bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3533: Zebrafish her3 knockout impacts developmental and rhabdomyosarcoma-related gene signatures
Authors: Kent Matthew R. , Calderon Delia , Silvius Katherine M. , Kucinski Jack P. , LaVigne Collette A. , Cannon Matthew V. , Kendall Genevieve C. 
Published: April 2023

Abstract
HES3 is a basic helix-loop-helix transcription factor that regulates neural stem cell renewal during development. HES3 overexpression is predictive of reduced overall survival in patients with fusion-positive rhabdomyosarcoma, a pediatric cancer that resembles immature and undifferentiated skeletal muscle and is most commonly driven by the fusion-oncogene PAX3-FOXO1. However, the mechanisms of HES3 cooperation in PAX3-FOXO1 driven rhabdomyosarcoma are unclear and are likely related to her3/HES3’s role in neurogenesis. To investigate HES3’s function during development, we generated a zebrafish CRISPR/Cas9 null mutation of her3, the zebrafish ortholog of HES3. Phenotypic characterization revealed that her3 null mutation is not embryonic lethal as they are present at expected Mendelian ratios. We observed a temporary growth delay her3 zebrafish null mutants and, rarely, eye defects in adults. Transcriptomic analysis of her3 null mutant embryos showed early dysregulation of a known downstream target, neurog1 and downregulation of genes involved in organ development, such as pctp and grinab, while genes pointing toward a terminal differentiation state, such as tmod, are upregulated. Differentially expressed genes in her3 null mutant embryos are enriched for HOX and SOX10 motifs, suggesting these may be key genes involved during HES3 dysregulation. Several cancer-related gene pathways are impacted, including the inhibition of matrix metalloproteinases and the tumor microenvironment pathway. To complement our zebrafish model, we are developing a double-inducible cell line model consisting of a tetracycline-inducible HES3 and a cumate-inducible PAX3-FOXO1 to further investigate the hypothesized cooperation between her3/HES3 and PAX3-FOXO1 in fusion-positive rhabdomyosarcoma. These two systems will allow us to elucidate conserved mechanisms of cooperation between HES3 and PAX3-FOXO1, and identify new therapeutic opportunities for children with fusion-driven rhabdomyosarcoma.
Citation Format: Matthew R. Kent, Delia Calderon, Katherine M. Silvius, Jack P. Kucinski, Collette A. LaVigne, Matthew V. Cannon, Genevieve C. Kendall. Zebrafish her3 knockout impacts developmental and rhabdomyosarcoma-related gene signatures. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3533.
Abstract 3555: Proteomic characterization of rhabdomyosarcoma-derived extracellular vesicles reveals a fusion-positive protein signature
Authors: Quaglietta Paula R. , Kissoondoyal Ashby , Malkin Ethan , Malkin David , Baertschiger Reto M. 
Published: April 2023

Abstract
Rhabdomyosarcoma is the most common pediatric soft tissue sarcoma. Current diagnostic methods involve imaging and tissue biopsy for staging, histology and fusion status assessment. There are presently no serum biomarkers for rhabdomyosarcoma diagnosis or surveillance. Novel approaches and identification of biomarkers are warranted for minimally invasive diagnostic/surveillance techniques. Extracellular vesicles (EVs) are recognized as critical mediators of intercellular communication and the pathophysiology of carcinogenesis and metastasis. This study aimed to characterize the proteome of rhabdomyosarcoma-derived EVs and identify proteomic signatures that correlate with clinical characteristics such as histological subtype, cancer stage, age and metastasis, using patient-derived cell lines. EVs were isolated from cell culture conditioned media by differential ultracentrifugation from four rhabdomyosarcoma cell lines (RH4, RH18, RH30, RD), each in triplicate. Liquid chromatography-tandem mass spectrometry identified EV protein cargo. Proteins identified in our samples were compared to all available EV data in the Vesiclepedia database, using the FunRich software. We identified 1,527 total proteins in our rhabdomyosarcoma-derived EVs. When comparing to Vesiclepedia, 96 unique proteins were identified from our EVs that were not previously annotated in EVs within the database. Expression of these 96 proteins was further evaluated based on clinical characteristics of the tumors the cell lines were derived from. We found a four-protein signature in the EV secretome of rhabdomyosarcoma cell lines correlated with FOXO1 fusion-positive status. This signature includes proteins involved in regulating extracellular matrix organization, regulating gene expression, protein biosynthesis and translation. Rhabdomyosarcoma cells secrete EVs with unique protein cargo based on clinical characteristics of the parent tumor such as histological subtype and FOXO1 fusion status. Further validation of these secreted EV proteins in biological fluids could prove an effective liquid biopsy technique for novel diagnostic approaches or surveillance in pediatric rhabdomyosarcoma.
Citation Format: Paula R. Quaglietta, Ashby Kissoondoyal, Ethan Malkin, David Malkin, Reto M. Baertschiger. Proteomic characterization of rhabdomyosarcoma-derived extracellular vesicles reveals a fusion-positive protein signature. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3555.
Abstract 3614: 3D model to study migration and invasion of lung cancer
Authors: Hardt Melina , Zatloukal Kurt , Popper Helmut H. 
Published: April 2023

Abstract
Background: In 2D cell cultures, migration of tumor cells is characterized by epithelial-to-mesenchymal transition (EMT), whereby tumor cells loose epithelial and express mesenchymal markers and very often change into a spindle cell phenotype. In contrast to 2D cultures, in 3D cultures tumor cells do not lose their differentiation. Small cell lung cancer usually moves as single cells or in small clusters, squamous cell and adenocarcinomas preferentially move in large clusters of cells. In tissue specimen, all of these carcinomas migrate in a hybrid EMT, not loosing their cytokeratin and E-cadherin expression. In AC and SCC also polarity and specification do occur, as some tumor cells act as leaders, providing orientation for the followers. However, 3D culture system might better depict real migration and come closer to the in vivo system as 2D cell culture systems.
Material and Method: A 3D bioprinter (TissueLabsR) was used to spot an alveolar-mimicking lung-specific matrix on cell culture wells. Microscopic channels out of pluronicsR are created within the matrix. Pluronics is removed by cooling, leaving empty channels within the protein matrix. These channels can be filled with different interleukins, such as IL23, which will direct migration of the tumor cells. Cultured cell lines from an adenocarcinoma (A549) and small cell carcinoma (NCI-H82) are layered on top of the matrix and migration into the matrix is studied using different time points.
Results and Discussion: A matrix simulating an alveolar structure was created with MatrigelR. The A549 cells were seeded on the matrix and grown for 2,5, 10, and 14 days. The cells formed cell clusters on the matrix and after one week started to invade the matrix in cell complexes. By immunohistochemistry we could show, that the A549 cells retained their epithelial differentiation. The H82 cells showed a different behavior. They did not form large clusters, but started to invade in small groups. The use of the bioprinter enabled us not only to create a 3D tumor culture system to simulate the in vivo conditions but also to create matrix structures for studying invasion and migration and to stimulate carcinoma cells with migration-activating substances.
Citation Format: Melina Hardt, Kurt Zatloukal, Helmut H. Popper. 3D model to study migration and invasion of lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3614.
Abstract 3636: Liver endothelium secreted LRG1 promotes metastatic colorectal cancer growth through the HER3/RSK/EIF4B AXIS
Authors: Rathore Moeez Ghani , Wright Michelle , Huang Wei , Taylor Derek , Li Yamu , Winter Jordan , Wang Zhenghe , Greenwood John , Moss Stephen , Wang Rui 
Published: April 2023

Abstract
Background: 25% of patients diagnosed with colorectal cancer (CRC) have liver metastasis at presentation, and ~80% of all metastatic CRC are developed in the liver. We previously reported that liver endothelial cells (ECs), a key component of the liver microenvironment, secrete LRG1 to promote CRC growth via activating human epidermal growth factor receptor (ERbB3, also known as HER3). However, we found that LRG1-induced HER3 activation is distinct from the canonical neuregulin 1 (NRG1)-induced HER3 pathway. The present study further validated LRG1 as a new HER3 ligand for promoting mCRC growth and elucidated the novel downstream signaling pathway induced by LRG1-HER3.
Methods: We first measured the binding affinity between HER3 and LRG1 by Biolayer interferometry (BLI). We then used in vitro and in vivo xenograft approaches to determine the effect of LRG1 monoclonal antibody (15C4) on HER3 activation and CRC growth. To further determine the role of LRG1 in promoting CRC growth in the liver, we used murine CRC cells in a syngeneic orthotropic liver injection model to establish CRC allografts in the liver of LRG1−/− mice with systemic LRG1 knockout and wild-type siblings (LRG1+/+). We also performed unbiased phospho-MS analysis and subsequent validations to determine the downstream signaling pathway activated by LRG1-HER3.
Results: We identified that LRG1 binds to HER3 with the affinity at ~100nM. The LRG1 antibody 15C4 completely attenuated LRG1-induced HER3 activation and in vitro and xenograft growth in vivo. Moreover, LRG1−/− mice with CRC allografts in the liver had 2 times longer overall survival than tumor-bearing LRG1+/+ mice. Lastly, unbiased -omics analysis identified eIF4-protein synthesis is significantly activated by LRG1. With target-specific inhibitors, we further determined that LRG1-HER3 activates the PI3K-PDK1-RSK1/3-eIF4 axis independent of AKT.
Conclusions: We identified LRG1 as a novel HER3 ligand and demonstrated that the liver microenvironment-derived LRG1 plays a key oncogenic role in mCRC, by activating a novel RSK-eIF4 survival pathway. Our findings highlighted the potential of blocking LRG1-HER3 and involved downstream pathways for treating patients with mCRC.
Citation Format: Moeez Ghani Rathore, Michelle Wright, Wei Huang, Derek Taylor, Yamu Li, Jordan Winter, Zhenghe Wang, John Greenwood, Stephen Moss, Rui Wang. Liver endothelium secreted LRG1 promotes metastatic colorectal cancer growth through the HER3/RSK/EIF4B AXIS. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3636.
Abstract 3639: Impact of KRAS oncogenic signaling in tumorigenesis and immune evasion driven by extracellular vesicles (EVs)
Authors: Vargas Zulaida M. Soto , Arteaga David C. , Sohal Ikjot S. , Hasan Humna , Kasinski Andrea L. 
Published: April 2023

Abstract
Mutations in Kristen rat sarcoma homolog (KRAS) occur in about 25% of cancers, making this protein one of the most frequently altered genes in cancer. KRAS mutations are not only associated with poor patient survival, but also with increased tumor aggressiveness and metastasis. Tumor development and metastasis are impacted by intercellular communication. Key mediators of cell-to-cell communication include vesicles secreted by cells, also known as extracellular vesicles (EVs). EVs released by cancer cells can mediate malignant transformation of non-cancerous cells as well as influence pre-metastatic niche formation. Conversely, inhibiting the secretion and release of EVs by cancer cells decreases tumor growth and metastasis. However, the mechanisms through which EVs drive tumorigenesis in KRAS mutant tumors are still understudied. Our lab recently reported that EVs from lung cancer cells drive invasion of non-tumorigenic lung epithelial cells. Here, we found that KRAS oncogenic signaling in the donor cells contributes to the EV-driven migration and invasion of non-tumorigenic lung epithelial cells. Previous studies have demonstrated that KRAS signaling drives immune evasion and promotes an “immune cold” tumor microenvironment. Our preliminary studies showed that EVs derived from KrasG12C lung cancer cells (H358) inhibit T cell proliferation and that blocking KRASG12C signaling, using ARS-1620 (KrasG12C inhibitor), impairs EV-driven T cell immunosuppression. We performed a comprehensive proteomic comparison between EVs isolated from H358 treated with ARS-1620 and those isolated from cells treated with DMSO (vehicle) and identified proteins involved in migration and invasion, such as LAMB-3, to be downregulated in EVs isolated from ARS-1620 treated cells. Ongoing studies aim to pinpoint the detailed mechanisms by which cancer cells utilize mutant KRAS signaling to promote loading of oncogenic cargo into EVs, and the resulting contributions to tumorigenesis and immune evasion.
Citation Format: Zulaida M. Soto Vargas, David C. Arteaga, Ikjot S. Sohal, Humna Hasan, Andrea L. Kasinski. Impact of KRAS oncogenic signaling in tumorigenesis and immune evasion driven by extracellular vesicles (EVs). [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3639.
Abstract 3641: Loss of Smad4 induces SPP1 secretion and immunosuppressive myeloid cell formation in pancreatic ductal adenocarcinoma
Authors: Murimwa Gilbert Z. , Rossi Francesca , Fleming Henry K. , Yazgan Zeynep , Alzhanova Dina , Huang Huocong , Brekken Rolf A. 
Published: April 2023

Abstract
Up to 50% of pancreatic ductal adenocarcinomas (PDA) harbor mutations in canonical TGFß pathway genes such as SMAD4 and TGFBR2. SMAD4 mutations have been shown to drive resistance to chemotherapy, metastasis, and margin positivity following surgical resection. To characterize the unique tumor microenvironment of SMAD4 mutant PDA, we developed isogenic, orthotopic mouse models using CRISPR mediated knockout of Smad4 in a cell line derived from KPfC (KrasLSL-G12D;Trp53fl/fl;Pdx1Cre) mice, a genetically engineered mouse model of pancreatic cancer. To characterize the alteration of the secretome caused by the loss of Smad4, we performed proteomic analysis with conditioned media collected from Smad4 wildtype and mutant cancer cell clones. Interestingly, the loss of Smad4 resulted in the production of neuroendocrine related factors. In addition, gene ontology analysis with the secretome suggested that these factors were associated with neuronal processes. These data indicate that loss of Smad4 in PDA cancer cells may lead to a neuroendocrine signature change. In particular, we found one of the most up-regulated secreted factors in the mutant cells was SPP1, which has been shown to promote tumor progression by affecting cancer cell stemness and macrophage polarization. To validate the proteomic results, we established orthotopic models with Smad4 wildtype and mutant clones. We found that loss of Smad4 resulted in larger tumor growth. Moreover, immunohistochemistry showed a significant up-regulation of SPP1 in the cancer cells of Smad4 mutant tumors. To further profile the alteration of immune landscape, we stained for markers of myeloid cells and lymphocytes. We found that Smad4 mutant tumors had an immunosuppressive microenvironment characterized by increased M2 macrophage, neutrophil and MDSC infiltration. Altogether, our study suggests that loss of SMAD4 in PDA may induce a neuroendocrine signature with increased SPP1 secretion in cancer cells. The production of this unique cancer cell secretome may result in a microenvironment marked by immunosuppressive myeloid cells.
Citation Format: Gilbert Z. Murimwa, Francesca Rossi, Henry K. Fleming, Zeynep Yazgan, Dina Alzhanova, Huocong Huang, Rolf A. Brekken. Loss of Smad4 induces SPP1 secretion and immunosuppressive myeloid cell formation in pancreatic ductal adenocarcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3641.
Abstract 3650: STAT1 suppresses KRAS-driven lung adenocarcinoma depending on the tumor microenvironment
Authors: Trenk Christoph , Sagmeister Rebecca , Horvath Jaqueline , Homolya Monika , Luca Andreea Corina , Eferl Robert , Moll Herwig , Casanova Emilio 
Published: April 2023

Abstract
Lung cancer is responsible for the majority of cancer-related deaths worldwide. Within this severe disease lung adenocarcinoma (LUAD) belongs to the most common form of lung cancer. LUAD is highly correlated with KRAS mutations. Although, the first KRAS inhibitors entered into clinics recently, therapy resistance arises. Since KRAS-mutant LUAD is an inflammation driven disease, we focus on the JAK-STAT pathway as an alternative target. Preliminary data suggests that human KRAS-mutant LUAD is correlated to enriched JAK-STAT signaling, as well as STAT1 upregulation. Accordingly, we aimed to explore the tumorigenic functions of STAT1 in this form of lung cancer. Genetically engineered C57BL6/N mice which develop autochthonous KRAS-driven and P53-deficient LUAD proficient (KP) and deficient for STAT1 (KPS) were used. In a second mouse model expression of ovalbumin was induced in tumor cells, mimicking a neoantigen to increase immune infiltration in the presence (KPO) or absence of STAT1 (KPOS). Kaplan-Meyer analysis was performed to assess the overall survival. Tumor burden, as well as immune infiltration was analyzed in lungs 6 and 10 weeks after tumor induction via H&E, IHC and IF staining. Furthermore, the lung fluid was collected by bronchoalveolar lavage and used to profile inflammatory cytokines. Deletion of STAT1 reduced the survival in KRAS-driven LUAD mice only when ovalbumin was expressed. Tumor burden and tumor grades were increased in tumors lacking STAT1 at 6 and 10 weeks after tumor initiation. Surprisingly, although tumors of KPOS mice showed a decreased infiltration by T cells and CD4 T cells, CD8 T cell infiltration did not change, suggesting a tumor suppressive function of STAT1 via T cell exhaustion. Moreover, the macrophage attracting cytokine CCL9 secretion was upregulated and the number of tumor infiltrating, immunosuppressive macrophages was increased in KPOS mice. This data implicates a tumor suppressive function of STAT1 by secretion of cytokines that can recruit suppressive myeloid immune cells to the TME leading to CD4 T cell exclusion and CD8 T cell exhaustion. Further studies are needed on the changes in the immune infiltrating cells to explain the mechanism behind these intriguing findings.
Citation Format: Christoph Trenk, Rebecca Sagmeister, Jaqueline Horvath, Monika Homolya, Andreea Corina Luca, Robert Eferl, Herwig Moll, Emilio Casanova. STAT1 suppresses KRAS-driven lung adenocarcinoma depending on the tumor microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3650.
Abstract 3673: Plk1 phosphorylation of PHGDH to regulate serine metabolism
Authors: Rao Xiongjian , Allison Derek B. , Flight Robert M. , Wu Yuanyuan , Scott Timothy , He Daheng , Wang RuiXing , Li Zhiguo , Li Chaohao , Kong Yifan , Mao Fengyi , Wang Chi , Higashi Richard M. , Lane Andrew N. , Moseley Hunter NB , Liu Xiaoqi 
Published: April 2023

Abstract
Polo-like kinase 1 (Plk1) has been reported to be highly expressed in most tumors, especially in advanced tumors, while how Plk1 elevation benefits tumor growth remains elusive. Metabolic reprogramming is one of the hallmarks of cancer, but how the tumors fully take advantage of deregulated metabolism is an enigma. Here, we found that Plk1 could divert serine metabolism from de novo synthesis to exogenous uptake by regulating PHGDH (phosphoglycerate dehydrogenase), the first rate-limiting enzyme of de novo serine biosynthesis. We show that PHGDH is phosphorylated by Plk1 at a cluster site (S512, S513, S517) and that Plk1-associated phosphorylation of PHGDH results in its protein degradation, thus reduced de novo serine biosynthesis. As a compensatory response, cells with an elevated level of Plk1 significantly increase the uptake of serine to produce more sphingosines, which are pivotal metabolites for the growth of cancer cells. Our finding may provide guidance on how to target de novo biosynthesis of serine, serine uptake or sphingosine metabolism to treat advanced prostate cancer.
Citation Format: Xiongjian Rao, Derek B. Allison, Robert M. Flight, Yuanyuan Wu, Timothy Scott, Daheng He, RuiXing Wang, Zhiguo Li, Chaohao Li, Yifan Kong, Fengyi Mao, Chi Wang, Richard M. Higashi, Andrew N. Lane, Hunter NB Moseley, Xiaoqi Liu. Plk1 phosphorylation of PHGDH to regulate serine metabolism. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3673.
Abstract 3680: Multiomics analysis of triple negative breast cancer identifies potential metabolic vulnerability for overcoming the resistance of neoadjuvant therapy
Authors: Ren Zuen , Kurmi Kiran , Morris Robert , Joshi Shakchhi , Zaniewski Eric , Kreuzer Johannes , Gioia Gabrielle Elena , Haas Wilhelm , Haigis Marcia C. , Ellisen Leif W. 
Published: April 2023

Abstract
Triple-negative breast cancer (TNBC) is a heterogeneous disease that remains currently medically incurable. We comprehensively analyzed clinical, proteomic, and metabolomic data of a cohort of 12 highly selected primary TNBCs. Among them, 7 patients after neoadjuvant chemotherapy had residual disease and micro- or macro-metastases that were non-pathologically complete response (non-pCR); 5 patients had pathologically complete response (pCR). Interestingly, both proteomic and metabolic profiling results revealed dramatically different molecular signatures in non-pCR relative to pCR. Non-pCR were enriched in metabolites involving aspartic acid, fumarate, nicotinamide, and adenosine that are critical intermediates for nitrogen recycling and tricarboxylic acid (TCA) cycle, which are essential fuels for tumor biomass. Moreover, non-pCR were enriched in urea cycle, oxidative phosphorylation, glycolysis, sirtuin signaling, nucleotide De Novo biosynthesis, which are resistance-associated. Putative therapeutic targets were identified in non-pCR patients. Collectively, our multiomics analysis demonstrated the heterogeneity of TNBCs at multi-levels and enabled the development of personalized therapies targeting unique tumor metabolic profiles.
Citation Format: Zuen Ren, Kiran Kurmi, Robert Morris, Shakchhi Joshi, Eric Zaniewski, Johannes Kreuzer, Gabrielle Elena Gioia, Wilhelm Haas, Marcia C. Haigis, Leif W. Ellisen. Multiomics analysis of triple negative breast cancer identifies potential metabolic vulnerability for overcoming the resistance of neoadjuvant therapy. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3680.