bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3583: Novel MRI contrast system using halloysite nanotubes embedded with iron-platinum nanoparticles for the regulation of the CHST11 expression for cancer treatment
Authors: Chan Ming-Hsien , Li Chien-Hsiu , Chang Yu-Chan , Hsiao Michael
Published: April 2023

Topic: TB Tumor Biology (TB03) In Vivo Imaging.

Multifunctional nanocomposites have been a popular research target in recent years. This research aims to produce magnetic nanocomposites with high saturation magnetization (Ms) and good biocompatibility to improve the disadvantage of iron-oxide was easy decomposition in the biological environment and overcome the unclear contrast problem of T2-weighted magnetic resonance imaging (MRI) of lung adenocarcinoma. In order to fit the suitable size of iron-based nanoparticles embedded into the halloysite nanotubes (HNT), the iron-platinum (FePt) nanoparticles were chosen as the MRI’s core materials. The FePt@HNT nanocomposites have low toxicity, and superparamagnetism is expected to be used in biomedical therapy and magnetic separation for disease treatment and environmental protection. Lung adenocarcinoma cancer cells grow in situ, usually invade, and are frequently recurrent after treatment. Since CHST11-mediated gene regulation may serve as a possible means of reducing tyrosine kinase inhibitor (TKI)-induced mitotic cell death, CHST11 modulation of integrin signaling may help lung cancer cells enhance their resistance to gefitinib. To enable the FePt@HNT nanocomposite to be effective in treating lung tumors, we designed miRNAs that inhibit CHST11 expression in an attempt to diagnose lung adenocarcinoma concurrently for treatment.

Keywords: Magnetic resonance imaging, halloysite nanotubes, FePt nanoparticles, CHST11, Lung adenocarcinoma

References

1. Zhou T, Jia L, Luo YF, Xu J, Chen RH, Ge ZJ, et al. Multifunctional nanocomposite based on halloysite nanotubes for efficient luminescent bioimaging and magnetic resonance imaging. Int. J. Nanomed. 2016;11:4765-76.

2. Abdullayev E, Joshi A, Wei WB, Zhao YF, Lvov Y. Enlargement of Halloysite Clay Nanotube Lumen by Selective Etching of Aluminum Oxide. ACS Nano 2012;6:7216-26.

Citation Format: Ming-Hsien Chan, Chien-Hsiu Li, Yu-Chan Chang, Michael Hsiao. Novel MRI contrast system using halloysite nanotubes embedded with iron-platinum nanoparticles for the regulation of the CHST11 expression for cancer treatment. [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 3583.
Abstract 3595: Epigenetic driven IL32 expression contributes to a JNK related cell stress response in breast cancer stem cells to promote cellular invasion
Authors: Wilson Megan A. , Benson Elayne M. , Gray Emma , Cairns Paige , Ouzounova Maria , Korkaya Hasan , Shull Austin Y.
Published: April 2023

Metastatic potential in basal-like breast cancers typically correspond with increased enrichment of EpCAM-/CD49f- cancer stem cells (CSC). With this premise in mind, it is important to better understand the mechanistic driver of these cell populations and their distinctive potential to interact with the tumor microenvironment (TME) for cancer promotion. Previous work from our lab has compared the 450K DNA methylation profile of EpCAM-/CD49f- poor breast cancer cell lines to that of EpCAM-/CD49f- enriched breast cancer cell lines and found the IL32 promoter to be hypomethylated in EpCAM-/CD49f- enriched cell lines, a result which corresponded basal-like patient samples in TCGA. By identifying IL32 being differentially regulated in CSC-enriched cell lines, we further sought to characterize IL32’s role in breast cancer aggressiveness. We first were able to identify several overarching mechanisms altered in siIL32 treated SUM15PT cells by RNAseq differential expression analysis (FDR p-value <0.01). Most notable from our RNAseq results was the significant enrichment of upregulated pathways involved in extracellular matrix (ECM) organization as well as significant enrichment of downregulated pathways involved in cellular and replicative stress responses. Particular examples of transcripts differentially expressed between control and siIL32-treated SUM159PT cells included COL6A1, ITGB3, and CD24 that were upregulated as well as NQO1, HMOX1, and CXCL2/CXCL3 that were downregulated. Furthermore, IL32 suppression decreased SUM159PT invasion in both an ECM-matrix cell invasion assay and a chick CAM xenograft/angiogenesis model. From our RNAseq results, we then performed a multi-pathway protein phosphorylation array to determine whether any key signaling events were affected by siIL32 knockdown in SUM159PT cells. Based on this approach, we were able observe a significant decrease in phosphorylated JNK and phosphorylated NFKB in siL32-treated cells when compared to control, both of which are well-established events that can coordinate both cell stress responses and cellular invasion. Collectively, our results reflect the notion that differential IL32 expression by promoter hypomethylation in breast CSCs plays a role to mitigating intracellular stress and subsequently allowing for breast cancer cell invasion and metastasis.

Citation Format: Megan A. Wilson, Elayne M. Benson, Emma Gray, Paige Cairns, Maria Ouzounova, Hasan Korkaya, Austin Y. Shull. Epigenetic driven IL32 expression contributes to a JNK related cell stress response in breast cancer stem cells to promote cellular invasion. [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 3595.
Abstract 3598: Cooperative NF-KappaB and Notch1 signaling promotes a macrophage-mediated metastatic phenotype in breast cancer
Authors: Duran Camille L. , Karagiannis George S. , Chen Xiaoming , Sharma Ved P. , Entenberg David , Condeelis John S. , Oktay Maja H.
Published: April 2023

Metastasis is a multistep process that leads to the formation of clinically detectable tumor foci at distant organs and, frequently, to patient demise. Only a subpopulation of breast cancer cells within the primary tumor can disseminate systemically and cause metastasis. To disseminate, cancer cells must express MenaINV, an isoform of the actin-regulatory protein, Mena, encoded by the ENAH gene that endows tumor cells with transendothelial migration activity allowing them to enter and exit the blood circulation. We have previously demonstrated that MenaINV mRNA and protein expression is induced in cancer cells by macrophage contact. Here, we report on the discovery of the precise mechanism by which macrophages induce MenaINV expression in tumor cells. We examined the promoter of the human and mouse ENAH gene and discovered a conserved NF-κB transcription factor binding site. Using live imaging of an NF-κB activity reporter and staining of fixed tissues from mouse and human breast cancer we further determined that, for maximal induction of MenaINV in cancer cells, NF-κB needs to cooperate with the Notch1 signaling pathway. Mechanistically, Notch1 signaling does not directly increase MenaINV expression, but instead enhances and sustains NF-κB signaling through retention of p65, an NF-κB transcription factor, in the nucleus of tumor cells, leading to increased MenaINV expression. In mice, these signals are augmented following chemotherapy treatment and abrogated upon macrophage depletion. Targeting Notch1 signaling in vivo decreased NF-κB signaling and MenaINV expression in the primary tumor and decreased metastasis. Altogether, these data uncover mechanistic targets for blocking MenaINV induction that should be explored clinically to decrease cancer cell dissemination and improve survival of patients with metastatic disease.

Citation Format: Camille L. Duran, George S. Karagiannis, Xiaoming Chen, Ved P. Sharma, David Entenberg, John S. Condeelis, Maja H. Oktay. Cooperative NF-KappaB and Notch1 signaling promotes a macrophage-mediated metastatic phenotype in breast 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 3598.
Abstract 3604: Epithelial Caveolin-1 regulates lung metastasis in advanced stages of breast cancer
Authors: Singh Dhirendra Pratap , Pathak Rashmi , Pandit Abhishek , Ebenezer Philip J. , Jambunathan Nithya , Kumar Sanjay , Plooy Alexander Du , Francis Joseph , White Mary E.
Published: April 2023

Organotypic metastasis is a well-orchestrated sequence of events that takes place in tumors. The interplay between secretory factors from the primary tumor and the microenvironment of a distant organ determines the fate of metastasis. A significant concern in effective triple-negative breast cancer (TNBC) treatment is lung metastasis. Caveolin-1 (Cav-1), a lipid raft protein, is an important component of the metastatic interplay and is reported to be upregulated in the advanced metastatic stage of breast cancer. Although Cav-1 has been shown to have oncogenic potential, its role in TNBC lung metastasis is not fully understood. Our data illustrates, knocking out Cav-1 in the 4T1 cells (a mouse breast cancer cell line) affects cell morphology, extracellular vesicle formation, and MMP release in vitro. Furthermore, a wound healing/cell migration assay showed reduced migration in Cav-1 KO 4T1 cells when compared to control 4T1 cells, and these findings were validated using Cav-1 siRNA. For investigating the metastatic role, we injected Cav-1 KO 4T1 and control 4T1 cells into the mammary fat pad of female BALB/c mice, a syngeneic mouse model of triple-negative breast cancer. We found no evidence of lung metastasis in Cav-1 KO 4T1 injected mice when compared to control 4T1 injected mice. This was further supported by mRNA profiling of the tumor. We observed 21 epithelial cell migration genes that are differentially expressed in Cav-1 KO tumors when compared to WT tumors. Correlation analysis from Breast Cancer Gene-Expression Miner v4.8 (bc-GenExMiner), a human database, showed a significant correlation between Cav-1 expression and several of the 21 epithelial cell migration genes, along with integrin 3 alpha (ITGα3). Interestingly, in silico protein docking predicted an interaction between Cav-1 and ITGα3, which was later confirmed by co-immunoprecipitation. The role of ITGα3 in metastasis was further corroborated by cell migration assay, which was affected when siRNA was used to silence ITGα3 gene expression. Collectively, these findings provide evidence that Cav-1 KO, along with ITGa3, plays an important role in distant lung metastasis in the 4T1 tumor-induced mice model.

Citation Format: Dhirendra Pratap Singh, Rashmi Pathak, Abhishek Pandit, Philip J. Ebenezer, Nithya Jambunathan, Sanjay Kumar, Alexander Du Plooy, Joseph Francis, Mary E. White. Epithelial Caveolin-1 regulates lung metastasis in advanced stages of breast 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 3604.
Abstract 3606: HDAC8-mediated inhibition of EP300 drives a neural crest-like transcriptional state that increases melanoma brain metastasis
Authors: Emmons Michael , Bennett Richard , Riva Alberto , Zhang Chao , Macaulay Robert , Dupéré-Richér Daphne , Fang Bin , Koomen John , Li Jiannong , Chen Ann , Seto Edward , Licht Jonathan , Smalley Keiran
Published: April 2023

Melanomas are heterogeneous and adopt multiple transcriptional states that can confer an invasive phenotype and resistance to therapy. Little is known about the epigenetic drivers of these cell states, limiting our ability to regulate melanoma heterogeneity and tumor progression. Here we identify stress-induced HDAC8 activity as the driver of a neural crest stem cell (NCSC)-like transcriptional state that increased the formation of melanoma brain metastases (MBM). Exposure of melanocytes and melanoma cells to multiple different stresses led to HDAC8 activation, a switch to a NCSC gene expression signature and the adoption of an amoeboid, invasive phenotype. This cell state enhanced the survival of melanoma cells under shear stress conditions and increased the formation of metastases in the brain. Single cell RNA-seq analyses showed that HDAC8 expression was correlated with the NCSC cell state in clinical MBM specimens. ATAC-Seq and ChIP-Seq analysis showed HDAC8 to alter chromatin structure by increasing H3K27ac and accessibility at c-Jun binding sites without changing global histone acetylation. The increased accessibility of Jun binding sites was paralleled by decreased H3K27ac and accessibility at MITF binding sites and loss of melanoma-lineage gene expression. Mass spectrometry-based acetylomics demonstrated that HDAC8 deacetylated the histone acetyltransferase (HAT) EP300 leading to its enzymatic inactivation. This, in turn, led to an increased binding of EP300 to Jun-transcriptional sites and decreased binding to MITF-transcriptional sites. Increased expression of EP300 decreased invasion and increased the sensitivity of melanoma cells to multiple stresses while inhibition of EP300 function increased invasion and resistance to stress. We identified HDAC8 as a novel mediator of transcriptional co-factor inactivation and chromatin accessibility that increases MBM development.

Citation Format: Michael Emmons, Richard Bennett, Alberto Riva, Chao Zhang, Robert Macaulay, Daphne Dupéré-Richér, Bin Fang, John Koomen, Jiannong Li, Ann Chen, Edward Seto, Jonathan Licht, Keiran Smalley. HDAC8-mediated inhibition of EP300 drives a neural crest-like transcriptional state that increases melanoma brain metastasis. [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 3606.
Abstract 3647: Anti-tumor actions of neutrophils are mediated via STAT5 inhibition
Authors: Rajgopal Sanjana , Alsamraae Massar , Costanzo-Garvey Diane , Cook Leah
Published: April 2023

The overall patient survival rate drops dramatically when localized prostate cancer (PCa) progresses to bone metastatic prostate cancer (BM-PCa). The treatment for BM-PCa is limited to palliative therapy, and it is currently incurable. Neutrophils are the most prevalent immune cell in the bone, and we have found that PCa enhances their recruitment and infiltration into the prostate tumor-bone microenvironment of patients with BM-PCa. Additionally, we observed that bone marrow neutrophils directly induce apoptosis of PCa cells both in vitro and in vivo and that this is mediated by the inhibition of STAT5, a transcription factor that promotes PCa progression. Moreover, STAT5 has also been found to contribute to the development of resistance to standard care androgen deprivation therapy. Neutrophils specifically target and kill only cells that express STAT5, as stable knockdown of STAT5 in BM-PCa cells makes them resistant to neutrophil-mediated cytotoxicity, while the overexpression of STAT5 in STAT5-negative PCa cells, which are generally resistant to killing by neutrophils, sensitizes them to neutrophil-mediated killing. Furthermore, transcriptomic analyses derived from the STAT5 knockdown and STAT5 overexpressing cells suggest that STAT5 expression in PCa induces pro-inflammatory signaling and that neutrophil-mediated cytotoxicity may be exerted via STAT5-induced IL-1 and/or IL-6. Preliminary data in our laboratory show that STAT5-induced production of IL-1 by PCa cells leads to their death by neutrophils, as knocking out the IL-1 receptor on neutrophils reduces their cytotoxic potential against STAT5-positive PCa cells. Interestingly, in addition to killing BM-PCa cells, we have demonstrated that bone marrow-derived neutrophils are also capable of killing pancreatic cancer cells. Further delineation of the role of neutrophils in STAT5 signaling in BM-PCa growth holds promise for enhancing neutrophil cytotoxicity in the bone which could lead to novel therapeutic options for bone metastatic prostate cancer.

Citation Format: Sanjana Rajgopal, Massar Alsamraae, Diane Costanzo-Garvey, Leah Cook. Anti-tumor actions of neutrophils are mediated via STAT5 inhibition. [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 3647.
Abstract 3648: MSU-42011, alone and in combination with selumetinib, reduces pERK levels in NF1 cancer cells and decreases CCL2 expression in THP-1 macrophages
Authors: Hung Pei-Yu , Lockwood Lizbeth , Liby Karen T.
Published: April 2023

Neurofibromatosis type 1 (NF1) is a common genetic disease that predisposes approximately 50% of affected individuals to develop plexiform neurofibromas (PNFs), which can progress to highly aggressive malignant peripheral nerve sheath tumors (MPNSTs) in approximately 10% of patients. NF1 is caused by mutations in the tumor suppressor gene NF1, which encodes for neurofibromin, a negative regulator of RAS activity. Selumetinib, a specific inhibitor of MEK1/2, is the only FDA-approved drug for NF1-associated PNFs. However, the anti-tumor effects of selumetinib are limited in MPNSTs and have dose-limiting side effects. Deficiency of the NF1 gene not only promotes tumorigenesis but also has broad effects on the immune cells and cytokine signaling driven by hyperactive RAS signaling. Because macrophages account for almost half of cells in NF1 lesions and their infiltration highly correlates with disease progression, we hypothesized that targeting tumor-promoting immune cells could be an alternative approach for NF1 treatment. The novel retinoid X receptor (RXR) agonist MSU-42011 reduces tumor growth in experimental Kras-driven cancers by decreasing pERK expression, reducing tumor-promoting immune cells like CD206+ macrophages and regulatory T cells, and increasing activated cytotoxic T cells. Here, we tested MSU-42011 and selumetinib, either alone or in combination, to evaluate their efficacy against NF1 cancer cells and macrophages using monoculture and conditioned media (CM) treatments. Treatment with 200 nM MSU-40211, 50 nM selumetinib, and the combination for 3 hours reduced pERK protein levels by approximately 40%, 70% and 90%, respectively vs. untreated controls. Treatment for 72 hours with selumetinib and the combination reduced the viability of PNF cells in a dose-dependent manner. Moreover, CM from human PNF cells increased monocyte chemoattractant CCL2 (C-C motif chemokine ligand 2) mRNA expression in human THP1 monocytes and THP1 macrophages differentiated by PMA. Notably, MSU-42011 and selumetinib alone similarly inhibited CCL2 mRNA expression by 25% in THP1 macrophages stimulated with CM from PNF cells, and the inhibition of CCL2 mRNA expression was enhanced to 50% with combination treatment. Taken together, our data suggest that MSU-42011 should be tested in relevant preclinical models of NF1.

Citation Format: Pei-Yu Hung, Lizbeth Lockwood, Karen T. Liby. MSU-42011, alone and in combination with selumetinib, reduces pERK levels in NF1 cancer cells and decreases CCL2 expression in THP-1 macrophages. [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 3648.
Abstract 3652: Circular RNAs repress interferon responses in the sarcoma cells and promote sarcoma growth by forging a pro-tumorigenic immune microenvironment
Authors: Guarnerio Jlenia
Published: April 2023

Circular RNAs (CircRNA) arise from events of non-conventional splicing in which the 3'-tail of an exon back-splices and joins the 5'-head of the upstream exon. Because of their structural conformation, these RNAs are incredibly stable in the cellular space, where they can regulate tumor-associated and anti-immunogenic pathways. Although circRNAs have been profiled in many tumors, their functional contribution to cancer progression is still poorly understood. In the current study, we profiled the most abundant circRNAs expressed in Soft-tissue sarcomas (STS) and characterized them functionally and mechanistically. Surgery and chemotherapy are the standard treatments for STS, but the 5-year survival rate is still below 20% for the advanced cases. Response to immunotherapies, such as immune checkpoint inhibitors, is also marginal for STS, likely because sarcoma cells are not sufficiently immunogenic to recruit a high volume of cytotoxic T cells. To implement discoveries on STS, we recently generated novel immunocompetent STS mouse models and characterized the sarcoma-elicited mechanisms to block immune-recruitment. Interestingly, we observed that two circRNAs, circCsnk1g3, and circAnkib1, promote sarcoma growth by shaping a pro-tumorigenic microenvironment, possibly due to their capabilities to regulate tumor-promoting elements extrinsic to the sarcoma cells. More specifically, we observed that circCsnk1g3 and circAnkib1 control the expression of interferon-related genes and pro-inflammatory factors in the sarcoma cells, thus directing immune cell recruitment into the tumor mass and hence their activation. Mechanistically, we observed that circRNAs might repress pro-inflammatory elements by buffering activation of the pathways mediated by RIG-I, the cytosolic viral RNA sensor. The current findings suggest that targeting specific circRNAs could augment the efficacy of sarcoma immune response to mainstay therapies.

Citation Format: Jlenia Guarnerio. Circular RNAs repress interferon responses in the sarcoma cells and promote sarcoma growth by forging a pro-tumorigenic immune 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 3652.
Abstract 3671: Targeting SLC2A1 and SLC38A2-mediated metabolic shift in SMARCA4/A2-dual deficient cancer
Authors: Zhu Xianbing , Fu Zheng , Chen Shary , Ong Dionzie , Aceto Giulio , Ho Rebecca , Steinberger Jutta , Li Eunice , Huntsman David , Foulkes William , Huang Sidong , Wang Yemin
Published: April 2023

SMARCA4 (BRG1) and SMARCA2 (BRM) are the two paralogous ATPases of the SWI/SNF chromatin remodelling complexes. These two ATPases are frequently inactivated in cancers and cancer cells deficient in either of them have been shown to depend on the remaining counterpart for survival. Contrary to this paralog synthetic lethality, concomitant loss of SMARCA4 and SMARCA2 occurs in a subset of cancers that are often associated with very poor patient outcomes. Here, we uncover that dual loss of SMARCA4/A2 represses the expression of SLC2A1, encoding the glucose transporter GLUT1, which results in reduced glucose uptake and glycolysis accompanied with increased dependency on oxidative phosphorylation (OXPHOS); adapting to this, these SMARCA4/A2-dual deficient cells rely on elevated SLC38A2, an amino acid transporter, to increase glutamine import for fueling OXPHOS. Consequently, SMARCA4/A2-dual deficient cells and tumours are highly sensitive to inhibitors targeting OXPHOS or glutamine metabolism. Furthermore, supplementation of alanine, also imported by SLC38A2, restricts glutamine uptake through competition and selectively induces death in SMARCA4/2-dual deficient cancer cells. At a clinically relevant dose, alanine supplementation synergizes with OXPHOS inhibition or conventional chemotherapy eliciting marked antitumor activities in SMARCA4/A2-dual deficient cancer cell lines and patient-derived xenografts. Our findings reveal multiple druggable vulnerabilities of SMARCA4/A2-dual loss exploiting a GLUT1/SLC38A2-mediated metabolic shift. Particularly, unlike dietary deprivation approaches, alanine supplementation can be readily applied to current regimens for better treatment of these aggressive cancers. * Authors contributed equally# Co-corresponding

Citation Format: Xianbing Zhu, Zheng Fu, Shary Chen, Dionzie Ong, Giulio Aceto, Rebecca Ho, Jutta Steinberger, Eunice Li, David Huntsman, William Foulkes, Sidong Huang, Yemin Wang. Targeting SLC2A1 and SLC38A2-mediated metabolic shift in SMARCA4/A2-dual deficient 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 3671.
Abstract 3675: Targeting the mevalonate pathway, a novel anti-ferroptosis pathway, in hepatocellular carcinoma (HCC) treatment
Authors: Chen Yiling , Lee Derek , Shuo Zhang Misty , Wing-Sum Cheu Jacinth , Chak-Lui Wong Carmen
Published: April 2023

Ferroptosis is a novel immunogenic and regulated cell death caused by iron-dependent lipid peroxidation. Glutathione peroxidase 4(GPX4)/glutathione and ferroptosis suppressor protein 1(FSP1)/Coenzyme Q10(CoQ10) are two major anti-ferroptosis systems that prevent lipid ROS-induced cell death. Our current study unprecedentedly uncovered that the Mevalonate pathway (MVA) protects HCC from ferroptosis by two parallel mechanisms: 1) by providing the lipophilic antioxidant CoQ10 for FSP1; 2) by generating isopentenyl pyrophosphate (IPP), a substrate for Selenocysteine (Sec)-tRNA modification for selenoprotein GPX4 translation. Selenoproteins are a class of proteins composed of selenocysteine, the 21st amino acid not found in the codon table. Selenocysteine is recoded by the stop codon UGA and synthesized on its tRNA. The Sec-tRNA incorporates Sec into selenoproteins such as GPX4. We showed that perturbation of the MVA pathway by genetic knockdown or pharmacologic inhibition repressed CoQ10 synthesis and the translation of selenoproteins, thereby leading to lipid-derived ROS and ferroptotic cell death. Furthermore, we showed that Mevalonate diphosphate decarboxylase (MVD), a MVA enzyme, was a transcriptional target of Nuclear Factor Erythroid 2- related factor 2 (NRF2), a master regulator of oxidative stress response. High expression of MVD was associated with poor prognosis in HCC. Strikingly, MVD inhibitor treatment profoundly suppressed HCC. Reduced tumor size and increased tumor-infiltrated effector T cells and macrophages were found in vivo. To conclude, our study has provided novel mechanistic insight about how NRF2/MVA pathway promotes HCC and suggested that MVD might be a good prognostic indicator and therapeutic target in human HCC.

Funding Support: RGC Postdoctoral Fellowship Scheme 2021/22 (Project #102010048)

Citation Format: Yiling Chen, Derek Lee, Misty Shuo Zhang, Jacinth Wing-Sum Cheu, Carmen Chak-Lui Wong. Targeting the mevalonate pathway, a novel anti-ferroptosis pathway, in hepatocellular carcinoma (HCC) treatment. [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 3675.
Abstract 3685: Targeting a novel cell surface serine protease TMPRSS11B for lung cancer therapy
Authors: Sunil Hari Shankar , Updegraff Barrett , Zhu Jingfei , Evers Bret M. , Minna John D. , Deberardinis Ralph J. , Oliver Trudy G. , O'Donnell Kathryn A.
Published: April 2023

Lung cancer is the leading cause of cancer-related deaths worldwide. Patients have few therapeutic options, and disease progression is inevitable. Anti-PD-1/PD-L1 antibodies can induce potent anti-tumor immunity and have been approved as a first-line therapy for non-small cell lung cancer (NSCLC). However, only ~20% of all NSCLCs benefit from checkpoint blockade, underscoring the critical need for the development of new and innovative approaches to identify actionable therapeutic targets to treat NSCLC. We 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 (Updegraff et al., Cell Reports, 2018). Our data show that TMPRSS11B promotes solubilization of Basigin, an obligate chaperone of the lactate monocarboxylate transporters MCT1 and MCT4, and that TMPRSS11B interacts and co-localizes with Basigin and MCT4. Moreover, Basigin release mediated by TMPRSS11B enhances lactate export and glycolytic metabolism, thereby promoting tumorigenesis. This is of significant interest because TMPRSS11B is frequently overexpressed in human lung squamous cell cancers (LSCC) and high expression is associated with poor survival of NSCLC patients. Moreover, as a cell surface protein and enzyme, TMPRSS11B represents an exciting putative target for therapeutic intervention. Our new studies demonstrate that the catalytic activity of TMPRSS11B may be important for the interaction with Basigin and enhanced lactate export. We also evaluated the effect of Tmprss11b depletion in the KLN205 syngeneic LSCC mouse model. CRISPR/Cas9 knockout of Tmprss11b in KLN205 LSCC cells significantly reduced tumor burden in immunocompetent DBA/2 WT mice. Moreover, mass spectrometry analysis revealed TMPRSS11B-interacting proteins that may represent novel substrates of this enzyme. Current studies are focused on elucidating the molecular mechanisms through which TMPRSS11B promotes tumorigenesis and investigating the efficacy of TMPRSS11B inhibition in syngeneic and autochthonous mouse models of LSCC, where the effects on immune cell function will be assessed.

Citation Format: Hari Shankar Sunil, Barrett Updegraff, Jingfei Zhu, Bret M. Evers, John D. Minna, Ralph J. Deberardinis, Trudy G. Oliver, Kathryn A. O'Donnell. Targeting a novel cell surface serine protease TMPRSS11B for lung cancer 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 3685.
Abstract 3703: Multimodal metabolic imaging and proteomics of radiation resistance in head and neck squamous cell carcinoma
Authors: Ivers Jesse D. , Dadgar Sina , Rajaram Narasimhan
Published: April 2023

Radiation resistance can halt a well-curated treatment plan in its tracks, but it may take weeks to realize this lack of efficacy resulting in lost time and unnecessary side effects. In this study, we sought to elucidate novel biomarkers of radiation resistance to predict treatment response in head and neck squamous cell carcinoma (HNSCC) at much earlier timepoints. We grew tumor xenografts from radiation-resistant UM-SCC-47 (n=25) and -sensitive UM-SCC-22B (n=37) cells in nude athymic mice. Each group was randomly divided into treatment and control groups, which were further subdivided into four timepoints for tumor excision: pre-treatment baseline, 1-, 24-, and 48-hours post-treatment. Prior to excision, we injected pimonidazole to quantify tumor hypoxia. We determined the two-photon excited fluorescence intensity and lifetime of NADH and FAD and quantified the optical redox ratio, ratio of free to bound NADH, and mean NADH lifetime of frozen sections of each tumor. We observed spatial and temporal heterogeneity in these high-resolution optical maps that indicated different metabolic programs in radiation-resistant and -sensitive tumors. A histogram-based heterogeneity analysis revealed time dependent shifts toward glycolysis in the radiation-resistant UM-SCC-47 treatment group, while the sensitive UM-SCC-22B treatment group showed the opposite trend. Proteomics performed on both cell lines identified multiple dysregulated proteins that are often implicated in HNSCC and associated with metabolic reprogramming — specifically, EGFR and SMAD4. We also found elevated levels of hypoxia and HIF-1 in the UM-SCC-47 tumors compared with the UM-SCC-22B tumors prior to and post-radiation. Using spatially co-registered optical imaging and immunohistochemical maps of the same field of view, we will present correlations between our optical markers and EGFR, SMAD4, and HIF-1. These studies will provide evidence of how protein dysregulation causes changes to optical metabolic imaging markers.

Citation Format: Jesse D. Ivers, Sina Dadgar, Narasimhan Rajaram. Multimodal metabolic imaging and proteomics of radiation resistance in head and neck squamous cell 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 3703.
Abstract 3706: The significance of cholesteryl ester profiles in hepatocellular carcinoma
Authors: Rahmanov Nazar , Liu Zhirong , Mao Jinghe , Kyle Patrick , Zhou Xinchun
Published: April 2023

Introduction: The association between cholesteryl ester (CE) profiles in the liver and oncogenesis, gender, and age disparities in hepatocellular carcinoma (HCC) has not been studied to date. This study aims to compare CE profiles between benign liver tissue (BLT) and HCC, and to correlate these findings with oncogenesis and disparities in patients with HCC.

Methods: Paired BLT and HCC samples obtained from 32 patients with the diagnosis of HCC were used for lipid extraction. Electrospray ionized mass spectrometry (ESI-MS) was used to determine CE profiles, including 22 individual CE species and the total levels of CE. The CE profiles in BLT and HCC were compared and stratified between subgroups based on the gender and age of the patients. The student T-test was used for statistical analysis and the significant p-value was set at 0.05.

Results: Total CE levels were higher in HCC than in BLT, but not statistically significant (1.8-fold, p=0.16). Interestingly, 5 saturated CE individual species and 2 mono-unsaturated individual CE species (C14:0, C16:0, C16:1, C18:0, C18:1, C19:0, and C20:0) were significantly higher in HCC than in BLT (2.4 to 14-fold higher). None of the individual poly-unsaturated CE species showed a statistical difference between HCC and BLT. In males, the levels of 3 saturated individual CE species (C16.0, 16:0, and C20:0) were significantly higher in HCC than in BLT (1.9 to 21.3-fold higher levels). In contrast, CE 16:1 was significantly lower in HCC than in BLT (0.9-fold, p=0.02) in males. In females, none of the CE profiles showed a significant difference between HCC and BLT. Total CE levels were 2.1-fold higher in males with HCC than in females with HCC (18.6nM vs. 8.92nM). In young patients (<66-year-old), individual CE species C14:0 was significantly higher in HCC than in BLT (19.1-fold, p=0.038). In old patients (≥66-year old), individual CE species C20:0 was significantly higher in HCC than in BLT. Total CE levels were 2.1-fold higher in young HCC patients than in old HCC patients (13.68nM vs. 6.45nM).

Conclusion: This study reveals that an increase in hepatic total CE levels and individual CE species are associated with the oncogenesis of HCC and show gender and age disparities. These findings could be helpful in prevention, treatment, exploring biomarkers, and understanding disparities in HCC.

Citation Format: Nazar Rahmanov, Zhirong Liu, Jinghe Mao, Patrick Kyle, Xinchun Zhou. The significance of cholesteryl ester profiles in hepatocellular 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 3706.
Abstract 3716: RNA N6-methyladenosine binding protein, YTHDF1 regulates DNA repair and cancer immunity in protein synthesis levels
Authors: Jang Dongjun , Jung Hae Rim , Oh Jaeik , Cho Sung-Yup
Published: April 2023

N6-methyladenosine RNA methylation is one of the most abundant post transcriptional modifications in eukaryotic messenger RNAs and plays crucial roles in cancer development and progression. YTHDF1 is an m6A reader protein and has been reported to increase the translation of mRNAs in cooperation with YTHDF3. In gastric cancer, YTHDF1 was reported to be overexpressed in cancer tissues, but detailed effects of YTHDF1 in cancer immunity have not been fully elucidated. In RNA sequencing data from a TGCA gastric cancer cohort, the expression of YTHDF1 was inversely correlated with the cytotoxic T cell markers, such as CD8A, GZMA, and GZMB. To investigate the effect of YTHDF1 overexpression on gene expression regulation in protein levels, we performed the proteomic analysis for newly synthesized proteins using non-canonical amino acid azidohomoalanine and click chemistry tagging reaction in the presence of interferon-γ (IFN-γ). Protein synthesis of genes in ‘DNA damage response’ and ‘DNA repair’ gene sets increased in YTHDF1-overexpressed samples. On the contrary, protein synthesis of genes in ‘Allograph rejection’, ‘NK cell mediated cytotoxicity’, and ‘NF-κB signaling’ gene sets decreased in YTHDF1-overexpressed samples. Especially, YTHDF1 reduced basal expression of DNA damage marker, γH2AX, suggesting that DNA damage repair was enhanced by YTHDF1 overexpression. Our study suggests that YTHDF1 plays pervasive roles in DNA repair and cancer immunity by regulating gene expression in protein levels.

Citation Format: Dongjun Jang, Hae Rim Jung, Jaeik Oh, Sung-Yup Cho. RNA N6-methyladenosine binding protein, YTHDF1 regulates DNA repair and cancer immunity in protein synthesis levels. [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 3716.
Abstract 3721: The C1 domain of capicua has functional importance in human CIC-DUX4
Authors: Luck Cuyler , Jacobs Kyle A. , Okimoto Ross A.
Published: April 2023

Genomic rearrangements between the HMG-box family transcriptional repressor capicua (CIC) and multiple partner genes result in fusion oncoproteins that activate transcription to drive sarcoma growth and progression. In the case of CIC-DUX4, the most common CIC-rearranged fusion oncoprotein, the fusion often retains over 90% of the wild type CIC protein structure including the HMG box and C-terminal C1 domain. Structure-function studies in Drosophila have suggested that the C1 domain of CIC, a non-HMG box DNA binding domain, is important for the transactivating capacity of a synthetic fly-human CIC-DUX4. Consistent with these findings, we performed a retrospective analysis of over 70 breakpoints described from CIC-DUX4 human derived tumors, which revealed that the C1 domain is structurally conserved within the context of the CIC-DUX4 fusion. To further understand the functional role of the C1 domain in human CIC-DUX4 we generated human CIC-DUX4 C1 domain and HMG-box deletion (alone or in combination) mutant constructs and performed a series of studies to test the functional impact on CIC-DUX4 protein levels, subcellular localization, and transcriptional activity. Through these analyses we have found that deletion of the C1 domain does not impact CIC-DUX4 oncoprotein expression or localization but does dramatically reduce target gene expression. In ongoing in vivo studies, we aim to evaluate the significance of the C1 domain in inducing CIC-DUX4 driven tumorigenesis. Collectively, our findings validate the functional importance of the C1 domain in human CIC-DUX4 and to our knowledge provide the first harmonized analysis of breakpoint locations in CIC-rearranged sarcoma.

Citation Format: Cuyler Luck, Kyle A. Jacobs, Ross A. Okimoto. The C1 domain of capicua has functional importance in human CIC-DUX4. [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 3721.
Abstract 3723: Identification and functional characterization of long noncoding RNAs that regulate TGF-<&946;>-Smad pathway as Smad co-factors
Authors: Kitagawa Masatoshi , Ota Kosuke , Ohhata Tatsuya , Sakai Satoshi
Published: April 2023

TGF-β-Smad pathway participates in various biological processes such as development, differentiation, growth regulation, and cancer progression including epithelial-mesenchymal transition (EMT). Long non-coding RNAs (lncRNAs) also involved in carcinogenesis and cancer malignancies. To identify lncRNAs regulating TGF-β-Smad pathway, we comprehensively identified lncRNAs induced by TGF-β and found and reported a lncRNA ELIT-1 (EMT-associated lncRNA induced by TGF-β-1) which was induced by TGF-β and promoted EMT by facilitating TGF-β-Smad signaling. ELIT-1 bound to Smad and participated in expression of TGF-β target genes including Snail. Prognosis of lung adenocarcinoma and gastric cancer patients with high expression of ELIT-1 was poor suggesting that ELIT-1 may be useful as a novel prognostic and therapeutic target. Recently, we found that lincNMR (long intergenic noncoding RNA-nucleotide metabolism regulation) was induced via TGF-β-Smad pathway in various cell lines. Moreover, RNA-seq analysis using lincNMRs-depleted cells indicated that lincNMRs was involved in expression of APOBEC3B, a cytidine deaminase, promoting C to U mutation and highly expressed in various human cancers. Although, regulatory mechanisms of APOBEC3B expression have not been fully elucidated, we proved that lincNMRs bound to Smad and promoted the activity of APOBEC3B promoter. These data suggest that lincNMRs participate in APOBEC3B expression by collaborating with TGF-β-Smad pathway. Additionally, high expression of lincNMRs was positively correlated with high expression of APOBEC3B in various cancer cell lines. The high expression of APOBEC3B as well as lincNMR was significantly associated with their poor prognosis in liver and lung cancer patients, suggesting that lincNMR may contribute to tumor malignancy by enhanced expression of APOBEC3B via TGF-β-Smad pathway.

Citation Format: Masatoshi Kitagawa, Kosuke Ota, Tatsuya Ohhata, Satoshi Sakai. Identification and functional characterization of long noncoding RNAs that regulate TGF-<&946;>-Smad pathway as Smad co-factors. [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 3723.
Abstract 3725: ABI1 regulates STAT3 transcription through a DNA binding activity
Authors: Li Xiang , Arya Neeru , Porter Baylee A. , Kemraj Allysa P. , Dong Xuesen , Frueh Dominique , Bah Alaji , Kotula Leszek
Published: April 2023

Prostate cancer (PCa) is characterized by the complexity of oncogenic signaling and heterogeneity of transcriptional landscapes. Adaptor protein ABI1 is a tumor suppressor in PCa, as evidenced by its loss or downregulation in high-grade and metastatic tumors. STAT3 activation is a hallmark of high-risk prostate tumors. ABI1 loss is associated with STAT3 activation leading to transcriptional reprogramming and epithelial-mesenchymal-transition (EMT) of prostate cancer cells (Nath, Li et al. Cell Commun Signal. 2019). EMT changes involve several homeobox transcription factors. The fact that ABI1 contains a homeobox homology region (HHR) suggests the possibility that it plays a role in EMT by directly regulating transcriptional activity by DNA binding. To examine this hypothesis, we set out to analyze ABI1-DNA binding. Structural NMR studies and in-vitro protein-DNA binding assays confirmed ABI1 binding to DNA regulated by alternative spliced ABI1 Exon 4 region located on the C-terminus of ABI1 HHR. RNA-sequencing data and PCa cell lines qPCR assays reveal the alternative ABI1 Exon 4 spliced-in is enriched in high Gleason-scored PCa samples. ABI1 and STAT3 co-regulate their nuclear vs. cytoplasm localization, and the subsequent functional studies using PCa lines expressing wild type or HHR Exon 4 deletion mutant of ABI1 demonstrated that HHR regulates the STAT3 DNA binding patterns, STAT3 mediated chromatin structure programming as well as its transcription activities. We propose that ABI1 is a critical regulator of STAT3 activity during prostate cancer progression.

Citation Format: Xiang Li, Neeru Arya, Baylee A. Porter, Allysa P. Kemraj, Xuesen Dong, Dominique Frueh, Alaji Bah, Leszek Kotula. ABI1 regulates STAT3 transcription through a DNA binding activity. [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 3725.
Abstract 3736: ZBTB46/FOXA2/HIF1A transcription activator complex promotes MCTP1-regulated neuroendocrine differentiation and epithelial-to-mesenchymal transition
Authors: Phan Vu Thuy Dung , Wen Yu-Ching , Chen Wei-Yu , Chen Wei-Hao , Jiang Kuo-Ching , Li Han-Ru , Tram Van Thi Ngoc , Chen Zi-Qing , Wang Wan-Hsin , Liu Yen-Nien
Published: April 2023

Androgen deprivation therapy (ADT)-induced neuroendocrine differentiation (NED) is a well-known lethal subtype of prostate cancer (PCa) with a median survival rate of less than one year. Despite the increasing research attention on this variant of PCa, the underlying mechanism orchestrating therapy-related neuroendocrine prostate cancer (NEPC) remains elusive. We found that ADT-induced hypoxia-associated ZBTB46/FOXA2/HIF1A signaling enhances the multiple C2 domain transmembrane protein 1 (MCTP1), which promotes NED and epithelial-to-mesenchymal transition (EMT) of PCa. Mechanistically, ZBTB46 physically interacts with the HIF1A/FOXA2 complex, in which ZBTB46 may be a co-activator of the hypoxia-related FOXA2 transcription factor. Interestingly, this ZBTB46/FOXA2/HIF1A complex accumulates after hypoxia and functions as a transcriptional activator of MCTP1. Hypoxia-upregulated MCTP1 subsequently leads to NED and the increase in EMT, whereas the opposite is true for the knockdown of MCTP1 in PCa cells. Consistent with previous results, MCTP1 is highly expressed in high-grade castration-resistance prostate cancer (CRPC) and small-cell PCa (SCPC) tissues and is associated with NE markers and ZBTB46/FOXA2/HIF1A abundance. In this study, we explored the direct interaction of ZBTB46 protein with hypoxia-related FOXA2/HIF1A complex in PCa cells under hypoxic conditions, which promote MCTP1-driven EMT and NED. Our finding suggests that MCTP1 could be used as a biomarker for diagnosing NEPC and as a therapeutic target in clinical applications.

Citation Format: Vu Thuy Dung Phan, Yu-Ching Wen, Wei-Yu Chen, Wei-Hao Chen, Kuo-Ching Jiang, Han-Ru Li, Van Thi Ngoc Tram, Zi-Qing Chen, Wan-Hsin Wang, Yen-Nien Liu. ZBTB46/FOXA2/HIF1A transcription activator complex promotes MCTP1-regulated neuroendocrine differentiation and epithelial-to-mesenchymal transition. [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 3736.