bluuebunny/crossref_metadata_2025_split huggingface dataset:
85 Detection of human angiotensin-converting enzyme 2 receptor (hACE2R) on human cancer cell lines
Authors: Moudgil Tarsem , Fox Bernard , Hu Hong-Ming 
Published: November 2021

BackgroundSARS-CoV-2 infections have delayed administration of treatments for some patients with cancer, increasing the number of avoidable deaths. However, we hypothesized that infection of cancer cells with SARS-CoV-2 might increase the immunogenicity of those cancer cells. Here we sought to determine whether non-small cell lung cancer (NSCLC) and head and neck squamous cell cancer (HNSCC) cell lines could be a potential target of SARS-CoV-2, which binds and infects host cells via interactions between the viral spike glycoprotein and the human angiotensin-converting enzyme 2 receptor (hACE2) receptor. Through an institutional research tissue protocol, our lab has established a panel of cancer cell lines of various histologies. Here we set out to identify whether HNSCC and NSCLC cell lines expressed the hACE2R. We also investigated the expression of neuropilin-1, a molecule reported to facilitate SARS-CoV-2 cell entry.MethodsEstablished cell lines were phenotyped by flow cytometric analysis utilizing the anti-hACE2R antibody from Novus Biologicals. Cell lines were also stained with mIgG1 and anti-CD3 antibodies as negative staining controls.ResultsWe identified that three of eight NSCLC cell lines expressed the hACE2R and two of these had strong expression of neuropilin-1. Evaluation of HNSCC cell lines identified seven of seven cell lines expressed detectable levels of hACE2R but only one of seven HNSCC celllines expressed substantial levels of neuropilin-1. Preliminary evaluation of a renal cell carcinoma (RCC) cell line revealed strong staining for hACE2R.ConclusionsOur study found that a majority of HNSCC cell lines (100% n=7) and approximately a third of the NSCLC cell lines (37.5%, n=8) tested express the hACE2R. Some cell lines express both hACE2R and neuropilin-1, potentially increasing their susceptibility for infection with SARS-C0V-2. While these studies were performed with cultured cell lines that may have modulated expression of hACE2R, it is possible that in vivo these tumors express the ACE2R and could serve as a target and possible reservoir for SARS-CoV-2.AcknowledgementsSupportThe Chiles foundation, Nancy Lematta
90 Unbiased proteomic profiling leads to the discovery of a novel non-invasive blood-based protein panel with significant positive predictive value in pancreatic and colorectal cancers
Authors: Beeler Kristina , Bruderer Roland , Tognetti Marco , Sklodowski Kamil , Mueller Sebastian , Kamber Dominique , Reiter Lukas 
Published: November 2021

BackgroundMass spectrometry-based discovery proteomics has recently emerged as a high-throughput method for the proteomic profiling in biofluid samples from large clinical and population screening cohorts. Despite this progress, a significant fraction of the plasma proteome is currently not covered by state-of-the-art discovery approaches and therefore not accessible for biomarker discovery. To close this analytical gap, we present a novel workflow combining automated plasma depletion and FAIMS-DIA-MS to bridge both sensitivity and scalability. We demonstrate the applicability of this workflow to support biomarker discovery and subject stratification in precision oncology in a case-control cohort.MethodsThe plasma samples were depleted in 96-well format using an automated MARS-14 depletion system. The depleted samples were processed to tryptic peptides and analyzed using a Thermo Scientific Orbitrap Exploris 480 equipped with a FAIMS Pro device. Data processing and analysis were performed using Biognosys’ SpectroMine and Spectronaut software.ResultsUsing the unbiased discovery workflow, we investigated a cohort comprising of 180 plasma samples from healthy donors and subjects diagnosed with pancreatic, breast, prostate, colorectal and lung (NSCLC) cancer at either early or late stage of the disease. Overall, the optimized FAIMS-DIA-MS quantified 2,741 proteins across all samples and 1,849 proteins on average per sample measurement. Based on estimated plasma protein concentrations (Human Protein Atlas), quantified proteins span across 8 orders of magnitude, down to single digit pg/mL. Within this dynamic range, we could interrogate the tissue leakage proteome, interleukins and signaling proteins. Using classification algorithms, we were able to select candidates to build protein panels which provide significant positive predictive values associated with different disease stages, especially in the sub-cohorts for pancreatic and colorectal cancer.ConclusionsWe demonstrate the capabilities of a novel discovery workflow for deep, quantitative profiling of plasma samples at large scale, providing a rich proteomic resource for precision oncology.
223 Potent inhibition of PI3Kδ promotes enhanced T cell stemness and mitochondrial fitness
Authors: RIvera Guillermo Rangel , Dwyer Connor , Knochelmann Hannah , Smith Aubrey , Cole Anna , Wyatt Megan , Ware Brandon , Paulos Chrystal 
Published: November 2021

BackgroundDurable responses have been observed with adoptive T cell therapy (ACT) in chemotherapy and immunotherapy refractory patients. However, current T cell products do not always lead to therapeutic responses. Therapy failure has been attributed to terminally-differentiated T cell profiles, diminished host engraftment and poor mitochondrial metabolism. T cell activation and a switch towards glycolysis is regulated by signals downstream of the PI3K signaling pathway. We hypothesized that by inhibiting the T cell specific PI3Kδ subunit, that we would prevent overt T cell differentiation, enhance mitochondrial metabolism and improve anti-tumor immunity of adoptively transferred T cells.MethodsTo test this, we primed melanoma-specific CD8+ pmel-1 T cells in the presence of increasing concentrations of CAL-101, a PI3Kδ specific inhibitor, and infused them into B16F10 tumor-bearing mice, following non-myeloablative total body irradiation. In vitro we tested how PI3Kδ inhibition altered T cell stemness features by flow cytometry and RNA sequencing. Further, we tested the ability of PI3Kδ inhibited T cells to survive against antigen rechallenge, and assessed how mitochondrial phenotypes changed using dyes indicating mass, membrane potential and reactive oxygen species. Moreover, we tested how real time mitochondrial respiration was changed with PI3Kδ inhibition via seahorse.ResultsPotent inhibition of PI3Kδ in vitro enhanced tumor immunity and survival in a dose dependent manner. High doses of CAL-101 enriched T cells with phenotypic and transcriptional signatures of stemness. We found that CAL-101 decreased glucose uptake and increased mitochondrial mass and membrane potential while reducing mitochondrial reactive oxygen species. Potent inhibition of PI3Kδ enhanced mitochondrial respiration, however gene expression of fatty acid oxidation was not enriched, suggesting an alternative pathway involved in the enhanced spare respiratory capacity of CAL-101 treated T cells.ConclusionsThese findings indicate that blocking PI3Kδ is sufficient to mediate lasting tumor immunity of adoptively transferred T cells by preserving stemness features and improving mitochondrial fitness. Our data suggest that addition of CAL-101 to ACT expansion protocols could greatly improve responses to solid tumors by rewiring T cell stemness and promoting mitochondrial fitness.AcknowledgementsWe would like to thank all the researchers and clinicians leading the way in translating immunological principles into effective therapies, as well as the patients whose support is integral for the furthering of medicine.Ethics ApprovalAll animal procedures performed at the Medical University of South Carolina or Emory University were approved by each university’s Institutional Animal Care & Use Committee, protocol number 0488 or 201900225, respectively
224 M2 macrophage-mediated immune suppression of chimeric antigen receptor T cells via PD-L1 signaling in prostate cancer
Authors: Yamaguchi Yukiko , Gibson Jackson , Ou Kevin , Priceman Saul 
Published: November 2021

BackgroundThe immune suppressive tumor microenvironment (TME) that inhibits T cell infiltration, survival, and anti-tumor activity has posed a major challenge for developing effective immunotherapies for solid tumors. Chimeric antigen receptor T cell therapy has shown unprecedented clinical response in treating patients with hematological malignancies, and intense investigation is underway to achieve similar responses with solid tumors. Immunologically cold tumors, including prostate cancers, are often infiltrated with abundant macrophages, and infiltration of M2 macrophages correlates with metastasis and poor prognosis.MethodsTo model this in vitro, we utilized a novel co-culture system with tumor cells, prostate stem cell antigen (PSCA)-directed CAR T cells, and polarized macrophages. To investigate the TME in vivo, we took advantage of ”humanized” MISTRG mice, which are immunocompromised mice with knocked-in human genes that support human hematopoiesis and efficient tumor-infiltration of myeloid cell populations. Humanized MISTRG mice were intratibially engrafted with LAPC9 tumor cells to model bone metastatic disease.ResultsWe observed significant hampering of PSCA-CAR T cell activity in vitro with the presence of M2 macrophages, but not M1 macrophages, coinciding with a robust induction of PD-L1 in both tumor cells and macrophages. We also observed PD-L1 expression in tumor-associated macrophages infiltrating tumors following PSCA-CAR T cell therapy in the humanized mice. Anti-PD-L1 monoclonal antibodies in combination with CAR-T cell therapy altered phenotype and survival of M2 macrophages, resulting in improved anti-tumor activity of PSCA-CAR T cells in the presence of M2 macrophages.ConclusionsRecently, immune checkpoint (IC) blockade (ICB) has been utilized in combination with chimeric antigen receptor (CAR) T cell therapy, with the notion that induction of immune responses with CAR T cells may instigate checkpoint pathways in immunologically cold tumors that would otherwise not respond to ICB. This study gives insights to a mechanism by which CAR T cells and ICB work in synergy to modulate immune landscape of immunologically cold tumors, and our ongoing studies will continue to elucidate the TME-mediated immunosuppression of CAR T cell therapy.
299 Broad T antigen-specific CD8+ T cell repertoire is associated with response to PD-1 blockade in virus positive merkel cell carcinoma
Authors: Hansen Ulla , Church Candice , Bentzen Amalie , Fling Steven , Ramchurren Nirasha , Topalian Suzanne , Nghiem Paul , Hadrup Sine 
Published: November 2021

BackgroundMerkel cell carcinoma (MCC) is an aggressive human skin cancer primarily induced by Merkel Cell Polyomavirus (MCPyV) driven by expression of the oncogenic T antigens (T-Ags): Large T and Small T antigen. Checkpoint inhibition therapy blocking the programmed cell death protein-1 (PD-1) pathway has proven effective with a clinical response rate up to 58%,1 highlighting the critical role of immune surveillance for tumor control. Yet, evidence for the impact of T-Ag-specific T cells following immunotherapy is still limited.MethodsPotential CD8+ T cell epitopes derived from the T-Ags and the Viral capsid protein 1 (VP1) were predicted with netMHCpan 4.0 as 9- and 10-mer peptides with a rank score <2 for binding to 33 different HLA class I types. Peripheral blood mononuclear cells (PBMC) were obtained from 24 MCPyV+ MCC patients prior and post anti-PD-1 therapy initiation (CITN-09/Keynote-017). T cell recognition of the MCPyV-derived peptides during therapy was evaluated using the high-throughput technology of DNA barcode labeled pMHC multimers. Phenotypic characteristics of multimer-binding T cells were evaluated for selected patients.ResultsAcross all patients, 40 T-Ag-specific CD8+ T-cell populations were detected recognizing 31 epitopes restriction to 14 different HLA class I types. T-Ag-specific CD8+ T cells were detected in responders (complete and partial response, n=17) during therapy with a trend of increased number of responses observed after therapy initiation. Whereas only a single T-Ag-specific population was detectable in non-responders (stable and progressive disease, n=7). Moreover, the T cell repertoire after therapy initiation was significantly increased in the responder group compared to non-responder with the T-Ag-specific T cells showing an activated effector memory phenotype.ConclusionsThe current study indicates that the T-Ag-specific T cells are associated with clinical benefit to checkpoint inhibitor therapy. Furthermore, the broad identification of novel T-Ag-derived T cell epitopes could potentially facilitate the use of targeted T cell therapy to enhance T cell recognition and clearance of MCC in combination with checkpoint inhibition.AcknowledgementsSupported by the Cancer Immunotherapy Trials Network (CITN)ReferenceNghiem P, Bhatia S, Lipson E. Three-year survival, correlates and salvage therapies in patients receiving first-line pembrolizumab for advanced Merkel cell carcinoma. J Immunother cancer 2021;9:e002478.
322 Melanoma-intrinsic hypoxia-inducible factor-1α results in diminished T cell accumulation within the tumor microenvironment
Authors: Higgs Emily , Gajewski Thomas , Trujillo Jonathan 
Published: November 2021

BackgroundThe hypoxia-inducible factor (HIF) system, consisting of the transcription factors HIF-1α and HIF-2α, mediates cellular adaptation to hypoxia, and can promote cancer progression, invasion, and metastasis. HIF pathway activation in the tumor microenvironment has been implicated in cancer immune evasion; however, a direct causal role for tumor cell-intrinsic HIF-1α and HIF-2α activation in mediating T cell exclusion and cancer cell resistance to immune checkpoint inhibitor therapy has not been demonstrated.MethodsWe performed gene expression analysis of melanoma tumors in the Cancer Genome Atlas (TCGA) data set to determine whether increased HIF-1α pathway activation correlated with reduced T cell-based inflammation. The magnitude of HIF-1α pathway activation across melanoma samples was determined by applying a quantitative scoring system on the expression of a melanocyte-specific hypoxia-induced, HIF-1α-target gene signature consisting of 81 genes. The Pearson correlation test was used to compare the HIF-1α activation score and our 160-gene T-cell-inflamed gene signature. To determine the impact of cancer cell-intrinsic HIF-1α or HIF-2α activation on the endogenous anti-tumor T cell response, we developed an inducible autochthonous mouse melanoma model driven by BRAFV600E expression and PTEN-deletion, with or without inducible expression of either a stabilized variant of HIF-1α or HIF-2α. These murine tumor models are being used to determine the impact of cancer cell-intrinsic HIF-1α or HIF-2α activation on tumor sensitivity to anti-PD-1/PD-L1 and anti-CTLA-4 treatment.ResultsGene expression analysis of human melanomas in the TCGA demonstrated a statistically significant inverse correlation between the HIF-1α activation score and T cell-inflammation score. Braf/PTEN murine melanomas with and without stabilized HIF-1α expression developed with comparable tumor onset and growth kinetics. Multiparameter immunofluorescence staining of melanoma tissue revealed a significant decrease in tumor-infiltrating T cells within Braf/PTEN melanoma tumors expressing stabilized HIF-1α compared to control Braf/PTEN melanomas.ConclusionsOur data demonstrate that tumor-cell intrinsic HIF-1α activation leads to diminished T cell accumulation within the tumor microenvironment, which has implications for cancer immunotherapy. The mechanism of this effect is being elucidated. These novel murine models will help elucidate the roles of cancer cell-intrinsic HIF-1α and HIF-2α activation in modulating the anti-tumor T cell response, providing mechanistic insight that will inform the evaluation of novel selective HIF inhibitors, which are showing promising anti-tumor activity in clinical trials in patients with advanced solid tumors.
382 The synthetic long peptide cancer vaccine UV1 in combination with ipilimumab induces a CD4+ Th1 anti-hTERT immune response and an inflammatory tumor microenvironment in patients with melanoma
Authors: Ellingsen Espen , Kerzeli Iliana , Anzar Irantzu , Mensali Nadia , Clancy Trevor , Mangsbo Sara , Aamdal Steinar , Aamdal Elin , Guren Tormod , Inderberg-Suso Else-Marit , Hovig Eivind , Gaudernack Gustav 
Published: November 2021

BackgroundCheckpoint inhibitors (CPIs) have revolutionized the treatment of malignant melanoma. Although melanoma patients may experience deep and durable clinical responses to CPI treatment, the majority develop disease progression requiring additional therapy. Combining CPIs with therapeutic cancer vaccines may augment the anti-tumor immune response and thus improve clinical outcomes. UV1 is a therapeutic cancer vaccine based on synthetic antigens derived from the tumor-associated antigen telomerase reverse transcriptase (hTERT). hTERT is activated in 85–90% of all cancers and leads to replication of telomeric DNA, an essential mechanism for increased proliferation, immortality, and invasiveness of cancer cells. In this phase I/IIa study combining UV1 with ipilimumab in patients with melanoma (NCT02275416), we hypothesized that UV1 and ipilimumab might provide synergy in the expansion of vaccine-induced T cells. Furthermore, an augmented CD4+ Th1 immune response targeting a shared tumor antigen may increase T cell infiltration in the tumor microenvironment, promoting immune-mediated cancer cell death.MethodsThe immune response dynamics were assessed by longitudinal immunomonitoring for up to 5 years using a standard proliferation assay. The phenotype and functionality of vaccine-induced T cells were assessed by patient-derived T cell cloning and subsequent in vitro characterization by flow cytometry, peptide stimulation, and cytokine release assays. On available biopsies harvested at baseline and 12 weeks after treatment initiation, the tumor microenvironment was assessed based on whole-exome sequencing, RNA sequencing, and immunohistochemistry.ResultsTwelve patients with metastatic melanoma were enrolled in the study and received up to 9 vaccine doses over a 20-week period. A persistent (up to 5 years) vaccine-induced immune responses were demonstrated in 91% of evaluable patients (10/11). Vaccine-specific T cell clones were polyfunctional CD4+ Th1 cells, producing both IFN-γ and TNF-α upon in vitro peptide stimulation. Differential gene expression analysis and immunohistochemical characterization of biopsies at baseline and 12 weeks showed induction of an inflammatory ”hot” tumor microenvironment in clinical responders with available paired biopsies.ConclusionsUV1 in combination with ipilimumab leads to robust and long-lasting CD4+ Th1 anti-hTERT immune responses sculpting the local tumor microenvironment.Trial RegistrationNCT02275416Ethics ApprovalThe study was approved by Regional Ethical Committee South East (ID number 25 165). The patients provided their written informed consent to participate in the study.
Virtue, Sanctity, and Charity of the Royal Women of Fez
Authors: Boloix-Gallardo Bárbara 
Published: November 2021

Abstract
The history of the Banū Marīn of the kingdom of Fez (seventh–ninth/thirteenth–fifteenth centuries) cannot and should not be reconstructed without a gender perspective which gives the women of this dynasty a place within its historic discourse. They played a key role in the political and religious legitimacy of the rulers, as reflected by the Banū Marīn historiography which, mirroring the idiosyncrasy of medieval Berber societies, afforded its women a visible space. However, as it was always subject to the clear interests of male political-religious legitimacy, this space for visibilisation is worthy of analysis. This study examines the different profoundly religious behaviour and capabilities which the chronicles assigned to different royal Merinid women, as related in the various anecdotes transmitted in their pages. Additional analysis is carried out on how these model characterisations aimed to increase the aura of spirituality of the amīrs, influenced by their close relationship and everyday contact with these women.
Church, State, and Pluralistic Society
Authors: Nikolajsen Jeppe Bach 
Published: November 2021

Abstract
This article demonstrates that Lutheran teaching on the two regiments can be drawn in different directions and how it was drawn in a particular direction for centuries so that it could provide a theoretical framework for mono-confessional Lutheran societies. It argues that the Lutheran two regiments theory can be developed along a different path, regaining some emphases in Luther’s early reflections: it can thereby contribute to an improved understanding of the role not only of the church but also of the state. While a number of Lutheran theologians believe that Lutheran teaching on the two regiments is particularly difficult to apply today, with some even contending that it should simply be abandoned, this article argues that Lutheran teaching on the two regiments could present a potential for a relevant understanding of the relationship between church, state, and society, and its ethical implications in a contemporary pluralistic society.
What We Talk About When We Talk About Early Release in International Criminal Law: The Sui Generis Nature of the Reduction of Sentence Under the Rome Statute
Authors: Fernández-Pacheco Estrada Cristina 
Published: November 2021

Abstract
Early release has been regularly granted by the ad hoc tribunals for over 20 years. However, it could be argued that some issues still remain contentious. In fact, in May 2020, the Practice Direction ruling early release in the Mechanism of the International Criminal Tribunals was amended. This was intended to clarify key matters, such as the time needed to be served before early release, the possibility of imposing conditions upon those released, and the unappealable character of the resulting decision. At a glance, it could be argued that the International Criminal Court is better equipped to confront the many challenges posed by early release. This is owing to its detailed regulation, which may consequently lead to a more reasoned and solid case law. After comparatively examining ten features key to the application of early release, however, this paper argues that the ultimate problem lies within the nature generally conferred to early release in the Rome Statute.
‘Jugendbewegung’: Gerhard Scholem as a Figure of the German Avant-Garde
Authors: Engel Amir 
Published: November 2021

Abstract
The purpose of this essay is to examine one of Gershom Scholem’s most obscure undertakings, a proto-Dadaist and antiwar text that he published, together with a few friends, during the first year of World War One. As is shown, this project has drawn heavily from the work of one of the leading avant-garde poets, later among the founding members of the Dada movement in Zurich, Hugo Ball. Discussed side by side, the works of Gershom Scholem and Hugo Ball offer a unique view onto the anti-war sentiment and the need for experimental language in an attempt to express a disdain so profound and fundamental that words and sentences seem unable to capture.