bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2473: Tissue factor-induced fibrinogenesis and cancer cell clustering contribute to polyclonal peritoneal metastasis in diffuse-type gastric carcinoma
Authors: Miyazaki Makoto , Nagamura Yuko , Yamaguchi Hideki 
Published: April 2023

Abstract
Diffuse-type gastric carcinoma is characterized by highly invasive growth, massive fibrosis, and peritoneal metastasis. Peritoneal metastasis is the leading cause of death in diffuse-type gastric carcinoma; however, the underlying molecular mechanisms remain largely unknown. Recent studies have established a concept of polyclonal metastasis, in which metastases arise from circulating tumor cell clusters with high metastatic potential. Polyclonal metastasis is a cause of tumor heterogeneity that contributes to tumor progression and therapeutic resistance. However, the clonality of peritoneally metastasized tumors and the evidence for polyclonal metastasis in diffuse-type gastric carcinoma have been poorly elucidated. In this study, we utilized multicolor fluorescent lineage tracking to examine the clonality of peritoneal metastasis in mouse xenograft models. When peritoneal metastasis was induced by intraperitoneal or orthotopic injection of multicolored cancer cells, each peritoneally metastasized tumor exhibited multicolor fluorescence, showing that it contains multiclonal cancer cell populations. Peritoneal cytology and in situ fluorescent imaging of micrometastases revealed that multicolored cancer cell clusters formed within the peritoneal cavity and collectively attached to the peritoneum. In vitro, peritoneal lavage fluid or cleared ascitic fluid derived from cancer patients elicited cancer cell clustering, which was inhibited by anticoagulants. Immunofluorescence and immunoblotting revealed that cancer cell clusters formed in vitro and in vivo were associated with fibrin formation. Moreover, tissue factor knockout in cancer cells significantly reduced fibrin formation, cell clustering, peritoneal attachment, and peritoneal metastasis. In an in vitro adhesion assay, fibrin formation facilitated the cancer cell-mesothelial cell adhesion. Thus, these results demonstrate that cancer cells activate the coagulation cascade through tissue factor to form fibrin-mediated cell clusters and accelerate peritoneal attachment, leading to the development of polyclonal peritoneal metastasis. Our findings identify a novel mechanism of peritoneal metastasis that may be a potential therapeutic target.
Citation Format: Makoto Miyazaki, Yuko Nagamura, Hideki Yamaguchi. Tissue factor-induced fibrinogenesis and cancer cell clustering contribute to polyclonal peritoneal metastasis in diffuse-type gastric carcinoma [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 2473.
Abstract 2560: Post-translational modification of focal adhesion kinase (FAK) protein increases in breast cancer cells upon exposure to conditioned medium derived from in vitro and in vivo adipocytes
Authors: Mubtasim Noshin , Boren Caleb , Barr Benjamin , Gollahon Lauren 
Published: April 2023

Abstract
Under obese conditions, dysregulation of the normal secretion of adipokines from fat cells has been shown to drive cancer progression. However, the role adipokine dysregulation plays in driving metastasis is still unclear. Recent studies have reported the activation of metastasis-associated cell signaling pathways. One such pathway involves Focal Adhesion Kinase (FAK), a non-receptor cytoplasmic protein that acts as a signal transducer between receptor activation by adipokines and downstream effector proteins that control the biological phenomenon of metastasis. Nevertheless, the association between obesity-induced changes in adipocyte secretory factors and the subsequent increase in the transactivation of FAK through phosphorylation is unknown. To elucidate the potential relationship of adipocyte-secreted factors towards activating FAK, breast cancer cells with low (MCF7) and high (MDA-MB-231) metastatic potential were exposed to in vitro and in vivo, murine adipocyte-derived, conditioned medium. For the in vitro adipocyte model, 3T3-L1 MBX cells differentiated to obesity, with fatty acids supplementation, were used. For the in vivo model, adipocyte-secreted factors from organ-cultured visceral fat harvested from C3H/HeJ female mice fed an either high fat (46%) or control (11%) diets, were collected in the medium. Adipocyte secretions from the in vitro and in vivo adipocyte samples were determined using a mouse adipokine proteomic profiler array and ELISA kit. Under obese conditions, both the 3T3-L1 MBX cells and C3H/HeJ visceral fat were found to have significant changes in their adipokine secretions in comparison to controls. Results showed that the breast cancer cells got increased FAK autophosphorylation when exposed to either in vitro or in vivo-derived obese adipocyte-conditioned medium in comparison to the untreated breast cancer cells.
Citation Format: Noshin Mubtasim, Caleb Boren, Benjamin Barr, Lauren Gollahon. Post-translational modification of focal adhesion kinase (FAK) protein increases in breast cancer cells upon exposure to conditioned medium derived from in vitro and in vivo adipocytes [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 2560.
Abstract 2563: Identifying BCLAF1 as a downstream effector of p85β in ovarian cancer
Authors: Wang Panpan , Rao Ling , Mak Victor CY , Cheung Lydia WT 
Published: April 2023

Abstract
The phosphatidylinositol 3-kinase (PI3K)/AKT pathway is dysregulated in a broad spectrum of human cancers. Genomic alterations of the pathway members can cause cell transformation and tumorigenesis. The class IA PI3K is a dimeric enzyme composed of a catalytic subunit and a regulatory subunit to initiate AKT activation, with the two major regulatory subunits being p85α and p85β. Apart from sharing a canonical function in binding to and stabilizing p110α, we and the others have demonstrated that p85α and p85β carry distinct activities. We have recently reported an oncogenic role of p85β in ovarian cancer cells. In this study, we analyzed multiple omics data to further explore the oncogenic mechanism of p85β. BCL2 Associated Transcription Factor 1 (BCLAF1) was identified as a downstream effector of p85β. We found that the protein level of BCLAF1 was consistently decreased upon PIK3R2 silencing in ovarian cancer cell lines, and that the decrease could be reversed after PIK3R2 overexpression. Importantly, knockdown of BCLAF1 reduced the oncogenicity of p85β, whereas BCLAF1 overexpression promoted cell viability, cell invasion and migration. Accompanied with the increase in total protein level was enhanced phosphorylation of multiple serine residues of BCLAF1. Functional analysis of the serine residues by expressing the phosphorylation-defective mutants suggested that Ser161, Ser512 and Ser531 are critical for BCLAF1 oncogenic functions in ovarian cancer. Together, the findings reveal a new downstream effector by which p85β exerts its oncogenic effects in ovarian cancer.
Citation Format: Panpan Wang, Ling Rao, Victor CY Mak, Lydia WT Cheung. Identifying BCLAF1 as a downstream effector of p85β in ovarian 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 2563.
Abstract 2573: A novel regulation of the Wnt/β-catenin pathway involving NLRP12 during colorectal tumorigenesis
Authors: Khan Shahanshah , Peng Lan , Lewis Cheryl M. , Gao Yunpeng , Mani Ram S. , Zaki Hasan 
Published: April 2023

Abstract
NLRP12, a member of the NOD-like receptor family, has been characterized as a negative regulator of NF-κB and MAPK pathways in immune cells. We previously demonstrated that Nlrp12-deficient mice are highly susceptible to azoxymethane plus dextran sodium sulfate induced colorectal tumorigenesis in mice. However, the precise mechanism of NLRP12-mediated protection against colorectal cancer (CRC) was poorly understood. RNA-seq analysis revealed that tumors of Nlrp12-deficient mice express higher levels of protooncogenes (cMyc, Ccnd1), Wnt target genes (Ctnnb1, Axin2, Yap1, Tcf3, Lgr5), matrix metalloproteinases, and epithelial to mesenchymal transition markers (Vim, Fn1, Foxc2, Zeb2, Ezh2) compared to wild-type tumor. Interestingly, higher expression of these genes in Nlrp12-deficient tumors was not linked to any inflammatory signaling pathways as there was no difference in the activation of NF-kB, ERK, JNK, AKT, and STAT3 signaling pathways between Nlrp12-/- and wild-type tumors. However, we observed higher activation of β-catenin in Nlrp12-deficient tumors. Using intestinal epithelial cell specific Nlrp12-conditional knockout mice we showed that Nlrp12 deficiency in intestinal epithelial cells leads to increased tumorigenesis, metastasis, and β-catenin activation. To further understand the role of NLRP12 in the regulation of the Wnt/β-catenin, we overexpressed NLRP12 in HEK293T cells and HCT116 cells followed by stimulation with Wnt ligand Wnt3A. NLRP12 overexpressed cells showed significantly reduced activation of β-catenin and expression of Wnt target genes. Consistently, knocking down of NLRP12 in HEK293T and HCT116 cells caused higher activation of the Wnt/β-catenin pathway. Further biochemical analysis revealed that NLRP12-mediated suppression of β-catenin activation is associated with reduced phosphorylation of GSK3β. Interestingly, human and mouse colorectal tumors showed reduced expression of NLRP12 along with increased activation of β-catenin and GSK3β phosphorylation. Altogether, these data suggest that NLRP12 is a potent negative regulator of the Wnt/β-catenin pathway in colorectal tumor cells, and the NLRP12-GSK3β axis could be a novel target for CRC treatment.
Citation Format: Shahanshah Khan, Lan Peng, Cheryl M. Lewis, Yunpeng Gao, Ram S. Mani, Hasan Zaki. A novel regulation of the Wnt/β-catenin pathway involving NLRP12 during colorectal tumorigenesis [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 2573.
Abstract 2575: A multi-omic screening approach identifies the E3-ubiquitin ligase DCAF13 as a novel vulnerability in lung adenocarcinoma
Authors: Wagner Vinona T. , Chang Yun-Chien , Giansanti Piero , Öllinger Rupert , Engleitner Thomas , Kuster Bernhard , Bassermann Florian 
Published: April 2023

Abstract
Lung adenocarcinoma (LuAD) is the most prevalent subtype of lung cancer but despite advances in targeted therapy approaches, LuAD patients still have a 5-year survival rate of less than 15%. This is largely owed to a high variability in cancer driver mutations, with only around 25% of patients being eligible for targeted therapy, highlighting the need for therapy approaches beyond the level of genetic modifications. In order to determine novel vulnerabilities of LuAD, we performed a CRISPR/Cas9-based genomic screen with a pooled sgRNA library targeting genes of the ubiquitin system. The screen identified the poorly characterized E3-ubiquitin ligase DCAF13 as an essential gene for LuAD cell survival. Data mining of LuAD patient data revealed that DCAF13 is significantly overexpressed in tumor versus normal tissue and that high DCAF13 expression correlates with adverse overall survival and progression free survival. Mechanistically, DCAF13 depletion leads to a decrease in cell proliferation and subsequent cell death, which can be rescued by apoptosis inhibition. Immunofluorescence revealed that DCAF13 localizes to the centrosome, cell-cell contacts and the nucleus of LuAD cells, indicating a potential role in transcription control. Functionally, DCAF13 is a predicted substrate recruiting unit of a CUL4-family ubiquitin ligase. Indeed, we show that DCAF13 binds to CUL4B and DDB1 to form an active ubiquitin ligase complex. In an effort to identify lung cancer specific relevant ubiquitylation substrates of DCAF13, we performed affinity-based interactome screens and non-affinity-based functional proteomics in LuAD cells. Cross-validation of these approaches revealed distinct tumor-suppressive transcription factors, which are known to play a role in tumorigenesis, embryogenesis and stem cell differentiation. Ongoing biochemical and transcriptomic analyses further determine the involvement of these transcription factors and DCAF13 in the establishment and maintenance of LuAD. Taken together, we nominate DCAF13 as a novel vulnerability in LuAD with potential implication in future targeted therapy strategies.
Citation Format: Vinona T. Wagner, Yun-Chien Chang, Piero Giansanti, Rupert Öllinger, Thomas Engleitner, Bernhard Kuster, Florian Bassermann. A multi-omic screening approach identifies the E3-ubiquitin ligase DCAF13 as a novel vulnerability in lung 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 2575.
Abstract 258: Whole genome analysis of hereditary leiomyomatosis and renal cell cancer identifies role of localized hypermutational events
Authors: Wu Selina , Boutros Paul C. 
Published: April 2023

Abstract
Hereditary leiomyomatosis and renal cell cancer (HLRCC) is a rare hereditary condition affecting over 1 in 200,000 people worldwide. Individuals with HLRCC are also at risk of developing an aggressive form of renal cell carcinoma (RCC), as 15-20% of patients develop type 2 papillary RCC associated with poor prognosis. Although RCC risk and cancer penetrance of this condition has been studied, whole genome sequencing of associated tumors has yet to be examined, and the landscape of structural variants of HLRCC is largely unknown. We have therefore performed whole genome sequencing of tumor/normal pairs of HLRCC patients to study the tumor molecular architecture in this form of RCC. Through germline variant and somatic mutation calling, we have identified specific somatic mutational drivers, including those at the structural variant level for the first time in HLRCC. We have also quantified both chromothripsis and kataegis in these tumors, identifying the specific role of localized hypermutational events. Similarly, using tumour subclonal reconstruction techniques, we have inferred the clonality of HLRCC-associated RCC tumors and elucidate the relative timing of key oncogenic events and mutational processes. We also compare each of these mutational features to their equivalents in other RCC tumour subtypes. These studies shed key light on the specific mutational processes and drivers of this rare hereditary malignancy and on its lethality.
Citation Format: Selina Wu, Paul C. Boutros. Whole genome analysis of hereditary leiomyomatosis and renal cell cancer identifies role of localized hypermutational events [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 258.
Abstract 2871: Spatial landscape of the tumor microenvironment in pancreatic ductal adenocarcinoma using multiplexed imaging and AI based analysis
Authors: Smith Michael J. , Huang Chi-Chou , Phan Tuan H. , Sasaki Hideki , White Katie O. , Heil-Chapdelaine Richard A. , Angus-Hill Melinda 
Published: April 2023

Abstract
Pancreatic tumors are highly heterogenous, often with more aggressive regions that are responsible for invasion and metastasis. These aggressive pancreatic ductal adenocarcinoma (PDAC) cells undergo epithelial to mesenchymal transition and create subregions in the tumor that evade treatments and provide a critical support niche for continued tumor growth and metastasis. Conventional chemotherapy plus radiation, or in advanced disease chemotherapy plus targeted drug therapy can lengthen patient survival. However, even in patients with local disease, the 5-year survival rate remains at around 40%, demonstrating that there is need for improved conventional and immune therapies for all pancreatic cancer patients. In this study, dozens of biomarkers have been used to probe tumor heterogeneity in pancreatic ductal adenocarcinoma (PDAC) tissue. We rely on the published finding that hypoxia gene expression is consistent in PDAC, regardless of tumor location, whether primary tumor or metastasis. With the spatial biology solution, Cell DIVETM and proprietary Spatial AI analytics software, dozens of biomarkers can be used to computationally reduce tumor heterogeneity, and to spatially define cells in the microenvironment within hypoxic and normoxic regions of PDAC, and in normal pancreas. We found that these hypoxic PDAC regions are devoid of immune cells in general, with the appearance of spatially co-localized populations of cell types consistent with tumor desmoplasia and inflammation. Taken together, multiplexed whole slide imaging and analysis enables spatially resolved whole tissue analysis of the tumor microenvironment, including new insights into spatial biology in the hypoxic PDAC environment.
Citation Format: Michael J. Smith, Chi-Chou Huang, Tuan H. Phan, Hideki Sasaki, Katie O. White, Richard A. Heil-Chapdelaine, Melinda Angus-Hill. Spatial landscape of the tumor microenvironment in pancreatic ductal adenocarcinoma using multiplexed imaging and AI based analysis [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 2871.
Abstract 2878: Identification of immunosuppressive and tumor-promoting neutrophils in human bone marrow and a novel mouse model that supports human neutrophil engraftment
Authors: Liu Wei , Che Keying , Lu Chun , Wei Jia , Li Yan 
Published: April 2023

Abstract
Immunosuppressive neutrophils are identified as a specialized heterogeneous population in cancer patients that accelerate tumor progression. However, the developmental stage and the source of this population remain elusive. To address this issue, we unified three reported neutrophil nomenclatures and demonstrated that immature neutrophils predominate tumor tissues of patients. On the quest to identify the origin of immature neutrophils, we unexpectedly discovered that immature neutrophils collected from the bone marrow of healthy people and mice reconstituted with human immune cells (HIS mice) are naturally immunosuppressive and tumor-promoting, whereas the mouse counterpart is not. Single-cell comparison of immature neutrophils isolated from human tumors, human bone marrow and HIS mouse bone marrow samples reveal FCN1 as a common marker for this heterogeneous population. To further investigate the link between peripheral and bone marrow immature neutrophils, we created a novel immunodeficient strain, to overcome the poor human neutrophil engraftment in the periphery. HIS mice based on this strain manifest similar kinetics of immature neutrophil expansion as cancer patients both in the bone marrow and periphery after tumor cell inoculation. Finally, we proved that eliminating bone marrow immature neutrophils delays tumor growth in HIS mice. In summary, human immunosuppressive neutrophils are predominantly immature neutrophils and targeting this population in bone marrow may represent a novel strategy for cancer immunotherapy.
Citation Format: Wei Liu, Keying Che, Chun Lu, Jia Wei, Yan Li. Identification of immunosuppressive and tumor-promoting neutrophils in human bone marrow and a novel mouse model that supports human neutrophil engraftment [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 2878.
Abstract 2881: Investigating cholesterol transfer between macrophages and prostate cancer cells
Authors: Osunmakinde Olabisi , Ruffell Brian 
Published: April 2023

Abstract
Background: Androgens stimulate androgen receptor (AR) signaling, driving prostate cancer (PC) cell proliferation and progression. Androgen deprivation therapy (ADT) effectively treats primary disease, but 20-30% of patients relapse after ~5 years, resulting in castration-resistant prostate cancer (CRPC), which has limited treatment options. We have previously shown in a murine model of CRPC that macrophages promote AR signaling in CRPC and that depleting tumor macrophages extends survival in response to ADT with Lupron. This correlates with reduced androgen levels in macrophage-depleted tumors and the ability of macrophages to transfer cholesterol, the anabolic precursor of androgens, to prostate cancer cells. However, the mechanism by which this transfer occurs, and whether cholesterol exchange mediates resistance to ADT, is currently unknown.
Methods: To assess cholesterol transfer from macrophages to cancer cells, we established a culture system involving prostate cancer cell lines and either the RAW264.7 macrophage cell line or bone marrow-derived macrophages (BMDMs). Macrophages were pre-loaded with cholesterol using fluorescently labeled low-density lipoprotein (LDL) and were co-cultured or placed in a transwell system with tumor cells for 24 hours. Cholesterol transfer was then measured by flow cytometry.
Results: Fluorescent signal from LDL was detected in 50-70% of tumor cells after 24 hours of co-culture. Cell contact was required for cholesterol transfer, as no fluorescence was detected in prostate cancer cells during transwell experiments. Macrophages were also able to internalize and transfer acetylated LDL, which is poorly taken up by tumor cells compared to unmodified LDL. Macrophage polarization with interleukin-4 did not impact cholesterol transfer and uptake of acetylated LDL occurred independently of the scavenger receptor CD36. The scavenger receptor SR-B1, which is often upregulated in prostate cancer cells, was dispensable for transfer, as knocking out SR-B1 via CRISPR/Cas9 in cancer cell lines did not reduce LDL uptake.
Conclusion: Cholesterol transfer between macrophages and prostate cancer cells occurs in a contact-dependent manner, independently of the scavenger receptors most commonly associated with cholesterol accumulation.
Citation Format: Olabisi Osunmakinde, Brian Ruffell. Investigating cholesterol transfer between macrophages and prostate cancer cells [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 2881.