bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3077: Integrin expression during ovarian cancer progression
Authors: Bano Nazia , Schmelz Eva M. 
Published: March 2024

Abstract
During peritoneal dissemination, ovarian cancer cells exfoliate from the primary tumor and aggregate in the peritoneal fluid. This elicits survival signals via changes in mitochondrial morphology and function. As spheroids adhere to secondary sites, mitochondrial function is restored in the form of recovery of mitochondrial morphology, increased mitochondrial membrane potential, glucose uptake and increased proliferation. These findings suggested a mitochondrial phenotypic switch from mitophagy in spheroids to upregulation of mitobiogenesis upon spheroids adhesion to secondary sites. Hence, our objective was to identify critical receptors that facilitate the adhesion of spheroids to secondary sites, focusing on integrins that facilitate cell-extracellular matrix (ECM) attachment and activate downstream signaling cascades. To investigate changes in integrin expression, we mimicked three major stages of ovarian cancer progression by developing adherent stage, spheroid stage and adherent spheroid stage using our mouse ovarian surface epithelial (MOSE) cell lines representing benign (MOSE-E) stage as well as slow-developing (MOSE-L) and fast-developing disease (MOSE-LTICv) in biologically relevant conditions. Using RT qPCR and Western blotting, we determined gene and protein expression levels of integrins and adhesion related genes. Confocal microscopy was used to determine protein localization during ovarian cancer progression. RT qPCR result indicated differential up-regulated expression of integrins in both spheroid and adherent spheroid stage, suggesting the role of different integrins during the stages of ovarian cancer progression. Specifically, we observed a decrease (integrin ɑ5, ɑ6) or increase (ɑ2𝛃;1) of integrins after aggregation and an upregulation of 𝛃;1, α2, αV, α3, α5 and immunomodulatory integrins such as αE and αM in adherent spheroids, indicating their role in mediating specific steps during progression of ovarian cancer from tumorigenesis to metastatic implantation. We also observed increased expression levels of extracellular matrix proteins and their modulators, and cell-to-cell adhesion genes during ovarian cancer progression that was more pronounced in the MOSE-LTICv cells which suggests their association with an aggressive cancer phenotype. Overall, this project will aid in the identification of adhesion targets to develop new anti-metastatic treatments that will reduce mortality in women with ovarian cancer.
Citation Format: Nazia Bano, Eva M. Schmelz. Integrin expression during ovarian cancer progression [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 3077.
Abstract 3079: AMPK-HIF-1α signaling enhances glucose-derived de novoserine biosynthesis to promote glioblastoma growth
Authors: Yun Hye Jin , Lee Jong-Ho 
Published: March 2024

Abstract
Cancer cells undergo cellular adaptation through metabolic reprogrammin to sustain survival and rapid growth under various stress conditions. However, it remains unknown how brain tumors modulate their metabolic flexibility in the naturally serine/glycine (S/G)-deficient brain microenvironment. We used a range of primary/stem-like and established glioblastoma (GBM) cell models in vitro and in vivo. To identify regulatory mechanisms of S/G deprivation induced metabolic flexibility, we employed high throughput RNA-sequencing, transcriptomic analysis, metabolic flux analysis, metabolites analysis, chromatin immunoprecipitation (ChIP), luciferase reporter, nuclear fractionation, cycloheximide-chase, and glucose consumption. The clinical significances were analyzed in the genomic database (GSE4290) and in human GBM specimens. The high throughput RNA-sequencing and transcriptomic analysis demonstrate that the de novo serine synthesis pathway (SSP) and glycolysis are highly activated in GBM cells under S/G deprivation conditions. Mechanistically, S/G deprivation rapidly induces reactive oxygen species (ROS)-mediated AMP-activated protein kinase (AMPK) activation and AMPK dependent hypoxia-inducible factor (HIF)-1α stabilization and transactivation. Activated HIF-1α in turn promotes expression of SSP enzymes phosphoglycerate dehydrogenase (PHGDH), phosphoserine aminotransferase 1 (PSAT1), and phosphoserine phosphatase (PSPH). In addition, HIF-1α-induced expression of glycolytic genes (GLUT1, GLUT3, HK2, and PFKFB2) promotes glucose uptake, glycolysis, and glycolytic flux to fuel SSP, leading to elevated de novo serine and glycine biosynthesis, NADPH/NADP+ ratio, and proliferation and survival of GBM cells. Analyses of human GBM specimens reveal that levels of overexpressed PHGDH, PSAT1, and PSPH are positively correlated with levels of AMPK T172 phosphorylation and HIF-1α expression and poor prognosis of GBM patients. Our findings reveal that metabolic stress-enhanced glucose-derived de novo serine biosynthesis is a critical metabolic feature of GBM cells and highlight the potential to target SSP for treating human GBM.
Citation Format: Hye Jin Yun, Jong-Ho Lee. AMPK-HIF-1α signaling enhances glucose-derived de novoserine biosynthesis to promote glioblastoma growth [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 3079.
Abstract 3082: The liver microenvironment promotes glycolysis in metastatic colorectal cancer by activating her3
Authors: Rathore Moeez Ghani , Curry Kimberly , Boutros Christina , Wang Zhenghe , Winter Jordan , Wang Rui 
Published: March 2024

Abstract
Background: Liver metastasis occurs in ~80% of all metastatic colorectal cancer (mCRC) and the liver has a unique endothelial cell (EC)-rich microenvironment that promotes cancer cell survival. We previously reported that liver ECs secrete soluble factors to promote CRC growth via activating CRC-associated the HER3 signaling pathway. The present study reports that ECs promote mCRC growth by shifting cancer metabolism towards glycolysis and elucidates the involved signaling pathway.
Methods: We performed mass spectrometry to profile the metabolic changes in mCRC treated with/without HER3 inhibition in a syngeneic, orthotopic mCRC model. We also performed Seahorse and lactate ELISA assays to determine the effect of liver EC-secreted factors on CRC metabolism. To further determine the role of HER3 in promoting glycolysis and CRC growth in the liver, we used siRNA silencing of HER3 and pharmacological inhibition of HER3 downstream signaling proteins to identify the key mediator(s) of HER3-induced metabolic shift in CRC.
Results: We determined that HER3 inhibition leads to increased oxidative phosphorylation metabolites (OXPHOS) and decreased glycolysis metabolites in CRC liver metastases. Conditioned medium containing EC-secreted factors decreased oxygen consumption (determined by Seahorse, a readout of OXPHOS) and increased lactate secretion (determined by ELISA, a readout of glycolysis) in CRC. As HER3 inhibition reversed the metabolic shift and increased OXPHOS as an alternative survival strategy, we found that simultaneous inhibition of HER3 and OXPHOS synergistically blocked CRC cell and tumor growth. Lastly, we found that PFK2 is activated by HER3 signaling, determined by phosphorylation, and determined that the HER3-AKT-PFK2 signaling axis is the key mediator of EC-induced metabolic shifts.
Conclusions: We identified that the liver microenvironment activates HER3 and the downstream AKT-PFK2 signaling to promote glycolysis and growth in CRC liver metastases by activating AKT-PFK2-dependent glycolysis. Our findings highlighted the potential of combining HER3 and OXPHOS inhibitions as a potential strategy for treating patients with mCRC.
Citation Format: Moeez Ghani Rathore, Kimberly Curry, Christina Boutros, Zhenghe Wang, Jordan Winter, Rui Wang. The liver microenvironment promotes glycolysis in metastatic colorectal cancer by activating her3 [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 3082.
Abstract 309: AHNAK nucleoprotein 2 tune oxidative stress and resistance to chemotherapy in triple negative breast cancer
Authors: Hao Na 
Published: March 2024

Abstract
Background: Resistance to chemotherapy is a major reason for the poorerprognosis in patients with triple negative breast cancer (TNBC). Growing evidence suggests that oxidative stress induced by reactive oxygen species (ROS) is an important factor in both tumourprogression and responses to chemotherapies, however, many signalling pathways and key molecular underlying the regulation of cancer cells to the ROS metabolism remains unclear.
Aim: (i) To investigate the mechanistic role of AHNAK nucleoprotein 2 (AHNAK2) in TNBC and its links with breast cancer mortality, particularly caused by chemotherapeutic resistance; and (ii) to test a hypothesis that AHNAK2 facilitates the abnormal activation of ROSscavenging system, result in the resistance to chemotherapy in TNBC.
Methods: The expression level of AHNAK2 was determined by immunohistochemical staining and analysed the mRNA expression profiles from The Cancer Genome Atlas of triple negative breast tumors. Roles of AHNAK2 in cancer cell growth, metastasis, chemotherapeutic resistanceand activation of oxygen scavenging systemwere determined by molecular and cell biology methods. A jetPEI nanocarrier was used as the vehicle for anti-AHNAK2 siRNAs in mouse models of cancer therapeutics.
Results: AHNAK2 is a novel triple negative breast cancer target, which is overexpressed in TNBC tissues and significantly associated with clinical pathology of chemotherapeutic resistance and poor prognosis of tumour patients. Mechanistically, AHNAK2 antagonizes the ROS accumulation induced by epirubicinvia activating JAK-STAT signaling pathway and transportingphosphorylated STAT1 & STAT4 to the nucleus to promote the expression of SOD2, a key gene in ROS metabolism.And epirubicin enhanced AHNAK2 expression by induced the accumulation of ROS,which hence the Sp1-driven transcription of AHNAK2,result inde novo resistance. Furthermore, AHNAK2-specific siRNAs encapsulated by jetPEI nanocarriers prominently inhibit tumor growth and epirubicin-induced resistance in vivo.
Conclusions: AHNAK2 plays a key role in resistance to chemotherapyby regulating the oxidative stressand ROS metabolism,representing a future biomarker and therapeutic target of TNBC.
Citation Format: Na Hao. AHNAK nucleoprotein 2 tune oxidative stress and resistance to chemotherapy in triple negative breast 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 309.
Abstract 3091: Discovery of highly selective ligands with unique binding modes for challenging oncology targets using DNA-encoded chemical libraries
Authors: Zhang Ying , Keefe Anthony , Clark Matthew A. 
Published: March 2024

Abstract
DNA-encoded chemical library (DEL) technology enables the efficient screening of large collections of encoded compounds and has emerged as a powerful tool for hit identification in recent years. The technology allows for the multiplexed interrogation of target-ligand binding interactions and results in DEL hits with unusual binding modes and desirable selectivity. This screening methodology is particularly effective for oncology targets. Examples of discovery of highly selective ligands for challenging oncology targets, including kinases, protein-protein interactions, and receptors, will be presented. We will illustrate how DEL campaign strategies directly aim at novel binding modes, and provide molecular details around the uniqueness of these interactions. Furthermore, we will show how these novel binding modes lead to unique pharmacology with direct benefit to patients. Lastly, we will demonstrate how X-Chem’s high-quality encoded libraries enhance the tractability and novelty of chemical equity discovered with the platform.
Citation Format: Ying Zhang, Anthony Keefe, Matthew A. Clark. Discovery of highly selective ligands with unique binding modes for challenging oncology targets using DNA-encoded chemical libraries [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 3091.
Abstract 3116: Bioluminescent 3D tumors with immune cell co-culture for high throughput screening (HTS)
Authors: Chou Fu-Ju , Kamali Samaneh , Gilardi Mara , Walling Brandon , Andar Abhay , Abarca-Heidemann Karin , Mancini Maria 
Published: March 2024

Abstract
In the past decade, cancer immunotherapy has revolutionized the cancer treatment strategy. Immune-oncology (IO) drugs play an important role in treating various malignancies. Immunotherapies work by re-educating and boosting patients’ immune systems to recognize and eliminate cancer cells. The immune co-culture assay provides invaluable information to understand the tumor microenvironment (TME) and observe the immune response after IO drug treatment. Recent advancements in the Champions TumorGraft3D (CTG3D) platform provides more consistent and accurate information for preclinical drug evaluation, but also efficiently simulate the tumor-Immune microenvironment (TIME) and interactions between cancer organoids and immunocyte. However, current analytic approaches in organoid-immune co-culture studies are still highly reliant on flow cytometry and high-resolution confocal imaging endpoints to provide the study outcome. There is a need for a fast and cost-effective co-culture platform to help set up experimental conditions. Easily traceable tags (e.g. bioluminescent and fluorescent) can quickly help establish reliable the effector (E) cell to tumor (T) cell conditions, thus enabling the user to test multiple immune-cell activator or engagers concentrations in a cost-effective and rapid manner. At Champions Oncology we provide stable Luc/GFP-tagged CTG3D models. We have built a reliable CTG3D-immune co-culture platform with various indications (including colon, prostate, gastric, non-small cell lung cancer, and leukemia) which can be used in co-culture with autologous TIL, allogenic TIL and NK cells, thus providing users a fast and cost-effective option to optimize the E:T ratio and monitor the drug performance.
Citation Format: Fu-Ju Chou, Samaneh Kamali, Mara Gilardi, Brandon Walling, Abhay Andar, Karin Abarca-Heidemann, Maria Mancini. Bioluminescent 3D tumors with immune cell co-culture for high throughput screening (HTS) [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 3116.
Abstract 3128: HRA00184-C004, preclinical characterization of a novel tissue factor-targeted antibody-drug conjugate
Authors: Ying Hanxiao , You Lingfeng , He Ning , Xu Shaoyu , Pu Pu , Lin Sophie , Feng Jun , Zhang Zhe , Ye Xin , Hu Min , He Feng 
Published: March 2024

Abstract
The physiological function of tissue factor (TF) is to initiate coagulation cascade by forming complex with plasma Factor VII after vascular injury. For Tisotumab vedotin (Tivdak), a TF targeting antibody-drug conjugate (ADC) approved for the treatment of cervical cancer, bleeding events were prespecified as adverse effects (AEs) of interest, besides ocular toxicity and peripheral neuropathy (known MMAE payload related AE). In order to mitigate the bleeding risk, a next generation of TF-targeting ADC is developed by Hengrui Pharmaceuticals. HRA00184-C004 is a novel ADC comprising a differentiated TF-targeted antibody (HRP07132) conjugated to a proprietary topoisomerase I inhibitor (Dxh, an exatecan derivative) via a protease-cleavable linker. In contrast to most TF-targeting antibodies, the naked HRP07132 does not interfere the interaction between TF and Factor VII or Factor X, has no impact on coagulation function in prothrombin time and thrombin generation assay, therefore reduced bleeding incidence is expected for HRA00184-C004. Meanwhile, HRA00184-C004 exhibited not only potent growth inhibition against multiple TF-expressing cancer cell lines but also bystander-killing effect in vitro. HRA00184-C004 also displayed robust and sustained anti-tumor activities in the PDAC HPAFII (TF high), TNBC MDA-MB-231 (TF moderate) and NSCLC H358 (TF low) TF-expressing xenograft models. Furthermore, in cynomolgus monkey, HRA00184-C004 is well-tolerated over a 6-week period with 12 mg/kg dosing every 3 weeks with acceptable PK profile. Notably, neither coagulation disorder nor skin rash was observed, which are superior to the historical NHP data reported by Tivdak. In conclusion, with the encouraging efficacy in preclinical study and promising safety profiles in NHPs, HRA00184-C004 has demonstrated the best-in-class potential and is scheduled to advance into clinical development.
Citation Format: Hanxiao Ying, Lingfeng You, Ning He, Shaoyu Xu, Pu Pu, Sophie Lin, Jun Feng, Zhe Zhang, Xin Ye, Min Hu, Feng He. HRA00184-C004, preclinical characterization of a novel tissue factor-targeted antibody-drug conjugate [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 3128.
Abstract 313: Benchmarking immune repertoire sequencing technologies
Authors: Maheshwari Shamoni , Morrison Carolyn , Taylor Sarah ,   BCR-SEQC consortium
Published: March 2024

Abstract
In the past decade, single cell gene expression technologies have transformed basic and translational research and are more frequently found in the toolbox of immunologists alongside the standard bulk B-cell receptor (BCR) sequencing and transcriptome profiling assays. While conventional bulk BCR assays capture the frequency of single chains, single-cell immune profiling reveals the frequency of paired heavy and light chains along with the cells transcriptional profile.
Here we present the results of a single-blind benchmarking experiment conducted in collaboration with the U.S. Food and Drug Administration (FDA) that compares the performance of standard bulk DNA and RNA BCR repertoire sequencing assays, with the 10x Genomics Single Cell Immune Profiling solution. The FDA selected nine unambiguously monoclonal BCR-expressing cell lines to generate a reference mixture that was shared with each study participant, without revealing the input proportions. This reference cell line mixture was also diluted to 10% and 1% with B-cells sorted from peripheral blood mononuclear cells (PBMC) to further test the limits of clonotype detection.
For each experiment, we targeted ~10,000 cells and in aggregate generated a paired gene expression and V(D)J dataset from ~100,000 cells. BCR clonotypes were detected in 78% of the captured cells across all samples profiled. Each of the 9 top unique clonotypes observed in our dataset corresponded to a cell line in the reference mixture, allowing us to decode the mixing percentages of each BCR-expression cell line. In addition, we were able to identify rare cell line clonotypes to as low as 0.01% in the dilution experiments. Furthermore, the multimodal nature of the single cell data also resolved the transcriptional profile of each cell line into its own cluster thus enabling independent verification of clonotype proportions.
In summary, we have generated a resource for the immunology community to evaluate and compare data generated with different repertoire sequencing technologies.
Citation Format: Shamoni Maheshwari, Carolyn Morrison, Sarah Taylor, BCR-SEQC consortium. Benchmarking immune repertoire sequencing technologies [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 313.
Abstract 3135: BSI-730, a bispecific antibody targeting HER2 and PD-L1 for the development of a first-in-class bispecific ADC
Authors: Hao Xiaoyao , Li Hongyan , Liu Xiaodong F. , Liu Jinyu , Dai Wenwen , Pan Ying , Hu Hui-Han , Li Jun , Xia Shukai , Lyu Qun , Davis Hugh M. , Chen Mingjiu , Peng Zeyu 
Published: March 2024

Abstract
Background: Antibody-drug conjugates (ADCs) have become a promising therapeutic option for patients with cancer. Bispecific ADCs targeting two tumor-associated antigens have potential differential anti-tumor advantages. HER2 and PD-L1 are co-expressed in some tumor cell types, such as breast cancer, gastric cancer, lung adenocarcinoma and urothelial carcinoma. A bispecific ADC targeting HER2 and PD-L1 should provide therapeutic benefit, particularly for patients with HER2 low cancer. Dual-target mediated cytotoxicity combined with PD-L1 mediated immunotherapy provides an innovative approach for cancer patients.
Experimental procedures: An anti-PD-L1 monoclonal antibody was identified from A/J mice immunized with PD-L1-ECD-Fc and screened by the Biosion proprietary SynTracer® High Throughput Endocytosis Platform. A “knob-in-hole” bispecific antibody, BSI-730, targeting HER2 and PD-L1 was created with trastuzumab and the humanized anti-PD-L1 scFv. Target binding specificity, binding activity, and affinity of BSI-730 were evaluated by protein-based ELISA, cell-based FACS and Biacore-based SPR, and the ligand blocking activity of PD-L1 portion was measured by competitive ELISA and cell-based FACS. The internalization activities on various cancer cell lines with different expression levels of HER2 and PD-L1 were evaluated by using the SynTracer® Platform.
Summary: BSI-730, a HER2/PD-L1 bispecific antibody, showed comparable binding affinity to HER2 as compared to trastuzumab, as well as comparable bioactivity to the parental anti-PD-L1 antibody regarding PD-L1 binding and PD-1/PD-L1 blocking. Based on a cell-based reporter assay, BSI-730 was able to reverse PD-L1 mediated T-cell suppression and exhibited comparable potency to the parental anti-PD-L1 antibody. The internalization of BSI-730 on HER2 low cancer cell lines was stronger than that of trastuzumab whereas the internalization of BSI-730 on HER2 high cancer cell lines was comparable to that of trastuzumab.
Conclusion: BSI-730 is a novel bispecific antibody for the development of a first-in-class bispecific ADC with the potential advantage of increased potency that can be further investigated to treat HER2 and PD-L1 co-expression tumors, particularly HER2 low tumors.
Citation Format: Xiaoyao Hao, Hongyan Li, Xiaodong F. Liu, Jinyu Liu, Wenwen Dai, Ying Pan, Hui-Han Hu, Jun Li, Shukai Xia, Qun Lyu, Hugh M. Davis, Mingjiu Chen, Zeyu Peng. BSI-730, a bispecific antibody targeting HER2 and PD-L1 for the development of a first-in-class bispecific ADC [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 3135.