bluuebunny/crossref_metadata_2025 huggingface dataset:
Induction of EBV latent membrane protein-2A (LMP2A)-specific T cells and construction of individualized TCR-engineered T cells for EBV-associated malignancies
Authors: Zhang Chaoting , Tan Qin , Li Shance , Shen Luyan , Zhang Jingtao , Liu Ying , Yang Wenjun , Lu Zheming
Published: July 2021

Background

Latent membrane protein-2A (LMP2A)-specific TCR-engineered T cells could be a promising treatment approach to Epstein–Barr virus-associated malignancies. However, previous studies mainly reported LMP2A-reactive TCRs only focusing on specific HLA subtypes and corresponding epitopes, and thus, they were only suitable for patients with specific HLA.

Methods

Due to hugely varied HLA subtypes and presented LMP2A epitopes in different individuals, our study attempted to develop an individualized approach, based on the weekly in vitro stimulation of peripheral T cells for 2 weeks with autologous dendritic cells (DCs) pulsed with a pool of LMP2A peptides covering LMP2A whole protein and combination analysis of high throughput TCRβ sequencing of prestimulated and poststimulated T cells and single-cell TCR sequencing of poststimulated T cells, and to identify LMP2A-specific TCRs of which poststimulated frequencies significantly increased than corresponding prestimulated frequencies.

Results

Following this approach, multiple LMP2A-reactive TCRs were identified, optimized and cloned into lentiviral vector, and then transduced into peripheral T cells. These engineerd T cells were demonstrated to specifically recognize the LMP2A presented by autologous DCs and lymphoblastoid cell lines in vitro and in vivo.

Conclusions

This approach provides an efficient procedure to isolate individualized LMP2A-specific TCRs for basic and translational research, as well as for clinical applications.
Photoluminescence of Ca4Ga2S7:Eu2+ in wide excitation intensity and temperature range
Authors: Urmanov B. D. , Leanenia M. S. , Yablonskii G. P. , Taghiyev O. B. , Taghiyev K. O. , Kazimova F. A. , Ibragimova T. Sh.
Published: July 2021

Photoluminescence properties of [Formula: see text] chalcogenide semiconductors have been studied under the impulse laser excitation in the range of 10–105 W/cm2 at room temperature. This study has shown that as a result of excitation, photoluminescence of [Formula: see text] is characterized by the emission in the interval of 450–575 nm with significant domination in the spectra line at 660 nm. Photoluminescence of [Formula: see text] quenches at wavelengths of 560 nm and 660 nm with constant time frames 258 ns and 326 ns, respectively. Moreover, the temperature measurements of photoluminescence were performed on the samples in the temperature range of 10–300 K.
Doing Fieldwork Without Knowing It
Authors: Rosaldo Renato
Published: July 2021

This perspective article explores the ways in which I was doing fieldwork without being aware of it from the time I began to speak and spoke English to my mother and Spanish to my father. This life experience taught me central concepts in anthropology, such as cultural relativity, and made me fascinated with the field before I was formally able to study it.
Ansa cervicalis stimulation increases pharyngeal patency in patients with obstructive sleep apnea
Authors: Kent David T. , Scott William C. , Zealear David , Schwartz Alan R.
Published: July 2021

Ansa cervicalis stimulation (ACS) is a recently proposed neurostimulation mechanism for generating caudal pharyngeal traction that may benefit patients with obstructive sleep apnea. Here, we document endoscopic findings with ACS during drug-induced sleep endoscopy and additionally detail the effects of ACS on expiratory airflow, when the pharynx is known to be most hypotonic.
Tricuspid Valve Replacement in a Patient with a Leadless Cardiac Pacemaker: Current Guidelines and Recommendations for Perioperative Management
Authors: Hames River , Hayanga J. W. Awori , Schmidt-Krings Diane , Goldhardt Timothy , Bozek John , Siddoway Donald , Schmidt Stanley , Lobban John , Hayanga Heather K.
Published: July 2021

Leadless cardiac pacemakers were developed to reduce complications associated with conventional transvenous pacemakers. While this technology is still relatively new, devices are increasingly being implanted. The perioperative management of patients with these devices has been underreported; we thus seek to add to the limited body of knowledge of perioperative management of patients with leadless cardiac pacemakers. An elderly female patient with a Micra VR transcatheter pacing system leadless cardiac pacemaker placed for tachycardia-bradycardia syndrome with intermittent complete heart block was scheduled for elective tricuspid valve replacement for severe tricuspid regurgitation. Pacemaker interrogation was performed several hours prior to the scheduled surgery based on the electrophysiologist’s availability; the device was kept in its programmed VVIR mode, and the base rate was increased from 60 to 80 beats per minute in anticipation of the upcoming surgery. Upon preoperative evaluation, the anesthesiologist asked that the electrophysiology team be placed on standby intraoperatively due to the concern that either oversensing in the setting of pacemaker dependence and/or undesirable tachycardia from rate-responsive pacing could occur. The surgeon used monopolar electrocautery for the duration of the cardiac surgery. Despite the patient having evidence of pacemaker dependence in the intensive care unit preoperatively, no electromagnetic interference leading to oversensing nor rate modulation was detected during intraoperative electrocardiographic and intraarterial invasive monitoring. Evidence-based guidelines regarding perioperative management specifically of leadless cardiac pacemakers do not exist. As these devices become more prevalent, further evaluation will be paramount to determine whether existing guidelines for perioperative management of conventional transvenous pacemakers apply.
Retrograde Balloon-Assisted Deep Enteroscopy in the Diagnosis of Metastatic Melanoma
Authors: Barge William , Albertini Mark R. , Cold Christopher , Abbott Daniel , Gopal Deepak
Published: July 2021

A 74-year-old male with a history of metastatic melanoma presents with persistently abnormal small bowel findings on PET-CT scan. The patient had persistent FDG uptake near the ileocolic junction on imaging, concerning for metastatic melanoma. Capsule endoscopy demonstrated ulcerated mucosa in the distal ileum. This area was biopsied and tattooed via retrograde double-balloon enteroscopy to confirm the diagnosis of metastatic melanoma and facilitate subsequent small bowel resection. The case illustrates a unique case of metastatic melanoma to the small bowel and the utility of capsule endoscopy and balloon-assisted enteroscopy to assist in diagnosis and management of metastatic disease.
A Case of Quadricuspid Aortic Valve Regurgitation: The Cardiac Clover
Authors: Dayco John S. , Cardozo Shaun
Published: July 2021

A quadricuspid aortic valve is a very rare congenital heart condition that can present as aortic regurgitation in the 5th and 6th decade of life. The following case report will describe a patient who presented with symptoms of severe aortic regurgitation and was found to have a quadricuspid aortic valve on echocardiography. The case will describe the clinical manifestations in which the patient presented and the subsequent diagnosis of the quadricuspid aortic valve. The rationale for the surgical approach will also be discussed, along with the patient’s clinical response.
12-Epi-Napelline Inhibits Leukemia Cell Proliferation via the PI3K/AKT Signaling Pathway In Vitro and In Vivo
Authors: Han Jia , Hou Wei , Cai Bi-qing , Zhang Fan , Tang Jian-cai
Published: July 2021

This study aimed to investigate the inhibitory effect of 12-epi-napelline on leukemia cells and its possible mechanisms. The inhibitory effects of 12-epi-napelline on K-562 and HL-60 cells were evaluated using the CCK-8 assay, cell cycle arrest and apoptosis were detected by flow cytometry, and the expression of related proteins was measured by western blot. A K-562 tumor model was established to evaluate the antitumor effect of 12-epi-napelline in vivo. A reduction in leukemia cell viability was observed after treatment with 12-epi-napelline. It was determined that the cell cycle was arrested in the G0/G1 phase, and the cell apoptosis rate was increased. Moreover, caspase-3 and Bcl-2 were downregulated, whereas cleaved caspase-3 and caspase-9 were upregulated. Further study revealed that 12-epi-napelline could suppress the expression of PI3K, AKT, p-AKT, and mTOR. Insulin-like growth factor 1 (IGF-1) attenuated 12-epi-napelline-induced apoptosis and ameliorated the repression of PI3K, AKT, p-AKT, and mTOR by 12-epi-napelline. Animal experiments clearly showed that 12-epi-napelline inhibited tumor growth. In conclusion, 12-epi-napelline restrained leukemia cell proliferation by suppressing the PI3K/AKT/mTOR pathway in vitro and in vivo.
A Combined Network Pharmacology and Molecular Docking Approach to Investigate Candidate Active Components and Multitarget Mechanisms of Hemerocallis Flowers on Antidepressant Effect
Authors: Ma Tiancheng , Sun Yu , Jiang Chang , Xiong Weilin , Yan Tingxu , Wu Bo , Jia Ying
Published: July 2021

Objective. The purpose of our research is to systematically explore the multiple mechanisms of Hemerocallis fulva Flowers (HF) on depressive disorder (DD). Methods. The components of HF were searched from the literature. The targets of components were obtained from PharmMapper. After that, Cytoscape software was used to build a component-target network. The targets of DD were collected from DisGeNET, PharmGKB, TTD, and OMIM. Protein-protein interactions (PPIs) among the DD targets were executed to screen the key targets. Afterward, the GO and KEGG pathway enrichment analysis were performed by the KOBAS database. A compound-target-KEGG pathway network was built to analyze the key compounds and targets. Finally, the potential active substances and targets were validated by molecular docking. Results. A total of 55 active compounds in HF, 646 compound-related targets, and 527 DD-related targets were identified from public databases. After treated with PPI, 219 key targets of DD were acquired. The gene enrichment analysis suggested that HF probably benefits DD patients by modulating pathways related to the nervous system, endocrine system, amino acid metabolism, and signal transduction. The network analysis showed the critical components and targets of HF on DD. Results of molecular docking increased the reliability of this study. Conclusions. It predicted and verified the pharmacological and molecular mechanism of HF against DD from a holistic perspective, which will also lay a foundation for further experimental research and rational clinical application of DD.
Abdominopelvic Mass Revealing Tuberculosis in a Young Woman
Authors: ELmajdoubi Hicham , EL Yahiyaoui Mariam , Baiss Marouane , Bouzroud Mohamed , Aboulfeth El Mehdi , Najih Mohammed , Bouchentouf Sidi Mohamed , ELkaoui Hakim , Bounaim Ahmed
Published: July 2021

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis and remains a health problem, especially in developing countries. Abdominal location represents 5 to 10% of all locations. The clinical symptoms are not very specific, and the discovery of an abdominal mass in a context of deterioration of general state may wrongly lead to the diagnosis of a tumor. Radiological explorations remain sensitive in the detection of abdominal masses but they cannot prejudge their etiology. Surgical exploration is sometimes the only recourse either for diagnostic purposes or complications, and the diagnosis can only be confirmed by bacteriological and histological examinations of the surgical specimen. We report the case of abdominopelvic mass and lymph nodes revealing tuberculosis.
Relapsed Brucellosis Manifested as Endocarditis with Vegetation on the Implantable Cardioverter Defibrillator Lead
Authors: Sawaf Bisher , Kanjo Wael , Alabbas Yasir , Hatim Ahmad , Bedardeen Unus K. , Dabdoob Wafer A.
Published: July 2021

Implantable cardioverter defibrillator lead endocarditis due to Brucella melitensis is a rare and life-threatening complication of brucellosis. Successful management requires a combination of medical treatment and device extraction. We present a case of relapsing brucellosis manifested as infective endocarditis colonizing the lead of the implantable cardioverter defibrillator with formation of vegetation on the lead. A 63-year-old male presented to the rehabilitation unit with hypotension. No other signs of infection were noted. The patient had a history of drinking unpasteurized milk since childhood and a previous episode of Brucella infective endocarditis. A transthoracic echocardiography showed an oscillating vegetation on the lead of the tip of the right atrial ICD, and the blood cultures were positive for Brucella melitensis. Surgical removal of the device was infeasible, and medical management was the only feasible option in this case.
DMTC: Optimize Energy Consumption in Dynamic Wireless Sensor Network Based on Fog Computing and Fuzzy Multiple Attribute Decision‐Making
Authors: Varmaghani Abbas , Matin Nazar Ali , Ahmadi Mohsen , Sharifi Abbas , Jafarzadeh Ghoushchi Saeid , Pourasad Yaghoub
Published: July 2021

Advances in wireless technologies and small computing devices, wireless sensor networks can be superior technology in many applications. Energy supply constraints are one of the most critical measures because they limit the operation of the sensor network; therefore, the optimal use of node energy has always been one of the biggest challenges in wireless sensor networks. Moreover, due to the limited lifespan of nodes in WSN and energy management, increasing network life is one of the most critical challenges in WSN. In this investigation, two computational distributions are presented for a dynamic wireless sensor network; in this fog‐based system, computing load was distributed using the optimistic and blind method between fog networks. The presented method with the main four steps is called Distribution‐Map‐Transfer‐Combination (DMTC) method. Also, Fuzzy Multiple Attribute Decision‐Making (Fuzzy MADM) is used for clustering and routing network based on the presented distribution methods. Results show that the optimistic method outperformed the blind one and reduced energy consumption, especially in extensive networks; however, in small WSNs, the blind scheme resulted in an energy efficiency network. Furthermore, network growth leads optimistic WSN to save higher energy in comparison with blinded ones. Based on the results of complexity analysis, the presented optimal and blind methods are improved by 28% and 48%, respectively.
Quality Evaluation of Randomized Controlled Trials of Rhodiola Species: A Systematic Review
Authors: Li Xiuzhu , Chen Weijie , Xu Yingqi , Liang Zuanji , Hu Hao , Wang Shengpeng , Wang Yitao
Published: July 2021

Background. Rhodiola is a worldwide used medicinal plant for its various medicinal functions, and the number of randomized controlled trials (RCTs) of Rhodiola is increasing in recent years. This study aims to evaluate the reporting quality and risk of bias of the current RCT reports of different Rhodiola species. Methods. Six databases including Embase, PubMed, Web of Science, the Cochrane Library, ClinicalTrial.gov, and China National Knowledge Infrastructure were searched to identify RCTs that used Rhodiola as a single intervention and were published in English or Chinese from inception to December 2020. The Consolidated Standards of Reporting Trials (CONSORT) 2010 statement was used as the checklist for assessment, and a scoring system was applied to the evaluation of RCTs. Score 0 represents no reporting or inadequate reporting, and score 1 represents adequate reporting. The risk of bias of the included studies was also assessed using the Cochrane Risk of Bias tool. Results. A total of 39 RCTs were included in this study, including 23 RCTs of Rhodiola rosea (R. rosea), 8 RCTs of Rhodiola crenulata (R. crenulata), and 8 RCTs of Rhodiola wallichiana (R. wallichiana). None of the included studies met all the CONSORT statement criteria, and the reporting quality of RCTs of the three Rhodiola species was all generally poor. Based on the risk of bias assessment, the majority of included studies were judged to have an unclear risk of bias in most domains due to inadequate reporting. Conclusions. There is inadequate reporting among the included RCTs of different Rhodiola species, and RCTs of Rhodiola with higher reporting quality and better methodological quality are needed.
Contemporary Lung Cancer Screening and the Promise of Blood-Based Biomarkers
Authors: von Itzstein Mitchell S. , Gerber David E. , Minna John D.
Published: July 2021

In this issue, the study by Dagnino and colleagues represents an important addition to the maturing field of blood-based biomarkers for lung cancer screening. Their comprehensive approach to analyzing circulating inflammatory proteins identified CDCP1 as a potential biomarker for distinguishing patients with or without lung cancer, a finding that was confirmed in a validation cohort. CDCP1 blood levels, when combined with smoking history, gave an AUC receiver operator characteristic of 0.75. Analysis of transcripts in peripheral blood cells suggested a Wnt/β-catenin signaling–based mechanism for CDCP1 in tumorigenesis providing biologic plausibility. CDCP1 now joins the ranks of other potential blood-based lung cancer screening biomarkers (including epithelial tumor marker proteins, tumor-associated miRNA, antitumor antibodies, and tumor-specific DNA methylation) that need validation in future clinical trials. Further exploration of how CDCP1 levels might be integrated into current lung cancer screening programs, including both detection of lung cancer, and evaluation of the need for invasive biopsies, as well as how CDCP1 performs in different racial populations, is warranted.

See related article by Dagnino et al., p. 3738
Artificial Intelligence in Surveillance of Barrett's Esophagus
Authors: Madabhushi Anant , Toro Paula , Willis Joseph E.
Published: July 2021

A study by Waterhouse and colleagues in a previous issue of Cancer Research describes the development and prospective validation of an artificial intelligence approach in conjunction with spectral imaging to enhance endoscopic detection of Barrett's esophagus-related neoplasia. The authors developed a novel spectral endoscope with external optics suitable for routine Barrett's esophagus surveillance with diffuse tissue reflectance to define multispectral data correlated with histopathology. A convolutional neural network was trained on the absis of the spectral signatures acquired as part of a small, prospective clinical trial to distinguish Barrett's esophagus from Barrett's esophagus neoplasia. The results from the study suggest the utility of artificial intelligence for diagnosis of Barrett's esophagus.

See related article by Waterhouse et al., Cancer Res 2021;81:3415–25
Standard-Dose Pembrolizumab Plus Alternate-Dose Ipilimumab in Advanced Melanoma: KEYNOTE-029 Cohort 1C, a Phase 2 Randomized Study of Two Dosing Schedules
Authors: Long Georgina V. , Robert Caroline , Butler Marcus O. , Couture Felix , Carlino Matteo S. , O'Day Steven , Atkinson Victoria , Cebon Jonathan S. , Brown Michael P. , Dalle Stéphane , Hill Andrew G. , Gibney Geoffrey T. , McCune Steven , Menzies Alexander M. , Niu Cuizhen , Ibrahim Nageatte , Moreno Blanca Homet , Diab Adi
Published: July 2021

Purpose:

Standard-dose pembrolizumab plus alternative-dose ipilimumab (1 mg/kg Q3W for 4 doses) were tolerable and had robust antitumor activity in advanced melanoma in cohort B of the phase 1 KEYNOTE-029 study. Cohort C evaluated standard-dose pembrolizumab with two other alternative ipilimumab regimens.

Patients and Methods:

Patients with treatment-naive unresectable stage III/IV melanoma were randomly assigned 1:1 to pembrolizumab 200 mg Q3W for ≤24 months plus ipilimumab 50 mg Q6W for 4 doses (PEM200+IPI50), or the same pembrolizumab regimen plus ipilimumab 100 mg Q12W for 4 doses (PEM200+IPI100). Primary end points were incidence of grade 3–5 treatment-related adverse events (TRAE) and objective response rate (ORR) per RECIST v1.1 by independent central review. Per protocol-defined thresholds, grade 3–5 TRAE incidence ≤26% indicated meaningful toxicity reduction and ORR ≥48% indicated no decrease in efficacy versus data reported for other PD-1 inhibitor/ipilimumab combinations.

Results:

Median follow-up on February 18, 2019, was 16.3 months in PEM200+IPI50 (N = 51) and 16.4 months in PEM200+IPI100 (N = 51). Grade 3–5 TRAEs occurred in 12 (24%) patients in PEM200+IPI50 and 20 (39%) in PEM200+IPI100. One patient in PEM200+IPI50 died from treatment-related autoimmune myocarditis. Immune-mediated AEs or infusion reactions occurred in 21 (42%) patients in PEM200+IPI50 and 28 (55%) in PEM200+IPI100. ORR was 55% in PEM200+IPI50; 61% in PEM200+IPI100.

Conclusions:

Pembrolizumab 200 mg Q3W plus ipilimumab 50 mg Q6W or 100 mg Q12W demonstrated antitumor activity above the predefined threshold; pembrolizumab plus ipilimumab 50 mg Q6W had lower incidence of grade 3–5 TRAEs than the predefined threshold, suggesting a reduction in toxicity.

See related commentary by Jameson-Lee and Luke, p. 5153
Organoid Platform in Preclinical Investigation of Personalized Immunotherapy Efficacy in Appendiceal Cancer: Feasibility Study
Authors: Forsythe Steven D. , Erali Richard A. , Sasikumar Shyama , Laney Preston , Shelkey Ethan , D'Agostino Ralph , Miller Lance D. , Shen Perry , Levine Edward A. , Soker Shay , Votanopoulos Konstantinos I.
Published: July 2021

Purpose:

Immunotherapy efficacy data on appendiceal cancer from clinical trials does not exist, due to appendiceal cancer incidence of 0.97 per 100,000. The goal of this study was to preclinically explore the application of immunotherapy in treating appendiceal cancer in a personalized organoid model.

Experimental Design:

Patient tumor organoids (PTO) were fabricated using unsorted tumor cells with and without enrichment with patient-matched immune components derived from peripheral blood leukocytes, spleen, or lymph nodes [immune-enhanced PTOs (iPTO)]. Organoids were cultured for 7 days, followed by treatment with immunotherapy (pembrolizumab, ipilimumab, nivolumab), and assessed for treatment efficacy.

Results:

Between September 2019 and May 2021, 26 patients were enrolled in the study. Successful testing was conducted in 19 of 26 (73.1%) patients, with 13 of 19 (68.4%) and 6 of 19 (31.6%) patients having low-grade appendiceal (LGA) and high-grade appendiceal (HGA) primaries, respectively. Immunotherapy response, with increased expression of granzyme B and cleaved caspase 3 and decreased expression of CK20 and ATP activity, was exhibited in 4 of 19 (21.1%) pembrolizumab-treated and 2 of 19 (10.5%) nivolumab-treated iPTOs. Post-immunotherapy cellular viability, in responding HGA organoids to pembrolizumab, decreased to less than 15% (P < 0.05). LGA iPTO treatment responses were observed in pembrolizumab and nivolumab, with an 8%–47.4% (P < 0.05) viability compared with controls. Ipilimumab showed no efficacy in the examined cohort.

Conclusions:

Immunotherapy shows measurable efficacy in appendiceal cancer organoids. Information derived from immunocompetent organoids may be applied in selecting patients for clinical trial enrollment in rare diseases where preclinical models of disease are lacking.
Abstract 1029: Dissecting the mechanism of action targeting TNFR2 with a lignd blocking anti-TNFR2 antibody that elicits potent anti-tumor activity
Authors: Zhao Xiaofeng , Liu Xiaoqing , Gao Zhenna , Wang Mengyu , Li Wenjing , Pang Ran , Yang Wenqing , Cao Zhuoxiao , Tang Renhong
Published: July 2021

Tumor necrosis factor receptor-2 (TNFR2) is a TNF receptor superfamily member and has been reported to have restricted expression in the immune system, including potent suppressive Tregs, myeloid suppressor cells and endothelial cells. The therapeutic potential of targeting TNFR2 for cancers has been previously demonstrated but with conflicting data reported by different groups. In order to gain further insight into the mechanism of action (MoA) of targeting TNFR2, we used an anti-murine TNFR2 surrogate antibody (suAb) and tested its functional activities both in vitro and in vivo. suAb was able to bind murine TNFR2 with high sub-nanomolar affinity, in the meantime, it can also block the interaction between murine TNFα and TNFR2 at both protein and cellular levels. CD4+ T cells were isolated from spleens of wildtype Balb/c mice, the co-incubation of which with murine TNFα and IL-2 rendering the significant proliferation of Treg within the CD4+ T cell population. Addition of suAb can potently reduce Treg proliferation while downregulating the expression of TNFR2 on these Tregs. Further detailed analysis by using isolated mouse FoxP3+ Treg confirmed that suAb could effectively suppress murine Treg proliferation induced by TNFα and IL-2. In vivo, suAb treatment resulted in significant tumor growth inhibition across various syngeneic tumor models (CT26, EMT6 and LL/2) as single agent or when combined with anti-mPD-1 (CT26). Tumor infiltrating lymphocytes (TILs) analysis revealed dramatic increase of the effective-memory pool of both CD4+ and CD8+ T cells together with an significantly improved CD8/Treg ratio. In addition, treatment with suAb led to decreased expression of PD-1 on tumor infiltrating CD8+ T cells suggesting a potential relieved exhaustion phenotype after the treatment. Based on this careful MoA characterization, corresponding human anti-TNFR2 lead antibodies have been identified and are currently under development as potential novel treatment for cancer patients.

Citation Format: Xiaofeng Zhao, Xiaoqing Liu, Zhenna Gao, Mengyu Wang, Wenjing Li, Ran Pang, Wenqing Yang, Zhuoxiao Cao, Renhong Tang. Dissecting the mechanism of action targeting TNFR2 with a lignd blocking anti-TNFR2 antibody that elicits potent anti-tumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1029.
Abstract 1047: Dual targeting of CK2 and MTOR as a novel therapeutic approach for high-risk B-cell acute lymphoblastic leukemia
Authors: Song Chunhua , Ge Zheng , Ding Yali , Tan Bihua , Dovat Elanora , Yang Yiping , Gowda Chandrika , Dovat Sinisa
Published: July 2021

Genomic defect and Casein Kinase II-mediated hyper-phosphorylation of Ikaros tumor suppressor cause the loss of function of Ikaros, and Ikaros dysfunction is characteristics of high-risk B-cell acute lymphoblastic leukemia (HR B-ALL) with poor prognosis. The increased activation of the PI3K/AKT/mTOR pathway is also observed in HR B-ALL and identifies mTOR as an attractive therapeutic target in HR B-ALL. Here, we report that Ikaros represses MTOR transcription and Ikaros' ability to repress MTOR in leukemia is impaired by oncogenic CK2 kinase. Treatment with the CK2 inhibitor, CX-4945, enhances Ikaros activity as a repressor of MTOR, resulting in reduced expression of MTOR in HR B-ALL. Thus, we designed a novel therapeutic approach that implements dual targeting of mTOR: direct inhibition of the mTOR protein (with rapamycin), in combination with Ikaros-mediated transcriptional repression of the MTOR gene (using the CK2 inhibitor, CX-4945). Combination treatment with rapamycin and CX-4945 shows synergistic therapeutic effects in vitro and in patient-derived xenografts from HR B-ALL. These data suggest that such therapy has the potential to reduce the death rate in HR B-ALL. The dual targeting of oncogene transcription, combined with inhibition of the corresponding oncoprotein provides a paradigm for a novel precision medicine approach for treating hematological malignancies.

Citation Format: Chunhua Song, Zheng Ge, Yali Ding, Bihua Tan, Elanora Dovat, Yiping Yang, Chandrika Gowda, Sinisa Dovat. Dual targeting of CK2 and MTOR as a novel therapeutic approach for high-risk B-cell acute lymphoblastic leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1047.
Abstract 1069: Determining the effect of bazedoxifene in combination with chemotherapy on colon cancer cells
Authors: Dmello Rhynelle S. , Thilakasiri Pathum , Duscio Belinda , Poon Ivan , Ernst Matthias , Chand Ashwini
Published: July 2021

Inflammatory cytokines, such as interleukin-6 (IL-6) and IL-11, activate the signal transducer and activator of transcription 3 (STAT3), which drives the gene transcription of cellular processes attributed as hallmarks of cancer. High levels of these cytokines, in combination with driver mutations, facilitate tumour growth and progression in preclinical models of colon cancer. Previous studies in our laboratory identified bazedoxifene (BZA), currently approved for the treatment of osteoporosis, as a small molecule inhibitor of GP130 (the receptor common to the IL-6 family of cytokines), selectively suppressing IL-6 and IL-11 signalling to reduce colon and gastric cancer growth in vivo (Thilakasiri et al. EMBO Mol Med 2019). The aim of this study is to determine the key cellular mechanisms influenced by GP130 inhibition to mediate anti-tumour effects in colon cancer cells. Additionally, we assessed the combined effects of BZA with standard of care chemotherapy drugs on cell proliferation and apoptosis. Using LIM2405 and HT29 colon cancer cells, the combined effect of BZA, fluorouracil and oxaliplatin on inducing apoptosis in LIM2405 and HT29 colon cancer cells was analysed using flow cytometry and confirmed using Western blotting and DIC microscopy. The effects of BZA was further analysed on HT29 xenograft growth and metastasis in vivo. Levels of STAT3 activation in response to IL-11/STAT3 signalling was characterised by Western blotting. Our results demonstrated that combination treatment with BZA, fluorouracil and oxaliplatin significantly increased apoptosis in LIM2405 cells and BZA significantly induced apoptosis in HT29 cells. Our data showed that BZA inhibited GP130-dependent STAT3 activity in the human colon cancer cell line LIM2405 and HT29. BZA treatment sensitized cells to chemotherapy leading to increased apoptosis. BZA treatment also reduced both primary tumour and metastatic burden in vivo demonstrating that gp130 mediated STAT3 inhibition is an attractive target for the treatment of early stage and advanced stage colon cancer.

Citation Format: Rhynelle S. Dmello, Pathum Thilakasiri, Belinda Duscio, Ivan Poon, Matthias Ernst, Ashwini Chand. Determining the effect of bazedoxifene in combination with chemotherapy on colon cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1069.
Abstract 1082: Regorafenib resistance is associated with senescence-like phenotype and EMT in colorectal cancer (CRC)
Authors: Kehagias Pashalina , Krayem Mohammad , Kindt Nadège , Najem Ahmad , Vandeputte Caroline , Journé Fabrice , Awada Ahmad , Ghanem Ghanem E. , Hendlisz Alain
Published: July 2021

The introduction of the multi-targeted tyrosine kinase inhibitor (TKI) regorafenib in clinical routine improve patients' survival with metastatic CRC who failed to respond to standard therapies, however at the cost of significant toxicities. Moreover, patients' tumor exhibit heterogeneous responses and become resistant. Its mechanism of action remains largely unknown, challenging the selection of patients through the identification of clinically useful predictive biomarkers. This study aimed to explore the effects on human CRC cell lines of short- and long-term exposure to regorafenib and investigate resistance-related mechanisms. Two representative CRC cell lines were used, HCT-116 and SW480. Short-term (3 days) and long-term exposure to IC50 values (HCT-116, 3µM and 6µM; SW480, 5µM and 6µM respectively) were intended to explore intrinsic and acquired resistance. The observation of early morphological changes (increased cell volume) in SW480 cells after regorafenib exposure led us to investigate a drug-initiated senescence-like phenotype commonly associated with resistance to treatment allowing cell death escape. These cells acquired stable senescent-like properties (slow-cycling cells, high β-galactosidase activity) and the majority of them were arrested in the G2/M cell cycle phase after long treatment exposure. A specific senescence-associated secretome (high sIL6Rα in cell supernatants) was also observed. In contrast, HCT-116 cells exposed to regorafenib presented early senescent features (high β-galactosidase activity) but developed over time an acquired resistance triggering EMT (high migration activity, upregulation of EMT-related factors) also known to contribute to TKI resistance. Moreover, the investigation of MAPK and PI3K/AKT pathways pointed to the latter as a significant player in acquired resistance of HCT-116 cells (increase of AKT phosphorylation) possibly related to the presence of a PI3KCA mutation in this cell line. Our findings provide new insights into the phenotypic plasticity of CRC cells under treatment pressure able to (1) either acquire stable senescent-like properties or (2) use early senescence state to undergo EMT. Our findings need further investigations of the major determinants involved in regorafenib-induced phenotype-switching and may help to develop new therapeutic strategies to overcome related resistances.

Citation Format: Pashalina Kehagias, Mohammad Krayem, Nadège Kindt, Ahmad Najem, Caroline Vandeputte, Fabrice Journé, Ahmad Awada, Ghanem E. Ghanem, Alain Hendlisz. Regorafenib resistance is associated with senescence-like phenotype and EMT in colorectal cancer (CRC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1082.
Abstract 1086: Pancreatic cancer musters HER2 signaling through DUSP6 to circumvent therapeutic MAPK inhibition
Authors: Bulle Ashenafi Shiferaw
Published: July 2021

Background: Oncogenic KRAS is present in almost all cases of pancreatic ductal adenocarcinoma (PDAC). The mitogen-activated protein kinase (MAPK) pathway is the most cardinal signaling cascade utilized by KRAS, but targeting RAF or MEK, the key signaling nodes within this pathway has been clincally unsuccesful, largely due to ERK reactivation. Targeting ERK kinases has recently emerged as a promising therapeutic strategy, with several ERK inhibitor combinations now ongoing in clinical trials. However, resistance to ERK inhibitionn is inevitable and should be investigated.

Aim: Identify adaptive mechanisms that can be targeted to potentiate ERK inhibitors in PDAC to facilitate development of novel therapeutic combinations.

Methods: Reverse-phase protein array (RPPA) was used in early-passage patient-derived cell lines (PDCLs) to identify potential resistance mechanisms. These were confirmed using RNA interference and overexpression in PDAC cell lines and PDCLs. The promising combinations were tested in heterotypic spheroid cultures and in 30 early-passaged PDAC patient-derived xenografts (PDXs).

Results: RPPA showed dramatic downregulation of DUSP4 and DUSP6 protein levels following MEK and ERK inhibition in PDAC cells, which coincided with phospho-activation of HER2 and HER3. Knockdown of DUSP6, but not DUSP4, was sufficient in activating HER2. Conversely, overexpression of DUSP6 curbed HER2 and ERK activation. Combined ERK inhibitor (ulixertinib) and bortezomib was able to prevent DUSP4 and DUSP6 downregulation in vitro but was ineffective in curbing in vivo tumor growth. On the other hand, ulixertinib plus PI3K or pan-HER inhibitor afatinib is effective in PDCLs in vivo. Addition of gemcitabine greatly augmented the therapeutic efficacy of these combinations and achieved prolonged tumor suppression in several PDX models.

Conclusions: Our study provided novel mechanisic insight on how PDAC cells evade MAPK inhibition via mustering HER2 signaling, as well as rational therapeutic combinations that can be readily tested in clinical trials.

Keys: DUSP6, HER2, KRAS, pancreatic ductal adenocarcinoma, therapeutic resistance

Citation Format: Ashenafi Shiferaw Bulle. Pancreatic cancer musters HER2 signaling through DUSP6 to circumvent therapeutic MAPK inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1086.
Abstract 1098: Activation of YAP1 confers ROS1 inhibitor resistance in ROS1-rearranged lung cancer
Authors: Yamazoe Masatoshi , Ozasa Hiroaki , Ohgimoto Tetsuya , Hosoya Kazutaka , Ajimizu Hitomi , Funazo Tomoko , Yasuda Yuto , Tsuji Takahiro , Yoshida Hironori , Itotani Ryo , Sakamori Yuichi , Kim Young Hak , Toyohiro Hirai
Published: July 2021

ROS1 gene arrangements occur in approximately 1% to 2% of non-small cell lung cancers. Crizotinib, the ROS1-tyrosine kinase inhibitor (TKI), has been approved for ROS1-rearranged lung cancer based on its dramatic therapeutic effect, but cures are usually not achieved. We have been focused on relationships between Yes associated protein1 (YAP1) and resistance to molecular target drugs. YAP1, a main mediator of the Hippo pathway, promotes cell proliferation and epithelial-mesenchymal transition and is associated with drug resistance in some cancer types. We previously demonstrated that YAP1 mediates survival against alectinib therapy in anaplastic lymphoma kinase-rearranged lung cancer cells. Then, in this study, we evaluated the role of YAP1 as an initial survival mechanism from lorlatinib in ROS1-rearranged lung cancer. We established a patient-derived EZR-ROS1-rearranged lung cancer cell line (KTOR71) from pleural effusion of a patient who acquired resistance to crizotinib. Subsequently, we conducted functional analysis of YAP1 involvement in drug-resistance, using this patient-derived cell line. DNA sequencing confirmed that KTOR71 cells harbored EZR-ROS1 fusion and ROS1 S1986F mutation, which is known crizotinib resistance mutation. KTOR71 cells were sensitive to lorlatinib, while resistant to crizotinib in cell viability assay (CVA). Nuclear localization of YAP1, which is an activation marker of YAP1, was assessed using immunofluorescence staining. In KTOR71 cells, YAP1 was activated after lorlatinib exposure. Then, to evaluate functional role of YAP1 in the initial survival against lorlatinib, the genetic inhibition of YAP1 in KTOR71 cells were performed using small interfering RNA (siRNA). Silencing of YAP1 by siRNA enhanced sensitivity to lorlatinib in KTOR71 cells in CVA. Furthermore, to confirm this functional role of YAP1 in vivo, we established the KTOR71 cell-derived xenograft model. The combination therapy with lorlatinib and verteporfin, an inhibitor of YAP1-TEAD interaction, sustained tumor remission significantly compared with lorlatinib monotherapy in vivo. Finally, we found that AKT was reactivated with activation of YAP1 and silencing of YAP1 by siRNA suppressed this AKT reactivation. These results suggest that YAP1 may be a potential drug target for ROS1-rearranged lung cancer.

Citation Format: Masatoshi Yamazoe, Hiroaki Ozasa, Tetsuya Ohgimoto, Kazutaka Hosoya, Hitomi Ajimizu, Tomoko Funazo, Yuto Yasuda, Takahiro Tsuji, Hironori Yoshida, Ryo Itotani, Yuichi Sakamori, Young Hak Kim, Hirai Toyohiro. Activation of YAP1 confers ROS1 inhibitor resistance in ROS1-rearranged lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1098.
Abstract 1102: Integrated approach towards defining mechanism based combinations to guide clinical development of glutaminase inhibitors
Authors: Kovacs Jeffrey J. , Spencer Nakia D. , Bristow Christopher A. , Carugo Alessandro , Giluiani Virginia , Miller Meredith A. , Harris Angela , Feng Ningping , Soth Michael L. , Le Kang , de Stanchina Elisa , Rudin Charles M. , Draetta Giulio , Yap Timothy A. , Jones Philip , Heffernan Timothy
Published: July 2021

Metabolic dysregulation is a hallmark of cancer, and recently, increasing evidence has shown a critical role for glutamine metabolism to support the bioenergetics and biosysnthetic needs of tumor and immune cells. We have previously disclosed the development of the GLS1 inhibitor, IPN60090, which is currently advancing through Phase 1 clinical studies (NCT03894540). Metabolic plasticity has been suggested to confer adaptive resistance to metabolic inhibitors and defines mechanisms that could be exploited to enhance therapeutic benefit. To address the clinical problems of innate drug resistance and adaptation, we interrogated metabolic and adaptive responses to IPN60090 in vitro and in vivo. We and others have previously identified KEAP1/NFE2L2 mutant non-small cell lung cancer models as sensitive to GLS1 inhibition. Through an integrated approach, including metabolic, transcriptomic, and proteomic analysis, we have identified molecular pathways that confer resistance to GLS1 inhibition. Nodes in these pathways that drive aquired resistance, may serve as additional patient stratification biomarkers in subsets on NSCLC or provide opportunites for further drug development. Additionally, unbiased in vivo functional genomics screening identified mulitple signaling pathways that act as critical nodes governing resistance to GLS1 inhibition. Drug combinations were tested in vitro and in vivo to identify those that are most synergistic with IPN60090. We found that PI3K/AKT/mTOR signaling is a major contributor to IPN60090 resistance, and we demonstrate that dual targeting of GLS1 and PI3K/AKT/mTOR signaling in tumors with KEAP1/NFE2L2 mutations results in synergistic anti-tumor efficacy. IPN60090, dosed in combination with inhibitors of these pathways yields regressions and off-treatment, durable responses in preclinical models of KEAP1-mutant NSCLC. Based on these data, combination strategies are being developed for Phase 1b expansion cohorts.

Citation Format: Jeffrey J. Kovacs, Nakia D. Spencer, Christopher A. Bristow, Alessandro Carugo, Virginia Giluiani, Meredith A. Miller, Angela Harris, Ningping Feng, Michael L. Soth, Kang Le, Elisa de Stanchina, Charles M. Rudin, Giulio Draetta, Timothy A. Yap, Philip Jones, Timothy Heffernan. Integrated approach towards defining mechanism based combinations to guide clinical development of glutaminase inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1102.
Abstract 1115: BET bromodomain inhibition synergizes with MEK inhibitors in uveal melanoma
Authors: Ambrosini Grazia , Musi Elgilda , Schwartz Gary K.
Published: July 2021

Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. Approximately 50% of the patients develop hepatic metastasis and the median survival rate is 12 months. Currently available drugs have shown limited clinical activity in patients with UM and there is an urgent need for new effective therapies. UM is characterized by activating mutations of G protein q/11, mutations in BAP1, and amplification of the oncogene MYC. Epigenetic dysregulation plays a critical role in UM pathogenesis, and we have demonstrated that Gnaq/11 mutant UM cells are addicted to BRD4 activity. BRD4 is a key regulator of transcriptional elongation by recruiting positive transcriptional elongation factor complex (PTEFb) to chromatin and activating RNA polymerase II-dependent transcription. While BET inhibitors have shown potent activity in pre-clinical studies, single-agent BET inhibitors exhibited limited efficacy in advanced UM clinical trials. MAPK signaling in constitutively active in Gnaq/11 mutant UM cells and previous studies have shown that the MAPK pathway mediates resistance to BET inhibition. Therefore, we sought to combine MEK and BET inhibitors in a panel of UM cells. We found synergistic activities with a variety of structurally distinct MEK inhibitors including trametinib and binimetinib in combination with BET inhibitors, such as I-BET and PLX2853. These combinations induced a greater reduction in cell viability and significant apoptosis compared to either monotherapy, in both parental and BET inhibitor-resistant cells. Western blot studies were performed to evaluate the mechanism of the combination treatment with BET and MEK inhibition and demonstrated a marked downregulation of p-ERK, MYC, Bcl-xL and upregulation of BIM with the combination therapy. Similarly, the combination treatment resulted in greater inhibition of tumor growth compared to either BET or MEK inhibition alone in a Gnaq mutant xenograft model. In conclusion, our studies suggest that co-targeting of MEK and BET proteins may be required to maximize the responses of UM cells to BET inhibitors. Clinical trials with the combination of BET and MEK inhibitors are warranted.

Citation Format: Grazia Ambrosini, Elgilda Musi, Gary K. Schwartz. BET bromodomain inhibition synergizes with MEK inhibitors in uveal melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1115.