tbuckley/cabot-search huggingface dataset:
Abstract 1798: Glyoxalase 1 (<i>GLO1</i>) as an oncometabolic enabler of prostate cancer progression: CRISPR/Cas9-based <i>GLO1</i>-deletion from human DU145-Luc2 cells blocks EMT and bone metastasis in SCID mice
Authors: Jana Jandova, Anne E. Cress, Georg T. Wondrak
Journal: Cancer Research (2024)

Abstract Glyoxalase 1 (encoded by GLO1) is a glutathione-dependent enzyme detoxifying the glycolytic byproduct methylglyoxal (MG), an oncometabolite involved in metabolic reprogramming. Recently, we have demonstrated that GLO1 is overexpressed in human prostate cancer cells and patient tumors. In order to inform the ongoing debate on the role of GLO1 as an oncometabolic enabler of tumor glucose metabolism and malignant progression, we performed CRISPR/Cas9-based GLO1-deletion in human DU145-Luc2 malignant prostate cancer cells. NanoString nCounterTM ('PanCancer-Progression-Panel') comparative gene expression profiling (GLO1_KO versus GLO1_wt) revealed a significant downregulation of EMT-related pathways in GLO1_KO cells; concordantly, phenotypical screening indicated a pronounced attenuation of matrigel invasiveness observable in GLO1_KO cells. Likewise, inclusion of MG or a small molecule GLO1 inhibitor (TLSC-702) blocked invasiveness of DU145 GLO1_wt cells. Downregulation of EMT-related genes (including MMP3, SPP1, CXCL8) was accompanied by increased expression of TXNIP (thioredoxin-interacting protein), a master regulator of cellular energy metabolism and redox homeostasis. In a bioluminescent SCID mouse bone metastasis model (intracardial injection of DU145-Luc2 GLO1_wt and GLO1_KO cells), GLO1 expression was necessary to cause mandibular bone metastases (as evidenced by the complete absence of bone metastases after GLO1 deletion). Given the availability of drug-like small molecule inhibitors of GLO1 enzymatic activity these data suggest that GLO1 represents a novel molecular target for the pharmacological suppression of prostate cancer bone metastasis. Citation Format: Jana Jandova, Anne E. Cress, Georg T. Wondrak. Glyoxalase 1 (GLO1) as an oncometabolic enabler of prostate cancer progression: CRISPR/Cas9-based GLO1-deletion from human DU145-Luc2 cells blocks EMT and bone metastasis in SCID mice [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 1798.
Abstract 4243: Revolutionizing cancer research, new drug screening approach with vascularized cancer organoid platform mimicking patient tumor microenvironment
Authors: Mingyun Lee, Yeo-Joon Yoon, Yong-Soo Lee, Jinguen Rheey
Journal: Cancer Research (2024)

Abstract Patient-derived organoids (PDOs) are improving our understanding of cancer heterogeneity and personalized medicine. In particular, patients' intrinsic and acquired treatment resistance is being evaluated with PDOs, which has become an essential factor in developing new anticancer drugs and designing successful clinical trials. However, there are obstacles to cancer heterogeneity and patient-specific cancer therapy due to lack of understanding of tumoral diversity resulting from the tumor microenvironment (TME). Complex direct and indirect interactions between various types of TME cells and the resulting tumoral diversity are challenges that must be addressed for customized cancer therapy. Standard cancer therapy targets tumor cells without considering possible damage on the tumor microenvironment that could impair therapy response. Therefore, the construction and analysis of a platform for co-culturing cancer organoids with other cell types that make up the TME over a long period of time is necessary. We generated and performed characterization of blood vessel organoid (BVO). The BVO expressed endothelial marker CD31 and pericyte marker PDGFRβ. We attempted to co-culture with patient-derived lung cancer organoid (LCO) on Matrigel by dissociating the constructed BVO, and it was confirmed that BVO and LCO were interconnected after 10 days. This new platform was treated with Gefitinib and Poziotinib, and after 5 days of drug treatment, the IC50 value increased compared to LCO-only culture. Additionally, flow cytometry analysis confirmed that co-culturing with BVO during drug treatment resulted in a lower percentage of dead LCO cells compared to LCO-only culture. Image-based analysis also verified that co-culturing with blood vessels enhances the viability of LCOs when treated with drugs. Taken together, the co-culture of cancer organoids and BVOs present a platform that closely mimics the in vivo TME compared to the existing cancer organoid culture method. This innovative platform demonstrates higher drug resistance than existing approaches, offering a novel method for drug screening. Citation Format: Mingyun Lee, Yeo-Joon Yoon, Yong-Soo Lee, Jinguen Rheey. Revolutionizing cancer research, new drug screening approach with vascularized cancer organoid platform mimicking patient tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4243.
Abstract 6808: Investigating the role of CXCR3+ monocytes on CAR T cell function
Authors: Claire Victoria Ong, Sabina Kaczanowska, Wei Ju, James Cronk, Sneha Ramakrishna, Rosandra Kaplan
Journal: Cancer Research (2024)

Abstract Chimeric antigen receptor T cells (CAR-Ts) are an emerging immunotherapy that have remarkable efficacy against leukemias and lymphomas but limited success against solid tumors. We conducted a phase I clinical trial (NCT 02107963) of GD2 CAR-T (GD2-CAR.OX40.28.z.ICD9) administration to children and young adults with neuroblastoma and, for the first time, osteosarcoma. While all patients progressed and were switched to alternative therapy, we observed a significant association between good CAR-T expanders and expression of C-X-C Motif Chemokine receptor 3 (CXCR3) on monocytes detected in pretreatment apheresis. This study seeks to uncover the possible interactions between CXCR3 and CAR-Ts that may alter CAR-T function and ultimately lead to better CAR-T expansion. We hypothesize that CXCR3+ monocytes induce functional change in CAR-Ts resulting in improved expansion. To recapitulate the interaction between tumor, CAR-Ts and monocytes, we took 143Bs (osteosarcoma cell line) and GD2 CAR-Ts and co cultured them with THP-1s (a monocytic leukemia cell line) that were either untransduced (UTD) or transduced via lentivirus to express CXCR3 (CXCR3+). The ratio of UTD or CXCR3+ THP-1s to CAR-Ts and 143Bs was varied between co cultures. Enzyme-linked immunoassay (ELISA) for interferon gamma (IFNy) expression was run as a proxy measurement for CAR-T cell activity. Flow cytometry was used to assess for expression of activation and exhaustion markers. Differences between UTD and CXCR3+ co cultures were evaluated for statistical significance via unpaired t-test. We observed that CXCR3 was elevated on GD2 CAR-Ts co cultured with CXCR3+ monocytes. This pattern was seen most prominently in CD4+ CAR-Ts, but also subtly in CD8+ T-cells. Exhaustion markers CD39 and LAG3 were elevated in CAR-Ts co cultured with CXCR3+ monocytes. Additionally, CAR-Ts co cultured with CXCR3+ monocytes displayed greater activity via elevated IFNy expression. We conclude that GD-2 CAR-T cells co cultured with CXCR3+ monocytes express elevated exhaustion markers due to possible increased activity when in the presence of CXCR3+ monocytes. Additionally, we suspect that elevated CXCR3 expression on CAR-Ts co cultured with CXCR3+ monocytes may be helping CAR-Ts better target tumor cells, thus leading to improved expansion. If CXCR3 can improve CAR-T cell function, this could lead to better optimization of CAR-Ts against solid tumors and create a viable treatment option for neuroblastoma and osteosarcoma patients. Citation Format: Claire Victoria Ong, Sabina Kaczanowska, Wei Ju, James Cronk, Sneha Ramakrishna, Rosandra Kaplan. Investigating the role of CXCR3+ monocytes on CAR T cell function [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 6808.
Abstract 3025: Germline p53 R181 variants and DNA binding cooperativity in tumorigenesis
Authors: Renyta Moses, Ryan Hausler, Gregory Kelly, Alexandra Indeglia, Sven Miller, John Karanicolas, Maureen Murphy, Kara Maxwell
Journal: Cancer Research (2024)

Abstract TP53 is the most frequently mutated gene in cancer, and its encoded protein p53 has many tumor-suppressive functions. There are several different classes of mutant p53 acquired in human cancer and inherited in cancer-prone families with Li-Fraumeni Syndrome, where individuals have an 80-90% increased risk of cancer. These include structural mutations that are generally (e.g R175H) or locally (e.g Y220C) misfolded, DNA contact mutations (e.g R273H); and oligomerization mutations (A347D). Cooperativity mutations affect binding of the p53 tetramer to DNA via disruption of a salt bridge formed by the negatively charged glutamic acid (E) 180 residue of one p53 monomer and the positively charged arginine (R) 181 residue of another. p53 “cooperativity” mutations at the R181 residue (R181H and R181C) have been increasingly identified in cancer-prone families undergoing genetic testing; however, the mechanism by which these variants disrupt p53 tumor suppression in humans is not understood. We show that the purified DNA binding domains of p53 variants R181H and R181C bind less cooperatively to the p53 binding site in the CDKN1A (p21) promoter, despite retaining wild-type levels of structural stability. RNA-sequencing of CRISPR knock-in colorectal and breast cancer cell lines shows reduced ability of R181H and R181C to transactivate a curated set of ~300 known p53 target genes. Upon treatment with p53 activating molecule Nutlin-3a, the R181-mutant cells fail to cell cycle arrest in the G1 phase and maintain high proliferative rates. Interestingly, we observe some residual apoptotic activity in R181H and R181C mutant cells treated with DNA-damaging agent 5-fluorouracil, despite losing the transactivation of proapoptotic p53 targets; this suggests that these mutants retain the p53 transcription-independent mechanism of apoptosis which is observed in other p53 variants such as P47S and A347D. These studies will define the pathogenicity of R181 variants and guide therapeutic intervention for patients that harbor these p53 mutations. Citation Format: Renyta Moses, Ryan Hausler, Gregory Kelly, Alexandra Indeglia, Sven Miller, John Karanicolas, Maureen Murphy, Kara Maxwell. Germline p53 R181 variants and DNA binding cooperativity in tumorigenesis [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 3025.
Abstract 1793: Transmembrane serine protease TMPRSS11B promotes tumorigenesis in lung squamous cell carcinoma through enhanced lactate export and modulation of the tumor microenvironment
Authors: Hari Shankar Sunil, Barrett L. Updegraff, Jingfei Zhu, Lisa Thomas, Bret M. Evers, Jean R. Clemenceau, Isabel Barnfather, John D. Minna, Ralph J. Deberardinis, Trudy G. Oliver, Tae Hyun Hwang, Kathryn A. O'Donnell
Journal: Cancer Research (2024)

Abstract Lung cancer is the leading cause of cancer-related deaths worldwide. Existing therapeutic options have limited efficacy, underscoring the critical need for the identification of new actionable therapeutic targets. We previously identified the Transmembrane Serine Protease TMPRSS11B as a novel gene that promotes the transformation of human bronchial epithelial cells in vitro and induces tumorigenesis in vivo. TMPRSS11B promotes extracellular release of Basigin, an obligate chaperone of the lactate monocarboxylate transporters MCT1/4, enhancing lactate export and glycolytic metabolism, and thereby promoting tumorigenesis. Importantly, TMPRSS11B is frequently overexpressed in human lung squamous cell cancers (LSCC) and high expression is associated with poor patient survival. Moreover, as a cell surface protein and enzyme, TMPRSS11B represents a tractable target for therapeutic intervention. To investigate whether TMPRSS11B activity impacts the host immune system and the tumor microenvironment (TME), we evaluated the effect of Tmprss11b depletion in a syngeneic LSCC mouse model, KLN205. Tmprss11b loss of function in KLN205 LSCC cells significantly reduced tumor burden in immunocompetent mice and triggered an accumulation of CD4+ T cells. We are currently extending these studies by investigating the effect of Tmprss11b loss of function using CRISPR/Cas9 editing in the Rosa26LSL-Sox2-IRES-GFP; Nkx2-1 fl/fl; Lkb1fl/fl (SNL) mouse model of LSCC. Tumor burden analysis and pH-sensitive fluorescent imaging are ongoing to assess changes in lactate accumulation. Moreover, RNA FISH analysis and spatial transcriptomics revealed that Tmprss11b expression is highly enriched in lung squamous tumors compared to lung adenocarcinomas and normal lung in SNL mice. Spatial analysis of the transcriptome is currently ongoing to assess the effects of Tmprss11b expression on the TME. Collectively, these studies will elucidate the molecular mechanisms through which TMPRSS11B promotes tumor growth and alters the tumor microenvironment in LSCC. Citation Format: Hari Shankar Sunil, Barrett L. Updegraff, Jingfei Zhu, Lisa Thomas, Bret M. Evers, Jean R. Clemenceau, Isabel Barnfather, John D. Minna, Ralph J. Deberardinis, Trudy G. Oliver, Tae Hyun Hwang, Kathryn A. O'Donnell. Transmembrane serine protease TMPRSS11B promotes tumorigenesis in lung squamous cell carcinoma through enhanced lactate export and modulation of the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1793.
Abstract 1830: Bergenin inhibits growth of human cervical cancer cells by decreasing galectin-3 and MMP-9 expression
Authors: Mayank Singh, Ravi Chauhan, Lakshay Malhotra, Ashna Gupta, Gunjan Dagar, Dayasagar Das, Shahab Uddin, Ethayathulla Abdul Samath, Muzafar A. Macha, Ammira Al shabeeb Akil, Ajaz A. Bhat
Journal: Cancer Research (2024)

Abstract Cervical cancer is still the leading of cancer mortality worldwide, even after introducing a vaccine against Human papillomavirus (HPV), due to low vaccine coverage, especially in the developing world. Cervical cancer is primarily treated by Chemo/Radiotherapy, depending on the disease stage, with Carboplatin/Cisplatin-based drug regime. These drugs being non-specific, target rapidly dividing cells, including normal cells, so safer options are needed for more targeted therapy and lower off-target toxicity. Natural products offer an attractive option compared to synthetic drugs due to their well-established safety profile and capacity to target multiple oncogenic hallmarks of cancer like inflammation, angiogenesis, etc. In the current study, we investigated the effect of Bergenin (C-glycoside of 4-O-methylgallic acid), a natural polyphenol compound that is isolated from medicinal plants such as Bergenia crassifolia, Caesalpinia digyna, and Flueggea leucopyrus. Bergenin has been shown to have anti-inflammatory, anti-ulcerogenic, and wound healing properties. Still, its anticancer potential has been realized only recently. We performed a proteomic analysis of cervical carcinoma cells treated with bergenin and found it to influence multiple hallmarks of cancers, including apoptosis, angiogenesis, and tumor suppressor proteins. It was also involved in many different cellular processes unrelated to cancer, as shown by our proteomic analysis. Further analysis showed bergenin to be a potent -angiogenic agent by reducing key angiogenic proteins like Galectin 3 and MMP-9 (Matrix Metalloprotease 9) in cervical carcinoma cells at cellular level as shown by western blotting assay. Further understanding of this interaction was carried out using molecular docking analysis, which indicated MMP-9 has more affinity for Bergenin as compared to Galectin-3. Cumulatively, our data provide novel insight into the anti-angiogenic mechanism of Bergenin in cervical carcinoma cells by modulation of multiple angiogenic proteins like Galectin-3 and MMP-9 which warrant its further development as an anticancer agent in cervical cancer. Citation Format: Mayank Singh, Ravi Chauhan, Lakshay Malhotra, Ashna Gupta, Gunjan Dagar, Dayasagar Das, Shahab Uddin, Ethayathulla Abdul Samath, Muzafar A. Macha, Ammira Al shabeeb Akil, Ajaz A. Bhat. Bergenin inhibits growth of human cervical cancer cells by decreasing galectin-3 and MMP-9 expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1830.
Abstract 1114: Evaluating strategies to harness metabolic vulnerabilities in cervical
Authors: Dr Julie K. Schwarz, Dr Nishanth Gabriel, Aunannya Banik
Journal: Cancer Research (2024)

Abstract Tumor cells frequently alter their metabolism to generate biomolecular intermediates for tumor maintenance and proliferation. Previous reports from Dr. Julie Schwarz’s laboratory have shown that cervical cancer cells with altered AKT/mTOR pathways show increased reliance on glucose and exogenous glutamine. The laboratory has introduced PIK3CA E545K mutation and PTEN null mutation in cervical cancer cell line SiHa. Further, to evaluate the role of glutamine metabolism in these genotypes, these cells were cultured in sequentially lower levels of glutamine until these cells were able to proliferate without exogenous glutamine supplementation. These isogenic long-term glutamine-deprived (QD) cells allow us to tease out the role of these specific mutations in cellular metabolism. In our study, long-term glutamine deprivation inhibits the proliferation of rapidly proliferating PI3K-AKT-altered cell lines. Even though glutamine is required for GSH synthesis, which quenches reactive oxygen species, QD cells were refractory to cisplatin and irradiation. Since PI3K-AKT altered were dependent on exogenous glutamine, it was expected that QD cells would have lower GLS levels to inhibit glutaminolysis. However, GLS levels in WT and QD were similar. Altogether, these data indicate that in PI3K-AKT altered cervical tumor glutamine antagonists can reduce tumor proliferation. Further glutamine reduction does not alter GLS levels and hence GLS can be targeted as well to further improve treatment outcomes. To evaluate the alternate metabolic dependencies in cervical cancer tumors, both cellular as well as secreted metabolites were evaluated. To identify alternative pathways that are activated in these glutamine-deprived cell lines, we cultured these cells in uniformly labeled 13C glucose. Mass spectrometric studies also evaluated the protein levels of various metabolic intermediates in glutamine-deprived cell lines compared to control cells. To confirm our observations, we used kit-based assays to evaluate metabolites in the cells. To further confirm the change in the metabolite pathway we evaluated changes in transcriptional changes of regulating proteins/enzymes. This pre-clinical study reinforced efforts that are anticipated to reveal new metabolic vulnerabilities of cancer cells that can be targeted by therapeutic interventions. This study also helped us evaluate druggable metabolic targets that can be targeted for effective tumor eradication along with conventional treatment. This understanding will help advance the pre-clinical trials persistent to the cure of cervical cancer. Citation Format: Dr Julie K. Schwarz, Dr Nishanth Gabriel, Aunannya Banik. Evaluating strategies to harness metabolic vulnerabilities in cervical [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 1114.
Abstract 424: FDX2-KO induces global down-regulation of iron-sulfur cluster-containing proteins and senescence-like growth arrest or death in ovarian cancer cells
Authors: Shuko Miyahara, Mai Ohuchi, Miyuki Nomura, Kayoko Hayashi, Muneaki Shimada, Nobuo Yaegashi, Nobuhiro Tanuma
Journal: Cancer Research (2024)

Abstract Iron-Sulfur clusters (Fe-S) are synthetized in mainly mitochondria and binds to various proteins (Fe-S proteins). Fe-S proteins play important roles in various cellular functions including energy metabolism, nucleic acid synthesis or redox homeostasis. However, roles of Fe-S clusters in cancer cells are not fully understood. Here, we report that loss of Fe-S biosynthesis causes proliferation arrest with DNA-damage response or cell death depend on TP53 status in ovarian cancer (OVC) cells. CRISPR-screening of all metabolism-related genes identified that Fe-S biosynthesis is essential for OVC proliferation. We chose one of the Fe-S biosynthesis factors, FDX2, and developed a cell line that can maintain FDX2 expression doxycycline (Dox) -dependently. In these cells, FDX2-loss induced irreversible growth arrest. FDX2-KO cells also showed enlarged and flatten morphology, up-regulation of SASP factors, and DNA damage through p53/p21 pathway activation. These were all senescence like phenotypes. To know these mechanisms, we performed proteome analysis and found that FDX2-loss caused global but differential post-transcriptional down-regulation of Fe-S proteins, in turn perturbing mitochondrial respiration, iron-regulation and redox homeostasis. These results all associated with DNA damage. Furthermore, we found that FDX2-KO induced caspase dependent cell death in TP53-KO condition instead of growth arrests. These results suggest that Fe-S cluster biosynthesis is significant in OVC proliferation and fate of FDX2-KO cells depends on TP53 status (senescence or cell death). FDX2 or Fe-S biosynthesis may be a new therapeutic target of OVC. Citation Format: Shuko Miyahara, Mai Ohuchi, Miyuki Nomura, Kayoko Hayashi, Muneaki Shimada, Nobuo Yaegashi, Nobuhiro Tanuma. FDX2-KO induces global down-regulation of iron-sulfur cluster-containing proteins and senescence-like growth arrest or death in ovarian 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 424.