bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 180: LINC00261 is functionally linked to tumor DNA mutational burden and tumor immune microenvironment composition in lung adenocarcinoma
Authors: Castillo Jonathan , Xue Tianchun , Norwood Mayela , Joseph Samantha , Epstein Alan L. , Wallace William D. , Kim Anthony W. , Marconett Crystal N.
Published: March 2024

Immunotherapy has emerged as a breakthrough in the improvement of survival outcomes for lung adenocarcinoma (LUAD), however effective response requires the combination of blocking inhibitory signals on the tumor surface and antigen presentation to the tumor surface for proper immune recognition. Several commercially available and robust methods exist for identification of tumors displaying immune-inhibitory surface receptors, such as PD-L1, however it is currently difficult to predict effectiveness of antigen presentation on the cell surface. To address this, we utilized clinical and next-generation sequencing data from The Cancer Genome Atlas (TCGA) to identify gene signatures that are correlated to tumor mutational burden (TMB) within cancers of epithelial origins as a surrogate for neoantigen signatures. We identified LINC00261 as a top gene correlated to TMB, whose expression activates DNA damage response pathways in vitro along with resistance to cisplatin. LINC00261 expression was also significantly correlated to MHC class I and II genes involved in endogenous neoantigen presentation expression within the TCGA-LUAD cohort. This relationship was confirmed in vitro through ectopic reintroduction of LINC00261 for key MHC class II presentation genes. Interferon gamma-induced MHC gene activation in vitro was also able to induce endogenous expression of LINC00261. Staining of primary human lung cancer sections suggested that loss of LINC00261 is associated with an immunosuppressive tumor microenvironment. Taken together, our results suggest there is a mechanistic relationship in the silencing of LINC00261 in LUAD and compromised DNA repair, accumulation of mutations, and reduced antitumor immune response.

Citation Format: Jonathan Castillo, Tianchun Xue, Mayela Norwood, Samantha Joseph, Alan L. Epstein, William D. Wallace, Anthony W. Kim, Crystal N. Marconett. LINC00261 is functionally linked to tumor DNA mutational burden and tumor immune microenvironment composition in lung adenocarcinoma [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 180.
Abstract 1820: Investigating the anticancer effects and pharmacological mechanisms of phenanthridine Amaryllidaceae alkaloid in bladder cancer
Authors: Su Yu-Chi , Chen Mei-Chuan
Published: March 2024

Bladder cancer (BC) ranks as the tenth most common malignancy worldwide. Despite the improved five-year survival rate of around 80% for bladder cancer patients, 25% of cases progress to muscle-invasive bladder cancer with associated metastases, leading to a notable five-year survival of 5%. This underscores the urgent clinical necessity for the development of novel therapeutic strategies in the management of bladder cancer. In this study, we demonstrate the anticancer potential of a phenanthridine Amaryllidaceae alkaloid (PAA), and investigate its underlying mechanisms in bladder cancer. The data demonstrates the selective cytotoxicity of PAA towards bladder cancer cells, revealing IC50 values of 2.47 μM in RT112 cells and 8.99 μM in normal uroepithelial SV-HUC-1 cells. Protein array analysis further showed that PAA attenuated phosphorylation of several kinases, which functions as critical regulators of cell survival and proliferation. PAA induces S phase retardation and increases the phosphorylation of ATM, potentially enhancing DNA damage capability. Furthermore, PAA induced apoptosis, as evidenced by the accumulation of sub-G1 cells and caspase activation. In summary, our findings demonstrate that PAA exerts diverse effects on cell cycle regulation and apoptosis. These results suggest that PAA holds promise as a potential candidate for the treatment of bladder cancer.

Citation Format: Yu-Chi Su, Mei-Chuan Chen. Investigating the anticancer effects and pharmacological mechanisms of phenanthridine Amaryllidaceae alkaloid in bladder 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 1820.
Abstract 1830: Bergenin inhibits growth of human cervical cancer cells by decreasing galectin-3 and MMP-9 expression
Authors: Singh Mayank , Chauhan Ravi , Malhotra Lakshay , Gupta Ashna , Dagar Gunjan , Das Dayasagar , Uddin Shahab , Samath Ethayathulla Abdul , Macha Muzafar A. , Akil Ammira Al shabeeb , Bhat Ajaz A.
Published: March 2024

Cervical cancer is still the leading of cancer mortality worldwide, even after introducing a vaccine against Human papillomavirus (HPV), due to low vaccine coverage, especially in the developing world. Cervical cancer is primarily treated by Chemo/Radiotherapy, depending on the disease stage, with Carboplatin/Cisplatin-based drug regime. These drugs being non-specific, target rapidly dividing cells, including normal cells, so safer options are needed for more targeted therapy and lower off-target toxicity. Natural products offer an attractive option compared to synthetic drugs due to their well-established safety profile and capacity to target multiple oncogenic hallmarks of cancer like inflammation, angiogenesis, etc. In the current study, we investigated the effect of Bergenin (C-glycoside of 4-O-methylgallic acid), a natural polyphenol compound that is isolated from medicinal plants such as Bergenia crassifolia, Caesalpinia digyna, and Flueggea leucopyrus. Bergenin has been shown to have anti-inflammatory, anti-ulcerogenic, and wound healing properties. Still, its anticancer potential has been realized only recently. We performed a proteomic analysis of cervical carcinoma cells treated with bergenin and found it to influence multiple hallmarks of cancers, including apoptosis, angiogenesis, and tumor suppressor proteins. It was also involved in many different cellular processes unrelated to cancer, as shown by our proteomic analysis. Further analysis showed bergenin to be a potent -angiogenic agent by reducing key angiogenic proteins like Galectin 3 and MMP-9 (Matrix Metalloprotease 9) in cervical carcinoma cells at cellular level as shown by western blotting assay. Further understanding of this interaction was carried out using molecular docking analysis, which indicated MMP-9 has more affinity for Bergenin as compared to Galectin-3. Cumulatively, our data provide novel insight into the anti-angiogenic mechanism of Bergenin in cervical carcinoma cells by modulation of multiple angiogenic proteins like Galectin-3 and MMP-9 which warrant its further development as an anticancer agent in cervical cancer.

Citation Format: Mayank Singh, Ravi Chauhan, Lakshay Malhotra, Ashna Gupta, Gunjan Dagar, Dayasagar Das, Shahab Uddin, Ethayathulla Abdul Samath, Muzafar A. Macha, Ammira Al shabeeb Akil, Ajaz A. Bhat. Bergenin inhibits growth of human cervical cancer cells by decreasing galectin-3 and MMP-9 expression [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 1830.
Abstract 1835: Discovery of HSN003839, a highly potent inhibitor of ubiquitin-specific protease USP21 for cancer therapy
Authors: Liu Xiaohui , Wang Ying , Xia Dongliang , Zhang Wei , He Yan , Tian Shijie , Feng Xuerong , Yang Xia , Zhang Qi , Liu Fei , Jiang Shiyi , Hu Ran , Tang Ming , Tang Xuelin , Liu Yuqing , Yuan Weijiao , You Xuedan
Published: March 2024

USP21 belongs to the ubiquitin-specific protease (USP) subfamily of deubiquitinating enzymes (DUBs), which catalyze the removal of ubiquitin chain from targeted proteins. Numerous efforts over the last decade have identified a wide range of USP21 substrates and revealed the momentous and multifaceted role of USP21 in physiological and pathological states, especially in tumorigenesis, indicating that USP21 is an appealing target for the therapy of many correlative diseases. Nevertheless, progress in the development of potent USP21 inhibitors remains limited. Herein, we present the discovery of HSN003839, a highly potent and selective USP21 inhibitor with excellent anticancer activity and drug-like properties. Following screening tool compounds to identify hits and subsequent optimization, we discovered a series of compounds with strong enzymatic inhibition for USP21 with IC50 < 100 nM. Among them, HSN003839 displayed low nanomolar cancer cells proliferation inhibitions and excellent ADMET properties. The mouse oral bioavailability (F) is 76.5% at 10 mg/kg. In a preliminary mouse CDX model, monotherapy of HSN003839 showed 105% tumor growth inhibition (TGI) at 30 mg/kg with no obvious body weight reduction. Further biological characterization, efficacy and toxicity studies are ongoing.

The structure of HSN003839 was not presented and will not be disclosed at the time of presentation at AACR meeting.

Citation Format: Xiaohui Liu, Ying Wang, Dongliang Xia, Wei Zhang, Yan He, Shijie Tian, Xuerong Feng, Xia Yang, Qi Zhang, Fei Liu, Shiyi Jiang, Ran Hu, Ming Tang, Xuelin Tang, Yuqing Liu, Weijiao Yuan, Xuedan You. Discovery of HSN003839, a highly potent inhibitor of ubiquitin-specific protease USP21 for cancer 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 1835.
Abstract 1851: Comprehensive multi-level proteomics characterization of gastric cancer
Authors: Wang Yuefan ,   Clinical Proteomic Tumor Analysis Consortium (CPTAC)
Published: March 2024

Gastric cancer is the leading cause of cancer related deaths worldwide, accounts for over 700,000 deaths each year. There is an urgent need for advancing therapeutic strategies for patients with gastric cancer. Addressing this need, the Clinical Proteomics Tumor Analysis Consortium (CPTAC) has conducted a comprehensive characterization of gastric cancer, aiming to elucidate the complex molecular mechanisms driving this cancer type, and to discover underlying cancer biology that will form the basis for developing new approaches for precision medicine. This comprehensive study includes an integrated multi-level proteomics analysis of 165 surgically resected gastric cancer and 41 normal adjacent tissues. We employed a multi-faceted approach, analyzing the proteome, phosphoproteome and glycoproteome, along with other protein modifications including ubiquitination, acetylation, and tyrosine phosphorylation. Furthermore, we extended our research to include 74 pairs of early-onset gastric cancer and normal adjacent tissues, analyzed using a proteomic approach. We can totally identify more than 12,000 protein groups with median identification of 9750 protein groups per sample. This expansive dataset helps constructing a comprehensive multi-omics atlas of gastric cancer, offering a deep understanding into the disease's molecular landscape. The depth of this study was enhanced by incorporating multi-level proteomics data across 6 tiers, facilitating an unprecedented multi-level proteomics analysis. Our integrated approach allows us to illustrate the changes in protein expression, protein modifications, signaling pathways, and protein networks during tumorigeneis. This analysis uncovers potential therapeutic targets for personalized treatments. Overall, our study provides a multi-omics atlas for gastric cancer, a profound insight into gastric cancer biology, contributing significantly to the understanding of its pathogenesis. It could pave the way for precise treatments for gastric cancer, underscoring the potential of multi-level proteomics analyses in advancing personalized medicine.

Citation Format: Yuefan Wang, Clinical Proteomic Tumor Analysis Consortium (CPTAC). Comprehensive multi-level proteomics characterization of 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 1851.
Abstract 1868: Target-dependent considerations for the design of bispecific T-cell engagers
Authors: Mai Matt , Tonikian Raffi , Bergqvist Peter , Amash Alaa , Blamey Nathalie , Conaghan Gabrielle , de Puyraimond Valentine , Farber Patrick , Goodman Allison , Lee Ahn , Scortecci Jessica Fernandes , Crichlow Cindy-Lee , Khodabandehloo Akram , Pinsky Tova , Caldwell Kate , Patterson Jessica , Pouliot Philippe , Tortora Davide , Urtatiz Oscar , Xiang Ping , Yu Irene , Brown Kirstin , Bullock Kelly , Chee Andrea , Masterman Stephanie K. , Aubuchon Neil , Devorkin Lindsay , Barnhart Bryan C. , Jacobs Tim
Published: March 2024

In this study, we present data from four T-cell engager (TCE) programs in which we leveraged our highly diverse CD3-binding antibodies to generate functional bispecifics for each tumor target. By aggregating tumor-cell killing and cytokine data across four tumor targets with different properties and target expression levels, we have gained novel insights into target-specific considerations for the development of CD3 TCEs. TCEs recapitulate a synapse between T cells and target cells. T cell function downstream of this synapse is determined by the complex interplay between multiple independent factors. These include properties of the CD3- and tumor-binding arms, such as binding kinetics and epitope, as well as target-dependent factors such as cell type and target density. We used our diverse CD3 antibodies to engineer bispecific TCEs against four solid tumor targets: PSMA, B7-H4, 5T4, and the peptide-MHC target MAGE-A4. For each target, we engineered and characterized hundreds of bispecific molecules using high-throughput assays, including T-cell dependent tumor-cell killing, cytokine release, developability assessments, and structural studies. Here, we compare data from across these programs to elucidate the impact of target-dependent parameters on TCE function. These data provide important insights for the selection of appropriate TCE targets and efficient design of bispecific antibodies with high therapeutic potential.

Citation Format: Matt Mai, Raffi Tonikian, Peter Bergqvist, Alaa Amash, Nathalie Blamey, Gabrielle Conaghan, Valentine de Puyraimond, Patrick Farber, Allison Goodman, Ahn Lee, Jessica Fernandes Scortecci, Cindy-Lee Crichlow, Akram Khodabandehloo, Tova Pinsky, Kate Caldwell, Jessica Patterson, Philippe Pouliot, Davide Tortora, Oscar Urtatiz, Ping Xiang, Irene Yu, Kirstin Brown, Kelly Bullock, Andrea Chee, Stephanie K. Masterman, Neil Aubuchon, Lindsay Devorkin, Bryan C. Barnhart, Tim Jacobs. Target-dependent considerations for the design of bispecific T-cell engagers [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 1868.
Abstract 2111: Impact of HER2 Expression on trastuzumab deruxtecan(T-DXd) efficacy
Authors: Chen Wanyi , Drago Joshua , Gupta Avantika , Liu Bo , Chandarlapaty Sarat
Published: March 2024

Background: T-DXd is an FDA-approved anti-HER2 Antibody Drug Conjugate (ADC), which has shown clinical efficacy in breast cancers with either overexpression or low-level expression of HER2. The benefit of T-DXd across the spectrum of HER2 expression has called into question the relevance of HER2 expression level as relevant parameter for T-DXd sensitivity. We sought to experimentally address the relevance of HER2 expression level upon the degree of T-DXd response.

Methods: A series of isogenic models was generated that expressed different levels of surface HER2 using a doxycycline-inducible promoter. These were analyzed for impact of differential expression level upon T-DXd internalization and drug response. Human breast cancer samples were analyzed for HER2 expression levels and association with clinical response to T-DXd under an IRB approved protocol.

Results: Induction of different levels of HER2 at specified doxycycline concentrations was verified in multiple models using immunoblotting and FACS. These different levels appeared to correspond to clinical breast cancer immunohistochemistry (IHC) scoring levels to 0, 1+, 2+, and 3+. The models were analyzed for T-DXd response and showed increasing levels of HER2 led to increase lysosomal uptake of T-DXd (p<0.0001). Moreover, elevation of HER2 levels led to lower concentrations of T-DXd needed to suppress tumor growth. Analyses of the effects of increasing levels of HER2 upon xenograft response to T-DXd are ongoing. Finally, retrospective analyses of a cohort of 50 breast cancer patients treated with T-DXd revealed that higher levels of HER2 corresponded to longer time on treatment (time to progression).

Conclusion: We observe a consistent relationship between elevated expression of HER2 and breast cancer response to T-DXd. The findings highlight the relevance of HER2 expression level as a biomarker of T-DXd response and support the potential value of therapeutic strategies that can induce HER2 surface expression levels to enhance response.

Citation Format: Wanyi Chen, Joshua Drago, Avantika Gupta, Bo Liu, Sarat Chandarlapaty. Impact of HER2 Expression on trastuzumab deruxtecan(T-DXd) efficacy [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 2111.
Abstract 2232: Association between sleep characteristics and cancer survival: Findings from the UK Biobank study
Authors: Che Jiajing , Lv Jiali , Pan Keyu , Sun Tiantian , Cao Chao , Li Shengxu , Zhang Tao , Xue Fuzhong , Yang Lin
Published: March 2024

Background

Sleep characteristics have been associated with patient outcomes after cancer. We aimed to appraise the causal relationship of chronotype, sleep duration, insomnia, and snoring with cancer survival.

Methods

This large prospective cohort study enrolled 38350 cancer survivors aged ≥40 years from the UK Biobank between 2004 and 2022. Habitual sleep characteristics were obtained from the self-reported baseline questionnaire, including chronotype, sleep duration, insomnia, and snoring. Cox proportional hazards regression models were applied to evaluate the association between sleep characteristics and cancer survival, then one sample Mendelian randomization (MR) analyses were further used to validate these causal relationships. In particular, restricted cubic splines (RCS) and non-linear MR analyses were applied to assess the possible nonlinear association between sleep duration and cancer survival.

Results

Among 38350 study participants (mean [SD] age, 59.8 [7.3] years; 24150 [63.0%] males; 35534 [92.7%] White individuals), 12671 (33.1%) reported evening preference chronotype; 9050 (23.6%) reported short and 3773 (9.8%) reported long sleep duration;12274 (32.0%) reported snoring; and 30426 (79.4%) reported experiencing insomnia sometimes or usually. During up to 18 years of follow-up, 6274 deaths occurred. Multivariable regression models showed that evening preference (HR = 1.09, 95% CI: 1.03 to 1.15), short (HR = 1.14, 95% CI: 1.07 to 1.21) and long sleep duration (HR = 1.37, 95% CI: 1.28 to 1.48), insomnia (HR = 1.14, 95% CI: 1.09 to 1.21) and snoring (HR = 0.83, 95% CI: 0.78 to 0.87) were associated with death among pan to cancer survivors. Restricted cubic splines suggested nonlinear associations between sleep duration and death after cancer (nonlinear P < 0.001). In the one-sample MR, genetically predicted sleep duration (HR = 0.71, 95% CI: 0.56 to 0.90) and snoring (HR = 1.18, 95% CI: 1.05 to 1.34) demonstrated a reversed cancer survival pattern compared to multivariable regressions. A nonlinear U-shape association between sleep duration and cancer survival was observed in restricted cubic spline analyses (P < 0.001) but not evident when evaluating by nonlinear MR (P > 0.05).

Conclusions

Evening preference chronotype, insomnia, snoring, and short sleep duration are likely to be causally associated with a higher risk of death among cancer survivors, whilst long sleep duration does not appear to be a causal factor.

Citation Format: Jiajing Che, Jiali Lv, Keyu Pan, Tiantian Sun, Chao Cao, Shengxu Li, Tao Zhang, Fuzhong Xue, Lin Yang. Association between sleep characteristics and cancer survival: Findings from the UK Biobank study [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 2232.
Abstract 2327: MADVAR: An algorithm that improves the relevance of computational biology output, while reducing compute time and space requirements
Authors: Silberberg Gilad , Ritchie Michael
Published: March 2024

High-throughput methods implemented in biology research produce a continuously growing array of data input that are used to produce data output with an increasing abundance of features. While growth in the volume and diversity of data input can be highly valuable for studying biological systems, it presents the challenge of managing enormous quantities of features, many of which are not relevant to the specific research question being asked. This excess data input burdens storage and computation of downstream clustering and machine learning tasks. A common approach used to manage this data input relies on filters applied to the features by their variance across the sample set, while applying random cutoffs. Our proprietary algorithm (“MADVAR”) enables the prioritization of variable features from high-throughput continuous data, by automatically finding an optimal cutoff for the distribution of the data. Based on the right-skew nature of biological data distribution, MADVAR finds and excludes the "0 variance peak" using the median of the distributions and the median absolute deviation (MAD). MADVAR enables a faster analysis with a reduced memory requirement, and dramatically improves clustering results with minimal loss of relevant features.

Citation Format: Gilad Silberberg, Michael Ritchie. MADVAR: An algorithm that improves the relevance of computational biology output, while reducing compute time and space requirements [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 2327.
Abstract 2333: Methylation based phylogeny and timing in cancer
Authors: Guteneva Nataliya , Lichter Eliezer , Leshchiner Ignaty
Published: March 2024

Knowing the chronological sequence of genetic alterations during cancer development and progression is crucial for understanding the process of transformation of normal tissue into malignant tumors. This can offer insights into the etiology and intrinsic cellular mechanisms of the process, presenting opportunities for early detection and treatment of initial disease stages, as well as highlight key events as potential therapeutic. Current methods of establishing progression histories of cancers rely either on direct sequencing of premalignant lesions, which are seldom available for tumors or on computational inference timing methods capable of reconstructing the history from primary tumor data. These methods require genome or exome sequencing data from a relatively large cohort of tumors and provide limited resolution due to overall scarcity of somatic mutations. We utilize a different source of genetic information, genome methylation, that also “records” historic information about acquisition and development of mutations. Over 28 million methylated CpG sites exist in the genome with several percent of them differentially methylated (DMS) between tissue types and tumor vs adjacent normal tissue. The several orders of magnitude higher density of distinctly methylated sites provides unprecedented resolution, needs fewer tumors, can be merged with driver mutation data and co-analyzed with “methylation clock” information that allows to establish early development, phylogeny and progression at unprecedented resolution. We developed methods for whole genome bisulfite sequencing (WGBS), reduced representation bisulfite sequencing (RRBS) and Oxford Nanopore Technologies native methylation sequencing data. Somatic DMS sites have distinct properties from somatic point mutations, can appear and disappear in the process of tumor evolution and development allowing for unique configuration of methylated islands. Our tools perform single sample and cohort level “timing”, multi-sample phylogeny reconstruction and cancer population reconstruction by integrating methylation and mutation information from the same sample. These new methods will significantly contribute to our understanding of cancer initiation and progression at a resolution not available previously.

Citation Format: Nataliya Guteneva, Eliezer Lichter, Ignaty Leshchiner. Methylation based phylogeny and timing in 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 2333.
Abstract 2608: Hu2G10 antibody-drug conjugates selectively target cleaved-activated Trop-2 in cancer cells and show therapeutic efficacy against multiple human cancers
Authors: Alberti Saverio , Trerotola Marco , Moschella Antonino , Giavazzi Raffaella , Guerra Emanuela
Published: March 2024

Trop-2 is a transmembrane calcium signal transducer which activates growth-signaling networks that converge on Akt, ERK, Cyclin D1, NFkB (1). Trop-2 cleavage by ADAM10 was shown to be an activator switch for induction of cancer growth and metastatic diffusion (2). We devised a strategy to obtain monoclonal antibodies (mAb) capable of recognizing ADAM10-exposed sites in cancer cells. We succeeded at obtaining mAb that efficiently bound Trop-2 expressing cancer cells and were able to inhibit cell growth in vitro. We humanized the 2G10 mAb as the IgG1/k Hu2G10 antibody. Hu2G10 binds cancer-specific, cleaved/activated Trop-2 with Kd 10−12 M. On the other hand, it binds uncleaved/wtTrop-2 in normal cells with Kd 3.16 × 10−8 M, thus promising an unprecedented therapeutic index in patients. Antibody-drug conjugates (ADC) of Hu2G10 were obtained using stable linkers, multiple conjugation chemistries and different payloads, among them topoisomerase I inhibitors, bleomycin, RNA polymerase inhibitor, MMAE and calicheamicin. Hu2G10-ADC selectively killed Trop-2-expressing cells in vitro. Dose-response curves were obtained for the BxPC-3 pancreatic cancer, DU-145 prostate cancer, HT29 colon cancer cell lines and KM12SM/Trop2 colon cancer transfectants, reaching EC50 values as low as 5.05 × 10−10 M. Hu2G10-ADC drove high anticancer effectiveness on cell- and patient-derived xenografts from colon, pancreas, prostate and ovarian cancer. Effectiveness followed a gradient of payload potency, with correspondingly high therapeutic indexes. Consistently, neither detectable toxic effects nor weight loss were observed in any subgroup of treated xenotransplant-bearing mice. Taken together these findings candidate anti-Trop-2 Hu2G10-ADC for best-in-class anticancer therapy. (1) E. Guerra et al., The Trop-2 signaling network in cancer growth. Oncogene 32, 1594-1600 (2013).(2) M. Trerotola et al., Trop-2 cleavage by ADAM10 is an activator switch for cancer growth and metastasis. Neoplasia 23, 415-428 (2021).

Citation Format: Saverio Alberti, Marco Trerotola, Antonino Moschella, Raffaella Giavazzi, Emanuela Guerra. Hu2G10 antibody-drug conjugates selectively target cleaved-activated Trop-2 in cancer cells and show therapeutic efficacy against multiple human cancers [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 2608.
Abstract 2698: TCF-1 is a critical regulator of intraepithelial lymphocytes in colorectal carcinoma
Authors: Yakou Marina H. , Ghilas Sonia , Tran Kelly , Liao Yang , Afshar-Sterle Shoukat , Kumari Anita , Schmid Kevin , Dijkstra Christine , Inguanti Chantelle , Ostrouska Simone , Wilcox Jordan , Smith Maxine , Parathan Pavitha , Allam Amr , Xue Hai-Hui , Belz Gabrielle T. , Mariadason John M. , Behren Andreas , Drummond Grant R. , Ruscher Roland , Williams David S. , Pal Bhupinder , Shi Wei , Ernst Matthias , Raghu Dinesh , Mielke Lisa A.
Published: March 2024

Tissue-resident immune cells reside within the gastrointestinal tract known as intraepithelial lymphocytes (T-IELs), including αβ and γδ T-IELs. This unique population of T cells constantly survey and are critical in maintaining the gastrointestinal epithelium. We show that T-IELs in various regions of the gastrointestinal tract have distinct features. T-IELs in the small intestine exhibit high expression of cytotoxic molecules important for cancer defense while colon T-IELs are distinctively regulated by the transcription factor, TCF-7/TCF-1, which suppresses their effector and cytotoxic properties, including reduced expression of granzymes, and their abundance is dependent on the microbiome. Targeted deletion of TCF-1 in γδ T-IELs using pre-clinical mouse models resulted in a distinct effector profile and reduced colon tumor burden. Furthermore, TCF-1 expression was significantly reduced in γδ T-IELs present in human colorectal cancers compared with normal healthy colon, strongly correlating with an enhanced γδ T-IEL effector phenotype and improved patient survival. Further investigation aims to elucidate diverse mechanisms by which TCF-1 controls tissue-specific anti-tumor immunity and uncover the important factors controlling TCF-1 expression. Our study underscores the necessity to consider tumor and organ microenvironment-specific factors in optimizing immunotherapy, advancing our understanding of T cell function in the colon to pave the way for innovative CRC treatments.

Citation Format: Marina H. Yakou, Sonia Ghilas, Kelly Tran, Yang Liao, Shoukat Afshar-Sterle, Anita Kumari, Kevin Schmid, Christine Dijkstra, Chantelle Inguanti, Simone Ostrouska, Jordan Wilcox, Maxine Smith, Pavitha Parathan, Amr Allam, Hai-Hui Xue, Gabrielle T. Belz, John M. Mariadason, Andreas Behren, Grant R. Drummond, Roland Ruscher, David S. Williams, Bhupinder Pal, Wei Shi, Matthias Ernst, Dinesh Raghu, Lisa A. Mielke. TCF-1 is a critical regulator of intraepithelial lymphocytes in colorectal carcinoma [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 2698.
Abstract 2700: Therapeutic IL-33 enhances the antigen presentation function of ILC2s to exert anti-tumor immune activity
Authors: Liu Jie , Feng Zhenchu , Wei Hongyan , Li Chang , Li Xiaoshi , Chen Wenlong , Ke Yixi , Wang Minjun , Liu Lantian , Zheng Cunni , Liu Quan
Published: March 2024

IL-33 regulates the function of ILC2s through ST2. However, the role of IL-33, especially exogenous IL-33, on ILC2s in tumors is controversial depending on the cytokines they secrete. Here we report that exogenous, but not endogenous, IL-33 significantly inhibits the progression of a variety of mouse solid tumors, including melanoma, colorectal cancer, lung adenoma, and breast cancer. By inoculating tumor cells in Il1rl1−/- and NSG mice, we found that the tumor-suppressive effect of IL-33 on tumors was dependent on the expression of ST2 receptors and the presence of lymphocytes. Regarding tumor-infiltrating immune cells, exogenous IL-33 significantly increases the proportions of ST2+ Tregs (1.10 ± 0.20% vs. 1.74 ± 0.36%), eosinophils (6.00 ± 1.33% vs. 22.73 ± 6.51%), Granzyme B+ CD8+ T cells (4.32 ± 2.77% vs. 11.53 ± 6.81%), and bulk ILC2s (0.08 ± 0.04% vs. 5.31 ± 1.33%) and IA/IE+ ILC2s (0.02 ± 0.01% vs. 3.67 ± 1.03%). In draining lymph nodes, IL-33 also significantly increases the proportions of ILC2s (0.03 ± 0.02‰ vs. 6.37 ± 1.71‰) and IA/IE+ ILC2s (0.003 ± 0.003‰ vs. 4.38 ± 1.21‰). Importantly, single-cell transcriptomic analysis on tumor-infiltrating ILC2s highlights the expression of genes related with antigen processing and presentation including H2-K1, H2-Q7, B2m, Hspa1a, Hspa1b, Creb1, Hspa4, Tap1 and Tap2. We testified this finding by DQ-OVA experiment and found that IL-33 could promote ILC2s to up-take and process exogenous antigens (17.78 ± 0.75 fold vs. 22.02 ± 0.75 fold). The antigen presentation function of ILC2s was further established by mixed lymphocyte culture where BALB/c ILC2s potently stimulated C57BL/6 CD8+ T cell proliferation, which was further augmented by IL-33 treatment. Taken together, our data demonstrate an anti-tumor function of therapeutic IL-33 that elevates the number and antigen presentation function of ILC2s.

Citation Format: Jie Liu, Zhenchu Feng, Hongyan Wei, Chang Li, Xiaoshi Li, Wenlong Chen, Yixi Ke, Minjun Wang, Lantian Liu, Cunni Zheng, Quan Liu. Therapeutic IL-33 enhances the antigen presentation function of ILC2s to exert anti-tumor immune activity [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 2700.
Abstract 2703: Targeting STING pathway as a therapeutic strategy against β-catenin driven immune escape and resistance to anti-PD1 therapy in hepatocellular carcinoma
Authors: Zhang Qingmei , Bi Jiacheng , Yeung Oscar Wai-Ho , Ding Tao , Chan Wing-Lim , Zhu Yueqin , Liu Jiang , Ng Kevin Tak-Pan , Man Kwan
Published: March 2024

β-catenin mutation occurs in around 40% of hepatocellular carcinoma(HCC); yet no precision therapies are available to treat this tumor subset. Immune checkpoint inhibitors (ICIs) are now widely used for the treatment of patients with advanced HCC. Unfortunately, preliminary evidence showed that β-catenin mutation drives resistance to anti-PD1 immunotherapy in HCC. In this study, we studied the molecular mechanisms underlying β-catenin mutation during HCC pathogenesis to develop novel therapeutics targeting β-Catenin-driven HCC. Firstly, three HCC cohorts were examined, including 70 untreated, 20 treated patients with nivolumab, and patients from a public TCGA database. By single-cell RNA sequencing analysis, we identified that anti-PD-1 treatment displayed effectiveness with the increase of CD27+CD8+ T-cell infiltration, correlating to improved prognosis with longer progression-free survival (PFS) and OS. Importantly, multiplex IHC staining showed that β-catenin activation is associated with significantly reduced CD8+CD27+ T cell infiltration in tumors (n=70), demonstrating that β-catenin promoted immune escape and resistance to anti-PD-1 treatment involved defective cytotoxic T-cell activity. Subsequently, we found that the interaction of β-Catenin with sirtuin 1 (SIRT1) mediated suppression of IRF3/IRF7 activity, resulting in inhibition of Type I interferon (IFN-I) production. However, by CRISPRi screening, we found that disruption of β-catenin-SIRT1 interaction via STING caused persistent high-level IFN-I response which increased the transcription of T-cell costimulatory molecule CD27 via IL10/STAT3/BATF signaling on infiltrated CD8+ T cells. This effect was also confirmed in the STING null mice, emphasizing that STING signaling is a therapeutic target to overcome β-catenin driven immune escape. Further, using hydrodynamic tail-vein injection murine HCC models, we found that the STING agonist inhibited β-catenin-SIRT1 activity, restored IFN signaling to rescue T-cell cytotoxic capacities, and enhanced the anti-PD-1 therapy efficacy. In conclusion, we demonstrated that STING reprogrammed β-Catenin mutated the HCC tumor microenvironment and a combination of STING and anti-PD-1 treatment represents an effective novel therapeutic strategy for HCC patients with β-Catenin mutation.

Citation Format: Qingmei Zhang, Jiacheng Bi, Oscar Wai-Ho Yeung, Tao Ding, Wing-Lim Chan, Yueqin Zhu, Jiang Liu, Kevin Tak-Pan Ng, Kwan Man. Targeting STING pathway as a therapeutic strategy against β-catenin driven immune escape and resistance to anti-PD1 therapy in hepatocellular carcinoma [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 2703.
Abstract 2759: Transcriptional intra-tumoral heterogeneity of putative therapeutic targets in colorectal cancer peritoneal metastases
Authors: Zhao Joseph J. , Ong Johnny CA , Chia Daryl Kai Ann , Teo Melissa Ching Ching , Tan Qiu Xuan , Ng Gillian , Tan Joey Wee-Shan , Ma Haoran , Ong Xuewen , Tay Su Ting , Tan Patrick , Sundar Raghav
Published: March 2024

Background: The poor prognosis of peritoneal metastasis (PM) in colorectal cancer (CRC) is typically attributed to, and characterized by, resistance to systemic chemotherapy and immunotherapy. The evolutionarily conserved plasma-peritoneal barrier as well as reduced tumor tissue vascularity contributes to poorer responses to systemically administered antineoplastic therapy. Beyond these, we sought to understand transcriptomic variations of important therapeutic targets implicated in peritoneal organotropism and by extension, resistance to systemic regimens in CRC PM.

Methodology: FFPE tissue samples from CRC patients were collected during oncological surgical resection of the synchronous PM (cytoreductive surgery [CRS] with or without hyperthermic intra-peritoneal chemotherapy [HIPEC]), or exploratory laparotomy for palliative surgery, with the majority of patients receiving systemic therapy prior to surgery. Samples were sequenced with whole transcriptomic sequencing. Paired and unpaired T-tests between PT and PM samples were undertaken with log2(fragments per kilobase of exon per million mapped [FPKM]) gene expression values.

Results: Fifty-six (22 primary tumor [PT], 34 peritoneal metastases [PM]) samples from 35 patients, comprising 21 paired PT-PM samples, were included in this analysis. TEAD1 (unpaired T-test, p=0.087; paired T-test, p=0.041), FGFR1 (unpaired T-test, p=0.003; paired T-test, p<0.001), PIK3CA (unpaired T-test, p=0.034; paired T-test, p=0.003), SMAD4 (unpaired T-test, p=0.095; paired T-test, p=0.010), and HAVCR2 (unpaired T-test, p=0.009; paired T-test, p=0.007) were found to be upregulated in the PM. Conversely, MET was found to be downregulated in PM samples (unpaired T-test, p=0.004; paired T-test, p=0.002). There were no significant differences in gene expression profiles of BRAF, RHOA, CD274, CTLA4, PDCD1 and TIGIT between PT-PM (all p>0.05).

Conclusion: Through CRC PT-PM comparisons, we identify potential therapeutic targets implicated in transcoelomic metastasis and provide rationale for development of immunotherapeutic biology and drug discovery for CRC PM. These findings underscore an exigent need to develop intra-peritoneal therapeutic strategies to overcome resistance to systemic regimens.

Citation Format: Joseph J. Zhao, Johnny CA Ong, Daryl Kai Ann Chia, Melissa Ching Ching Teo, Qiu Xuan Tan, Gillian Ng, Joey Wee-Shan Tan, Haoran Ma, Xuewen Ong, Su Ting Tay, Patrick Tan, Raghav Sundar. Transcriptional intra-tumoral heterogeneity of putative therapeutic targets in colorectal cancer peritoneal metastases [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 2759.
Abstract 2802: Enterococcus faecalis-derived X facilitates pancreatic tumor by regulating immune microenvironment
Authors: Wang Bin , Yang Chao , Xie Yongjie , Zhou Tianxing , Yu Jun , Hao Jihui
Published: March 2024

Previous research in human samples has found a link between the intratumoral microbiome and the development and progression of pancreatic cancer. Nevertheless, the bacterial strains that may have an effect, and the underlying mechanisms, are largely unknown. To identify potential bacterial species, microbiome sequencing was performed on fecal and tumor samples obtained from KPC mice and patients with pancreatic tumors in this study. To identify metabolites associated with bacteria, untargeted metabolomics were analyzed on serum and tumor samples. Furthermore, advanced and high-throughput techniques, such as single-cell RNA sequencing, tumor organoids, engineered bacteria and germ-free mouse models, were used to elucidate the specific influence on the immune microenvironment of pancreatic tumors. Our findings reveal an increased abundance of Enterococcus faecalis (E. faecalis) in the gut during the development of pancreatic cancer, accompanied by its accumulation in the tumor. Patient-derived E. faecalis strain and its metabolite X, facilitated pancreatic tumor growth, reduced the recruitment of CD8+T cells, and caused their dysfunction in the tumor, relying heavily on E. faecalis intratumoral colonization. We found that X inhibits the production of anti-tumor chemokines like CCL21 and CCL8 in tumor cells, reducing peripheral CD8+ T cell infiltration into the tumor. Furthermore, X was found to inhibit glycolysis and compromise the function of tumor-infiltrating CD8+ T cells by decreasing the activity of lactate dehydrogenase (LDH) in CD8+ T cells. Additionally, oral doxycycline, an antibiotic that targets intratumoral E. faecalis, effectively enhanced the therapeutic effect of gemcitabine in pancreatic cancer. Finally, we present evidence that X may predicts efficacy and survival in pancreatic cancer patients. Together, our findings provide new insights into the role of E. faecalis in mediating the immune environment of pancreatic tumors, highlighting E. faecalis and X as potential therapeutic targets for pancreatic cancer patients.

Citation Format: Bin Wang, Chao Yang, Yongjie Xie, Tianxing Zhou, Jun Yu, Jihui Hao. Enterococcus faecalis-derived X facilitates pancreatic tumor by regulating immune 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 2802.
Abstract 2835: Comparison of xenograft and syngeneic cancer cachexia models for preclinical drug testing
Authors: Draheim Kyle , Gietl Diana , Shinn Karyn , Durham William , Zhao Fei , Thompson Caitlin , Baldwin Alexis , Harris Glenn , Hall Chassidy , Fadden Patrick , Avrutskaya Anya
Published: March 2024

Cancer cachexia is a wasting syndrome characterized primarily by loss of skeletal muscle and adipose tissue mass and contributes greatly to overall cancer-related mortality (for recent review see Argiles 2023). An estimated 25-75% of cancer patients develop cachexia which leads to impacts on therapeutic tolerability, well-being, and eventually overall survival (Bossi 2021). Some preclinical mouse and rat cancer models manifest cachexia as they progress. In this study we established a syngeneic (immune competent) and a xenograft (immune incompetent) tumor model of cachexia with the intention of comparing the models with respect to typical study endpoints and to provide a platform for testing novel therapeutic agents aimed at inhibiting or reversing the effects of cachexia. The Colon26 syngeneic mouse colon adenocarcinoma model was reported as an experimental cancer cachexia model by Tanaka, et. al. in 1990. The model was developed in male BALB/c × DBA/2 F1 (first generation cross) mice. We typically implant Colon26 in female BALB/c mice for preclinical drug testing of anticancer agents and noticed a strong tendency for mice bearing Colon26 tumors to develop signs of cachexia on study, but not always consistently. A review of the literature showed there are clear gender differences with male cancer patients having higher prevalence of cachexia and greater muscle loss compared with female patients (Zhong 2020). Based on this we implanted Colon26 tumors in male BALB/c mice to establish the cachexia model. The HT1080 human fibrosarcoma model HT1080 was characterized as a cachexic model by Bernardo, et. al. in female severe combined immunodeficiency disease (SCID) mice. We developed the HT1080 model in male SCID mice based on the literature and previous experience. Both models were characterized by multiple endpoints including overall body weight loss, skeletal muscle weight loss, reduction of adipose tissue, and cardiac muscle loss, as well as tracking a broad panel of relevant cytokine levels at study endpoint. Both models generate robust wasting of skeletal muscle and adipose tissue providing an opportunity to assess novel anti-cachexia therapies.

Citation Format: Kyle Draheim, Diana Gietl, Karyn Shinn, William Durham, Fei Zhao, Caitlin Thompson, Alexis Baldwin, Glenn Harris, Chassidy Hall, Patrick Fadden, Anya Avrutskaya. Comparison of xenograft and syngeneic cancer cachexia models for preclinical drug testing [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 2835.