bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5466: In vivo characterization of neuroblastoma intratumoral heterogeneity
Authors: Squires Willow R. , Weiss Alex , Cohen-Gogo Sarah , Shlien Adam , Kaplan David , Irwin Meredith S. , Hayes Madeline N. 
Published: March 2024

Abstract
Background: Neuroblastoma is a pediatric solid tumor that arises from neural crest-derived progenitor cells of the peripheral sympathetic nervous system (PSNS). Neuroblastoma patients display a significant level of genetic and phenotypic heterogeneity, with amplification of the MYCN oncogene associated with poor clinical outcomes. To better understand underlying genetic mechanisms associated with aggressive neuroblastoma, our group incorporated patient-relevant loss-of-function mutations into the established MYCN-driven zebrafish model of neuroblastoma (Tg(dbh:EGFP-MYCN)), which resulted in increased tumor penetrance and a highly metastatic phenotype.
Aims & Methods: We aimed to investigate metastases-associated intratumoral heterogeneity in vivo using our patient-relevant metastatic zebrafish models, with functional validations performed using human neuroblastoma cell lines.
Results: Single-cell RNA sequencing of metastatic zebrafish neuroblastoma tumors revealed intra-tumor cell heterogeneity, with a subset of cells highly expressing markers of more differentiated adrenergic-like cell fates (th, dbh, phox2a) while others enriched for markers of less differentiated mesenchymal-like progenitor cells (prrx1a, vim, fn1a). Differential cell labeling in zebrafish neuroblastoma suggests distinct tumor cell populations for follow-up validation and characterization throughout metastatic progression.
Conclusion: Single-cell RNA sequencing of zebrafish metastatic neuroblastoma showed evidence of intratumoral heterogeneity that warrants further investigation. We are characterizing neuroblastoma intratumoral heterogeneity in our in vivo metastatic zebrafish models to examine the association between neuroblastoma tumor composition and metastatic potential. Altogether, this work aims to improve our understanding of the evolution of tumor cell heterogeneity associated with neuroblastoma metastasis and uncover novel therapeutic opportunities to improve patient outcomes.
Citation Format: Willow R. Squires, Alex Weiss, Sarah Cohen-Gogo, Adam Shlien, David Kaplan, Meredith S. Irwin, Madeline N. Hayes. In vivo characterization of neuroblastoma intratumoral heterogeneity [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 5466.
Abstract 548: Exploiting pronounced selection pressure from therapeutic bottlenecks to identify orthogonal immunotherapy targets
Authors: Fisk J. Nic , Townsend Jeffrey P. 
Published: March 2024

Abstract
Evolutionary processes play a crucial role in the development of cancer, where tumor cells continuously adapt, leading to the dominance of cell populations over their non-tumor counterparts, evading treatment and ultimately resulting in death. Effective cancer therapies induce significant and targeted selection pressures. During and in response to the induced bottleneck by therapy, the heterogeneous cancer cell population either diminishes or evolves, causing a subsequent selective sweep and the emergence of cells with resistance traits. However, this bottleneck phase presents a unique opportunity to predictably reduce the diversity of tumor cells. In this context, we propose a framework to quantify the selection on variants before, during, and after the therapeutic bottleneck, enhancing our understanding of the mechanisms underlying the acquisition of therapeutic resistance. Furthermore, we combine measures of selection intensity with the prediction of neoantigens in a patient cohort to identify potential alternative targets for immunotherapy. The outcome is a strategy that exploits the predictability introduced by the primary therapeutic bottleneck to eliminate the remaining tumor cell population. Finally, we illustrate the efficacy of this strategy in the context of several cancer types and empirically identify potential cancer type specific and pan-cancer immunotherapy targets orthogonal to primary, first-line therapies.
Citation Format: J. Nic Fisk, Jeffrey P. Townsend. Exploiting pronounced selection pressure from therapeutic bottlenecks to identify orthogonal immunotherapy targets [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 548.
Abstract 551: Investigation of the antipoliferative effect of the Xylopia Aethiopicaextract in triple-negative breast cancer cells
Authors: Soni Shailvi , Modi Shail Rakesh , Andey Terrick 
Published: March 2024

Abstract
Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer and has very limited treatment options. Xylopia Aethiopica (XA) is a common spice that is widely used in the regions of central and western Africa. It is known to have various pharmacological properties, one of which is anti-inflammatory activity. The anti-inflammatory effect may portend antiproliferative activity. We hypothesized that the extract of XA will exert antiproliferative activity against TNBC cells. The study aimed to establish the molecular mechanism underlying the antiproliferative activity of XA extract (XAE) against TNBC. The dried fruits of XA were crushed in a mixer and macerated using diethyl ether for 72 h. The macerate was subjected to vacuum and rotary evaporation to remove the organic solvent, and a stock solution was prepared in dimethyl sulfoxide (DMSO). Assessment of the efficacy of XAE was performed using serial dilutions (100 µg/ml, 50 µg/ml, 10 µg/ml, 5 µg/ml, and 1µg/ml) of the XAE stock solution, followed by in vitro testing against two TNBC cell lines (MDA-MB-231, and MDA-MB-468 cells). The cytotoxic effect of the XAE was then measured using the resazurin dye assay. XAE resulted in a concentration-dependent cell death in both cell lines. At the highest concentration (100 µg/ml), XAE induced cytotoxic cell death of 89% and 91% in MDAMB231 and MDAMB468 cells, respectively. This was consistent with IC50 values of 32.95 ± 1.32 µg/ml (MDAMB231) and 20.83 ± 1.05 µg/ml (MDAMB468). At the molecular level, western blot analysis of mitogenic markers associated with MAPK oncogenic signaling pathway demonstrated the inhibition of EGFR and AKT protein levels after a 48-h treatment. Acridine orange-ethidium bromide (AO/EB) staining showed a concentration-dependent induction of apoptosis in both cell lines. Flow cytometric analysis was performed to quantify the apoptotic cells. A significant increase of apoptotic cells was observed in treatment groups as compared to control after 72 h. Apoptotic induction was associated with increased p53 protein levels after 48 h treatment compared to control. The study results underscore the anticancer effects of XAE in TNBC cells mediated through induction of apoptosis.
Citation Format: Shailvi Soni, Shail Rakesh Modi, Terrick Andey. Investigation of the antipoliferative effect of the Xylopia Aethiopicaextract in triple-negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 551.
Abstract 5548: Cannabidiol enhances efficacy of atezolizumab via upregulation of PD-L1 expression by cGAS-STING pathway in triple negative breast cancer cells
Authors: Kim Bu Gyeom , Kim Bo Ram , Kim Dae Yeong , Yun Hyemi , Kim Dayoung , Kim Woo Young , Kang Sanghee , Lee Sun Il , Oh Sang Cheul 
Published: March 2024

Abstract
The current treatment for patients with triple negative breast cancer (TNBC) relies on cytotoxic therapy. Currently, atezolizumab and chemotherapy can be combined in patients with TNBC, but this approach is not effective for all patients with low reactivity to atezolizumab. As there is a lack of alternative treatment options, new anti-cancer drugs are urgently needed to enhance atezolizumab reactivity against TNBC. Recent strategies have focused on regulating the expression of programmed death-ligand 1 (PD-L1) or enhancing immune response activation by combining anti-cancer drugs with immune checkpoint inhibitors (ICIs). Cannabidiol (CBD), a cannabinoid component derived from the cannabis plant, has been reported to have anti-cancer therapeutic potential because of its capacity to induce apoptotic cell death in tumor cells while avoiding cytotoxicity in normal cells. Previous studies have demonstrated the effects of CBD on apoptosis in various types of cancer cells. However, the potential role of CBD as an immune modulator in the regulation of PD-L1 expression and the anti-cancer immune response remains to be explored. In this study, we found that CBD stimulated PD-L1 expression in TNBC cells, which significantly induced CBD-mediated cGAS-STING pathway activation. Taken together, we demonstrated that the combination of CBD and anti-PD-L1 antibody enhances the anti-cancer immune response in vitro and in vivo experiments. Our findings identified the mechanism of PD-L1 regulation by CBD in TNBC cells and suggested that CBD could be a potential candidate for the development of new combinatorial strategies with ICIs in TNBC patients.
Citation Format: Bu Gyeom Kim, Bo Ram Kim, Dae Yeong Kim, Hyemi Yun, Dayoung Kim, Woo Young Kim, Sanghee Kang, Sun Il Lee, Sang Cheul Oh. Cannabidiol enhances efficacy of atezolizumab via upregulation of PD-L1 expression by cGAS-STING pathway in triple negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5548.
Abstract 5561: Whole slide Imaging Mass Cytometry allows the rapid profiling of the immune landscape of histopathologically aggressive prostate tumors
Authors: Gorman Jennifer L. , Liu Lydia Y. , Hartig Jordan P. , Parsotam Nikesh , Khoo Amanda , Ignatchenko Vladimir , Asbury Sarah , Afiuni Somi , Gonzalez Ricardo , Geuenich Michael J. , Harrigan Caitlin F. , Lee Yuju , Chen Jianan , Lim Liang , Raza Qanber , Angel Peggi M. , Campbell Kieran , Liu Stanley K. , Downes Michelle R. , Drake Richard R. , Kislinger Thomas , King David , Jackson Hartland W. 
Published: March 2024

Abstract
Patients with Gleason pattern 4 or 5 prostate tumors have a high risk of biochemical recurrence and worse survival. Affected by their local environment, dynamic immune cell phenotypes can both hinder or drive cancer progression. To understand the proteomic environments that facilitate specific immune cell responses and interactions in advanced prostate cancer, we have spatially resolved the immune cell content and metabolic features of Gleason pattern 4 and 5 and subpatterns of prostate tumors. To scale quantitative multiplexed histopathology measurements to human disease, we have applied a new high-throughput whole slide modality of high-plex Imaging Mass CytometryTM to simultaneously quantify immune cells, metabolic states, prostate specific pathways, and biomarkers in 125 whole slide sections from 42 patients over 100 cm2 of tissue. It is essential to image whole slide sections in order to fully evaluate the complete heterogeneity of these cancer samples. In parallel, multi-region macrodissection shotgun proteomics and both glycan and extracellular matrix MALDI imaging mass spectrometry have quantified the proteome associated with specific immune cell environments. Together, we identify localized proteomic microenvironments associated with specific tumor supportive or inhibitory immune cell content in human tumors that may help differentiate which high-risk patients will have a rapid biochemical recurrence following radical prostatectomy.
Citation Format: Jennifer L. Gorman, Lydia Y. Liu, Jordan P. Hartig, Nikesh Parsotam, Amanda Khoo, Vladimir Ignatchenko, Sarah Asbury, Somi Afiuni, Ricardo Gonzalez, Michael J. Geuenich, Caitlin F. Harrigan, Yuju Lee, Jianan Chen, Liang Lim, Qanber Raza, Peggi M. Angel, Kieran Campbell, Stanley K. Liu, Michelle R. Downes, Richard R. Drake, Thomas Kislinger, David King, Hartland W. Jackson. Whole slide Imaging Mass Cytometry allows the rapid profiling of the immune landscape of histopathologically aggressive prostate tumors [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 5561.
Abstract 5566: Streamlining tissue sample preparation: A comparative study on the efficiency and reproducibility of the Omni-Stainer™ dewaxing and HIER protocol
Authors: Samusik Nikolay , Goltsev Yury , Kedei Noemi 
Published: March 2024

Abstract
The evolution of life sciences research has entailed a shift toward understanding cellular functions within tissue microenvironments, with an emphasis on cell- and tissue-specific relationships. A key challenge in these endeavors is the need for accurate data acquisition, which can be impeded by variability in the multistep manual assay preparations. In the quest for developing reliable and efficient tissue sample preparation workflows, two protocols using multiple dewaxing agents have been compared: the Omni-Stainer™ Dewaxing and HIER antigen retrieval protocol from Parhelia Biosciences, Inc. and the analogous one on Leica BOND Rx. The objective of this research was to assess the efficiency and reproducibility of these methods in the context of a four-color morphology immunostaining used for GeoMx DSP RNA and protein detection assays (Nanostring). We examined the efficiency of automated dewaxing using various non-toxic xylene alternatives, including C12-15 alkyl Benzoate, diethylene glycol, methyl octanoate or methyl decanoate, and compared them to the proprietary Leica Bond Dewaxing solution. Then, we compared the quality of post-HIER immunostaining using two different four-antibody panels on human jejunum samples. After several rounds of optimization, we found that the V4 of the open-source Omni-Stainer™ dewax-HIER protocol resulted in identical quality of morphology immunostaining compared to proprietary Leica Bond protocol. This study validates the Omni-Stainer™ Dewaxing and HIER protocol as a viable and efficient approach for automation of sample prep for GeoMx DSP studies.
Citation Format: Nikolay Samusik, Yury Goltsev, Noemi Kedei. Streamlining tissue sample preparation: A comparative study on the efficiency and reproducibility of the Omni-Stainer™ dewaxing and HIER protocol [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 5566.
Abstract 5586: Hijacking mutant E3 ligase for targeted protein degradation: Discovery of cancer-specific PROTACs for the treatment of FBXW7 R465C driven cancers
Authors: Zhang Hailong 
Published: March 2024

Abstract
Many cancers carry LOF mutations of tumor suppressor genes and drug discovery against these cancers are challenging. Fbxw7 is a major tumor suppressor E3 ligase and is frequently mutated (LOF) in cancer. Fbxw7 R465C mutation correlates with worse survival and resistance to both chemo- and targeted therapies. Blueray have discovered Fbxw7 R465C-selective ligands and developed PROTAC molecules selectively targeting cancers carrying Fbxw7 R465C mutation. The Fbxw7 R465C-based PROTACs show superior degradation activity for several target proteins and inhibit proliferation of cancer cells carrying Fbxw7 R465C mutations. These PROTACs show tumor growth inhibition in Fbxw7 R465C mutant cell derived xenograft models with clear PK/PD correlations. Development of bi-functional molecules such as PROTACs bring new options to target cancers with LOF mutations.
Citation Format: Hailong Zhang. Hijacking mutant E3 ligase for targeted protein degradation: Discovery of cancer-specific PROTACs for the treatment of FBXW7 R465C driven 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 5586.
Abstract 5597: Clinical significance of RNA polymerase I inhibitor in pancreatic cancer therapy
Authors: Ahmad Mudassier 
Published: March 2024

Abstract
Pancreatic cancer (PanCa) is the third leading cause of cancer-related deaths in the United States with limited therapeutic options available. Gemcitabine (GEM), a deoxycytidine nucleoside analog is currently considered the most effective therapy for PanCa. However, it shows only a marginal survival benefit of six months. Aberrant ribosome biogenesis occurs in most tumor types. We observed that PanCa cells are addicted to ribosome biogenesis (RiBi), which supports their highly aggressive metastatic phenotypes. Thus, strategically targeting RiBi process could be one of the ideal strategies for the prevention and treatment of PanCa. In this study, we elucidated the molecular mechanisms of POLR1A (RPA194) overexpression and how its targeting along with p53 status impacts RNA polymerase I inhibitor therapy against PanCa. The expression level of RPA194 was significantly elevated in pancreatic tumor tissues when compared with adjacent normal pancreatic tissues. BMH-21 is a potent pharmacological inhibitor of RNA Pol I, which is known to degrade RPA194 protein. Our results demonstrated that BMH-21 can selectively induce apoptosis in various PanCa cells but not in HPNE cells. We also found that the cytotoxic effect of BMH-21 was dependent on the expression pattern of RPA-194 and p53 status. We further examined the therapeutic efficacy of BMH-21 in orthotopic xenograft mouse models by using two different PanCa cells, AsPC1 which contains non-functional p53 and MIA PaCa-2 which contains functional mutant p53. We observed that BMH-21 significantly inhibited the growth of tumors derived from both cancer cell lines. Interestingly, BMH-21-mediated inhibition of tumor growth was more significant in tumors derived from MIA PaCa-2 cells compared to AsPC1 cells. Further examination revealed that the inhibition of tumor growth was correlated with RPA194 degradation followed by inhibition of cell proliferation. Overall, our results strongly suggest that BMH-21 is a promising non-toxic agent for the treatment of advanced PanCa and its therapeutic potential depends on RPA194 expression and p53 status in PanCa cells.
Citation Format: Mudassier Ahmad. Clinical significance of RNA polymerase I inhibitor in pancreatic 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 5597.
Abstract 5606: The role of KMT2C in the homologous recombination repair in epithelial ovarian cancer
Authors: Ma Mirielle C. , Lin Z. Ping , Ratner Elena S. 
Published: March 2024

Abstract
Background: Epithelial ovarian cancer (EOC) with homologous recombination (HR) deficiency is susceptible to PARP inhibitor therapy due to synthetic lethality. Identification of the patient population for PARP inhibitor therapy is imperative to reduce recurrence of EOC and improve the survival outcomes of patients. Current biomarkers for PARP inhibitor therapy consist of an array of genes involved in the HR repair pathway in EOC, including BRCA1, BRCA2 and PALB2. Through TCGA analysis, we found that the lysine (K)-specific methyltransferase 2C (KMT2C) gene is mutated in 10.3% of all EOC patients and considerably enriched in BRCA1 mutated EOC patients. However, the role of KMT2C in HR repair and PARP inhibitor sensitivity of EOC remains largely undefined.
Methods: BRCA2-mutated PEO1 and BRCA2-wild type PEO1-NR cells were transfected with KMT2C siRNA and assayed for KMT2C protein expression by western blotting. DR-GFP (HR) and EJ5-GFP (NHEJ)-SKOV3 cell lines were co-transfected with the ISceI-expressing plasmid pCBA-ISceI and KMT2C siRNA. GFP-positive cells were analyzed by flow cytometry. SKOV3 cells were transfected with KMT2C siRNA and the expression of BRCA2 and Rad51 mRNA was determined using quantitative RT-PCR analysis.
Results: siRNA-mediated knockdown of KMT2C resulted in marked down-regulation of KMT2C protein in PEO1 and PEO1-NR cells. KMT2C knockdown caused a significant decrease in ISceI-induced HR activity in DR-GFP-SKOV3 cells (p &lt; 0.05). In contrast, KMT2C knockdown had no effects on ISceI-induced NHEJ activity in EJ5-GFP-SKOV3 cells. As a control, Rad51 knockdown caused complete suppression of HR activity and had no effects on NHEJ activity. Furthermore, quantitative RT-PCR analysis of key HR genes showed that KMT2C knockdown resulted in a moderate but significant down-regulation of BRCA2 (p &lt; 0.05) and Rad51 (p &lt; 0.01) expression in SKOV3 cells.
Conclusion: Our results suggest that KMT2C partakes in the HR repair pathway in EOC. However, the contribution of KMT2C to HR activity is not rate-limiting. Based on the known function of KMT2C in histone modification, we speculate that KMT2C serves to positively regulate the transcription of HR genes and therefore promote HR activity. Future investigation into the exact roles of KMT2C in HR repair, etiology of EOC, and diagnostic biomarkers for the PARP inhibitor sensitivity of EOC is warranted.
Citation Format: Mirielle C. Ma, Z. Ping Lin, Elena S. Ratner. The role of KMT2C in the homologous recombination repair in epithelial ovarian 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 5606.