bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4861: Incidence rates of bladder and kidney cancers among U.S. military servicemen: Comparison with the rates in the general U.S. population
Authors: Bytnar Julie A. , McGlynn Katherine A. , Kern Sean Q. , Shriver Craig D. , Zhu Kangmin
Published: March 2024

Background: The military population may differ from the general population in factors related to the incidence of bladder and kidney cancers. However, incidence rates of these cancers have not been systematically compared between the two populations. This study compared incidence rates of bladder and kidney cancers and their trends between active-duty servicemen and men in the general U.S. population.

Methods: Data were obtained from the Department of Defense’s (DoD) Automated Central Tumor Registry (ACTUR) and the National Cancer Institute’s Surveillance, Epidemiology and End Results (SEER) database. Patients were active-duty servicemen in ACTUR and men in SEER who were diagnosed with malignant bladder and kidney cancers and 18-59 years at diagnosis from 1990-2013. Age-adjusted rates, incidence rate ratios (IRR), and their 95% confidence intervals (95% CI) were compared between the two populations by age, race, and cancer stage. Trends in the incidence were also compared.

Results: Incidence rates were lower in ACTUR than SEER for bladder cancer overall (IRR=0.55, 95% CI=0.48-0.62) and by age, race, or tumor stage. However, for 50-59-year-old men, the rates did not differ between the populations. Trend analysis showed bladder cancer incidence decreased more in ACTUR than SEER over the study period. Kidney cancer incidence rates were marginally lower overall (IRR=0.92, 95% CI=0.84-1.00) and significantly lower for 30-49-year-old Black men in the military than the general population. For kidney cancer, the trends were similar between the two populations with increased incidence from 1999 to 2013.

Conclusion: Lower bladder and kidney cancer incidence in ACTUR, notably in younger individuals, may be primarily associated with healthier status and better health care. The lack of differences in bladder or kidney cancer incidence among 50-59-year-old men between the populations might result from more cumulative military-related exposures in this age group, which effects were offset by healthier status and better medical care. Disclaimer: The contents of this publication are the sole responsibility of the authors and do not necessarily reflect the views, opinions or policies of NCI, USUHS, HJF, the DoD or the Departments of the Army, Navy or Air Force. Mention of trade names, commercial products or organizations does not imply endorsement by the US Government.

Citation Format: Julie A. Bytnar, Katherine A. McGlynn, Sean Q. Kern, Craig D. Shriver, Kangmin Zhu. Incidence rates of bladder and kidney cancers among U.S. military servicemen: Comparison with the rates in the general U.S. population [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4861.
Abstract 4965: Detection of status of cancer in radiology notes using artificial intelligence
Authors: Arya Ankur , Niederhausern Andrew , Bahadur Nadia S. , Shah Neil J. , Nichols Chelsea , Chatterjee Avijit , Philip John
Published: March 2024

Introductory Statement: The goal is to use an AI model to replicate the human curator’s response on cancer status in radiology notes of ~20,000 cancer patients.

Introduction: MSKCC currently has ~100,000 patients with genomic testing (IMPACT) and continues to accrue more. Clinicians use this genomic data for research but lack clinical structured data to analyze alongside the genomic data. We use a vendor called VASTA Global to manually curate unstructured paragraph text. Depending on the data model, we found that a patient’s full cancer history can take up to 8 hours to curate. Therefore, we want to implement AI to automate the curation of this data to save time and cost.

We hope to achieve a faster curation process that allows us to accomplish more than 1 patient a day to catch up to the 100,000 MSK-IMPACT cohort. To decrease the average curation time per patient, we investigated the use of an NLP model to replicate manual curation of the PRISSMM™ ontology field for change in cancer status from radiology reports. Manual curation of radiology reports is time intensive due to curators understanding their cancer status and the volume of scans patients undergo.

Methods:A pre-trained Bidirectional Encoder Representations from Transformers (BERT) model is fine-tuned using training data using GPUs in IBM Cloud Pak for Data (CPD) platform. The hyperparameters were adjusted using accuracy and F1 metric of evaluation data. Using one vs. rest approach the model is evaluated on held out test data with results shown in table 1.Table 1 includes class 1-5: Progressing/worsening/enlarging, stable/no change, improving/responding, not stated/indeterminate or mixed.

Summary:The weighted AUCROC ~ 0.97, F1 ~ 0.85 and accuracy ~ 93%. These metric scores improve by choosing notes with higher class probabilities only.

Conclusion:Due to the above 0.9 calculated accuracy this NLP model is successful in replicating the curated results of cancer status and the next steps will be to run this model on a new cohort of patients not yet curated.

Cancer Status Metrics Class Prevalence Precision Recall F1 Score AUCROC Accuracy 1 38.9% 88.7% 87.0% 87.9% 96.5% 90.7% 2 24.7% 89.9% 88.2% 89.1% 98.3% 94.6% 3 15.9% 87.6% 90.7% 89.1% 98.7% 96.5% 4 15.7% 77.0% 72.6% 74.7% 94.4% 92.3% 5 4.8% 68.1% 74.4% 71.1% 97.9% 97.1% Weighted Average 85% 86% 85% 97% 93%

Citation Format: Ankur Arya, Andrew Niederhausern, Nadia S. Bahadur, Neil J. Shah, Chelsea Nichols, Avijit Chatterjee, John Philip. Detection of status of cancer in radiology notes using artificial intelligence [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4965.
Abstract 497: Plant viruses against cancer
Authors: Steinmetz Nicole F. , Chung Young Hun , Moreno-Gonzalez Miguel , Zhao Zhongchao
Published: March 2024

Immunotherapy has significantly transformed the landscape of cancer treatment; however, concerns about safety, efficacy, and long-term outcomes across various tumor types persist. We have developed an intratumoral immunotherapy strategy with an engineered plant virus nanotechnology and demonstrated efficacy in tumor mouse models and canine cancer patients. This method is designed to elicit a systemic (abscopal effect), safe, and enduring anti-tumor immune response that is not limited by tumor type. The power of immunotherapy lies in synergistic combination treatments and toward this goal, we are engineering the next-generation plant virus nanotechnologies. We demonstrate synergistic combination with chemotherapy and immunization strategies the hone in on tumor-associated antigens and key players of inflammation. The combinatorial immunotherapy strategies prevent recurrence post-surgery in mice and canine patients. We will discuss the engineering design principles leading toward synergetic combination immunotherapies to treat and prevent cancer; efficacy studies and underlying mechanism is elucidated through immunomics studies.

Selected references: Steinmetz N.F. et al (2023) Viral nanoparticle vaccines against S100A9 reduce lung tumor seeding and metastasis. Proc. Natl. Acad. Sci. U.S.A. 120 (43) e2221859120. Wang C. and Steinmetz N.F. (2020) A Combination of Cowpea mosaic virus and Immune Checkpoint Therapy Synergistically Improves Therapeutic Efficacy in Three Tumor Models. Advanced Functional Materials, 2002299.

Citation Format: Nicole F. Steinmetz, Young Hun Chung, Miguel Moreno-Gonzalez, Zhongchao Zhao. Plant viruses against cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 497.
Abstract 4973: Impact of technology on quality of thoracic multidisciplinary cancer conferences
Authors: Ibekwe Opuruiche , Gaudioso Carmelo , Attwood Kristopher M. , Pokharel Saraswati , Roche Charles L. , Nwogu Chukwumere E.
Published: March 2024

Background: Complex cancer cases require discussion at Multidisciplinary Cancer Conferences (MCCs) to determine the best management. The study aim was to assess the impact of navify® tumor board solution (NTBS) technology on the quality of information presented, case discussions, and care plans at thoracic MCCs.

Methods: Between September 2020 and February 2022, using a before-after study design, we prospectively collected data through direct observation of thoracic MCCs at an academic cancer center. The NTBS technology was introduced in March of 2021. Additionally, we reviewed medical records to assess several factors including the rate of change in care plans as decided by the MCC, compliance of all care plans with national guidelines, and concordance of treatment received with MCC recommendations. Observational data was collected via a validated tool “Metric for the Observation of Decision-Making". We used SAS v9.4 (Cary, NC) for statistical analyses.

Results: We identified 151 and 166 thoracic cancer cases before and after implementation of NTBS, respectively. The overall quality of case presentation and discussion, represented by a mean composite score (the summation of individual variables scored on a 1-5 scale, poor to excellent), increased from 56.8 to 82.0 (p<.001). This improvement was also observed across multiple sub-components of the composite score including case history (3.4 vs 4.6), reason for presentation (3.2 vs 4.6), comorbidities/performance status/psychosocial information (2.8 vs 4.2), patient views (1.2 vs 3.3), radiology and pathology presentations (3.5 vs 4.8 and 3.2 vs 4.4, respectively), and contribution from surgical, medical, and radiation oncologists (3.9 vs 4.3, 3.3 vs 3.8, and 3.5 vs 3.8, respectively), as well as the moderation by the MCC chair (4.9 vs 5.0). All these results were statistically significant with p<.001. There was no statistically significant difference between the two cohorts in rate of change in care plans by the MCC, care plan compliance with national guidelines, and concordance of treatment received with MCC recommendations.

Conclusion: Technology improves the quality of information and discussion at tumor boards. However, this study did not demonstrate an impact on compliance with practice guidelines. Practitioners should explore the available tumor board technology platforms to optimize the conduct of MCCs in their respective institutions.

Citation Format: Opuruiche Ibekwe, Carmelo Gaudioso, Kristopher M. Attwood, Saraswati Pokharel, Charles L. Roche, Chukwumere E. Nwogu. Impact of technology on quality of thoracic multidisciplinary cancer conferences [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4973.
Abstract 4978: Genomic characterization of metastatic pancreatic cancer according to tumor sample sites
Authors: Goksu Suleyman Y. , Ozer Muhammet , Goksu Busra Bacik , Sanford Nina N. , Jones Amy , Verma Nilesh , Kazmi Syed
Published: March 2024

Background: Our research aimed to evaluate and compare the genetic variations in the primary and metastatic sites of patients with metastatic pancreatic cancer.

Methods: In this study, we selected individuals with metastatic pancreatic cancer from the data provided by the American Association for Cancer Research Project GENIE (version 14.1). The comparison of genetic mutations between primary and metastatic tumor sites was conducted using the Fisher exact test, augmented with the Benjamini-Hochberg adjustment technique.

Results: In a study of 3,108 patients with metastatic pancreatic cancer, 38.9% had metastatic site tumor samples, while 61.1% had samples from the primary tumor. The most frequent mutations were KRAS (89.4%), TP53 (72.9%), SMAD4 (21.2%), and CDKN2A (17.4%). Metastatic samples showed higher rates of CDKN2A, CDKN2B, and MTAP mutations compared to primary site samples (40.1% vs. 29.1%; 20.3% vs. 10.4%; and 20.8% vs. 8.9%, respectively, all with p<0.001 and q<0.001). In patients with young-onset pancreatic cancer (younger than 50 years), there was no notable difference in gene mutations between the two sample groups.

Conclusion: Our findings corroborate the notion that identifying the precise genes that contribute to the development of metastatic pancreatic cancer is vital for gaining a more comprehensive understanding of the biological mechanisms. Moreover, genomic profiling could be a key in pinpointing potential biomarkers for diagnosis and therapy.

Citation Format: Suleyman Y. Goksu, Muhammet Ozer, Busra Bacik Goksu, Nina N. Sanford, Amy Jones, Nilesh Verma, Syed Kazmi. Genomic characterization of metastatic pancreatic cancer according to tumor sample sites [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4978.
Abstract 4985: Pivotal roles for cancer cell-intrinsic mPGES-1 and autocrine EP4 signaling in suppressing anti-tumor immunity
Authors: Markosyan Nune , Kim Ilkyu , Arora Charu , Cheng Noah , Schechter Emma , Tobias John , Stanger Ben , Vonderheide Robert
Published: March 2024

Tumor cell-derived prostaglandin E2 (PGE2) is a tumor cell-intrinsic factor that supports immunosuppression in the tumor microenvironment (TME). Using a genetically engineered mouse model of pancreatic ductal adenocarcinoma, we demonstrate that deleting the terminal PGE2 synthesis enzyme or disrupting autocrine PGE2 signaling through cognate EP4 receptors on tumor cells reverses the T cell-low, myeloid cell-rich TME to activate T cell immunity and suppress tumor growth. Knockout (KO) of Ptges (the gene encoding PGE2 synthesis enzyme mPGES-1) or the EP4 receptor gene (Ptger4) in KPCY (KrasG12D/P53R172H/Yfp/CrePd) pancreatic tumor cells abolished growth of implanted tumors in a T cell-dependent manner (KO vs control p<0.0001 for both). Blockade of the EP4 receptor in combination with immunotherapy, but not immunotherapy alone, induced complete tumor regressions and immunological memory. Mechanistically, Ptges and Ptger4 KO tumor cells exhibited altered T and myeloid cell attractant chemokines, became more susceptible to TNF-α killing, and exhibited reduced adenosine synthesis. In hosts treated with an adenosine deaminase inhibitor, Ptger4 KO tumor cells accumulated adenosine and gave rise to tumors. These studies reveal an unexpected role for autocrine mPGES1-PGE2-EP4 signaling axis in pancreatic cancer cells, nominating mPGES-1 inhibition and EP4 blockade as immune-sensitizing approaches in cancer therapy.

Citation Format: Nune Markosyan, Ilkyu Kim, Charu Arora, Noah Cheng, Emma Schechter, John Tobias, Ben Stanger, Robert Vonderheide. Pivotal roles for cancer cell-intrinsic mPGES-1 and autocrine EP4 signaling in suppressing anti-tumor immunity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4985.
Abstract 50: ALK inhibitors increase ALK expression and sensitize neuroblastoma cells to ALK.CAR-T cells
Authors: Bergaggio Elisa , Tai Wei-Tien , Aroldi Andrea , Mecca Carmen , Landoni Elisa , Nüesch Manuel , Mota Ines , Metovic Jasna , Molinaro Luca , Ma Leyuan , Alvarado Diego , Ambrogio Chiara , Voena Claudia , Blasco Rafael B. , Li Tongqing , Klein Daryl , Irvine Darrell J. , Papotti Mauro , Savolodo Barbara , Dotti Gianpietro , Chiarle Roberto
Published: March 2024

Neuroblastoma is the most common extracranial solid tumor of childhood, with limited success in treating refractory or relapsed cases using current therapies. Chimeric antigen receptor (CAR) T cell therapy targeting GD2 has shown promise in neuroblastoma treatment, but relapses are associated with loss of antigen expression. The selection of the best tumor antigen is critical for the therapeutic success of CAR-T cells in hematologic malignancies and solid tumors. The Anaplastic Lymphoma Kinase (ALK) receptor is expressed by most neuroblastomas while virtually absent in most normal tissues. ALK is an oncogenic driver in neuroblastoma and ALK inhibitors show promising clinical activity. All these features make ALK an attractive target for CAR-T cell therapy. We developed ALK.CAR-T cells that show potent efficacy in monotherapy against neuroblastoma with high ALK expression without toxicity. For neuroblastoma with low ALK expression, combination with ALK inhibitors specifically potentiates ALK.CAR-T cells but not GD2.CAR-T cells. In neuroblastoma cell lines and in a patient-derived xenograft (PDX), the combination of ALK inhibitors and ALK.CAR-T cells significantly reduced tumor growth and extended mice survival. Mechanistically, ALK inhibitors impair tumor growth and upregulate the expression of ALK, thereby facilitating the activity of ALK.CAR-T cells against neuroblastoma. Thus, while neither ALK inhibitors nor ALK.CAR-T cells will likely be sufficient as monotherapy in neuroblastoma with low ALK density, their combination specifically enhances therapeutic efficacy. These findings provide insights into the potential of ALK.CAR-T cells as a novel therapeutic strategy for neuroblastoma. A Phase I clinical trial to test ALK.CAR-T cells in combination with ALK TKIs in children with refractory/relapsed neuroblastoma is being implemented based on these results.

Citation Format: Elisa Bergaggio, Wei-Tien Tai, Andrea Aroldi, Carmen Mecca, Elisa Landoni, Manuel Nüesch, Ines Mota, Jasna Metovic, Luca Molinaro, Leyuan Ma, Diego Alvarado, Chiara Ambrogio, Claudia Voena, Rafael B. Blasco, Tongqing Li, Daryl Klein, Darrell J. Irvine, Mauro Papotti, Barbara Savolodo, Gianpietro Dotti, Roberto Chiarle. ALK inhibitors increase ALK expression and sensitize neuroblastoma cells to ALK.CAR-T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 50.
Abstract 5053: Clinicopathologic and molecular genetic features of STAT3-mutated myeloid neoplasms
Authors: Ye Matthew T. , Zuo Zhuang , Calin Steliana , Ye Fengxi , He Hua , Kamata Wataru , You M. James
Published: March 2024

STAT3 is a key transcription factor that can mediate cancer progression through phosphorylation or gain-of-function mutations. STAT3 mutations have been mainly associated with large granular lymphocytic T-cell leukemia. Whereas STAT3 activation is common in myeloid neoplasms, it is primarily mediated through STAT3 phosphorylation. STAT3 mutation has only rarely been reported in myeloid neoplasms. We assessed the clinicopathologic and molecular genetic features of 32 STAT3-mutated myeloid neoplasms. Twenty (62.5%) cases were classified as acute myeloid leukemia (AML), 7 (21.9%) as myelodysplastic syndrome (MDS), and 5 (15.6%) as chronic myelomonocytic leukemia (CMML). It is of note that STAT3 mutation was not detected in myeloproliferative neoplasms (MPN). STAT3 mutations were detected at the initial diagnosis in 22 of 25 (88%) cases, suggesting STAT3 mutation is an early event in pathogenesis. However, STAT3 mutations represented the dominant clone in only 6 (18.8%) cases, all classified as AML. Most are missense mutations, located at the SH2 domain. The median variant allele frequencies were 6.8%, 4.6% and 3.1% for AML, MDS and CMML, respectively, and in a case of CMML, the VAF increased from 2.4% to 34.8% upon leukemic transformation. STAT3 mutation was accompanied by co-existing mutations in all cases, with SRSF2 (40.6%), TET2 (40.6%), ASXL1 (37.5%), and SETBP1 (34.4%) being most common. Cases with STAT3 mutations were usually associated with morphologic dysplasia and increased blasts. Monosomy 7/del7q was the most common cytogenetic abnormality, seen in 11 (34.4%) cases. With a median follow-up of 24.5 months, 21 patients died, 6 had persistent disease, and 5 achieved complete remission. We summarize that STAT3 mutation is present in a variety of myeloid neoplasms, but not seen in MPN. It is often an early events and always with concurrent mutations. It may play an important role in the pathogenesis and progression of myeloid neoplasms by activating JAK-STAT pathway, and may serve as a potential therapeutic target.

Citation Format: Matthew T. Ye, Zhuang Zuo, Steliana Calin, Fengxi Ye, Hua He, Wataru Kamata, M. James You. Clinicopathologic and molecular genetic features of STAT3-mutated myeloid neoplasms [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5053.
Abstract 5098: Second-line treatment outcomes in Extensive Stage Small Cell Lung Cancer (ES-SCLC) patients after first-line immuno-chemotherapy
Authors: Wan Rui , Fan Linjie , Zhong Jia , Duan Jianchun , Wang Zhijie , Hao Xuezhi , Wang Jie
Published: March 2024

Background: First-line immune checkpoint inhibitors (ICIs) has significantly improved overall survival (OS) of ES-SCLC. However, second-line treatment options are still controversial with limited overall survival. This study aims to evaluate the efficacy of different second-line regimens for relapsed ES-SCLC after first-line immuno-chemotherapy.

Patients and Methods: We retrospectively reviewed 96 ES-SCLC patients receiving first-line immuno-chemotherapy in Cancer Hospital Chinese Academy of Medical Sciences, from Jan 2019 to Dec 2022. Clinical data were collected from electronic medical records. The clinical outcomes, including objective response rate (ORR), progression-free survival (PFS) and overall survival (OS) were assessed by Kaplan-Meier method and standard log-rank test.

Results: From January 4, 2019 to December 18, 2022, a total of 96 ES-SCLC patients received first-line immuno-chemotherapy and 49 of them went through second-line treatment in our center. Patient characteristics were listed as follows : median age 63 (31-79), male 76 (79.2%) , heavy smoker 59 (61.5%), liver metastasis 40 (41.7%), brain metastasis 33 (34.4%) and refractory recurrence 58 (60.4%). Second-line treatments included immunotherapy crossover and combination with chemotherapy (n=40), anlotinib with or without chemotherapy (n=5), chemotherapy alone (n=4). ORR of the whole group in the second-line setting was 22.4%. Median PFS was 3.23 months (95% CI 1.96-4.50). Cross-line immunotherapy failed to improved survival for relapsed ES-SCLC compared with those without ICIs ( mPFS 3.23 vs 3.13m, p=0.829). Anlotinib based treatment showed numerically superior PFS for relapsed SCLC compared with those without anlotinib therapy (mPFS 6.00 vs 3.13m, p=0.089).

Conclusion: This retrospective study indicated cross-line ICIs failure to improve clinical outcome for relapsed ES-SCLC, and implied the potential benefits of anlotinib in second-line setting.

Citation Format: Rui Wan, Linjie Fan, Jia Zhong, Jianchun Duan, Zhijie Wang, Xuezhi Hao, Jie Wang. Second-line treatment outcomes in Extensive Stage Small Cell Lung Cancer (ES-SCLC) patients after first-line immuno-chemotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5098.
Abstract 5160: Evaluation of circulating microRNAs as predictive and prognostic biomarkers for advanced melanoma patients
Authors: Durante Giorgio , Comito Francesca , Naddeo Maria , De Matteis Serena , Bonafè Massimiliano , Dika Emi , Ardizzoni Andrea , Ferracin Manuela
Published: March 2024

Introduction Metastatic melanoma is the deadliest form of skin cancer whose incidence has been rising dramatically over the last few decades. The advent of Immune checkpoint inhibitors (ICIs) has strongly improved the survival of melanoma patients. However, a high fraction of patients shows primary resistance or will develop secondary resistance during therapy. For these reasons, it is of utmost importance to develop biomarkers that could predict response or resistance to therapies. Among the most promising biomarkers analyzed in peripheral blood there are circulating microRNAs (miRNAs). The aim of this study is to identify a miRNA signature able to predict prognosis and clinical relapse in advanced melanoma patients treated with ICIs.

Methodology We collected longitudinal blood draws from metastatic melanoma patients treated with ICIs at Bologna University-Hospital (Italy). We separated plasma from whole blood, extracted RNA and determined the absolute levels of miR-155-5p, miR-424-5p and miR-320a using EvaGreen-based droplet digital PCR. Moreover, in the subset of first-line anti-PD-1 treated patients, we performed cytofluorimetric leukocyte analysis.

Results and discussion We found that patients with pre-therapy (baseline) high expression of circulating miR-155-5p, miR-320a and mir-424-5p had longer progression-free survival (PFS), and overall-survival (OS). In patients treated with anti PD-1 as first-line treatment, we found that baseline miR-155-5p levels were significantly higher in patients who responded to therapy, and therefore miR-155-5p could have a predictive value. Moreover, using a flow cytometric approach, we noted that baseline miR-155-5p levels correlated with CD4+Naïve (r:-0.91, p:0.027) and CD4+PD1+ T-cells (r:0.94, p:0.015) in patients treated with first-line anti-PD-1 treatment.

Conclusion These preliminary findings propose baseline circulating miRNAs as potential predictive and prognostic biomarkers in the setting of advanced melanoma patients treated with immunotherapy.

Citation Format: Giorgio Durante, Francesca Comito, Maria Naddeo, Serena De Matteis, Massimiliano Bonafè, Emi Dika, Andrea Ardizzoni, Manuela Ferracin. Evaluation of circulating microRNAs as predictive and prognostic biomarkers for advanced melanoma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5160.
Abstract 5187: VistaPlex immunofluorescence assay panels provide reliable deep spatial phenotyping for diverse immuno-oncology applications
Authors: Christians Arne , Jackson Charles E. , Boog Jannik , Ingalls Matthew H. , Northcutt Adam , Cashion Kayla E. , Rauer Jonah , Kwarta Karen , Schwarz J Spencer , Braubach Oliver
Published: March 2024

Multiplex immunofluorescence (mIF) is a powerful tool that enables deep phenotyping of immune and cancer cells across a wide variety of tissue types. The initial effort and cost required to develop high-plex mIF panels is significant and poses a considerable hurdle for adoption of spatial biology. To streamline high-plex assay development, we developed a process for creating and validating multiplex antibody panels for use in immuno-oncology applications that leverage CellScape™, an automated end-to-end platform for Precise Spatial Multiplexing. Our multiplex assay development was a rigorous, multi-stage, iterative process to evaluate antibodies for suitability, specificity, and reproducibility. These assay panels are formulated as ready-to-use VistaPlex™ assay kits and are intended to enable rapid, reliable spatial phenotyping of key immune populations.

To confirm the utility of VistaPlex, we deployed the Human FFPE Spatial Immune Profiling Panel to stain and image various FFPE tissue samples on the CellScape platform. With CellScape, tissues are mounted in enclosed fluidic chambers while on-instrument staining, imaging and signal erasure are conducted in iterative rounds. The panel presented here contains antibodies against 16 common biomarkers used in immune-oncology applications. Among these are CD3, CD4, CD8, CD20, and CD45 to detect cell lineages, PD-1 and PD-L1, to interrogate immune checkpoints, and markers that inform about cell state and activation. Spatial immune profiling data were obtained from human tonsil, colon, breast, and lung tissue samples. Deep immune phenotyping was performed using unsupervised clustering analysis and AI-assisted cluster labelling, and biomarker distributions were studied in the context of cellular neighborhoods and other proximity analyses. Overall, we identified more than 19 cell phenotypes with varying frequencies and spatial arrangements across tissue samples.

Our data are robust and demonstrate that VistaPlex Assay kits for spatial profiling present an efficient and cost-effective solution for users that want to break into spatial biology.

Citation Format: Arne Christians, Charles E. Jackson, Jannik Boog, Matthew H. Ingalls, Adam Northcutt, Kayla E. Cashion, Jonah Rauer, Karen Kwarta, J Spencer Schwarz, Oliver Braubach. VistaPlex immunofluorescence assay panels provide reliable deep spatial phenotyping for diverse immuno-oncology applications [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5187.
Abstract 5194: Pre-operative markers extracted from extracellular vesicles related with lung cancer relapse
Authors: Koba Hayato , Nanjo Shigeki , Yano Seiji
Published: March 2024

Background: Pre- and post-operative immunotherapy have been a new standard of care for lung cancer patients. On the other hand, post-operative Osimertinib; EGFR-TKI therapy had obtained new evidence to the EGFR mutant lung cancer patients. Both treatments revealed to make patient survival prolonged. However, there are quite a lot patients who had experienced disease relapse, despite of multimodal therapy. Extracellular vesicles (EVs) have reported to have potent for searching a new target to cancers.

Purpose: Exploration of novel risk factors related with post-operative recurrence of lung cancer.

Methods: We retrospectively examined patient background, disease-free survival, overall survival, and preoperative liquid biopsy analysis in lung cancer patients who underwent surgery at Kanazawa University Hospital between January 2017 and May 2020, in whom preoperative chemotherapy was not administered. EVs were extracted from 500 μl serum within 7 days before surgery using MagCapture Exosome isolation Kit PS Ver.2 (FUJIFILM). First, we focused the six cases that had the 3 phase samples: preoperative, disease free and relapse phase. According to the EV concentrations calculated by BCA method, comprehensive protein assessment of these subjects was performed using shotgun mass spectrometry. Secondly, all cases’ prognosis were analyzed in connection with the detected marker using Dot blotting.

Results: All EV samples from primary 6 cases were able to be detected and median concentration of those was 27 μg/ml (9-82). We focused on three factors (LCEV1-3) among the 6 cases analysis, when we disease fluctuation. Among LCEV1-3 examination to all preoperative samples, LCEV2 had been related with disease recurrence.

Discussions: EVs contain amount of information such as cancer biomarker. Not only that, these factors may have a potency for therapeutic target. Further investigations are needed.

Citation Format: Hayato Koba, Shigeki Nanjo, Seiji Yano. Pre-operative markers extracted from extracellular vesicles related with lung cancer relapse [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5194.
Abstract 52: CRISPR-mediated knock-out of A20 lowers the activation threshold and induces metabolic rewiring of CAR-T cells
Authors: Chu Nina Jia , Nava-Barbero Carina , Merlino Ashley , Lee Ping-Hsien , Baker David , Taylor Ben , Moody Gordon , Giardino Letizia
Published: March 2024

Background: Common failures of chimeric antigen receptor (CAR)-T cell therapy regimens stem from immune escape of target antigen low tumors. Strategies to improve CAR-T cell therapy efficacy include enhancing intrinsic effector function while preventing T cell exhaustion. The ubiquitin-modifying enzyme A20, is a key negative regulator of the NF-kB pathway downstream of T cell receptor activation in T cells. Dysfunctional CD8 T cells in tumors have high expression of A20, and deletion of A20 enhances antitumor activity of CD8 T cells in murine tumor models. We tested whether CRISPR-mediated knock-out (KO) of tumor necrosis factor α-induced protein 3 (TNFAIP3), the gene for A20, increases the effector function and alters the metabolic state of CAR-T cells.

Results: A20 deletion led to upregulation of NF-κB target genes and resulted in higher levels of activation marker expression and effector cytokine secretion compared to unarmored CAR-T upon incubation with target cells. In addition, A20 KO CAR-T cells maintained prolonged expression of activation markers upon antigen clearance in the absence of enhanced differentiation, and were more resistant to dysfunction in a repeat challenge serial kill assay. In vivo, A20 deficient CAR-T exhibited increased anti-tumor efficacy against low, medium, and high target expressing tumor models. Consistent with NF-κB upregulation, A20 KO cells significantly upregulated key glycolytic intermediates suggesting that amino acid anabolism was fueled by glycolysis and the pentose phosphate pathway. Moreover, the preferential increase of glycolysis over oxidative phosphorylation confirmed that A20 KO CAR-T cells were skewed towards an effector-like phenotype. Notably, glutathione and its cysteine-glycine bi-products were upregulated upon activation to a higher level in A20 KO CAR-T suggesting enhanced resistance to oxidative stress, in line with their prolonged persistence.

Conclusions: CRISPR-mediated knock-out of A20 lowers the activation threshold of CAR-T cells producing enhanced in vitro and in vivo anti-tumor efficacy particularly against tumors with very low levels of target expression. A20 KO induces metabolic rewiring of CAR-T cells by promoting glycolysis and amino acid anabolism to sustain their superior effector functions. This intrinsic armoring strategy can be applied to any CAR-T in any indication to increase tumor control and reduce immune escape.

Citation Format: Nina Jia Chu, Carina Nava-Barbero, Ashley Merlino, Ping-Hsien Lee, David Baker, Ben Taylor, Gordon Moody, Letizia Giardino. CRISPR-mediated knock-out of A20 lowers the activation threshold and induces metabolic rewiring of CAR-T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 52.
Abstract 5271: Pseudohypoxia by HIF-PHD inhibitors activates tumor immune response for MSS colorectal cancer
Authors: Ohara Toshiaki , Chen Yuehua , Hamada Yusuke , Nishimura Seitaro , Kawai Hotaka , Noma Kazuhiro , Tazawa Hiroshi , Fujiwara Toshiyoshi , Matsukawa Akihiro
Published: March 2024

Purpose: Microsatellite-stable (MSS) colorectal cancer is resistant for immune therapy, necessitating the exploration of novel modalities for its conquest. Recent revelations have revealed the genetic upregulation of HIF-1α as a potential activator of CD8+ T-cells, enhancing an anti-tumor effect. Pseudohypoxia, characterized by normoxic upregulation of Hypoxia-Inducible Factor (HIF-1α), is known to be induced by iron chelators, including HIF-PHD inhibitors. In this study, we aimed to examine whether pseudohypoxia by HIF-PHD inhibitor can enhance immune response and elucidate the underlying mechanism.

Method: Colon 26, SW480 and HT29 were used as murine and human MSS colorectal cancer model. Human and murine CD8+ T-cells were represented by Jurkat cells and isolated splenic CD8+ T-cells, respectively. HIF-PHD inhibitors Roxadustat and Vadadustat were used in the study, and a subcutaneous Colon 26 tumor model was established in BALB/c WT and nude mice. Hypoxic culture conditions were maintained for 48 hours at 2% O2.

Results: Roxadustat and Vadadustat did not suppress the proliferation of cancer cells under conditions marked by HIF-1α upregulation. Roxadustat and Vadadustat suppress the colon 26 subcutaneous tumor growth in WT mice, the effect was cancelled in the nude mice. The treatment regimen was associated with an augmentation in the total number and effector-like subtypes of CD8+ T-cells (Tex) infiltrating the tumor. Combination therapy involving Roxadustat and PD-1 antibody synergistically inhibited tumor growth and was accompanied by an increase in PD-1lo, Ly108-, TIM-3+ CD8+ T-cells within the tumor microenvironment. Immunohistochemistry revealed that Interleukin-2 (IL-2) expression of murine spleen was stronger in Roxadustat than control group. Interestingly, HIF-PHD inhibitors induced the secretion of IL-2 from isolated CD3+ T-cells of murine spleens in ELISA, a phenomenon uniquely observed under pseudo-hypoxic conditions created by HIF-PH inhibitors, rather than actual hypoxia. This result indicates that pseudo-hypoxia may induce more robust hypoxic response than real hypoxia.

Conclusion: The induction of pseudohypoxia by HIF-PHD inhibitors activates the tumor immune response for MSS colorectal cancer via induction of splenic IL-2.

Citation Format: Toshiaki Ohara, Yuehua Chen, Yusuke Hamada, Seitaro Nishimura, Hotaka Kawai, Kazuhiro Noma, Hiroshi Tazawa, Toshiyoshi Fujiwara, Akihiro Matsukawa. Pseudohypoxia by HIF-PHD inhibitors activates tumor immune response for MSS colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5271.
Abstract 5273: A novel immunogenic cell death that exploits the cancer-specific overexpression of Rad51
Authors: Soloveychik Maria , Chahwan Charly
Published: March 2024

Overexpression of the Homologous Recombination (HR) DNA repair pathway is often correlated with poor prognosis and metastatic disease. SyntheX developed STX100, a small, stabilized peptide that is able to disrupt the function of Rad51 - a crucial component of HR. STX100 exhibits low nanomolar affinity to the target, cell permeability, and proteolytic stability. STX100 has acute mono-agent activity that is selective towards HR-overexpressing cancer cells. Interestingly, STX100-mediated cell killing is independent of canonical cell death mechanisms (apoptosis, necroptosis, pyroptosis, ferroptosis, etc.). Rather, the elevated abundance of Rad51 in cancer cells relative to healthy tissues underlies the selective cytotoxicity resulting from an acute calcium surge upon STX100-mediated target engagement. The mechanism translates to in vivo models, where a single dose can achieve sustained survival when administered locally as a mono-therapy or in combination with an immune checkpoint blockade agent.

Citation Format: Maria Soloveychik, Charly Chahwan. A novel immunogenic cell death that exploits the cancer-specific overexpression of Rad51 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5273.
Abstract 5343: Depletion of gut microbiota with broad spectrum antibiotics drives Pten-null prostate cancer growth in mice
Authors: De Velasco Marco A. , Kura Yurie , Sakai Kazuko , Nishimoto Mitsuhisa , Mori Yasunori , Yoshimura Kazuhiro , Fujita Kazutoshi , Nishio Kazuto , Uemura Hirotsugu
Published: March 2024

Gut microbes have the potential to be either promoters or inhibitors of cancer. Use of antibiotics during cancer therapy can lead to dysbiosis, which may affect treatment outcomes. Here, we examine the effects of broad-spectrum antibiotics on cancer progression in mice with Pten-null castration resistant prostate cancer (CRPC). After five or eight weeks, orchiectomized conditional Pten-null knockout (KO) mice aged 50 and 90 weeks were given two biweekly cycles of a broad-spectrum antibody cocktail (BSAbx), respectively. The effects of BSAbx on tumor burden were compared, and molecular and immunological responses were analyzed by quantitative IHC (qIHC) and qRt-PCR at week four after antibiotic treatment. BSAbx treatment resulted in tumor burdens of 1.2 and 2.4-fold higher than in control mice in mice aged 50 and 90 weeks, respectively. qIHC analysis of KI67 showed increased proliferation in stromal and cancer cells. Additionally, androgen receptor (AR) protein and gene expression levels of Ar and Ar target genes (Fkbp5, Tmprss2, and Timp4) were elevated in tumors of BSAbx-treated mice. Restoring gut microbiota by fecal microbiota transplantation (FMT) from KO mice CRPC after BSAbx restored AR transcriptional activity to basal levels and lessened tumor growth. FMT from non-castrated KO with castration-naïve prostate cancer or heathy wildtype mice further reduced AR Transcriptional activity. Moreover, altering the gut microbiota of CRPC mice with FMT from heathy mice resulted in a tendency for lower tumor burden, 21% reduction versus control mice. Immunologically, BSAbx treatment resulted in decreased CD11b+/GR1+ cells in tumors and were associated with decreased interferon-gamma (IFN-gamma), dendritic cell functions and T cell-mediated cytotoxicity gene signatures. Mice that received FMT form health mice showed enhanced T cell mediated immunity as demonstrated by higher IFN-gamma signatures and tumor abundances of CD8+ T cells expression IFN-gamma, TNF-alpha and the cytotoxic granule granzyme B. Our study shows that broad spectrum antibiotics caused dysbiosis and promoted prostate cancer growth in mice with Pten-null prostate cancer. The effects were partly mediated through the upregulation of AR signaling and the impairment of neutrophil and T cell-mediated immune responses in tumors.

Citation Format: Marco A. De Velasco, Yurie Kura, Kazuko Sakai, Mitsuhisa Nishimoto, Yasunori Mori, Kazuhiro Yoshimura, Kazutoshi Fujita, Kazuto Nishio, Hirotsugu Uemura. Depletion of gut microbiota with broad spectrum antibiotics drives Pten-null prostate cancer growth in mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5343.
Abstract 5346: A novel humanized CD16A model to evaluate efficacy of human antibody therapeutics in triggering ADCC
Authors: Liu Xiao , Niu Zhenlan , Lv Ruili , Lin Qingcong , Kuraguchi Mari , Xu Qingqing
Published: March 2024

Antibody-dependent cellular cytotoxicity (ADCC) is a mechanism by which immune effector cells engage the Fc portion of antibodies bound to antigen on the surface of another cell, thereby targeting that cell for destruction. In mice, the Fcgr4 receptor is primarily responsible for triggering ADCC, while the FCGR3A receptor (CD16A) mediates this function in human NK cells, which are an attractive target cell type for cancer immunotherapy. Given these differences, we sought to generate humanized CD16A preclinical models to evaluate the efficacy of antibodies in triggering human CD16A-mediated ADCC in vivo. Previously, we established humanized CD16A mice on immunocompetent backgrounds for evaluating efficacy of novel antibodies in preventing growth of syngeneic tumor models. Here, we introduce a novel humanized CD16A mouse on the CB-17-SCID background, in order to inoculate human tumor cells for therapeutic efficacy testing. We confirmed expression of CD16A in human NK cells and monocytes/macrophages, but not granulocytes in the model; expression of murine Fcgr4 was no longer detectable, indicating successful generation of the model. Assessment of leukocyte populations in the humanized model indicated no significant alterations in the percentages of NK cells, dendritic cells, granulocytes, monocytes and macrophages in the spleen or blood of the humanized mice compared with wild-type CB-17 SCID mice. Subsequently, the ability of these mice to grow human tumor cells was evaluated: B-hHep3B-Luc, HC116, K-562, and NCI-N87 tumors were all successfully established. Finally, efficacy of an anti-human CLDN18.2 antibody was evaluated against B-hCLDN18.2 BxPC-3 tumors in the humanized mice, which showed modest anti-tumor efficacy at 30 mg/kg when compared to isotype control-treated mice. In summary, humanized B-hCD16A mice (CB-17-SCID) provide a suitable model for evaluating the ability of antibody candidates to induce ADCC.

Citation Format: Xiao Liu, Zhenlan Niu, Ruili Lv, Qingcong Lin, Mari Kuraguchi, Qingqing Xu. A novel humanized CD16A model to evaluate efficacy of human antibody therapeutics in triggering ADCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5346.
Abstract 5355: A novel subset of Treg-like MDSCs expressing LAG-3 found in advanced gastric cancer
Authors: Iwasawa Takumi , Yamauchi Suguru , Fukunaga Tetsu , Orita Hajime , Ito Tomoaki , Kazunori Kato
Published: March 2024

The development and progression of cancers are frequently promoted by immunosuppressive cells, such as Treg and myeloid-derived suppressor cells (MDSCs) mediated by various immune checkpoint molecules. Therefore, MDSCs are potential diagnostic and therapeutic targets in cancer. However, the definitive cell surface markers for defining MDSC and the subset that correlates with cancer progression are not clear. In our previous studies, there was no difference in the amount of total granulocyte MDSCs (G-MDSCs) between early and advanced gastric cancer. We inferred that there is a novel subset of G-MDSCs involved in cancer progression rather than the previous marker-based classification. The purpose of this study is to analyze MDSCs in gastric cancer patients at different stages of progression and to identify subsets of G-MDSCs that are involved in cancer progression. As a result, we identified a subset of novel G-MDSCs that correlate with cancer progression by expressing LAG-3. LAG-3 is a molecule that is expressed in Tregs and causes strong immunosuppression, suggesting that it is involved in the immunosuppressive potential of MDSCs.

Citation Format: Takumi Iwasawa, Suguru Yamauchi, Tetsu Fukunaga, Hajime Orita, Tomoaki Ito, Kato Kazunori. A novel subset of Treg-like MDSCs expressing LAG-3 found in advanced gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5355.