bluuebunny/crossref_metadata_2025_split huggingface dataset:
An engineered Trop2/CD3 bispecific antibody for treatment of gastric, pancreatic, and colorectal cancers.
Authors: O Diana , Chow Kronos , Iyer Deepak , Wong Monarca , Wong Tony K , Wong Dennis A , Luk John Moon 
Published: June 2023

e14521 Background: Given its over-expression and poor prognosis in a variety of solid tumors, trophoblast cell surface antigen 2 (Trop2) is regarded as a potential therapeutic target. Trop2-directed antibody-drug conjugates have been developed for treatment of solid tumors; however, their clinical benefits can be limited by the associated undesirable effects which is likely attributed to the on-target off-tumor effect arising from the low basal Trop2 expression on the surface of multiple normal epithelial tissues. The present study aims to develop a humanized Trop2/CD3 bispecific T-cell engager antibody that can promote immunological synapse formation with tumor cells overexpressing Trop2, redirecting T cell-mediated killing specifically to cancers with reduced risk of on-target off-tumor effects. Methods: A Trop2/CD3 bispecific antibody was generated by linking a humanized anti-Trop2 monoclonal antibody with a CD3-binding scFv. To tune the affinity of the bispecific antibody for tumor target, in silico paratope prediction was firstly done using ProABC-2 (Ambrosetti F et al., 2020), following by alanine substitution of the identified paratope residues individually. A total of 6 CDR variants were generated with their binding to Trop2 examined using biolayer interferometry. Selected candidates were further characterized for their binding to Trop2-positive cancer cells and activation of T cells upon target engagement. Redirection of T cell-mediated killing on cancer cells expressing high Trop2 level was also studied in vitro. Results: Substitution of a complementarity-determining residue (CDR) on the heavy chain of anti-Trop2 antibody with an alanine significantly reduced binding affinity to Trop2. This CDR variant was selected for further functional characterization. Apart from weaker binding to Trop2-positive tumor cells, the variant also demonstrated a slower cytokine induction, a less acute T-cell activation, and a cytotoxic effect which was more restricted to Trop2-high cancer cells. Conclusions: Our new Trop2/CD3 bispecific antibody demonstrates potent anti-tumor activity, while at the same time shows a reduced risk of on-target off-tumor toxicity. The drug candidate warrants further investigations including toxicity study in monkeys. [Table: see text]
Tolerability and response of palliative radiotherapy in patients receiving tumor-infiltrating lymphocyte therapy.
Authors: Rogers Max , McKenzie Grant , Rattani Abbas , Yusuf Mehran , Chesney Jason Alan , Dunlap Neal E. 
Published: June 2023

e14527 Background: Tumor-infiltrating lymphocyte (TIL) therapy is an emerging therapeutic option for patients with metastatic tumors having progressed on multiple lines of systemic therapy. It is unknown how TIL therapy impacts the tolerability and effectiveness of palliative radiotherapy (RT). We herein present a case series of patients having received both TIL therapy and palliative RT along with the associated local responses and toxicity profiles. Methods: A single institution retrospective database review identified patients treated with TIL therapy following progression on ≥2 standard treatment(s) for metastatic disease. Patients were eligible for inclusion if they received palliative radiotherapy treatment within 1 year of TIL delivery as pre- or post-TIL therapy. Patient demographic data was collected and descriptive statistics reported. Radiographic imaging response was assigned in accordance with Response Evaluation Criteria in Solid Tumors (RECIST). Toxicities were graded in accordance with Common Terminology Criteria for Adverse Events (CTCAE) Version 5.0. Results: Between 2017–2021, a total of 10 patients receiving TIL therapy treated across 12 courses of palliative RT were included for analysis. 8 of 10 patients (80%) had a primary diagnosis of malignant melanoma and 2 (20%) had non-small cell lung cancer. RT was administered to 7 patients pre- and 3 patients post-TIL therapy. Within the pre-TIL cohort, 2 patients received RT between TIL harvest and infusion. Patients received an average total dose of 26 Gy (range, 8-45 Gy). Median patient follow-up after RT was 6 months. Of the patients who received RT prior to TIL therapy, 6 of 9 sites demonstrated at-least stable disease at the site of RT versus 1 of 3 sites receiving post-TIL RT. Across the cohort, 2 patients experienced Grade 1 toxicities associated with RT treatment. No Grade 2+ events were reported. Conclusions: Palliative RT appears to be safe and tolerable when delivered both prior to and post-TIL therapy. RT may be an option for bridging therapy between TIL harvest and infusion. Local response rates to RT were greater in patients receiving RT prior to TIL infusion. Further investigations are warranted to determine the optimal dose, fractionation, and timing of RT when integrating it into future TIL therapy protocols. [Table: see text]
Weight loss (WL) as a predictive factor for immune oncology (IO).
Authors: Fugere Brad , Chen Jim Zhongning , Mazahreh Farah , Fugere Tyler , Mazahreh Liyan , Safar A. Mazin 
Published: June 2023

e14528 Background: WL is an adverse prognostic factor/symptom reported in ~40% cancer (ca) patients (pts) at diagnosis agnostic to site or histologic grade. The mechanism of WL in ca, however, remains elusive. The role of energetic waste in ca WL is doubtful (it neglects likely dietary compensation). On the other hand, lymphocytic activation, a clonal, energetically demanding process is accompanied with anorexia limiting caloric intake which leads frequently to WL if protracted over months as seen in lymphoma (B-symptom) or indecisive immune engagement (Eng) with tuberculosis. WL of > 5% of body weight, was an adverse prognosticator and exclusion criterion in non-small cell lung cancer (NSCLC) chemotherapy (CT) trials, likely due to CT or steroids' abrogation of the putative, limited immune ca containment. We hypothesized that WL in ca reflects months of immune Eng, ineffective as it may be at containing the tumor, but nevertheless representing immune Eng with the ca. Such cases stand the best chance for IO benefit. Notably, IO impact in ca mirrors the 40% of WL in ca pts. Methods: Tumor registry retrieved data from all stage 4 cancers treated with monotherapy IO in our institution. We dichotomized pts to beneficiaries (B) or non-beneficiaries (NB). Bs survive longer than median survival for the respective stage/line of therapy in each ca derived from published trials. To account for missing WL data prior to IO, we reasoned that weight gain of ≥10 lbs (reliably charted over 1 year from IO infusions) is acceptable surrogate for WL and met our criteria. Results: WL in our cohort (n = 53) of Melanoma, NSCLC, renal cell carcinoma, and bladder ca was 43% of B; 10% in NB (ratio > 4:1) rejecting the null hypothesis (p < 0.05). PDL-1, Tumor Mutation Burden, both inconclusive in identifying durable benefit, were limited in our set precluding further analysis. Conclusions: Our results support, though do not prove, that WL identifies the appropriate IO pts. Improved outcome despite a historically-adverse prognosticator (WL); the fact that IO was the only ca therapy used, both support our interpretation. We hope to stimulate others to examine independent larger datasets to confirm this notion and to allow comprehensive subset analyses. For WL to be utilized properly the relevant clinical context is crucial, accounting for GI obstructing ca (esophageal ca), enzymatic anomalies (pancreatic ca), volume fluctuations (cirrhosis), catabolism (prolonged hospitalization), or dental issues, to name a few. Additionally, factors such as tumor bulk, perhaps discerned from the T of TNM staging, deserve examination since more vigorous immune activation against larger targets could produce more impact on appetite and caloric intake. Finally, available clinical trial datasets, not tumor registries, might provide a more crisp, reliable source as we continue this investigation. WL can, with confirmatory work, be clinically incorporated as a predictive factor for IO benefit.
"Armed" CEA CAR-T with a SIRPγ-CD28 chimeric co-receptor to exhibit the enhanced antitumor activity in preclinical study of colorectal cancer.
Authors: Zhao Yongchun , Zhu Xiuxiu , Shen Junjie , Xu Yanmin , Yang Zhi , Qi Yanan , Chen Jun , Hong Juan , Wang Linling , Qin Lei , Liu Ge , Huang Xia , Hu Jingwen , Zhao Wenxu , Chen Yunfan , Jiang Xiaorui , Jiao Liurong , Chen Nan , Qian Cheng 
Published: June 2023

e14530 Background: Nowadays, CAR-T cell therapy still faces limited therapeutic efficacy for solid tumors. SIRPγ is known as a ligand for a well-known immune checkpoint molecule CD47. It is distinguished from other members of its family in that it has a very short intracytoplasmic tail and is incapable of transducing signals on its own. Of note, the role of SIRPγ still largely unknown to date. Preliminary data indicated SIRPγ plays a key role in T-cell transendothelial migration and promotes antigen-specific T-cell proliferation. We thus designed a novel SIRPγ-CD28 chimeric receptor comprising the extracellular part of SIRPγ, the transmembrane and intracellular domains of CD28 and used as the co-receptor for a CEA-targeting CAR. Here, we report on the activity of this "armed" CAR-T with the SIRPγ-CD28 co-receptor in colorectal tumor (CRC) and its anti-tumor efficacy in mice xenograft model. Methods: In vitro luciferase-based cytotoxicity assay were used by coculturing CAR-T and DLD1-CEA CRC cells to assess the activities of CAR-T w/wo the co-receptor. The incubation supernatants were collected for detecting the release of cytokines. In vivo evaluation of the anti-tumor efficacy of CAR-T with the SIRPγ-CD28 co-receptor versus control T cell, and control CAR-T was performed in NOG mice CRC xenograft model. DLD1-CEA cells expressing luciferase reporter were s.c. implanted and T cells were i.v. injected. Anti-tumor efficacy was assessed by in vivo system. Results: CAR-T with the SIRPγ-CD28 co-receptor exhibited specific cytotoxicity only to CEA+ CRC cells, and the efficacies of CAR-T w/wo the co-receptor were comparable (88.6% vs 87.8%, p > 0.05). However, cytokines were significantly higher in the CAR-T group with the co-receptor. The levels of IFNγ, IL2 and TNFα in groups of CAR-T w/wo the co-receptor were 72,793.33 vs 19,013.33 pg/ml (p < 0.05), 1834.67 vs 25.15 pg/ml (p < 0.05) and 79.65 vs 0 pg/ml (p < 0.05), respectively. In mice CRC xenografts model, the tumors were totally eliminated in the CAR-T group with the co-receptor at 21 days and lasted until the end of experiment, while the tumor growth was suppressed in the control CAR-T group at first and out-of-control in the end (Table). Conclusions: These preclinical studies demonstrated the potential of SIRPγ-CD28 co-receptor as a novel T cellular activation signals for CAR-T. The "armed" CAR-T exhibits a fabulous anti-tumor efficacy in CRC and has great potential for applications in other solid tumors. [Table: see text]
Innovative platforms for development of TCR-T cell therapies against novel tumor targets: Triple negative breast cancer.
Authors: Abbott Rachel Judith Mary , Hammerl Dora , Kortleve Dian , van Brakel Mandy , Wijers Rebecca , Roelofs Daphne , Badoux Madelon , Debets Reno 
Published: June 2023

e14536 Background: Adoptive T cells therapies have shown great potential over the past decade, particularly with the use of CAR-T cells to treat haematological tumors. However, their effect in the treatment of solid cancers is limited, due to a lack of relevant therapeutic targets and suppressive mechanisms of the tumor microenvironment. To counteract these challenges, we have developed Pan Cancer T (PCT) platforms to yield novel TCR-T cell therapies against untapped (intracellular) tumor targets exclusively expressed in 10 hard-to-treat solid tumors (including breast, melanoma, colorectal, glioma, lung cancer). Furthermore, we are developing engineering strategies aiming to improve the longevity of T cells within the inhibitory solid tumor microenvironment. Methods: The platforms we have developed to enable generation of genetically engineered TCR-T cells to recognize tumor-specific (intracellular) antigens include: 1) PCT Target Discovery (identifying and selecting safe, immunogenic tumor targets by combining advanced in silico and laboratory techniques); 2) PCT TCR Discovery (identifying, selecting, and validating potent TCR-T cells binding with high specificity and avidity); 3) PCT Next Gen Engineering (identifying drivers of immune evasion and implementing TCR-T cell-based gene-engineering approaches to overcome suppression and/or improve durability). Results: Using our PCT Target Discovery platform, we have validated a set of 30 unique tumor targets which are safe, immunogenic and highly shared. Our first showcase target is PCT-1, which is of particular interest in triple negative breast cancer (TNBC), where it is expressed by >85% of patients, with high homogeneity in >80% of all tumor cells. PCT-1 is also expressed in other tumor types, including >90% melanoma and >45% multiple myeloma patients. The PCT TCR Discovery platform has enabled development of a highly specific and active TCR against PCT-1, which outperformed standard of care TNBC treatments (ie, cis-platinum and sacituzumab govitecan) in advanced preclinical efficacy models, including in vitro killing of patient-derived 3D organoids and an in vivo model in NSG mice. Conclusions: The PCT platforms have enabled development of a promising TCR-T against a novel target antigen expressed in TNBC, an aggressive form of breast cancer that affects younger women with limited treatment options. A portfolio of 29 additional untapped targets has also been identified, providing potential to generate TCR-based therapies for treatment against a much larger range of solid tumors. Furthermore, we are developing next generation technologies, aiming to enhance the durability of responses within the hostile tumor microenvironment, which should be broadly applicable to TCR-T therapies.
Development of allogenic nonviral RNA-based CAR-NK therapy targeting CDH17 in relapsed/refractory gastrointestinal cancer.
Authors: Roy Jaydeep , Lin Vivian , Rahmani Mehran , Chow Kronos , Lin James Chieh-Liang , Chang Alarng , Luk John Moon , Chan Anthony Chun Fung 
Published: June 2023

e14539 Background: Gastrointestinal (GI) cancers are commonly fatal after metastasis. The incidence and mortality of GI tumors are high, and the benefits of existing therapies are limited. Cadherin-17 (CDH17) is highly prevalent and specific to GI cancers which has potential for the development of novel treatments for GI cancers. Cell-based immunotherapy using genetically engineered autologous Chimeric Antigen Receptors T (CAR-T) cells has been clinically efficacious for treating hematological malignancies. However, its manufacturing is extremely expensive and lengthy with ~10% failure production rate. Equipping NK cells with CAR may represent a safer and equally effective approach for use in solid tumors as an alternative to autologous CAR-T-cell therapy. Nonviral RNA-based expression of CAR vector in NK cells will provide immediate and multiple accessibility to cancer patients, lower risk of abnormal genetic alteration mediated by viral integration, and potentially limit side effects. This paper demonstrates the development of RNA-CAR-NK targeting CDH17-positive GI cancers. Methods: Ex vivo expanded human peripheral blood NK (Magicell-NK, Medigen, Taiwan) cells were used in this study. To generate anti-CDH17 (C7) CAR-NK cells, mRNA containing a novel CDH17-specific single-chain variable fragment and the 4-1BB costimulatory domain was electroporated to Magicell-NK cell. Flow cytometry analysis was performed to evaluate the cell viability and the expression level of CAR and cytotoxic surface markers of the CAR-NK cells. CDH17 specific cytolytic activity against AGS gastric cancer cell line and the concentration of cytokines released by CAR-NK cells were measured by in vitro cytotoxicity assay and cytokine release assay, respectively. Results: Our results demonstrated efficient electroporation with high-level C7 CAR expression ( > 80%) and optimal CAR-NK cell viability (~60%). Interestingly, 12 h co-culture of C7 CAR-NK cells with AGS gastric cancer cells at 1:1 and 2:1 effector/target (E:T) ratio showed a significantly enhanced antitumor response of C7 CAR-NK cells on AGS-CDH17+ cells but not on AGS-CDH17- cells, indicating CDH17 target-specific cytolytic activity. Moreover, we observed an elevated level of cytokines such as INF-γ and TNF-α released by C7 CAR-NK cells only in co-culture with AGS-CDH17+ cells, confirming the mechanisms behind the CAR-mediated cytotoxicity. Furthermore, these results were supported by an increased level of surface CD107a or LAMP-1 expression in C7 CAR-NK cells during the 12 h of co-culture with AGS-CDH17+ cells at 1:1 and 2:1 E:T ratio. Conclusions: Our findings suggest that a non-viral based CDH17 CAR-NK cell therapy could be a potential allogeneic, off-the-shelf CAR-NK therapy candidate in GI cancer and support the rationale of further investigating it in vivo and clinical trials.
Prognostic impact of baseline circulating extracellular vesicles (BCEVs) in the setting of patients with advanced non-small cell lung cancer (ANSCLC) eligible for first line treatment with osimertinib.
Authors: Gristina Valerio , Barraco Nadia , Taverna Simona , Manno Mauro , Raccosta Samuele , Lamantia Maria , Carreca Anna Paola , Incorvaia Lorena , Badalamenti Giuseppe , Galvano Antonio , Bazan Viviana , Russo Antonio , Carreca Ignazio Ugo 
Published: June 2023

e14553 Background: In the liquid biopsy era, the interrogation of extracellular vesicles (EVs) content might be complementary for cancer prognostication and response assessment. In this explorative study, we aimed to describe whether the serial characterization of BCEVs plasma tumor-derived EVs could longitudinally reflect response and resistance to available first-line treatments, investigating the potential to predict clinical outcomes in patients with advanced NSCLC (aNSCLC). Methods: From 2020/02 to 2022/05, 27 treatment-naïve patients with aNSCLC (stageIIIB/IV), eligible for first-line treatment with an EGFRTKI (Osimertinib) or ICIs (e.g. Pembrolizumab ± platinum‐based chemotherapy) based on the predictive molecular pathology, were enrolled. Blood samples (∼5 mL) were collected into K2EDTA tubes early in the morning with fasting condition. We isolated EVs from 2 ml of plasma using exo-Easy Maxi Kit (Qiagen-USA). EVs’s size and distribution was determine by use of Dynamic Light Scattering method (DLS). Besides a Bradford assay (cfEV) for the indirect quantification of EV amount based on EVs proteins content was performed. According to the radiologic response, EV analyses were evaluated in patients with available plasma samples from baseline (T0) to the first radiologic evaluation of disease within 12 weeks (T1). Results: From 2020/02 to 2022/05, 27treatment-naïve patients with aNSCLC (stageIIIB/IV), eligible for first-line treatment with an EGFRTKI (Osimertinib) or ICIs (e.g. Pembrolizumab ± platinum‐based chemotherapy) based on the predictive molecular pathology, were enrolled. Blood samples (∼5 mL) were collected into K2EDTA tubes early in the morning with fasting condition. We isolated EVs from 2 ml of plasma using exo-Easy Maxi Kit (Qiagen-USA). EVs’s size and distribution was determine by use of Dynamic Light Scattering method (DLS). Besides a Bradford assay (cfEV) for the indirect quantification of EV amount based on EVs proteins content was performed. According to the radiologic response, EV analyses were evaluated in patients with available plasma samples from baseline (T0) to the first radiologic evaluation of disease within 12 weeks (T1). Conclusions: This real-world study confirmed the in vivo feasibility of isolating and characterizing BCEVs via DLS and Bradford assay from the plasma of patients with ANSCLC. The quantitative changes of the plasma EVs correlated with survival in the real-time monitoring of such first-line patients, mostly in the oncogene-addicted population receiving osimertinib.
The clinical impact of soluble natural killer cell group 2-member D (NKG2D) receptor ligands on tumor tumorigenicity and anti-tumor immunity.
Authors: Serritella Anthony V. , Saenz-Lopez Pablo , Dhar Payal , Liu Sizhe , Wu Jennifer 
Published: June 2023

e14557 Background: NKG2D is an activating receptor expressed on all human natural killer (NK) cells and CD8 T-cells. It is widely accepted that human tumors shed soluble NKG2D ligands, such as soluble major histocompatibility complex-I chain-related molecule (sMIC), to evade immune activation. Harnessing the NKG2D/NKG2D-ligand axis has emerged as a promising avenue for immunotherapy. However, controversy exists over whether soluble NKG2D ligands are immunosuppressive or immunostimulatory. Methods: Cell lines for melanoma (B16F10), Lewis lung carcinoma (LLC1), and prostate cancer (TRAMP-C2, T-C2) were used as parental cell lines in this study. Derivative cell lines over-expressing the soluble human NKG2D ligand sMICB (B16F10-sMICB, TC2-sMICB and LLC-sMICB) were created. We inoculated these 3 syngeneic transplantable tumor models and their respective parental lines into mice and compared tumor incidence/growth over time. We also compared surface markers representative of cell stemness (SCA-1, CD166 and CD133) in mouse cancer cells between the sMICB-expressing tumor cells and their respective parental cell lines. We then characterized the tumor immune microenvironment by flow cytometry. Results: B16F10-sMICB mice demonstrated a significant delay in tumor development compared to control mice (23 vs. 6 days, p < 0.05). Over longer follow-up, rapid tumorigenesis still occurred in the experimental group, with a 50% tumor incidence by day 26. By contrast, the TRAMP-C2-sMICB mice demonstrated higher tumorigenic ability (p < 0.05). LLC-sMICB mice demonstrated no difference in tumorigenicity. Time to tumor onset was more strongly associated with expression of stemness-like markers rather than sMIC. Once tumors were established, they grew significantly more aggressively in the TRAMP-C2-sMICB, LLC-sMICB and B16F10-sMICB lines vs. their parental counterparts. sMICB expressing tumors contained significantly lower percentages of NK cells and CD8 T-cells in tumor infiltrates than their respective parental lines. NK cells and CD8 T Cells from sMICB-expressing tumors had an impaired capacity to produce interferon gamma. Conclusions: Expression of a soluble NKG2D ligand had minimal impact on tumor establishment but facilitated more aggressive growth of established tumors due to immune suppression. Soluble NKG2D ligands imprint negative anti-tumor immunity onto established tumors. Our data differentiated the impact of NKG2D ligands on tumor onset vs. tumor progression and reinforced the translational concept that soluble NKG2D ligands are strong therapeutic targets for future study.