bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3748: Evaluation of immune cells from patient-derived tumor tissue and peritoneal washing cytology in correlation with immunohistochemical markers in endometrial cancer
Authors: Shin Changmin , Lee Seungmee , Chung Hyewon , Kim Jin Young , Ha Eunyoung , Seo Ji Hae , Shin Sojin
Published: March 2024

Background This study aims to evaluate the characteristics and clinical correlations of tumor-infiltrating lymphocytes (TILs) in endometrial cancer, derived from fresh tumor tissue and peritoneal washing cytology. Furthermore, it seeks to explore the correlation between TILs and immunohistochemical markers in endometrial cancer.

Methods In this study, we investigated the nature of TILs extracted from endometrial tissue and peritoneal washing cytology, along with their correlation with immunohistochemical markers. Samples collected from endometrial cancer patients during surgery, with informed consent, were analyzed for immune cell composition using flow cytometry. We examined the association between pathological findings, a range of immunohistochemical markers (including LVSI, PD-L1, ER, P53, HER2, MLH1, MSH2, MSH6, PMS2, MSI), and the status of tumor-infiltrating immune cells (such as CD3-CD19+ B Cells, CD3-CD16+CD56+ NK Cells, CD4+CD25+ regulatory T cells (Treg), and CD3+CD8+ cytotoxic T cells).

Results Our analysis encompassed 25 endometrial cancer-derived samples. Notable observations include an increase in CD3-CD19+ B cells in the LVSI-positive group and a decrease in the ER-positive group (p<0.05, respectively). CD3-CD16+CD56+ NK Cells decreased in the ER-positive group, while CD3+CD8+ cytotoxic T cells were decreased in the HER2-positive group (p<0.001, p<0.05, respectively). Other immunohistochemical markers showed no significant differences.

Conclusion The study suggests that tumor-infiltrating immune cells from patient-derived tissue and peritoneal washing cytology may reflect the molecular and clinical characteristics of endometrial cancer. However, further research with an expanded sample size and correlation with oncologic outcomes is warranted to validate these findings.

Citation Format: Changmin Shin, Seungmee Lee, Hyewon Chung, Jin Young Kim, Eunyoung Ha, Ji Hae Seo, Sojin Shin. Evaluation of immune cells from patient-derived tumor tissue and peritoneal washing cytology in correlation with immunohistochemical markers in endometrial 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 3748.
Abstract 3765: Single cell spatial proteomics analysis and computational evaluation pipeline
Authors: Bozorgui Behnaz , Dereli Zeynep , Thibault Guillaume , Weinstein John N. , Korkut Anil
Published: March 2024

Resolving tissue and proteomic heterogeneity is critical to decoding the structure and function of tumor-immune microenvironment (TIME). Such understanding requires profiling of tumor and immune cell proteomic features with spatial resolution at the single-cell level. Although such spatially resolved methods and data sets are becoming increasingly available, analytical and computational methods that can extract the highly complex features and interactions within TIME are lacking. To address that problem, we have developed a computational pipeline we call the Spatial Proteomics Analysis and Computational Evaluation Pipeline (SPACE). The SPACE pipeline is composed of many analysis modules for processing and mining highly multiplexed imaging-based data types to explore TIME composition, organization, and heterogeneity. The pipeline generates and interprets biomarker expression and positional information from multiplexed images using algorithms for image indexing, image registration, quality control, segmentation, identification and removal of non-specific signals, data normalization, automatic identification of missing data, and adjustment for left-over signals. The accurate intensity measurements at single cell level are then used to calculate the proposed spatial features that represent cellular interactions in TIME. A hierarchical decision tree of cell markers is used to annotate types and identities for individual cells. The SPACE enables statistical and differential analyses of complex spatial features as well as cell types/identities with respect to clinical annotations and genomic alterations. The visualization of spatial and imaging data is made possible through an open-source OMERO image repository and spatial maps that integrate diverse markers in a single representation. Here, we demonstrate the applications of our pipeline in diverse gastrointestinal tumor types (e.g., small bowel adenocarcinoma) and validate the importance of integrating tissue heterogeneity at spatial and single-cell level. The framework is applicable to nearly all highly multiplexed imaging data platforms, including but not limited to, CycIF, CODEX, and imaging mass cytometry.

Citation Format: Behnaz Bozorgui, Zeynep Dereli, Guillaume Thibault, John N. Weinstein, Anil Korkut. Single cell spatial proteomics analysis and computational evaluation pipeline [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 3765.
Abstract 3822: Oncotype DX testing and differences in use of chemotherapy by age in an integrated system
Authors: Aller Ashley , Lyon Liisa , Habel Laurel , Shirazi Aida , Liu Raymond
Published: March 2024

Introduction: Oncotype DX (ODX) identifies breast cancer patients who are less likely to benefit from chemotherapy. We investigated patient factors associated with clinical application of ODX testing and chemotherapy use.

Methods: We identified a cohort of adult breast cancer patients with stage I-IIA, estrogen receptor positive, human epidermal growth factor receptor 2 negative, lymph node negative disease diagnosed within Kaiser Permanente Northern California between 2015 - 2021 eligible for ODX testing. Chi-square tests were used for bivariate analysis of categorical variables. Multivariable logistic regression was used to identify independent predictors of chemotherapy use in female patients with high risk ODX scores (>25).

Results: 8017 patients were eligible for ODX testing; of which 5,136 received testing (64%) and 576 patients (7%) had high risk ODX. ODX testing increased over the years (45% of eligible patients in 2015; 79% of eligible patients in 2021). Testing was frequent in larger lesions (74%), poorly differentiated tumors (73%), and younger patients (83% in patients <40 years at diagnosis; 12% in patients > 80 years). Chemotherapy was frequently administered in younger patient, and for larger, poorly differentiated tumors. For patients who possessed a high risk ODX score, chemotherapy administration differed by age. In comparison with the 90% of high risk patients <45 years who received chemotherapy, 58% of patients 60+ years at diagnosis with high risk ODX received chemotherapy. Comorbidity index and chemotherapy administration was not statistically significant (Table 1).

Conclusion: Oncologists frequently use ODX testing to guide chemotherapy management. Despite high risk ODX scores, older patients received less chemotherapy compared to their younger counterparts. Further investigation is needed to determine how age impacts clinician decision making and clinical outcomes following ODX testing.

Table 1. Multivariable Logistic Regression Analysis of Factors Associated with Chemotherapy Women > 60 years with high risk oncotype DX scores (N=467) Adjusted Odds Ratio (95% Confidence Interval) P-values (Calculated using the Wald χ2 Statistic) Age at Cancer Diagnosis (Reference: 60 - 69 years old) 70-79 0.62 (0.40-0.96) 0.03 >79 0.11 (0.02-0.55) 0.01 Charlson Comorbidity Index (Reference = 0) 1 0.76 (0.48-1.22) 0.26 2 0.62 (0.30-1.29) 0.20 3 0.65 (0.26-1.60) 0.35 4 0.54 (0.20-1.44) 0.22 5+ 0.37 (0.10-1.35) 0.13

Citation Format: Ashley Aller, Liisa Lyon, Laurel Habel, Aida Shirazi, Raymond Liu. Oncotype DX testing and differences in use of chemotherapy by age in an integrated system [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 3822.
Abstract 3824: Efficacy of trastuzumab in combination with capecitabine and cisplatin vs 5-FU and cisplatin therapy for Korean patients with HER2-positive advanced gastric cancer Health Insurance Review and Assessment service (HIRA) data retrospective analysis
Authors: Lee hyun woo , Kim Jang-Hee , Kang Seok Yun , Park Tae Jun , Choi Yong Won
Published: March 2024

The combinations of oral fluoropyrimidines and cisplatin such as capecitabine and cisplatin (XP) or 5-FU and cisplatin (FP) are regarded as a standard therapy against HER-2 positive advanced gastric cancer (AGC). Trastuzumab-XP(HXP) and trastuzumab-FP(HFP) is the most common regimen against AGC in Korea. The aim of the present study is to evaluate the effectiveness of trastuzumab-XP vs trastuzumab-FP.

Patients with HER2-positive AGC were assigned to the trastuzumabXP or trastuzumabFP group. We then retrospectively compared the efficacy and safety between both groups. As a first-line chemotherapy, trastuzumab in combination with XP or FP was administered to 2,399 patients: 1,913 with HXP and 486 with HFP. In the HXP group vs HFP group, median progression-free survival (mPFS), and median overall survival (mOS) were 8.02 vs 6.08 months (p <0.0001) and 13.27 vs 9.89 months (p <0.0001). In HFP group, mPFS and mOS were 10.15 vs 5.36 months (p <0.0001) and 15.15 vs 8.90 months (p <0.0001) in comparison HFP-H maintenance to HFP group. In HXP group, mPFS and mOS were 10.64 vs 13.44 vs 5.03 months (p <0.0001) and 20.14 vs 22.18 vs 9.10 months (p <0.0001) in comparison HXP-H maintenance, HXP-HX maintenance, and HXP group.

Trastuzumab in combination with XP and HX maintenance is a potential first-line therapeutic option for patients with HER2-positive AGC.

Citation Format: hyun woo Lee, Jang-Hee Kim, Seok Yun Kang, Tae Jun Park, Yong Won Choi. Efficacy of trastuzumab in combination with capecitabine and cisplatin vs 5-FU and cisplatin therapy for Korean patients with HER2-positive advanced gastric cancer Health Insurance Review and Assessment service (HIRA) data retrospective analysis [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 3824.
Abstract 3833: Primary cutaneous diffuse large B-cell lymphoma-leg type and clinicopathologic determinants of survival: Analysis of a pooled database
Authors: Haddad Philip A. , Amankwah Millicent
Published: March 2024

Background: Primary Cutaneous Diffuse Large B-cell Lymphoma-Leg Type (PCDLBCL-LT) is a rare but non-indolent subtype of B-cell lymphoma. While most PCDLBCL-LT starts over the lower extremities, some cases may present at other cutaneous sites. The determinants of survival outcomes in this disease are not very well defined. Therefore, we conducted this pooled database analysis to identify the prognostic factors, clinicopathological characteristics, and therapeutic strategies that influence survival in this disease.

Methods: To study the demographic characteristics, molecular and immunohistochemical signatures, therapeutic interventions, survival, and prognostic factors, we compiled a pooled database of 122 cases that fit the diagnostic criteria for PCDLBCL-LT. Kaplan-Meier survival curves were constructed. Cox proportional hazards model and Log-rank tests were used to assess the influence of demographic and clinicopathologic factors on overall survival (OS) and disease-free survival (DFS).

Results: A total of 122 patients with confirmed PCDLBCL-LT were identified. The median age was 76. There was a male preponderance with M:F of 2.4. Seventy-three percent involved the extremities. The median duration of symptoms before diagnosis was 3.5 months. Involvement of lymph nodes, bone marrow, bone, and CNS occurred in 11%, 3%, 4%, and 3%, respectively. Constitutional symptoms were reported in 5%. The median OS and DFS of the whole group were 39 and 32 months, respectively. Patients younger than 70 had better median DFS (NR vs. 24 months, p=0.02) with a non-significant better median OS. Lymphadenopathy (45 vs. 18 months, p=0.006) and stage T3 compared to T1/T2 (16 vs 41 months, p=0.006) were associated with worse OS. Sex, constitutional symptoms, IHC phenotype, extra-lymphatic/extra-cutaneous organ involvement, and histologic cutaneous patterns did not impact OS. Compared to no treatment, chemotherapy, XRT+/-surgery, and stem cell transplant had numerically better OS without reaching statistical significance (5 vs. 39 vs. 41 vs. NR months, p=0.15). Patients who attained CR as their best response also had a superior median OS compared to responses less than CR and none (61 vs. 23 vs. 11 months, p<0.0001).

Conclusion: This study presents updated clinicopathologic data from a large, pooled cohort of patients with PCDLBCL-LT. It identifies age, lymph node involvement, T stage, type of therapy, and quality of response to treatment as critical determinants of OS.

Citation Format: Philip A. Haddad, Millicent Amankwah. Primary cutaneous diffuse large B-cell lymphoma-leg type and clinicopathologic determinants of survival: Analysis of a pooled database [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 3833.
Abstract 3852: Comprehensive genomic profiling of recurrent or metastatic head and neck cancer presenting with combined positive score 90 or more
Authors: Saito Yuki , Hidenori Kage , Kobayashi Kenya , Fukuoka Osamu , Yamamura Koji , Shinozaki-Ushiku Aya , Oda Katsutoshi , Kondo Kenji
Published: March 2024

Background. Biomarkers that predict response to immune checkpoint inhibitor (ICI) in recurrent metastatic squamous cell carcinoma of the head and neck (RMHNSCC) are not well known. Previously, we have found that Combined positive score (CPS) ≥90 and albumin >3.5 respond well to ICI treatment in patients with RMHNSCC treated with ICI irrespective of addition of chemotherapy. In the present study, we investigated whether RMHNSCC with CPS ≥90 has a characteristic genomic mutational profile.

Methods. CPS measurements were performed on a single tumor specimen at the time of diagnosis of RMHNSCC, as approved by Japanese health insurance. Tumor specimens were obtained from the primary tumor as much as possible, and sections that best reflected anti-tumor immunity were decided by a pathologist. We retrospectively reviewed the results obtained from comprehensive genomic profiling (CGP) testing of RMHNSCC cases with CPS ≥90 who were treated by ICI experienced at our institution.

Results. There were 117 eligible RMHNSCC cases. The number of primary lesions were oral cavity in 28 cases (23.9%), nasopharynx in 10 cases (8.5%), oropharynx in 21 cases (17.9%), hypopharynx in 33 cases (28.2%), and others in 18 cases (15.4%). Of 117 patients, 28 patients (23.9%) had a CPS ≥90. Of the 28 cases, the number of primary lesions were oral cavity in 12 cases (42.9%), nasopharynx in 4 cases (14.3%), oropharynx in 0 cases (0%), hypopharynx in 8 cases (28.6%), and others in 3 cases (10.7%). When ICI was administered, CPS ≥90 had a statistically significantly better prognosis than CPS <90 for OS and PFS. Of the 28 cases with CPS ≥90, CGP test was performed in 6 cases. The primary lesions in the 6 cases consisted of 3 oral cavity, 2 nasopharynx, 1 hypopharynx, and 1 maxillary sinus. All three oral primary cases had co-mutations in the TERT promoter and HRAS. Two of the nasopharyngeal primary cases had mutations in JAK3 and STAT, respectively. One of the hypopharyngeal primary cases had mutations in FGFR3.

Conclusion. RMHNSCC with CPS ≥90 had unique mutations that could account the enhanced PD-L1 expression, such as TERT/HRAS co-mutation, JAK-STAT pathway mutation, and FGFR pathway mutation. The above findings may be useful for predicting the efficacy of ICI for RMHNSCC.

Citation Format: Yuki Saito, Kage Hidenori, Kenya Kobayashi, Osamu Fukuoka, Koji Yamamura, Aya Shinozaki-Ushiku, Katsutoshi Oda, Kenji Kondo. Comprehensive genomic profiling of recurrent or metastatic head and neck cancer presenting with combined positive score 90 or more [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 3852.
Abstract 3879: Development of potent, highly selective and efficacious SMARCA2 degraders
Authors: Yang Lin , Tu Wenbin , Huang Liyue , Leng Lingying , Jiang Wei , Wang Meilin , Wen Bo , Sun Duxin , Stuckey Jeanne , Wang Shaomeng
Published: March 2024

In human lung, melanoma and other types of human cancers, the mammalian SWItch/Sucrose Non-Fermentable (SWI/SNF) helicase SMARCA4 is frequently mutated, which leads to inactivation of its functions. SMARCA2, a close homologous protein of SMARCA4, is an attractive synthetic lethality target for human cancers with SMARCA4 deficiency. Herein, we report the discovery and biological evaluation of potent, highly selective and efficacious SMARCA2 PROTAC degraders exemplified by UM-SMD-3236. UM-SMD-3236 has a DC50 value of <1 nM in inducing degradation of SMARCA2 in cells and demonstrates >400-fold selectivity over SMARCA4 protein. Of significance, while UM-SMD-3236 achieves a Dmax of >95% against SMARCA2, it shows a Dmax of 50% against SMARCA4 in cells. In vivo, a single intravenous dose of UM-SMD-3236 attains 85-93% of SMARCA2 depletion in tumor tissues for 7 days, while showing no reduction of the SMARCA4 protein. Weekly intravenous administration of UM-SMD-3236 is highly effective in inhibition of tumor growth in SMARCA4 deficient xenograft models of human cancer. Importantly, UM-SMD-3236 shows no signs of toxicity in mice at highly efficacious doses. UM-SMD-3236 represents a highly promising SMARCA2 degrader for extensive evaluation as a potential new therapy for the treatment of SMARCA4-deficient human cancers.

References 1. Oike, T.; Ogiwara, H.; Tominaga, Y.; Ito, K.; Ando, O.; Tsuta, K.; Mizukami, T.; Shimada, Y.; Isomura, H.; Komachi, M.; Furuta, K.; Watanabe, S.-I.; Nakano, T.; Yokota, J.; Kohno, T. A Synthetic Lethality-Based Strategy to Treat Cancers Harboring a Genetic Deficiency in the Chromatin Remodeling Factor BRG1. Cancer Res. 2013, 73, 5508-5518. 2. Yang, L.; Tu, W.; Huang, L.; Miao, B.; Kaneshige, A.; Jiang, W.; Leng, L.; Wang, M.; Wen, B.; Sun, D.; Wang, S. Discovery of SMD-3040 as a Potent and Selective SMARCA2 PROTAC Degrader with Strong in vivo Antitumor Activity. J. Med. Chem. 2023, 66, 10761-10781.

Citation Format: Lin Yang, Wenbin Tu, Liyue Huang, Lingying Leng, Wei Jiang, Meilin Wang, Bo Wen, Duxin Sun, Jeanne Stuckey, Shaomeng Wang. Development of potent, highly selective and efficacious SMARCA2 degraders [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 3879.
Abstract 3885: A ubiquitin-specific proximity labeling method to study drug-induced ubiquitylation
Authors: Huang Hai-Tsang , Lumpkin Ryan J. , Tsai Ryan W. , Donovan Katherine A. , Su Shuyao , Zhao Xu , Chen James , Fischer Eric S. , Sellers William R.
Published: March 2024

Targeted protein degradation is a promising therapeutic paradigm that offers new ways to disrupt protein functions. Currently, most studies that characterize the pharmacological effects of protein degraders utilize target-specific assays and global proteomics. However, information on the direct molecular consequence of protein degraders, i.e., induced protein ubiquitylation, is often lacking. Here we report the development of E-STUB (E3-substrate tagging by ubiquitin biotinylation), a ubiquitin-specific proximity labeling method that identifies ubiquitylated substrates in proximity to an E3 ligase of interest. We demonstrated that E-STUB can accurately identify the ubiquitylated targets induced by protein degraders in both CRBN and VHL systems—two of the most utilized E3 ligases by heterobifunctional degraders—and is likely generalizable to other E3 ligases. While we observed a high level of consistency between ubiquitylation and degradation for the induced substrates, E-STUB also detected ubiquitylation of non-degraded targets, providing novel and complementary insights to global proteomics. We also envision that E-STUB could be specifically used for the development of chemical inducers of proximity that catalyze non-degradative ubiquitylation events. In summary, we expect that the E-STUB approach can be widely useful for studying the principles of chemically induced ubiquitylation and the advancement of targeted protein degradation.

Citation Format: Hai-Tsang Huang, Ryan J. Lumpkin, Ryan W. Tsai, Katherine A. Donovan, Shuyao Su, Xu Zhao, James Chen, Eric S. Fischer, William R. Sellers. A ubiquitin-specific proximity labeling method to study drug-induced ubiquitylation [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 3885.
Abstract 3937: Investigating the role of fibulin 3 in pancreatic tumorigenesis
Authors: Song Hyemin , Hsu Jasper , Lin Xiaoxue , Ogawa Satoshi , Pantazopoulou Vasiliki , Peck Kristina , Bottomley Chelsea , Okhovat Shira , Stamp McKenna , Curtis Kassidy , Esko Jeffrey D. , Engle Dannielle D.
Published: March 2024

Pancreatic ductal adenocarcinoma (PDA) is a highly lethal malignancy with a five-year survival rate of less than 11%, making it one of the deadliest cancers. Notably, 95% of PDA cases exhibit KRAS mutations, contributing to highly dysregulated cell signaling networks. Despite extensive research into the interplay of various oncogenic pathways in pancreatic tumorigenesis, targeted therapies against these signaling networks have displayed modest efficacy, with most patients rapidly developing therapeutic resistance. Recent studies highlight the significance of the carbohydrate antigen 19-9 (CA19-9) as a functional biomarker linked to disease progression and resistance. However, understanding the detailed functions of CA19-9-modified proteins in PDA has been limited due to the lack of adequate models for synthesizing CA19-9. To address this challenge, we have developed unique mouse and 3D organoid culture models capable of producing CA19-9. CA19-9 elevation in mice with KRAS-mutations results in an aggressive PDA phenotype through increased tumor proliferation and microenvironment (TME) remodeling. Furthermore, our research has identified Fibulin 3 (FBLN3), an extracellular matrix glycoprotein, as a secreted, CA19-9-modified protein that stimulates tumorigenesis. In this study, we found that FBLN3 not only facilitates tumor proliferation but also drives epithelial-mesenchymal transition (EMT) and promotes STAT3 pathways activation in the CA19-9pos, KRAS-mutant PDA organoids. Additionally, in both CA19-9pos, KRAS-mutant PDA mouse models and human PDA, FBLN3 is expressed by cancer-associated fibroblasts (CAFs), suggesting its role in TME remodeling. These findings have significant implications for providing novel insights into PDA biology and paracrine signaling mechanisms, identifying FBLN3 as a potential therapeutic target for the improved treatment of this devastating disease.

Citation Format: Hyemin Song, Jasper Hsu, Xiaoxue Lin, Satoshi Ogawa, Vasiliki Pantazopoulou, Kristina Peck, Chelsea Bottomley, Shira Okhovat, McKenna Stamp, Kassidy Curtis, Jeffrey D. Esko, Dannielle D. Engle. Investigating the role of fibulin 3 in pancreatic tumorigenesis [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 3937.
Abstract 3961: Impact of CD8 resident memory T cells on antitumor immunity and response to cancer immunotherapy
Authors: Corgnac Stephanie , DAMEI Isabelle , Mami-Chouaib Fathia
Published: March 2024

CD8+ resident memory T (TRM) cells expressing the CD103 integrin accumulate in human lung tumors and are associated with a favorable prognosis. We have previously demonstrated that a high density of CD103+CD8+ T cells in human lung tumors is a biomarker of response to immune checkpoint blockade (ICB) immunotherapy. However, a large fraction of human tumors is only weakly infiltrated by CD8+ TRM cells, and the reason of this TRM desert is not explained. Therefore, defining the molecular signals that give rise and maintain TRM cells within the tumor microenvironment (TME) and the mechanisms that potentiate antitumor TRM functions are important challenges. Data obtained in pre-clinical mouse colorectal cancer and melanoma models demonstrate that knocking-out CD103 impairs antitumor T-cell immunity and response to ICB and therapeutic peptide vaccine. Moreover, immunization of melanoma-bearing mice with the cancer vaccine results in a dramatic decrease in the percentage of CD103+ TRM cells among CD8+ tumor-infiltrating T lymphocytes (TIL). Algorithm analyses applied to spectral cytometry data of TIL support the conclusion that upon activation with the peptide vaccine a TRM subset gives rise to tumor-specific effector T cells. The mechanisms associated with the decrease in CD103 expression on CD8+ T cells and lead to a TRM differentiation shift will be presented as well as the key role of TGF-b in CD8+CD103+ TRM formation and persistence in the TME. Our data demonstrate that tumor CD8+CD103+ TRM are capable of mounting a potent antitumor immunity and suggest that a fine balance between diverse signals in the tumor ecosystem is required to induce their differentiation and thereby optimize response to cancer immunotherapy.

Citation Format: Stephanie Corgnac, Isabelle DAMEI, Fathia Mami-Chouaib. Impact of CD8 resident memory T cells on antitumor immunity and response to cancer immunotherapy [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 3961.
Abstract 3968: Class I HLA-independent lysis of cancer cells by tumor-infiltrating CD8 T cells
Authors: Xie Hongyan , Jiang Aiping , Perera Jonathan , Dey Aonkon , Smith Neal , Sade-Feldman Moshe , Manguso Robert T. , Hacohen Nir , Jenkins Russell W.
Published: March 2024

T cell-based therapies have transformed cancer treatment, although intrinsic or acquired resistance develops in nearly half of the patients. Tumor-infiltrating CD8+ T lymphocytes (TILs) are key determinants of anti-tumor immunity by secretion of cytotoxic granules, such as granzymes and perforin, and effector cytokines, such as TNFa and IFNg. A deep understanding of how CD8+ TILs eliminate cancer cells will help identify resistance mechanisms and develop new therapeutic strategies. Here, we demonstrate that TILs eliminated matched melanoma cells in a time and dose-dependent manner but in a class I HLA-independent fashion. Specifically, blocking class I HLA-TCR interaction either by the deletion of B2M (resulting in loss of class I HLA surface expression) or TCR genes did not alter the activity of these TILs. We further confirmed that TIL-mediated killing is contact-dependent. After excluding the potential roles of CD1d and MR1-restrictive T cells, gamma delta T cells, and NKG2D in TIL-mediated killing, a whole-genome CRISPR/Cas9 screen was performed in cancer cells. We identified that genes involved in JAK-STAT and cell-extrinsic apoptosis pathways are necessary for TIL-mediated killing. Together, these findings demonstrate that expanded CD8 TILs from melanoma patients are capable of eliminating melanoma cells via a novel, class I MHC-independent mechanism by coactivation of both JAK-STAT and cell-extrinsic apoptosis pathways.

Citation Format: Hongyan Xie, Aiping Jiang, Jonathan Perera, Aonkon Dey, Neal Smith, Moshe Sade-Feldman, Robert T. Manguso, Nir Hacohen, Russell W. Jenkins. Class I HLA-independent lysis of cancer cells by tumor-infiltrating CD8 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 3968.
Abstract 402: Aneuploidy-associated SQLE gain promotes prostate cancer aggressiveness by altering lipid metabolism
Authors: Janas Thomas Walter , Su Xiaofeng A. , Stopsack Konrad H. , Schmidt Daniel R. , Ma Duanduan , Li Zhe , Penny Kathryn L. , Lotan Tamara L. , Mucci Lorelei A. , Vander Heiden Matthew G. , DeArment Elise , Amon Angelika , Scheet Paul A.
Published: March 2024

Prostate cancer (PCa) is the second leading cause of cancer-related death in men in the US. Epidemiology studies on primary PCa cohorts in Physicians' Health Study and Health Professionals Follow-up Study (PHS and HPFS) have shown that high levels of whole-genome aneuploidy, featured by imbalanced chromosome numbers, correlate with lethal progression in PCa. However, details of the mechanisms of how aneuploidy drives PCa aggressiveness are still unclear. Here, we used the case of chromosome 8q (chr 8q, the long arm of chr 8) gain to study aneuploidy-associated prostatic malignancies. Chr 8q gains are the most frequent gain events that occur in approximately 23% of PCa cases. By using the PHS and HPFS cohorts, we modeled the increased expression of each gene located on chr 8q, for predicting the risks for lethal progression, and obtained each corresponding gene’s odds ratio (OR). By ranking the ORs, we revealed that a cholesterol biosynthesis gene, squalene monooxygenase (SQLE), is one of the top associators with lethal progression, amongst all chr 8q genes. SQLE plays a pivotal role in cholesterol synthesis. Previous lymphoma studies have shown that loss of SQLE contributes to cholesterol auxotrophy, and squalene build-up protects against oxidative cell death. In our experimental study, we have used normal and cancerous TMPRSS2-ERG-driven organoid models and found that over-expression of SQLE promotes formation of invasive structures and proliferation in cancer organoids. Interestingly, overexpression of SQLE decreased the protein levels of TMPRSS2-ERG, which appeared to be independent of androgen receptor levels. We also utilized the TMPRSS2-ERG positive VCaP cell line, which harbors gains of SQLE gene copies. We found that knocking down SQLE expression significantly upregulated ERG protein levels. Our recent study has shown that ERG (or other ETS) positive prostate cancers have a strong correlation with downregulation of fatty acid metabolism signature. We speculate that gain of SQLE can drive aggressiveness of prostate cancer by modulating lipid metabolism for growth and migration. Inhibition of SQLE could translate to better clinical outcomes regarding prostate cancer lethality.

Citation Format: Thomas Walter Janas, Xiaofeng A. Su, Konrad H. Stopsack, Daniel R. Schmidt, Duanduan Ma, Zhe Li, Kathryn L. Penny, Tamara L. Lotan, Lorelei A. Mucci, Matthew G. Vander Heiden, Elise DeArment, Angelika Amon, Paul A. Scheet. Aneuploidy-associated SQLE gain promotes prostate cancer aggressiveness by altering lipid metabolism [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 402.
Abstract 4049: The beta-carboline Harmine sensitives microsatellite stable colorectal cancer mouse model to anti-PD-1 through upregulating MHC-I dependent antigen presentation machinery and improving the infiltration of CD8 T cells into the tumor microenvironment
Authors: Gao Ruize , Van Moer Kris , Pulido Coralie , Oudin Anaïs , Murera Diane , Ramos Teresa L. , Poussard Margaux , Schläpfer Andreas , Ives Annette , Auclair Christian , Janji Bassam
Published: March 2024

Colorectal cancer (CRC) is the most common gastrointestinal cancer and the third leading cause of cancer-related death in both sexes. Mismatch repair deficient/microsatellite instability-high (MSI) colorectal cancer patients benefit from immune checkpoint blockades (ICBs)-based immunotherapy; however, this benefit has not been translated into microsatellite stable (MSS) colorectal cancer. It has been proposed that the loss of major histocompatibility complex class I (MHC-I) in tumor cells limits the use of ICB in colorectal cancer. Harmine displays a number of biological and pharmacological properties affecting the immune response and tumor phenotype. Here, we assessed the impact of combining harmine, also referred to as ACB1801 molecule, on improving the responsiveness of ICB in the mouse MSS CRC model. Our results showed that combining ACB1801 significantly improves the therapeutic benefit of anti-PD-1 in the CT26 CRC mouse model by decreasing tumor growth and improving the survival of tumor-bearing mice. The improvement of anti-PD-1-based therapy by combining ACB-1801 was associated with a modification of the immune landscape of tumors as evidenced by an increase in the CD8+ T cells and a decrease in the Treg infiltrations into the tumor microenvironment. By profiling the cytokine/chemokine network, we showed that ACB1801 induced the expression and the release of the proinflammatory chemokine CXCL10 by CRC tumor cells in vitro, which could be responsible for driving CD8+ T cells to the tumor microenvironment. Mechanistically, we showed that ACB1801 increased the expression of MHC-I genes, including TAP1 and TAPASIN, and improved the antigen loading on MHC-I in CRC cells in vitro. Overall, our study highlights the value of combining ACB1801 and anti-PD-1 as a therapeutic opportunity to convert MSS colorectal cancer into an “immune hot” cancer, which may define the future treatment paradigm of colorectal cancer for which there is a great unmet need.

Citation Format: Ruize Gao, Kris Van Moer, Coralie Pulido, Anaïs Oudin, Diane Murera, Teresa L. Ramos, Margaux Poussard, Andreas Schläpfer, Annette Ives, Christian Auclair, Bassam Janji. The beta-carboline Harmine sensitives microsatellite stable colorectal cancer mouse model to anti-PD-1 through upregulating MHC-I dependent antigen presentation machinery and improving the infiltration of CD8 T cells into the tumor microenvironment [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 4049.
Abstract 4076: Interleukin-18 engineered for resistance to IL-18 binding protein (IL-18BP) and half-life extension to enhance its therapeutic potential
Authors: Chamoun Jean , DiLea Clifford , Dreaden Kristiana , Gurel Pinar , Heiber Joshua , Newman Robert G. , Pearson Su-Ping , Wang Chunhua , Zhao Yanchun , Whitmore Mark
Published: March 2024

Interleukin-18 (IL-18) possesses a unique combination of innate and acquired immune functions with high potential to be transformative in cancer immunotherapy of solid tumors. In clinical studies, recombinant wild-type IL-18 was limited by rapid upregulation of IL-18 binding protein (IL-18BP), which serves as a negative feedback checkpoint of the IL-18 pathway. As its name implies, IL-18BP tightly binds to IL-18, thereby blocking its ability to activate the IL-18 receptor. We have focused our protein engineering efforts to obtain variants of IL-18 that are resistant to IL-18BP suppression while retaining IL-18 biological activity. To further capitalize on the immunological effects of IL-18, the IL-18BP resistant variants were fused to half-life extension protein scaffolds for enhanced in vivo exposure. Rational protein design and combinatorial approaches were used to generate a pool of IL-18 variants that were screened for potency and resistance to IL-18BP. Several IL-18 variants showed undetectable binding to IL-18BP (up to 1 mM IL-18BP) and displayed a spectrum of potencies relative to recombinant wild-type IL-18. Importantly, these variants retained their full biological activity in the presence of super-physiological levels of IL-18BP, as tested with in vitro cell-based IFN-γ release assays. In vivo pharmacokinetic studies in mice compared several IL-18BP resistant variants fused to a half-life extension scaffold with a non-half-life enhanced IL-18BP resistant variant. The half-lives for the fusion proteins ranged from 15 - 30h, while the non-fused variant showed a half-live of 0.6h. Furthermore, the IL-18 variants displayed a more durable, Th1 immune response compared to a non-half-life enhanced variant. Thus, IL-18 variants with strong resistance to IL-18BP that are fused to a half-life extension protein scaffold displayed enhanced pharmacokinetic and pharmacodynamic properties in preclinical mouse models. These protein engineering modifications will enable us to select an appropriate development candidate with enhanced pharmaceutical properties to achieve the full therapeutic potential of IL-18 to assess in patients with cancer.

Citation Format: Jean Chamoun, Clifford DiLea, Kristiana Dreaden, Pinar Gurel, Joshua Heiber, Robert G. Newman, Su-Ping Pearson, Chunhua Wang, Yanchun Zhao, Mark Whitmore. Interleukin-18 engineered for resistance to IL-18 binding protein (IL-18BP) and half-life extension to enhance its therapeutic potential [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 4076.
Abstract 4139: Prolyl 4-hydroxylase subunit alpha 1 acts as a novel target for lung adenocarcinoma brain metastasis
Authors: He Yan , Luo Zhenyu , Dong Yucheng , Wan Rui , Zhang Xue , Bai Hua , Wang Jie
Published: March 2024

Lung cancer is the most lethal cancer worldwide, with a high incidence of distant metastasis. Many lung adenocarcinoma (LUAD) patients experience brain metastasis during the disease process with unfavorable prognosis. Our previous study has discovered that P4HA1 was exclusively upregulated in brain metastatic cells. Through the analysis of single-cell RNA sequencing of LUAD samples, we found that the expression of P4HA1 in brain metastatic tumor cells is higher than that in primary tumor cells. This finding was further confirmed by immunohistochemical staining. Significantly, a strong presence of P4HA1 in primary tumors is associated with a reduced duration of disease-free survival in individuals with LUAD. Overexpression of P4HA1 in A549 and PC9 cell lines significantly increased the incidence of brain metastasis in BALB/c nude mice, whereas knockdown of P4HA1 in high brain metastatic cells significantly decreased the incidence of brain metastasis and reduced the tumor foci in brain with improved survival. Utilization of DHB, a P4HA1 inhibitor, exerts both therapeutic and preventive effects in relieving brain metastases in nude mice. Furthermore, remarkable inhibition of tumor growth by DHB was observed in patient derived xenograft (PDX) models. RNA sequencing analysis demonstrated that the reduction of P4HA1 expression had a substantial impact on cell adhesion and extracellular matrix (ECM)-receptors interactions. Western blotting analysis verified that the decrease in P4HA1 expression led to a drop in the expression of adhesion molecules ICAM1, VCAM1, NCAM1, as well as integrin molecules ITGA2 and ITGA3. Furthermore, when endothelial cells were co-cultured with conditioned medium from tumor cells, it was observed that the expression of the tight junction molecule ZO1 was reduced after co-culturing with conditioned medium from P4HA1 knockdown A549 cells. The results from the in vitro blood-brain barrier (BBB) model and in vivo biofluorescence imaging indicate that P4HA1 promoted brain metastasis of LUAD via modulating the ability of tumor cells to cross the blood-brain barrier. In conclusion, our study reveals that overexpression of P4HA1 promotes metastatic spread to the brain, and targeting P4HA1 could be a promising therapeutic strategy for the treatment of lung adenocarcinoma brain metastasis.

Citation Format: Yan He, Zhenyu Luo, Yucheng Dong, Rui Wan, Xue Zhang, Hua Bai, Jie Wang. Prolyl 4-hydroxylase subunit alpha 1 acts as a novel target for lung adenocarcinoma brain metastasis [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 4139.
Abstract 414: Accelerating DDR related drug discovery through a customized cell panel
Authors: Huang Cong , Li Xiaoqing , Li Zhengtai , Tan Yan , Pan Huawei , Zhou Lizhen , Mao Kejun , Qin Yanyan , An Zhida , Song Wanxiao , Li Yifan , Jin Shuyu , Bing Tiejun
Published: March 2024

DNA damage is a major threat to cell survival. Not properly repaired DNA damage can lead to cell senescence, apoptosis or tumor development. DDR (DNA Damage Repair) is a collection of bioprocesses which cells utilize in order to identify and correct the DNA damage of the genome. The usage of various compounds for targeting DNA damage responses as well as attenuating DNA repair is also an approach for cancer therapy that has been widely studied in recent years. Despite the success of drug discovery of DDR related inhibitor, drug resistance has become an emerging issue of the field. Here we have constructed multiple cancer cell lines which are either DDR gene, such as XRCC1 and FANCD2, knock-out or resistant to certain DDR inhibitors. Together with the corresponding parental cell lines, we have generated a DDR cell panel which covers 11 different cancer types. We have first validated those DDR inhibitor sensitive or resistant cell lines with functional cell proliferation assays. Additionally, RNAseq has been performed for all constructed resistant cell lines. In combination with a thorough bioinformatic analysis using our in-house generated algorithms, we can provide the genetic background information of the resistant cell lines and top features that potentially contribute to the resistance mechanism. The representative data in this study has shown that LINC02709, BDKRB1, and PRKCQ are the top featured genes in A375 Vemurafenib resistant cell. Meanwhile, cell cycle, ECM-receptor interaction, and DNA replication are the most enriched KEGG terms. Lastly, we have validated this cell panel against multiple inhibitors targeting different DDR vital proteins such as ATM, PARP, POLQ etc. The result has shown that our unique DDR cell panel is capable of providing fast and comprehensive evaluation of DDR related inhibitors, thus facilitate faster and more efficient discovery of DDR inhibitors in the cancer therapy field.

Citation Format: Cong Huang, Xiaoqing Li, Zhengtai Li, Yan Tan, Huawei Pan, Lizhen Zhou, Kejun Mao, Yanyan Qin, Zhida An, Wanxiao Song, Yifan Li, Shuyu Jin, Tiejun Bing. Accelerating DDR related drug discovery through a customized cell panel [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 414.
Abstract 4189: BALB/c-hIL12RB1/hIL12RB2: A robust model for preclinical research in human IL-12 therapy
Authors: Fang Yuan , Wang Huiyi , Xing Jun , Yang Lu , Zhao Jing , Gao Xiang , Ju Cunxiang
Published: March 2024

Interleukin-12 (IL-12) is a multifaceted cytokine primarily produced by antigen-activated dendritic cells, macrophages, and neutrophils. Its receptor is a composite of IL12RB1 and IL12RB2, and the union of these two subunits forms a high-affinity receptor complex proficient in mediating IL-12 signaling. As a pivotal regulator of innate and adaptive immunity, IL-12 plays a crucial role in steering the differentiation of naïve T cells into Th1 cells. It is renowned as a T cell stimulatory factor, fostering T cell proliferation. Furthermore, IL-12 has the capacity to augment the activation of cytotoxic lymphocytes and natural killer cells (NK), thereby enhancing interferon (IFN-γ) production. Empirical evidence consistently demonstrates that IL-12 synergistically exerts anti-tumor effects in conjunction with various immune cells, such as T cells and NK cells. Its effectiveness in preclinical studies has solidified IL-12 as an appealing drug target, leading to the development of activating antibodies, therapeutic IL-12 proteins, and tumor vaccines as potential treatments.

To delve deeper into the functionality and therapeutic potential of IL-12, we have engineered a humanized mouse model, known as BALB/c-hIL12RB1/hIL12RB2. Notably, this model fully preserves the intracellular domains of mouse IL12RB1 and IL12RB2 proteins, ensuring the normal propagation of intracellular signaling. It also effectively expresses human IL12RB1 and IL12RB2. In vitro functional validation experiments have verified that BALB/c-hIL12RB1/hIL12RB2 mice can proficiently generate IFN-γ when induced by human IL-12.

Furthermore, in vivo efficacy experiments have yielded compelling results. Intratumoral administration of human IL-12 mRNA in BALB/c-hIL12RB1/hIL12RB2 mice led to a significant suppression in the growth of CT26 tumors, culminating in complete tumor regression. Subsequent rechallenge experiments conducted after the discontinuation of treatment revealed no tumor recurrence, signifying the enduring inhibitory effect of hIL12 mRNA on mouse tumors.

In conclusion, the development of BALB/c-hIL12RB1/hIL12RB2 mice stands as a robust model for advancing preclinical research in the realm of human IL-12 therapy.

Citation Format: Yuan Fang, Huiyi Wang, Jun Xing, Lu Yang, Jing Zhao, Xiang Gao, Cunxiang Ju. BALB/c-hIL12RB1/hIL12RB2: A robust model for preclinical research in human IL-12 therapy [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 4189.