bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5112: Immune biomarkers of clinical response after NSAID treatment in combination with vaccination targeting EGFR-COX2-CDC25B in the APC(Min/+) mouse model
Authors: Cecil Denise L. , Liu Ying , Corulli Lauren , Disis Mary L. 
Published: April 2023

Abstract
Studies from our group have shown that administration of NSAIDs in mouse models of Familial Adenomatous Polyposis (FAP) results in a decrease in the development in small intestinal polyps with a significant increase in CD8 tumor infiltrating lymphocytes (TIL) and downregulation of PD-L1 on tumors. Furthermore, we have demonstrated that NSAIDs could be used in combination with vaccination to even further reduce the development of intestinal polyps. We questioned what tissue specific biomarkers could define the efficacy of response with the combination treatment. APC(Min/+) male and female mice (4-6 weeks old) were immunized with a multi-antigen (EGFR, COX2, CDC25B) peptide vaccine with CFA/IFA as an adjuvant. Control mice received immune adjuvant alone. Regular chow or chow containing 400 ppm naproxen was provided intermittently (3 weeks on, 3 weeks off for 3 cycles) or continuously (18 weeks on) at the initiation of the vaccine regimen. Animals were euthanized at 22 weeks and tumors quantified. TIL were evaluated by flow cytometry. Since intermittent naproxen treatment was just as effective at inhibiting tumor development as continuous naproxen treatment, with or without vaccine, we did not differentiate between NSAID regimens. We observed an increase in CD8+ (83%; p&lt;0.0001), Tbet+CD4+ (94%; p&lt;0.001), effector memory (EM) CD4+ (76%; p&lt;0.0001) and EM CD8+ (94%; p&lt;0.0001) TIL in the tumors that did develop in the mice treated with the combination NSAID and vaccination as compared to control. There was increased CD8+ (Pearson correlation coefficient: −0.506, p&lt;0001), Tbet+CD4+ (Pearson correlation coefficient: -0.6222, p&lt;0.001), EM CD4+ (Pearson correlation coefficient: 0.-6074, p&lt;0.001) and EM CD8+ (Pearson correlation coefficient: -0.7155, p&lt;0.001) in mice with greater tumor inhibition than those with less protection. Receiver operator curves revealed that CD8+ (AUC: 0.7623; p=0.002), Tbet+CD4+ (AUC 0.7273; p=0.008), EM CD4+ (AUC 0.6782; p=0.04) and EM CD8 (AUC 0.7897; p&lt;0.001) TIL can differentiate mice receiving the combination of NSAID and vaccination as compared to either monotherapy alone. These data define potential biomarkers of response that can guide human clinical trials of NSAIDs +/- vaccination in patients with FAP.
Citation Format: Denise L. Cecil, Ying Liu, Lauren Corulli, Mary L. Disis. Immune biomarkers of clinical response after NSAID treatment in combination with vaccination targeting EGFR-COX2-CDC25B in the APC(Min/+) mouse model. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5112.
Abstract 5135: Standardized off-the-shelf engineered NK cell therapy with improved ADCC properties to treat malignancies
Authors: Lin Yanan , Zhang Shengbang , Ran Jie , Song Can , Zhu Shengcang , Shi Hui , Guo Ping , Tan Shengjiang , Gu Yuchun , Wu Lida 
Published: April 2023

Abstract
In the treatment of solid tumors, chimeric antigen receptor(CAR)-engineered NK (CAR-NK) cells have distinct advantages over CAR-T cells, such as a lack graft-versus-host disease in allogenic setting to make “off the shelf” medicine; much safer because they are less likely to cause cytokine storms; and NK cells have their own activated receptors that recognize tumor surface antigens and thus have a natural ability to kill a wide range of tumors. In addition, NK cells are the main performers of antibody-dependent cell-mediated cytotoxicity (ADCC), which binds to the Fc-terminus of antibodies through their surface Fc receptor CD16A, thereby killing antibody-targeted tumor cells. Enhancing the ADCC action of NK cells themselves enhances the therapeutic efficacy of monoclonal antibodies targeting tumor surface antigens and is therefore of great importance in the treatment of tumors, especially solid tumors. We screened two optimal structures among nine different Fc chimeric receptors, and NK cells overexpressing these two receptors killed K562 and Daudi six times more intensely than normal NK cells when combined with rituximab, while the killing ability was comparable to that of normal NK without rituximab, indicating that NK cells expressing chimeric Fc receptors have stronger ADCC effects. Moreover, these NK cells can kill target cells in multiple rounds. In vivo experiments in mice demonstrated that NK cells expressing chimeric Fc receptors in combination with EGFR monoclonal antibodies had a stronger inhibitory effect on tumor growth than monoclonal antibodies or NK cells alone. Here we provide a novel broad “off the shelf” NK cells that can significantly enhance the killing ability of hematological and solid tumors in combination with different monoclonal antibodies.
Citation Format: Yanan Lin, Shengbang Zhang, Jie Ran, Can Song, Shengcang Zhu, Hui Shi, Ping Guo, Shengjiang Tan, Yuchun Gu, Lida Wu. Standardized off-the-shelf engineered NK cell therapy with improved ADCC properties to treat malignancies. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5135.
Abstract 5138: Systemic agonist anti-CD40 treatment delivers precise melanoma immunotherapy within established autoimmune landscapes
Authors: Valencia Julio C. , Sanford Michael , Young Howard A. 
Published: April 2023

Abstract
The overlapping signatures of type 1 interferons (IFN) and IFN-gamma (IFNg) has proven challenging in human autoimmunity and cancer immunotherapy, particularly for resistance to anti-programmed death 1 (anti-PD1) monotherapy. By contrast, agonist abs targeting of CD40 (anti-CD40) antibodies (abs) has clinically elicit anti-tumor immunity with limited autoimmune complications. Here, we explore the relationships between anti-PD1 or agonist CD40 responses in melanoma tumors from type 1 IFN alpha receptor (IFNAR)-sufficient and type 1 IFNAR-deficient lupus-prone mice that overexpress IFNg. To address this, RNA-sequencing (RNA-seq) data on tumors treated with either anti-PD1 or agonist anti-CD40 was evaluated for tumor objective responses compared to untreated controls. Cellular and canonical pathway identification from RNA-seq data were interrogated from the IPA library for significantly differential expressed genes (DEG). Variance patterns of the anti-PD1 and anti-CD40 monotherapies identified known resistance and response pathways highly concordant with IFN co-expression patterns in the RNA-sequencing data, respectively. IPA biomarker analysis for uniquely regulated genes showed that anti-PD1 induced only 59 DEG; while anti-CD40 induced 494 DEG including Il-27, and PR/SET Domain 1 (PRDM1), a transcription factor regulated by IL-27. Specifically, signatures of MDSC development (irf8, il10); and adaptive resistance (pdcd1, pdcd1lg2, and TNFRSF5/CD40)) were negatively associated with response to anti-P1 monotherapy. By contrast, signatures of IFN activation (stat1, Ifng, and Ifngr1), chemotaxis (Cxcl10, Il12b and Il-27), attractants (Sema3g and Sema4a), and antigen presentation MHC II (h2-Aa, h2-Ab1, and ciita) were associated positively with response to agonist anti-CD40 monotherapy. Importantly, anti-CD40 alone delivered anti-tumor immune responses in type 1 IFN receptor-deficient lupus-prone mice that overexpress IFNg (Ifnar-/-ARE-/-) mice suggesting synergy with therapies blocking type 1 IFN signaling. Thus, these findings indicate that the rational use of CD40 agonists abs provide a better therapeutic platform to deliver precise anti-melanoma responses in complex autoimmune landscapes.
Citation Format: Julio C. Valencia, Michael Sanford, Howard A. Young. Systemic agonist anti-CD40 treatment delivers precise melanoma immunotherapy within established autoimmune landscapes. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5138.
Abstract 5148: Interrogating immuno-oncological interactions in the tumor microenvironment
Authors: Patterson Ben , Chen g-Yi , Fernandez Nicolas , Colbert Leiam , He Jiang 
Published: April 2023

Abstract
Single-cell RNA sequencing is a powerful and innovative tool for assessing individual cancer prognoses and outcomes; however, the process of generating this data typically destroys the spatial complexity of the tumor microenvironment. Understanding the spatial complexities within cancer provides crucial information for evaluating individual tumor development and progression. Vizgen’s MERSCOPETM Platform, built on Multiplexed Error-Robust Fluorescence in situ Hybridization (MERFISH) technology, enables the direct profiling of the spatial organization of intact tissue with subcellular resolution. Here, we present a Pan-Cancer approach to demonstrate the MERSCOPE’s ability to characterize breast, colon, prostate, ovarian, lung, and skin cancers from human clinical samples. Specifically, using a 500-gene panel to assess the canonical signaling pathways of cancer, cancer type-specific genes, select immune genes, proto-oncogenes, and tumor-suppressor genes, we show that the MERSCOPE was able to spatially profile gene expression across multiple tumor types. We utilized the Vizgen Cell Boundary Stain kit to generate a molecular and cellular atlas of individual patient tumors by clustering cells based on gene expression and mapping their spatial organization. To gather insight into immuno-oncological interactions and their effect on gene expression across these tumor types, we performed a neighborhood analysis to determine how proximity to immune cells altered gene expression in tumor cells, and vice versa. We found significant gene expression changes in MHC activation and downstream pathways in immune cells located nearest to tumor cells; in tumor cells nearest to immune cells, we identified increased expression of genes related to immune response mediation and inflammation. Furthermore, we compared the annotated immune cell populations across the various cancer types to observe differences in gene expression. Here, we used publicly available single-cell RNA-sequencing data to impute gene expression for these immune cell populations. This analysis demonstrated the transcriptional variation possible in identical immune cell types that are acting in different tissue contexts. Notably, we were able to map and catalogue different patterns of gene expression relating to inflammation, immune signaling, and immune cell exhaustion. These findings demonstrate how the MERSCOPE platform can provide deep insights into the complex heterogeneity observed between different tumor types and tissues.
Citation Format: Ben Patterson, g-Yi Chen, Nicolas Fernandez, Leiam Colbert, Jiang He. Interrogating immuno-oncological interactions in the tumor microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5148.
Abstract 5190: Systematic immune-profiling of immune-deficient mouse models: A rational to select ideal host for tumor implantation
Authors: Luche Hervé , Hadjem Lillia , Mello Marielle , Canavese Priscilla , Angelis Fabien , Joachim Anais , Bouilly Sylvie , Guinut Frederic , Fiore Frederic , Malissen Bernard , Zarubica Ana , Corcuff Erwan 
Published: April 2023

Abstract
Preclinical tumor oncology research relies historically on the analysis of mouse or human tumor cell lines implanted onto severely immune-deficient mouse models carrying nude, SCID or Rag mutations on different genetical background (C57Bl/6N, BalbC, NMRI, CB17 or NOD). These lines harbor defects in several leukocyte lineages among which Tc, Bc and NKc might be affected. Humanized mice are emerging models that have been transplanted with human cells or tissues (and/or equipped with human transgenes). Currently, the most advanced strains are the nonobese diabetic, severe combined immunodeficiency (NOD-SCID) mouse with complete disruptions in the interleukin-2 (IL-2) common γ-chain (IL2R γ null) receptor (NSG) and BALB/c Rag2−/− IL2R γ null SirpaNOD mice (BRGS).1 Juvenile chimera NSG or BRGS are reconstituted with hematopoietic progenitor cell (HPCs) from human cord blood yielding robust engraftment of a human immune system (HIS).. Recent work showing the binding of human IgG4 to mouse FcgR receptor reducing clinical efficacy of therapeutic antibodies highlights the need to characterize extensively the residual immune system present in these models prior humanization in order to precisely evaluate immunotherapies. In other to fulfill this task, we genetically modified the mouse genome introducing mutations involved in Tc, Bc and NKc development. We then immuno-phenotyped over 20 lines of mouse mutants produced from the same animal house to minimize the impact of microbiota on innate immune cell populations. High content cytometry analysis of BM, spleen, thymus and peripheral blood was performed in a standardized manner. The resource obtained of these immunotypes was used to highlight similarities and differences among these lines across several genetical background. This work has shown instrumental in selecting the most appropriate model to use for humanization followed before tumor implantation.
Citation Format: Hervé Luche, Lillia Hadjem, Marielle Mello, Priscilla Canavese, Fabien Angelis, Anais Joachim, Sylvie Bouilly, Frederic Guinut, Frederic Fiore, Bernard Malissen, Ana Zarubica, Erwan Corcuff. Systematic immune-profiling of immune-deficient mouse models: A rational to select ideal host for tumor implantation. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5190.
Abstract 5326: Comparison of CAR-T cell-mediated cytotoxicity assays with suspension tumor cells using high-throughput plate-based image cytometry method
Authors: Chan Leo Li-Ying , Sun Yu-Jun , Chen Yi-Chun , Hua Wei-Kai , Wu Sariena Chiung-Yuan 
Published: April 2023

Abstract
In the recent decade, chimeric antigen receptor (CAR)-T cell therapy has revolutionized strategies for cancer treatments due to its highly effective clinical efficacy and response for B cell malignancies. Currently, there are six CAR-T cell products available in the market including Kymriah, Yescarta, Tecartus, Breyanzi, Abecma, and Carvykt. The success of CAR-T cell therapy has stimulated the increase in the research and development of various CAR constructs to target different tumor types. Therefore, a robust and efficient in vitro potency assay is needed to quickly identify potential CAR gene design from a library of construct candidates. Traditionally, in vitro CAR-T cell-mediated cytotoxicity is assessed using release assays such as 51Cr (radioactivity), calcein (fluorescence), and LDH (enzymatic). However, release assays indirectly measure cell death via molecules released in supernatant and typically limited to only endpoint assays. In addition, handling and disposing of 51Cr hazardous materials is less preferred. Luciferase reporter assay, although highly sensitive, has similar drawbacks as the release assays. Finally, flow cytometry method can directly measure cell death and viability, but can be time-consuming and requires a large number of CAR-T cells when the effector-to-target (E:T) ratio is high. Furthermore, additional steps are required for adherent cells that require trypsinization. Image cytometry methodologies have been utilized for various CAR-T cell-mediated cytotoxicity assay using different fluorescent labeling methods, mainly due to their ease-of-use, ability to capture cell images for verification, and higher throughput performance. In this work, we employed the Celigo high-throughput plate-based Image Cytometer to evaluate and compare two CAR-T cell-mediated cytotoxicity assays using GFP-expressing or fluorescent dye-labeled myeloma and plasmacytoma cells. Performing time- and E:T ratio-dependent CAR-T cell-mediated cytotoxicity assays, the GFP-based method demonstrated higher sensitivity in detecting the level of cytotoxicity when compared to the CMFDA/DAPI viability method. We have established the criteria and considerations for the selection of cytotoxicity assays that are fit-for-purpose to ensure the results produced are meaningful for the specific testing conditions.
Citation Format: Leo Li-Ying Chan, Yu-Jun Sun, Yi-Chun Chen, Wei-Kai Hua, Sariena Chiung-Yuan Wu. Comparison of CAR-T cell-mediated cytotoxicity assays with suspension tumor cells using high-throughput plate-based image cytometry method. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5326.
Abstract 5536: Landscape and analysis of ERBB2 amplification and short variant mutations in large-scale Chinese patients with colorectal cancer
Authors: Chen Zhiqin , Yu Shijun , Wu Yin , Zhang Bei , Zhang Ding , Gao Yong , Quan Ming 
Published: April 2023

Abstract
Background: Human epidermal growth factor receptor 2 (ERBB2) amplification is a rapidly emerging therapeutic target in CRC, in spite of a relatively low incidence. While ERBB2 amplification has been considered as a critical biomarker to predict the efficacy and safety of anti-ERBB2 therapy of CRC, its point mutation status is largely ambiguous.Methods: Blood or tissues samples from 2454 Chinese patients with CRC were collected, and the genomic landscape, clinical characteristics and genetic features of ERBB2 in these samples were retrospectively investigated using next-generation sequencing (NGS) analysis. Results: Of the 2454 colorectal cancer patients, 3.46% (85) carried ERBB2 amplification, 2.24% (55) carried ERBB2 mutation, and some patients harbored both ERBB2 amplification and point mutation at the same time. Microsatellite instability high (MSI-H) was found in approximately 32.7% (18/55) of patients with ERBB2-mutated CRC, but almost negative in ERBB2-amplified CRC. The median fold change of ERBB2 amplification was 30 (Range 4 to 245). Among the 16 mutation sites detected from 55 patients with ERBB2 point mutations, p.R678Q was the most common mutation site (28%), followed by p.V842I (24%) and p.S310F/Y (12%), which appeared to be slightly different from previous reports. ERBB2 mutation was more frequently found in CRC patients aged &lt; 50 years. 50.9% of CRC patients with wild-type ERBB2 carried KRAS mutation, which was significantly higher than those with mutated ERBB2 (25.6%). A higher incidence of BRAF mutation was observed in patients with mutated-ERBB2 (8.5% vs. 2.3%). In addition, CRC patients with ERBB2 CNV showed a higher prevalence of TP53 mutation comparing with those with ERBB2 SNV (92.3% vs. 58.3%), while KRAS mutation exhibited a contrary trend (14.1% vs. 45.8%)Conclusions: The overall variation rate of ERBB2 is 5.4% in Chinese patients with CRC, which is accompanied by significantly different molecular pathological characteristics compared to patients with wild-type ERBB2, and different mutation types (CNV or SNV) display different molecular pathological characteristics.
Citation Format: Zhiqin Chen, Shijun Yu, Yin Wu, Bei Zhang, Ding Zhang, Yong Gao, Ming Quan. Landscape and analysis of ERBB2 amplification and short variant mutations in large-scale Chinese patients with colorectal cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5536.
Abstract 554: The role of Mycobacterium w as a novel treatment for osteosarcoma
Authors: Stefan Kirsten , Dheeraj Arpit , Tailor Dhanir , Kumar Sushil , Li Wenqi , Modi Rajiv , Joshipura Manjul , Khamar Bakulesh , Davis Lara E. , Malhotra Sanjay V. 
Published: April 2023

Abstract
Osteosarcoma (OS) is the most common malignant primary tumor of bone and occurs in adolescents and young adults as well as in the elderly. Despite a number of large clinical trial efforts, metastatic sarcoma remains a lethal disease, and there have been no meaningful advances in therapy or improvements in patient outcomes for decades. Mycobacterium w (Mw, also known as Mycobacterium indicus pranii) is a non-pathogenic strain of mycobacterium which has been used as an immunomodulator against leprosy, tuberculosis and several cancers and is approved for treatment of NSCLC along with chemotherapy in India. Mw promotes a Th1 immune response and its administration in tumor bearing animals is associated with an increase in total number of tumor infiltrating immune cells (TIIC) with decrease in FOXP3 and PD-1 expressing TIIC in pancreatic cancer and melanoma. Previous studies in our lab have shown that Mw promotes mature dendritic cell and activated CD8+ T cell accumulation within the tumor microenvironment in a mouse model of pancreatic cancer. We compared the effects Mw with standard of care doxorubicin on OS tumor progression in a subcutaneous K7M2 syngeneic mouse model. Mw treatment was more effective at tumor inhibition than doxorubicin. Immunohistochemistry analyses of OS tumors revealed an increase in macrophages following Mw treatment. We aim to further analyze the tumor immune microenvironment using a panel of 28 antibodies to determine alterations due to Mw treatment. With these results we hope to show that Mw is a promising candidate for further development as a therapy for OS.
Citation Format: Kirsten Stefan, Arpit Dheeraj, Dhanir Tailor, Sushil Kumar, Wenqi Li, Rajiv Modi, Manjul Joshipura, Bakulesh Khamar, Lara E. Davis, Sanjay V. Malhotra. The role of Mycobacterium w as a novel treatment for osteosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 554.