially designed skip‐connected subnetworks. Then, multiple residual shrinkage modules are stacked together to learn hierarchical features from raw input. Furthermore, a novel class rebalancing strategy is further designed to improve learning performance from imbalanced data. The experiments are conducted on Japanese classical literature datasets. The results demonstrate that the proposed method obtains more than 89.01% testing accuracy on Kuzushiji‐MNIST dataset, which outperforms the existing deep learning‐based character image recognition methods. Moreover, without additional data augmentation techniques, the proposed approach also achieves 97.16% balanced accuracy on Kuzushiji‐49 dataset, which is the best one compared with existing published results. When considering the computational performance, the number of trainable parameters of the proposed method is competitive with that of the typical ResNet‐18.
Supplementary Methods from RETRACTED: Variable Inhibition of Thrombospondin 1 against Liver and Lung Metastases through Differential Activation of Metalloproteinase ADAMTS1
Authors: Lee Yoon-Jin , Koch Moritz , Karl Daniel , Torres-Collado Antoni X. , Fernando Namali T. , Rothrock Courtney , Kuruppu Darshini , Ryeom Sandra , Iruela-Arispe M. Luisa , Yoon Sam S.
Published: April 2023

Supplementary Methods from RETRACTED: Variable Inhibition of Thrombospondin 1 against Liver and Lung Metastases through Differential Activation of Metalloproteinase ADAMTS1
Data Supplement from RETRACTED: CD44 Expression Denotes a Subpopulation of Gastric Cancer Cells in Which Hedgehog Signaling Promotes Chemotherapy Resistance
Authors: Yoon Changhwan , Park Do Joong , Schmidt Benjamin , Thomas Nicholas J. , Lee Hae-June , Kim Teresa S. , Janjigian Yelena Y. , Cohen Deirdre J. , Yoon Sam S.
Published: April 2023

Supplementary Figure 1 - Expression of CD44, other stem cell markers, and Hedgehog pathway proteins in gastric cancer cells.
Suppl. Figure 7 from Integrated Genomic and Immunophenotypic Classification of Pancreatic Cancer Reveals Three Distinct Subtypes with Prognostic/Predictive Significance
Authors: Wartenberg Martin , Cibin Silvia , Zlobec Inti , Vassella Erik , Eppenberger-Castori Serenella , Terracciano Luigi , Eichmann Micha David , Worni Mathias , Gloor Beat , Perren Aurel , Karamitopoulou Eva
Published: April 2023

Oncoprint
Data from Predominance of mTORC1 over mTORC2 in the Regulation of Proliferation of Ovarian Cancer Cells: Therapeutic Implications
Authors: Montero Juan Carlos , Chen Xi , Ocaña Alberto , Pandiella Atanasio
Published: April 2023

Abstract
mTOR is a serine/threonine kinase that acts by binding different sets of proteins forming two complexes, termed mTORC1 and mTORC2. mTOR is deregulated in a substantial proportion of ovarian tumors. Despite the use of drugs directed to mTOR in ongoing clinical trials, the functional relevance of the individual mTORC branches in ovarian cancer is not known. Here, we show that mTORC1 and mTORC2 were constitutively active in ovarian cancer cell lines. Knockdown of raptor or rictor, proteins required for the function of mTORC1 or mTORC2, respectively, resulted in profound inhibition of ovarian cancer cell proliferation. The knockdown of raptor had a more important inhibitory effect than the knockdown of rictor, indicating mTORC1 had a predominant role over mTORC2 in the control of ovarian cancer cell proliferation. Rapamycin decreased the proliferation of ovarian cancer cells, and this was accompanied by inhibition of the phosphorylation of S6, a protein used as readout of mTORC1 function. However, rapamycin had only a marginal effect on the phosphorylation status of 4E-BP1, another mTORC1 substrate. Therefore, mTORC1 probably controls p4E-BP1 along two distinct pathways, one of them sensitive to rapamycin and another insensitive. The dual PI3K/mTOR inhibitor BEZ235 was more efficient than rapamycin in its inhibitory action on ovarian cancer cell proliferation. Biochemically, BEZ235 completely inhibited pS6, p4E-BP1, and pAkt. Our results suggest that broad-spectrum mTOR inhibitors that block mTORC1 and mTORC2 are more desirable for their clinical development in ovarian cancer than agents exclusively targeting one of the mTOR branches. Mol Cancer Ther; 11(6); 1342–52. ©2012 AACR.
Data from Ovarian Cancer Stem Cell–Like Side Populations Are Enriched Following Chemotherapy and Overexpress EZH2
Authors: Rizzo Siân , Hersey Jenny M. , Mellor Paul , Dai Wei , Santos-Silva Alessandra , Liber Daniel , Luk Louisa , Titley Ian , Carden Craig P , Box Garry , Hudson David L. , Kaye Stanley B. , Brown Robert
Published: April 2023

Abstract
Platinum-based chemotherapy, with cytoreductive surgery, is the cornerstone of treatment of advanced ovarian cancer; however, acquired drug resistance is a major clinical obstacle. It has been proposed that subpopulations of tumor cells with stem cell–like properties, such as so-called side populations (SP) that overexpress ABC drug transporters, can sustain the growth of drug-resistant tumor cells, leading to tumor recurrence following chemotherapy. The histone methyltransferase EZH2 is a key component of the polycomb-repressive complex 2 required for maintenance of a stem cell state, and overexpression has been implicated in drug resistance and shorter survival of ovarian cancer patients. We observed higher percentage SP in ascites from patients that have relapsed following chemotherapy compared with chemonaive patients, consistent with selection for this subpopulation during platinum-based chemotherapy. Furthermore, ABCB1 (P-glycoprotein) and EZH2 are consistently overexpressed in SP compared with non-SP from patients' tumor cells. The siRNA knockdown of EZH2 leads to loss of SP in ovarian tumor models, reduced anchorage-independent growth, and reduced tumor growth in vivo. Together, these data support a key role for EZH2 in the maintenance of a drug-resistant, tumor-sustaining subpopulation of cells in ovarian cancers undergoing chemotherapy. As such, EZH2 is an important target for anticancer drug development. Mol Cancer Ther; 10(2); 325–35. ©2010 AACR.
Data from Knockdown of Oncogenic KRAS in Non–Small Cell Lung Cancers Suppresses Tumor Growth and Sensitizes Tumor Cells to Targeted Therapy
Authors: Sunaga Noriaki , Shames David S. , Girard Luc , Peyton Michael , Larsen Jill E. , Imai Hisao , Soh Junichi , Sato Mitsuo , Yanagitani Noriko , Kaira Kyoichi , Xie Yang , Gazdar Adi F. , Mori Masatomo , Minna John D.
Published: April 2023

Abstract
Oncogenic KRAS is found in more than 25% of lung adenocarcinomas, the major histologic subtype of non–small cell lung cancer (NSCLC), and is an important target for drug development. To this end, we generated four NSCLC lines with stable knockdown selective for oncogenic KRAS. As expected, stable knockdown of oncogenic KRAS led to inhibition of in vitro and in vivo tumor growth in the KRAS-mutant NSCLC cells, but not in NSCLC cells that have wild-type KRAS (but mutant NRAS). Surprisingly, we did not see large-scale induction of cell death and the growth inhibitory effect was not complete. To further understand the ability of NSCLCs to grow despite selective removal of mutant KRAS expression, we conducted microarray expression profiling of NSCLC cell lines with or without mutant KRAS knockdown and isogenic human bronchial epithelial cell lines with and without oncogenic KRAS. We found that although the mitogen-activated protein kinase pathway is significantly downregulated after mutant KRAS knockdown, these NSCLCs showed increased levels of phospho-STAT3 and phospho–epidermal growth factor receptor, and variable changes in phospho-Akt. In addition, mutant KRAS knockdown sensitized the NSCLCs to p38 and EGFR inhibitors. Our findings suggest that targeting oncogenic KRAS by itself will not be sufficient treatment, but may offer possibilities of combining anti-KRAS strategies with other targeted drugs. Mol Cancer Ther; 10(2); 336–46. ©2011 AACR.
Data from Cdk4/6 Inhibition Induces Epithelial–Mesenchymal Transition and Enhances Invasiveness in Pancreatic Cancer Cells
Authors: Liu Fang , Korc Murray
Published: April 2023

Abstract
Aberrant activation of Cyclin D-Cdk4/6 signaling pathway is commonly found in pancreatic ductal adenocarcinoma (PDAC). Here, we show that PD-0332991, a highly specific inhibitor for Cdk4 and Cdk6, exerted growth inhibitory effects on three human PDAC cell lines. Microarray analysis revealed that PD-0332991 downregulated cell-cycle–related genes, but upregulated genes implicated in extracellular matrix (ECM) remodeling and pancreatic cancer cell invasion and metastasis. Moreover, PD-0332991 enhanced invasion in TGF-β–responsive PDAC cell lines that harbor a wild-type SMAD4 gene (COLO-357, PANC-1), but not in TGF-β–resistant AsPC-1 cells that harbor a mutated SMAD4. PD-0332991 also induced epithelial–mesenchymal transition (EMT) in COLO-357 and PANC-1, but not in AsPC-1 cells. Inhibition of CDK4/6 using shRNA mimicked the effects of PD-0332991 on EMT induction. Furthermore, PD-0332991 increased Smad transcriptional activity in luciferase readout assays and activated TGF-β signaling. SB-505124, an inhibitor of the type-I TGF-β receptor (TβRI) kinase, completely blocked EMT induction by PD-0332991. When combined with PD-0332991, SB-505124 inhibited the growth of COLO-357 and PANC-1 cells. Taken together, these data suggest that anti-Cdk4/6 therapy could induce EMT and enhance pancreatic cancer cell invasion by activating Smad-dependent TGF-β signaling, and that combining PD-0332991 and SB-505124 may represent a novel therapeutic strategy in PDAC. Mol Cancer Ther; 11(10); 2138–48. ©2012 AACR.
Data from sFLT01: A Novel Fusion Protein with Antiangiogenic Activity
Authors: Bagley Rebecca G. , Kurtzberg Leslie , Weber William , Nguyen Tri-Hung , Roth Stephanie , Krumbholz Roy , Yao Min , Richards Brenda , Zhang Mindy , Pechan Peter , Schmid Steve , Scaria Abraham , Kaplan Johanne , Teicher Beverly A.
Published: April 2023

Abstract
sFLT01 is a novel fusion protein that consists of the VEGF/PlGF (placental growth factor) binding domain of human VEGFR1/Flt-1 (hVEGFR1) fused to the Fc portion of human IgG1 through a polyglycine linker. It binds to both human VEGF (hVEGF) and human PlGF (hPlGF) and to mouse VEGF (mVEGF) and mouse PlGF (mPlGF). In vitro, sFLT01 inhibited the proliferation of human umbilical vein endothelial cells and pericytes stimulated by either hVEGF or hPlGF. In vivo, sFLT01 had robust and significant antitumor activity in numerous preclinical subcutaneous tumor models including H460 non–small cell lung carcinoma, HT29 colon carcinoma, Karpas 299 lymphoma, MOLM-13 AML (acute myeloid leukemia), 786-O, and RENCA renal cell carcinoma (RCC). sFLT01 also increased median survival in the orthotopic RENCA RCC model. sFLT01 had strong antiangiogenic activity and altered intratumoral microvessel density, blood vessel lumen size and perimeter, and vascular and vessel areas in RCC models. sFLT01 treatment resulted in fewer endothelial cells and pericytes within the tumor microenvironment. sFLT01 in combination with cyclophosphamide resulted in greater inhibition of tumor growth than either agent used alone as a monotherapy in the A673 Ewing's sarcoma model. Gene expression profiling indicated that the molecular changes in the A673 sarcoma tumors are similar to changes observed under hypoxic conditions. sFLT01 is an innovative fusion protein that possessed robust antitumor and antiangiogenic activities in preclinical cancer models. It is a dual targeting agent that neutralizes both VEGF and PlGF and, therefore, has potential as a next generation antiangiogenic therapeutic for oncology. Mol Cancer Ther; 10(3); 404–15. ©2011 AACR.
Data from Exon 11 Skipping of E-Cadherin RNA Downregulates Its Expression in Head and Neck Cancer Cells
Authors: Sharma Sanjai , Liao Wei , Zhou Xiaofeng , Wong David T.W. , Lichtenstein Alan
Published: April 2023

Abstract
E-cadherin is an important tumor suppressor gene whose expression is lost when cells acquire a metastatic phenotype. We analyzed the role of E-cadherin missplicing as a mechanism of its downregulation by analyzing a misspliced E-cadherin transcript that lacks exon 11 of this gene. This results in a frameshift and a premature termination codon that targets this transcript for degradation. Tumor tissues, including breast (20%, n = 9), prostate (30%, n = 9) and head and neck (75%, n = 8) cancer, express the exon 11-skipped transcripts (vs. nonmalignant controls) and its levels inversely correlate with E-cadherin expression. This is a novel mechanism of E-cadherin downregulation by missplicing in tumor cells, which is observed in highly prevalent human tumors. In the head and neck cancer model, nontumorigenic keratinocytes express exon 11–skipped splice product two- to sixfold lower than the head and neck tumor cell lines. Mechanistic studies reveal that SFRS2 (SC35), a splicing factor, as one of the regulators that increases missplicing and downregulates E-cadherin expression. Furthermore, this splicing factor was found to be overexpressed in 5 of 7 head and neck cell lines and primary head and neck tumors. Also, methylation of E-cadherin gene acts as a regulator of this aberrant splicing process. In 2 head and neck cell lines, wild-type transcript expression increased 16- to 25-folds, whereas the percentage of exon 11-skipped transcripts in both the cell lines decreased five- to 30-folds when cells were treated with a hypomethylating agent, azacytidine. Our findings reveal that promoter methylation and an upregulated splicing factor (SFRS2) are involved in the E-cadherin missplicing in tumors. Mol Cancer Ther; 10(9); 1751–9. ©2011 AACR.
Data from Cetuximab Reverses the Warburg Effect by Inhibiting HIF-1–Regulated LDH-A
Authors: Lu Haiquan , Li Xinqun , Luo Zhongguang , Liu Jie , Fan Zhen
Published: April 2023

Abstract
Hypoxia-inducible factor-1 (HIF-1) plays a critical role in reprogramming cancer metabolism toward aerobic glycolysis (i.e., the Warburg effect), which is critical to supplying cancer cells with the biomass needed for proliferation. Previous studies have shown that cetuximab, an EGF receptor–blocking monoclonal antibody, downregulates the alpha subunit of HIF-1 (HIF-1α) through the inhibition of EGF receptor downstream cell signaling and that downregulation of HIF-1α is required for cetuximab-induced antiproliferative effects. However, the mechanism underlying these actions has yet to be identified. In this study, we used the Seahorse XF96 extracellular flux analyzer to assess the effect of cetuximab treatment on changes in glycolysis and mitochondrial respiration, the two major energy-producing pathways, in live cells. We found that cetuximab downregulated lactate dehydrogenase A (LDH-A) and inhibited glycolysis in cetuximab-sensitive head and neck squamous cell carcinoma (HNSCC) cells in an HIF-1α downregulation–dependent manner. HNSCC cells with acquired cetuximab resistance expressed a high level of HIF-1α and were highly glycolytic. Overexpression of a HIF-1α mutant (HIF-1α/ΔODD) conferred resistance to cetuximab-induced G1 phase cell-cycle arrest, which could be overcome by knockdown of LDH-A expression. Inhibition of LDH-A activity with oxamate enhanced the response of cetuximab-resistant cells to cetuximab. Cetuximab had no noticeable inhibitory effect on glycolysis in nontransformed cells. These findings provide novel mechanistic insights into cetuximab-induced cell-cycle arrest from the perspective of cancer metabolism and suggest novel strategies for enhancing cetuximab response. Mol Cancer Ther; 12(10); 2187–99. ©2013 AACR.
Data from Combinations of BRAF, MEK, and PI3K/mTOR Inhibitors Overcome Acquired Resistance to the BRAF Inhibitor GSK2118436 Dabrafenib, Mediated by NRAS or MEK Mutations
Authors: Greger James G. , Eastman Stephen D. , Zhang Vivian , Bleam Maureen R. , Hughes Ashley M. , Smitheman Kimberly N. , Dickerson Scott H. , Laquerre Sylvie G. , Liu Li , Gilmer Tona M.
Published: April 2023

Abstract
Recent results from clinical trials with the BRAF inhibitors GSK2118436 (dabrafenib) and PLX4032 (vemurafenib) have shown encouraging response rates; however, the duration of response has been limited. To identify determinants of acquired resistance to GSK2118436 and strategies to overcome the resistance, we isolated GSK2118436 drug-resistant clones from the A375 BRAFV600E and the YUSIT1 BRAFV600K melanoma cell lines. These clones also showed reduced sensitivity to the allosteric mitogen-activated protein/extracellular signal–regulated kinase (MEK) inhibitor GSK1120212 (trametinib). Genetic characterization of these clones identified an in-frame deletion in MEK1 (MEK1K59del) or NRAS mutation (NRASQ61K and/or NRASA146T) with and without MEK1P387S in the BRAFV600E background and NRASQ61K in the BRAFV600K background. Stable knockdown of NRAS with short hairpin RNA partially restored GSK2118436 sensitivity in mutant NRAS clones, whereas expression of NRASQ61K or NRASA146T in the A375 parental cells decreased sensitivity to GSK2118436. Similarly, expression of MEK1K59del, but not MEK1P387S, decreased sensitivity of A375 cells to GSK2118436. The combination of GSK2118436 and GSK1120212 effectively inhibited cell growth, decreased ERK phosphorylation, decreased cyclin D1 protein, and increased p27kip1 protein in the resistant clones. Moreover, the combination of GSK2118436 or GSK1120212 with the phosphoinositide 3-kinase/mTOR inhibitor GSK2126458 enhanced cell growth inhibition and decreased S6 ribosomal protein phosphorylation in these clones. Our results show that NRAS and/or MEK mutations contribute to BRAF inhibitor resistance in vitro, and the combination of GSK2118436 and GSK1120212 overcomes this resistance. In addition, these resistant clones respond to the combination of GSK2126458 with GSK2118436 or GSK1120212. Clinical trials are ongoing or planned to test these combinations. Mol Cancer Ther; 11(4); 909–20. ©2012 AACR.
Data from Nonclinical Evaluation of the Serum Pharmacodynamic Biomarkers HGF and Shed MET following Dosing with the Anti-MET Monovalent Monoclonal Antibody Onartuzumab
Authors: Mai Elaine , Zheng Zhong , Chen Youjun , Peng Jing , Severin Christophe , Filvaroff Ellen , Romero Mally , Mallet William , Kaur Surinder , Gelzleichter Thomas , Nijem Ihsan , Merchant Mark , Young Judy C.
Published: April 2023

Abstract
Onartuzumab, a humanized, monovalent monoclonal anti-MET antibody, antagonizes MET signaling by inhibiting binding of its ligand, hepatocyte growth factor (HGF). We investigated the effects of onartuzumab on cell-associated and circulating (shed) MET (sMET) and circulating HGF in vitro and nonclinically to determine their utility as pharmacodynamic biomarkers for onartuzumab. Effects of onartuzumab on cell-associated MET were assessed by flow cytometry and immunofluorescence. sMET and HGF were measured in cell supernatants and in serum or plasma from multiple species (mouse, cynomolgus monkey, and human) using plate-based immunoassays. Unlike bivalent anti-MET antibodies, onartuzumab stably associates with MET on the surface of cells without inducing MET internalization or shedding. Onartuzumab delayed the clearance of human xenograft tumor-produced sMET from the circulation of mice, and endogenous sMET in cynomolgus monkeys. In mice harboring MET-expressing xenograft tumors, in the absence of onartuzumab, levels of human sMET correlated with tumor size, and may be predictive of MET-expressing tumor burden. Because binding of sMET to onartuzumab in circulation resulted in increasing sMET serum concentrations due to reduced clearance, this likely renders sMET unsuitable as a pharmacodynamic biomarker for onartuzumab. There was no observed effect of onartuzumab on circulating HGF levels in xenograft tumor-bearing mice or endogenous HGF in cynomolgus monkeys. Although sMET and HGF may serve as predictive biomarkers for MET therapeutics, these data do not support their use as pharmacodynamic biomarkers for onartuzumab. Mol Cancer Ther; 13(2); 540–52. ©2013 AACR.
Data from Pharmacologic Activation of PKM2 Slows Lung Tumor Xenograft Growth
Authors: Parnell K. Mark , Foulks Jason M. , Nix Rebecca N. , Clifford Adrianne , Bullough Jeremy , Luo Bai , Senina Anna , Vollmer David , Liu Jihua , McCarthy Virgil , Xu Yong , Saunders Michael , Liu Xiao-Hui , Pearce Scott , Wright Kevin , O'Reilly Marc , McCullar Michael V. , Ho Koc-Kan , Kanner Steven B.
Published: April 2023

Abstract
Inactivation of the M2 form of pyruvate kinase (PKM2) in cancer cells is associated with increased tumorigenicity. To test the hypothesis that tumor growth may be inhibited through the PKM2 pathway, we generated a series of small-molecule PKM2 activators. The compounds exhibited low nanomolar activity in both biochemical and cell-based PKM2 activity assays. These compounds did not affect the growth of cancer cell lines under normal conditions in vitro, but strongly inhibited the proliferation of multiple lung cancer cell lines when serine was absent from the cell culture media. In addition, PKM2 activators inhibited the growth of an aggressive lung adenocarcinoma xenograft. These findings show that PKM2 activation by small molecules influences the growth of cancer cells in vitro and in vivo, and suggest that such compounds may augment cancer therapies. Mol Cancer Ther; 12(8); 1453–60. ©2013 AACR.
Data from Activation of AR Sensitizes Breast Carcinomas to NVP-BEZ235's Therapeutic Effect Mediated by PTEN and KLLN Upregulation
Authors: Wang Yu , Yu Qi , He Xin , Romigh Todd , Altemus Jessica , Eng Charis
Published: April 2023

Abstract
NVP-BEZ235 is a newly developed dual PI3K/mTOR inhibitor, being tested in multiple clinical trials, including breast cancer. NVP-BEZ235 selectively induces cell growth inhibition in a subset, but not all, breast cancer cell lines. However, it remains a challenge to distinguish between sensitive and resistant tumors, particularly in the pretreatment setting. Here, we used ten breast cancer cell lines to compare NVP-BEZ235 sensitivity and in the context of androgen receptor (AR) activation during NVP-BEZ235 treatment. We also used female SCID mice bearing breast tumor xenografts to investigate the beneficial effect of dihydrotestosterone/NVP-BEZ235 combination treatment compared with each alone. We found that AR-positive breast cancer cell lines are much more sensitive to NVP-BEZ235 compared with AR-negative cells, regardless of PTEN or PI3KCA status. Reintroducing AR expression in NVP-BEZ235 nonresponsive AR-negative cells restored the response. DHT/NVP-BEZ235 combination not only resulted in a more significant growth inhibition than either drug alone, but also achieved tumor regression and complete responses for AR+/ER+ tumors. This beneficial effect was mediated by dihydrotestosterone (DHT)-induced PTEN and KLLN expression. Furthermore, DHT could also reverse NVP-BEZ235–induced side effects such as skin rash and weight loss. Our data suggest that AR expression may be an independent predictive biomarker for response to NVP-BEZ235. AR induction could add benefit during NVP-BEZ235 treatment in patients, especially with AR+/ER+ breast carcinomas. Mol Cancer Ther; 13(2); 517–27. ©2013 AACR.
Data from Preclinical Evaluation of AMG 925, a FLT3/CDK4 Dual Kinase Inhibitor for Treating Acute Myeloid Leukemia
Authors: Keegan Kathleen , Li Cong , Li Zhihong , Ma Ji , Ragains Mark , Coberly Suzanne , Hollenback David , Eksterowicz John , Liang Lingming , Weidner Margaret , Huard Justin , Wang Xianghong , Alba Grace , Orf Jessica , Lo Mei-Chu , Zhao Sharon , Ngo Rachel , Chen Ada , Liu Lily , Carlson Timothy , Quéva Christophe , McGee Lawrence R. , Medina Julio , Kamb Alexander , Wickramasinghe Dineli , Dai Kang
Published: April 2023

Abstract
Acute myeloid leukemia (AML) remains a serious unmet medical need. Despite high remission rates with chemotherapy standard-of-care treatment, the disease eventually relapses in a major proportion of patients. Activating Fms-like tyrosine kinase 3 (FLT3) mutations are found in approximately 30% of patients with AML. Targeting FLT3 receptor tyrosine kinase has shown encouraging results in treating FLT3-mutated AML. Responses, however, are not sustained and acquired resistance has been a clinical challenge. Treatment options to overcome resistance are currently the focus of research. We report here the preclinical evaluation of AMG 925, a potent, selective, and bioavailable FLT3/cyclin-dependent kinase 4 (CDK4) dual kinase inhibitor. AMG 925 inhibited AML xenograft tumor growth by 96% to 99% without significant body weight loss. The antitumor activity of AMG 925 correlated with the inhibition of STAT5 and RB phosphorylation, the pharmacodynamic markers for inhibition of FLT3 and CDK4, respectively. In addition, AMG 925 was also found to inhibit FLT3 mutants (e.g., D835Y) that are resistant to the current FLT3 inhibitors (e.g., AC220 and sorafenib). CDK4 is a cyclin D–dependent kinase that plays an essential central role in regulating cell proliferation in response to external growth signals. A critical role of the CDK4–RB pathway in cancer development has been well established. CDK4-specific inhibitors are being developed for treating RB-positive cancer. AMG 925, which combines inhibition of two kinases essential for proliferation and survival of FLT3-mutated AML cells, may improve and prolong clinical responses. Mol Cancer Ther; 13(4); 880–9. ©2014 AACR.
Data from NF1 Deletion Generates Multiple Subtypes of Soft-Tissue Sarcoma That Respond to MEK Inhibition
Authors: Dodd Rebecca D. , Mito Jeffrey K. , Eward William C. , Chitalia Rhea , Sachdeva Mohit , Ma Yan , Barretina Jordi , Dodd Leslie , Kirsch David G.
Published: April 2023

Abstract
Soft-tissue sarcomas are a heterogeneous group of tumors arising from connective tissue. Recently, mutations in the neurofibromin 1 (NF1) tumor suppressor gene were identified in multiple subtypes of human soft-tissue sarcomas. To study the effect of NF1 inactivation in the initiation and progression of distinct sarcoma subtypes, we have developed a novel mouse model of temporally and spatially restricted NF1-deleted sarcoma. To generate primary sarcomas, we inject adenovirus containing Cre recombinase into NF1flox/flox; Ink4a/Arfflox/flox mice at two distinct orthotopic sites: intramuscularly or in the sciatic nerve. The mice develop either high-grade myogenic sarcomas or malignant peripheral nerve sheath tumor (MPNST)-like tumors, respectively. These tumors reflect the histologic properties and spectrum of sarcomas found in patients. To explore the use of this model for preclinical studies, we conducted a study of mitogen-activated protein kinase (MAPK) pathway inhibition with the MEK inhibitor PD325901. Treatment with PD325901 delays tumor growth through decreased cyclin D1 mRNA and cell proliferation. We also examined the effects of MEK inhibition on the native tumor stroma and find that PD325901 decreases VEGFα expression in tumor cells with a corresponding decrease in microvessel density. Taken together, our results use a primary tumor model to show that sarcomas can be generated by loss of NF1 and Ink4a/Arf, and that these tumors are sensitive to MEK inhibition by direct effects on tumor cells and the surrounding microenvironment. These studies suggest that MEK inhibitors should be further explored as potential sarcoma therapies in patients with tumors containing NF1 deletion. Mol Cancer Ther; 12(9); 1906–17. ©2013 AACR.
Data from Characterization of LY2228820 Dimesylate, a Potent and Selective Inhibitor of p38 MAPK with Antitumor Activity
Authors: Campbell Robert M. , Anderson Bryan D. , Brooks Nathan A. , Brooks Harold B. , Chan Edward M. , De Dios Alfonso , Gilmour Raymond , Graff Jeremy R. , Jambrina Enrique , Mader Mary , McCann Denis , Na Songqing , Parsons Stephen H. , Pratt Susan E. , Shih Chuan , Stancato Louis F. , Starling James J. , Tate Courtney , Velasco Juan A. , Wang Yong , Ye Xiang S.
Published: April 2023

Abstract
p38α mitogen-activated protein kinase (MAPK) is activated in cancer cells in response to environmental factors, oncogenic stress, radiation, and chemotherapy. p38α MAPK phosphorylates a number of substrates, including MAPKAP-K2 (MK2), and regulates the production of cytokines in the tumor microenvironment, such as TNF-α, interleukin-1β (IL-1β), IL-6, and CXCL8 (IL-8). p38α MAPK is highly expressed in human cancers and may play a role in tumor growth, invasion, metastasis, and drug resistance. LY2228820 dimesylate (hereafter LY2228820), a trisubstituted imidazole derivative, is a potent and selective, ATP-competitive inhibitor of the α- and β-isoforms of p38 MAPK in vitro (IC50 = 5.3 and 3.2 nmol/L, respectively). In cell-based assays, LY2228820 potently and selectively inhibited phosphorylation of MK2 (Thr334) in anisomycin-stimulated HeLa cells (at 9.8 nmol/L by Western blot analysis) and anisomycin-induced mouse RAW264.7 macrophages (IC50 = 35.3 nmol/L) with no changes in phosphorylation of p38α MAPK, JNK, ERK1/2, c-Jun, ATF2, or c-Myc ≤ 10 μmol/L. LY2228820 also reduced TNF-α secretion by lipopolysaccharide/IFN-γ–stimulated macrophages (IC50 = 6.3 nmol/L). In mice transplanted with B16-F10 melanoma, tumor phospho-MK2 (p-MK2) was inhibited by LY2228820 in a dose-dependent manner [threshold effective dose (TED)70 = 11.2 mg/kg]. Significant target inhibition (>40% reduction in p-MK2) was maintained for 4 to 8 hours following a single 10 mg/kg oral dose. LY2228820 produced significant tumor growth delay in multiple in vivo cancer models (melanoma, non–small cell lung cancer, ovarian, glioma, myeloma, breast). In summary, LY2228820 is a p38 MAPK inhibitor, which has been optimized for potency, selectivity, drug-like properties (such as oral bioavailability), and efficacy in animal models of human cancer. Mol Cancer Ther; 13(2); 364–74. ©2013 AACR.