bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intracellular retention of TCRαβ/CD3 and CD7 to generate novel allogeneic CAR-T cells targeting CD7 for treating T-cell malignancies.
Authors: Li Jun , Liu Kaichun , Fu Jianmin , Chen Shanshan , Zhang Yinhang , Xu Guanggui 
Published: June 2024

e14531 Background: The treatment for patients with T-ALL/LBL who relapse after hematopoietic stem cell transplantation (HSCT) are currently restricted. CD7 is highly expressed on the surface of T-ALL/LBL T cells and CAR-T cell therapy provides a promising avenue towards remission. However, CD7-targeting immunotherapies may be compromised by T-cell fratricide. Furthermore, in addition to the long production time, high cost, and high risk of production failure, the biggest challenges of autologous CAR-T cell therapy are how to isolate a sufficient number of healthy T cells from malignant T cells. Thus, allogeneic CAR-T cells can overcome these hurdles. Methods: A novel non-gene-editing allogeneic CAR-T platform was developed on the base of intracellular retention of membrane proteins, and named ThisCART (TCR and/or HLA-I intracellular sequestered). CD7-directed ThisCART cells are readily produced with a single lentiviral vector, encoding both a CD7-targeting CAR, an anti-CD7and anti-CD3 single chain antibody with the KDEL peptide fused to its C-termini. The surface expression of CAR, TCRαβ/CD3 and CD7 was measured by flow cytometer. Specific tumor cytotoxicity was evaluated both in vitro and in vivo. Furthermore, the safety of donor derived ThisCART cells were tested in a xenogeneic GvHD model. Results: The manufacturing platform of CD7 ThisCART was able to achieve over 130-folds of ex vivo CAR-T expansion in all batches, with the purity of products (CAR-positive/CD7-negative/TCRαβ-negative) above 99%. In preclinical models, CD7 ThisCART did not induce GvHD, and exhibited superior antitumor function compared to Control T. CD7 ThisCART cells were readily produced to a quantity and quality supporting clinical investigation. Conclusions: CD7 ThisCART cells based on intracellular retention of TCRαβ/CD3 complex and CD7 demonstrated superior CD7-dependent cytotoxicity and no xenogeneic GvHD in murine models. With streamlined single-vector-based production and potent cytotoxicity and favored preclinical safety profile, CD7 ThisCART warrant clinical investigation.
Predicting immunotherapy toxicity through single-cell sequencing of peripheral immune cells: A pilot study.
Authors: Lim Jennifer Wen Ying , Chin Venessa T. , Keith Patricia , McCloy Rachael , Neavin Drew , Xue Angli , Kaczorowski Dominik , Spenceley Ellie , Arora Himanshi , Joshua Anthony M. , Powell Joseph 
Published: June 2024

e14538 Background: Immune checkpoint inhibitors are widely used in oncology. However, immune-related adverse events (iRAEs) are idiosyncratic and can be severe. The overall incidence is 27%, with severe iRAEs occurring in 6% of patients. We hypothesised that distinct immune cell phenotypes (gene expression) correlate with immune-related toxicity from checkpoint inhibitor immunotherapy. Single-cell RNA sequencing (scRNA-seq) is a powerful tool that can assay the gene expression of thousands of individual cells. RPS26 expression mediates CD4/CD8 T cell activation through a known trans-expressive quantitative trait locus at rs1131017 and is linked with autoimmunity. FOS and JUN encode for c-Fos and c-Jun, which dimerise to create the activator protein-1 complex, a well-established transcriptional factor of the immune system. c-Fos is an immunoregulator known to suppress pro-inflammatory cytokines, suggesting that c-FOS deficiency plays a crucial role in developing autoimmunity. We aimed to analyse the transcriptomic differences of peripheral immune cells between patients who experienced severe checkpoint inhibitor therapy iRAE and those who did not. Methods: We identified a pilot cohort of eight patients who developed grade three or above toxicity due to immune checkpoint inhibitors and eight controls who were matched for age, cancer and treatment type. Peripheral blood mononuclear cells were collected prospectively at baseline and two subsequent treatment cycles. Cells were captured on the 10x Chromium platform, and libraries were sequenced to 30,000 reads per cell. Samples were demultiplexed, and the data was analysed with the Seurat package on R statistical software (version 4.3.0). Results: Five patients had baseline samples only, two patients had two samples from two cycles, and one patient had samples from three cycles. After quality control filtering, we identified 36,288 cells from 16 individuals and 24 samples. Immune cells were annotated with the Azimuth package into 29 subtypes. We identified differences in cell proportions of immune cell subsets in the toxicity vs control group, including B naive cells (5.57% vs 3.71% p=0.01) and reduced cytotoxic CD4 T lymphocyte cells (0.66% vs 2.26%, p=0.005). Differential gene expression analysis identified a significantly high expression of ribosomal genes, particularly RPS26, in multiple cell types at baseline (p<0.01) and reduced expression of FOS and JUN-related genes in CD14 and CD16 monocytes, CD4 and CD8 T cells in the toxicity cohort (p<0.01). Conclusions: Overexpression of ribosomal genes and reduced expression of FOS/ JUN genes amongst PBMCs at baseline are potentially predictive biomarkers of severe checkpoint inhibitor immunotherapy toxicity. Validation of these findings in a larger cohort is in progress.
Analysis of long-lasting tumor-specific antibodies from murine tumors treated with Aliya pulsed electric fields.
Authors: Pastori Chiara , Nafie Ebtesam , Neal Robert Evans 
Published: June 2024

e14543 Background: Pulsed electric field (PEF) tissue destruction has demonstrated meaningful systemic immune responses in mice that are superior to thermal ablation, and synergize effectively with systemic chemotherapy and checkpoint blockade (CPB) immunotherapy. Immune cell populations are increased by PEF, including tumor specific activated CD8 T-cells and plasma B-cells. Antitumor immunity can include the presence of circulating anti-cancer antibodies. However, the presence and role of circulating tumor specific antibodies induced by PEF have not been explored. Serological antibodies are often detected with the ELISA assay, where tumor-associated antigens are coated to a microtiter plate to attract the corresponding serum antibodies. Here, we quantify IgG tumor specific antibodies after PEF using an in-house ELISA. Methods: Mice bearing immune-warm (EMT6) and immune-cold (4T1) orthotopic murine breast tumors were treated with a biphasic PEF dose comparable to Aliya titrated for mouse tumors. To detect tumor specific circulating IgG1 immunoglobulin in serum, an ELISA plate was coated with EMT6 and 4T1 protein lysate, as well as with a known 4T1-expressed tumor-specific antigen (gp70). Serum draws from the treated, untreated, and naïve mice were exposed to the coated wells for 1 hour. Bound IgG were detected by anti-mouse IgG (HRP tagged) antibody added to the wells and incubated 1 hour. A TMB substrate was added and color development was allowed for 15 minutes. The optical density was quantified using a microtiter plate reader. Results: ELISA quantification of serum IgG at multiple timepoints show that 4T1 tumor-specific IgG antibodies were increased 14.3- and 2.6-fold by day 20 in the PEF-treated and untreated group survivors relative to Naïve mice, respectively, (OD=0.86 and 0.15 v. 0.06). Furthermore, IgG antibodies specific to the gp70 antigen of the 4T1 cell line were increased by 44.9- and 2.7-fold in the PEF-treated and untreated groups relative to naïve mice (OD=3.14 and 0.19 v. 0.07). In the EMT6 model, at 3 months post-treatment, only the PEF-ablated group mice survived, and showed 15.5-fold higher antibodies against EMT6 tumor antigens compared to naive mice (OD=0.62 v. 0.04). Conclusions: The specific form of PEF in this study generated tumor-specific IgG long-lived antibodies in mice for immune-warm and immune-cold tumor models, corroborating previous findings of increased murine B-cell and plasma cell populations. This effect has not been well-reported for other ablation technologies. Increases were evident by Day 20 post-PEF and persisted at least 3 months. These murine data suggest the PEF used may invoke potent, long-lived immune responses and anti-tumor immune surveillance, offering a potential approach for in situ tumor vaccination to improve metastatic disease outcomes. Future studies should determine response durability and its role in attaining distant tumor clearance.
Effect of SLAMF8 on the phagocytosis of tumor-associated macrophages in colorectal cancer.
Authors: Zhang Qun , Liu Zhihao , Han Xingzhi , Qian Xiaoping 
Published: June 2024

e14571 Background: Phagocytosis plays an essential role in innate immunity and is key in bridging innate immunity and adaptive immunity. Phagocytosis checkpoints promote tumor immune escape by inhibiting phagocytosis by tumor-associated macrophages (TAMs). SLAMF8, which is mainly expressed on the surface of TAMs, may be involved in regulating the tumor immune microenvironment. However, its regulation of the phagocytosis of TAMs remains unclear. The present study aims to investigate the effect of SLAMF8 on the phagocytosis of TAMs and to provide a new target for tumor immunotherapy. Methods: The dataset including 458 colorectal cancer (CRC) tissues from the TCGA database was used to analyze the correlation between the expression of SLAMF8 and several classical phagocytosis checkpoint molecules, and the above correlation was further verified by the RNA-sequencing data of 20 CRC tissues in our center. In vitro, the effects of SLAMF8 expression level on tumor cell proliferation and macrophage phagocytosis were also evaluated. RNA sequencing was used to analyze the effect of SLAMF8 on the expression of genes related to promoting macrophage phagocytosis. The effect of SLAMF8 on the expression levels of proteins related to FcR-mediated phagocytosis was detected by flow cytometry. Results: The results from the TCGA dataset and our validation dataset both showed that the SLAMF8 expression was significantly positively correlated with the expression levels of classical phagocytosis checkpoint molecules such as SIGLEC11, SIGLEC10, SIGLEC9, CD274and LILRB1. In vitro, functional tests showed that the phagocytosis of CT26 CRC cells by macrophages was significantly enhanced after SLAMF8 knockdown. RNA sequencing results suggested that the expression levels of molecules related to FcR-mediated phagocytosis were significantly upregulated after the decreased expression of SLAMF8.Flow cytometry analysis confirmed that SLAMF8 inhibited the expression levels of FcR-mediated phagocytosis related proteins such as CD16/32, CD64, and CD11b. Conclusions: SLAMF8 may release the “don’t-eat me” signal to inhibit FcR-mediated phagocytosis of tumor cells by macrophage. Further studies are needed to investigate the underlying mechanism of SLAMF8 on the regulation of macrophage phagocytosis and to test its possibility as a new immunotherapy target.
A new modality of anti-cancer drug: Ferritin-drug conjugates.
Authors: Lee Jeewon , Lee Bo Ram , Seol Yong Hwan , Hwang Hyun Ju 
Published: June 2024

e14590 Background: Success criteria of ADCs is the target overexpression in tumor, i.e. in general ~ 14-fold in tumor vs. normal cells, which is the reason why it is hardly possible to develop clinically potent ADCs against several cancers with low target expression, which remains a major challenge yet, although fam-trastuzumab deruxtecan-nxki (Enhertu; AZN) and sacituzumab govitecan-hziy (Trodelvy; Gilead) recently exhibit an exceptionally enhanced efficacy for TNBC. Methods: Payloads were conjugated to a human apoferritin-derived, cancer-targeting scaffold (a self-assembled 24-mer protein with a diameter of ~ 12 nm, showing a high avidity for cancer cells) via a particular payload-releasing linker based on thiol-maleimide/imidazole-metal ion/EDC-NHS chemistry. Results: A new modality of anti-cancer drug - ferritin-drug conjugates (FDCs) - was developed, demonstrating a notably enhanced performance in cancer treatment as compared to current, clinically available anti-cancer drugs such as fam-trastuzumab deruxtecan-nxki, which is due to the important advantages such as far lowered EC50, high cancer-targeting efficiency, excellent structural stability (even in DMSO (up to 30%)), high loading capacity (high drug to ferritin ratio (DFR > 20)) and consistent homogeneity in drug conjugation, no immune side effects, reduced drug resistance, and/or reduced off-target toxicity. Conclusions: FDCs are believed to open up a novel, attractive clinical route for both potent and safe therapy of a broad range of cancers.
Strong single-agent and PD-1/PD-L1 combination antitumor activity of antisense oligonucleotides suppressing Neuropilin 1 expression.
Authors: Petropoulos Konstantin , Maaske André , Kirchhammer Nicole , Festag Julia , Buchi Mélanie , Michel Sven , Zippelius Alfred , Klar Richard , Jaschinski Frank 
Published: June 2024

e14592 Background: While the introduction of immune checkpoint inhibitors (ICIs) is considered as a breakthrough in cancer therapy, large proportions of patients still do not respond or develop resistance, indicating a need for new targets and therapies to provide long-term therapeutic benefits. Neuropilin-1 (NRP1) is a membrane-bound multi domain protein expressed by different cell types in the tumor microenvironment. NRP1 promotes cell migration, survival, and angiogenesis via distinct domains. Moreover, it mediates immunosuppression by stabilization of regulatory T cells, attraction of immunosuppressive myeloid cells into the tumor microenvironment, and exhaustion of effector T cells. Therapeutic benefit is expected to be superior by simultaneous suppression of all functions and consequently all domains, which can be optimally achieved by suppression of its expression, for example mediated by antisense oligonucleotides (ASOs), instead of e.g. domain specific antibodies. Methods: Using our in-house bioinformatics and screening systems, we have identified potent and specific locked nucleic acid (LNA) modified ASOs specifically targeting NRP1. Mice bearing established syngeneic MC38 or EMT6 tumors were treated by intraperitoneal injection of unformulated ASO either as monotherapy or in combination with ICIs. Tumor growth and survival were determined as well as target engagement. Results: After systemic administration of different NRP1-specific ASOs we observed a potent downregulation of NRP1 expression in different cell types in tumors, including myeloid cells, T cells and non-immune cells as well as a strong reduction of soluble NRP1 plasma levels. Suppression of NRP1 correlated with both a delayed tumor growth or even complete tumor regression and prolonged survival as a monotherapy which was further improved in combination with ICIs. Particularly in an EMT6 breast cancer model, combination of NRP1 ASOs with PD-1 antibody resulted in tumor rejection and long-term survival in most animals. Rechallenge of long-term surviving mice with EMT6 tumor cells did not result in tumor growth. Conclusions: We could show that ASO-mediated suppression of NRP1 expression has strong potential as a novel therapeutic strategy, especially in combination with ICIs. Additional studies to further elucidate the mechanisms underlying the observed anti-tumor efficacy as well as IND-enabling activities are underway to progress the program towards clinical testing.
Pembrolizumab-induced myocarditis: A systematic review of case reports.
Authors: Hasan Walaa , Eysa Athar , Chauhan Darshita , El-daraysah Murad , Ezzelden Ahmed 
Published: June 2024

e14605 Background: Immune checkpoint inhibitors, such as pembrolizumab, have revolutionized cancer therapy but are associated with immune-related adverse events (irAEs), including myocarditis, which can have serious clinical consequences. This systematic review aims to comprehensively analyze case reports to better understand pembrolizumab-induced myocarditis. Methods: Using keywords of "pembrolizumab" or "Keytruda" and "myocarditis" in the PubMed/Medline, ScienceDirect and clinicaltrials.gov databases and manual searches on Google Scholar, we reviewed all cases reported up to the year 2023. In total, 296 studies received primary screening. Duplicated articles and those meeting the exclusion criteria (not myocarditis, not human, not English, trials done in vitro, combination immunotherapies) were excluded. Review articles, retrospective studies, systematic reviews, and meta-analyses were also excluded. Finally, 45 studies matching the inclusion criteria were included (total of 46 patients). Data on patient demographics, clinical presentation, diagnostic criteria, management strategies, and outcomes were extracted and analyzed. Results: In the included 46 patients, the mean age was 65.5 years, with a slight male predominance (56.5%). Predominant cancer types included lung cancer (30.4%), thymoma (15.2%), melanoma (13.0%), and genitourinary cancers (13.0%). 45.7% of patients developed myocarditis after their first cycle of Pembrolizumab while 41.3% developed myocarditis after two cycles. 47.8% of patients presented with dyspnea, 23.9% with fatigue, 10.9% with chest pain, while 13% of the patients were asymptomatic and were subsequently diagnosed with myocarditis while presenting with other immune-related adverse events. Troponin levels were assessed in 45 individuals, with only 2.2% (one patient) having normal levels. ECG changes included ST segment elevation in 21.7%, conduction abnormalities (RBBB 15.2%, LBBB 6.5%, AV block 26.1%, and one unspecified conduction abnormality) in 50%, and arrhythmias in 37% of cases. Treatment included steroids for all, with varying outcomes. Four patients each received one of the following treatments: batacept, ruxolitinib, alemtuzumab, and methotrexate. All four of these patients showed improvement. Two patients received infliximab (4.3%) and both of them died. Outcomes revealed 34.8% mortality, 63.0% improvement, and 2.2% deterioration, including one sudden cardiac death 35 days post-discharge. Conclusions: Pembrolizumab-induced myocarditis is a rare but potentially life-threatening irAE. This systematic review provides a comprehensive overview of the clinical characteristics, diagnostic challenges, treatment approaches, and outcomes of this condition. Further research is needed to elucidate the underlying mechanisms and identify risk factors for pembrolizumab-induced myocarditis, enabling early detection and prevention strategies.
Immune checkpoint inhibitors and immune mediated diabetes mellitus: Meta-review of case reports.
Authors: Van Loo Aiden Michael , Patel Manthan , Mohammed Adil , Kim Jun Hwan , Silvers Samantha , Ebrahim Rami , Bolton Tiara , Martin Chad , Li Hunter , Muslehuddin Zainab , Desai Rupak , Reddy S. Sethu , Haddad Nicholas 
Published: June 2024

e14606 Background: Use of Immune checkpoint inhibitors (ICIs) is an increasingly popular antitumor immunotherapy targeting immunoreceptors on T lymphocytes, enhancing the anti-neoplastic immune response. ICIs are becoming first line therapy options for non-small cell lung cancer and gastric adenocarcinoma and can be used as second line for variety of others. Various endocrinologic adverse events have been reported with use of ICIs. Of these, new onset Type-1 diabetes mellitus (T1DM) is of particular interest given its significant morbidity and noted frequency of 1-5% of patients who receive ICIs. There is limited consolidated information about symptomatic presentations, risk factors, and demographics of patients developing Type 1 DM in relation to ICI use. We performed a systematic review of published case reports to identify commonalities between patient disease presentation, ICI use, and development of immune-mediated diabetes mellitus. Methods: A comprehensive systemic literature search from PubMed Web Science, CINAHL, and Google Scholars was performed identifying case reports showing development of new onset diabetes mellitus in patients using immune checkpoint inhibitors. Descriptive analysis was performed to better identify characteristics and presentation of patients. Results: Based on our parameters, 130 reports with 149 cases were included. Primary tumors among the patients comprised lung cancer (34.9%) and melanoma (31.5%). Amongst all the cases, about 88% of the patients were treated with anti-PD-1 (program cell death) receptor, with nivolumab alone (34.9%) and pembrolizumab alone (26.2%) being the most common agents. Cases were more commonly found to be males (61.7%) and of Asian (36.6%) or North American (35.6%) background. Patient ages ranged from 14-95 with a mean of 62.3. Presentation most commonly started after the third cycle (13.4%) and was noted to occur between 1-50 doses. Diabetic Ketoacidosis was diagnosed in 69.8% of individuals, and the average presenting blood glucose was 611 mg/dl with an HbA1c of 8.2%. Autoantibody data was largely missing; however, anti-GAD antibodies were noted to be present in 29.5% of cases. 34.2% of patients had coexisting endocrinopathies with thyroiditis being the most common, occurring in 20% of patients. 1.3% of patients did not survive the initial hospitalizations, with the vast majority (95.3%) remaining on insulin and a select few (2%) were eventually able to discontinue insulin. Conclusions: New onset diabetes is a rare but a potential life-threatening metabolic urgency. Thus, it is paramount that clinicians maintain close monitor of blood glucose and endocrine function when a cancer patient who recently started ICIs gets admitted for hyperglycemia. Further research is warranted to understand onset of diabetes mellitus at molecular level.