bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1562: HMGA2 regulates GPX4 expression and oxidative stress
Authors: Dike Precious Elechi , Campbell Taaliah , Awolowo Mojisoluwa , Odero-Marah Valerie
Published: March 2024

Prostate cancer (PCa) is a leading cause of mortality, primarily due to its ability to metastasize to the bone. The High Mobility Group AT-Hook 2 (HMGA2) plays a crucial role in regulating gene expression and has been implicated in tumorigenesis and the metastatic process. Our recent study revealed that overexpression of the wild-type/full-length HMGA2 isoform in PCa cells promotes cancer progression by triggering epithelial mesenchymal transition (EMT), whereas truncated HMGA2 isoform promotes progression through oxidative stress signaling. We hypothesize that HMGA2 regulates GPX4 expression and oxidative stress in PCa. We studied the expression of HMGA2 and GPX4 in various PCa cell lines including enzalutamide resistant cell line (C4-2B MDVR), LNCaP cells overexpressing HMGA2 isoforms (wild type and truncated HMGA2). Our analysis of HMGA2 and GPX4 expression in diverse PCa cell lines, shows an inverse relationship between HMGA2 and GPX4 levels. Elevated HMGA2 expression coincides with reduced GPX4 expression, leading to heightened oxidative stress and susceptibility to ferroptosis. Moreover, treatment of enzalutamide resistant cell line C4-2B MDVR with ferroptosis inducer RSL3, further suppresses GPX4 expression, resulting in decreased cell proliferation. Our results unveils the intricate relationship between HMGA2, GPX4 regulation and oxidative stress in the context of prostate cancer progression. Exploiting this oxidative stress pathway offers a promising therapeutic approach for cancer treatment, especially for enzalutamide-resistant cancer cells.

Acknowledgements: These studies were supported by NIH/NIGMS/RISE SR25GM060414 and NIH/NIMHD 2U54MD007590; 5U54MD013376.

Citation Format: Precious Elechi Dike, Taaliah Campbell, Mojisoluwa Awolowo, Valerie Odero-Marah. HMGA2 regulates GPX4 expression and oxidative stress [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 1562.
Abstract 1569: Keratin X promotes cell metastasis and sorafenib resistance via the AMPKα/Snail axis in hepatocellular carcinoma
Authors: Lin Yang-Hsiang , Yeh Chau-Ting
Published: March 2024

A great majority of liver cancers are developed from cirrhotic livers. The key molecules involved in cirrhosis-mediated hepatocarcinogenesis have not been clearly defined. Understanding the oncogenic pathways and their associations with cancer growth, metastasis and drug resistance may help scientists devise new effective therapeutic agents. In the present study, we identified the intermediate filament-forming protein keratin X (KRTX) as a key factor in cirrhosis-mediated hepatocarcinogenesis. KRTX was highly expressed in HCC tissues and correlated with poor survival outcomes. Experimental verification revealed that overexpression of KRTX enhanced cell growth, tumor cell metastasis, glycolysis and sorafenib resistance. Conversely, knockdown of KRTX led to opposite effects. Mechanistically, KRTX inhibited AMPKα phosphorylation, thereby enhancing Snail protein stability. Additionally, TGFβ1 positively regulated KRTX expression, resulting in a KRTX/AMPKα (Thr172)/Snail axis-mediated increase in cell motility. Treating with 2-deoxyglucose reversed the KRTX-induced metastasis and sorafenib resistance. In clinic, KRTX expression is positively correlated with Snail overexpression and high KRTX/Snail expression had poor prognosis in HCC patients. Our data revealed a signaling pathway from TGFβ1, KRTX, through p-AMPKα (Thr172) to Snail. Targeting KRTX and its pathway may be effective for treating patients with metastatic HCC.

Citation Format: Yang-Hsiang Lin, Chau-Ting Yeh. Keratin X promotes cell metastasis and sorafenib resistance via the AMPKα/Snail axis in hepatocellular carcinoma [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 1569.
Abstract 1583: Neutrophil pro-tumorigenic role in progression and therapy resistance in pancreatic ductal adenocarcinoma
Authors: Sturgeon Reegan , Johnson Esther , Molczyk Caitlin , Goel Paran , Tinsley Parker , Abrahams Lauren , Maher Michael , Singh Rakesh K.
Published: March 2024

Pancreatic cancer (PC) continues to be a challenge for today’s therapeutics, with a meager 5-year survival rate of 12%. Due to the pancreas’ location, patients often receive diagnosis late, leaving them with limited treatment options. Often, they fail to respond to chemotherapy regimens or develop resistance to those therapies. Tumor-promoting inflammation is a hallmark of cancer, contributing to tumor cells’ survival and proliferation. Infiltrating leukocytes and pro-inflammatory cytokines released into the tumor microenvironment (TME) often cause this inflammation. My lab has previously linked increases in pancreatic ductal adenocarcinoma (PDAC) progression and chemotherapy resistance to inflammation and the CXCR2 pathway. In the present study, we examined the role of neutrophil recruitment in PDAC progression using patient samples and KRAS-dependent KC and KPC murine models. Our data suggest a positive association between tumor-associated neutrophils (TANs) and PDAC progression. In vitro, we observed neutrophil-PDAC interaction enhancing the survival of both neutrophils and PDAC cell lines. Moreover, this survival was mediated by cellular aggressiveness and therapy resistance. Concerning survival, we saw increased expression of anti-apoptotic genes in the neutrophils upon PDAC cancer cell conditioned media treatment. We also observed modulated expression of immunoregulatory molecules. Our data suggest that increased recruitment of TANs supports PDAC progression and survival, and PDAC cell-neutrophil interaction modulates neutrophil survival and immunoregulatory phenotype.

Citation Format: Reegan Sturgeon, Esther Johnson, Caitlin Molczyk, Paran Goel, Parker Tinsley, Lauren Abrahams, Michael Maher, Rakesh K. Singh. Neutrophil pro-tumorigenic role in progression and therapy resistance in pancreatic ductal adenocarcinoma [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 1583.
Abstract 1592: Single-cell transcriptomics reveals unique stromal and metabolic heterogeneity in liver metastatic pancreatic cancer
Authors: Singh Rima , Halbrook Christopher
Published: March 2024

Pancreatic ductal adenocarcinoma (PDA) is the 3rd leading cause of cancer related death in the US. This mortality is strongly linked to the complex PDA tumor microenvironment that drives resistance to chemotherapy and immune suppression. Most patients succumb to metastatic disease, with liver being the most frequently colonized organ. Despite this, most pre-clinical studies have focused on primary PDA models to study the disease. Accordingly, the extent that the liver metastatic niche promotes immune evasion and resistance to treatment remains unclear.To directly compare primary vs. metastatic tumors in PDA tumors, we have optimized a syngeneic experimental model to contrast primary vs. metastatic lesions. To define the similarities and differences between cell types present in these tumors, we employed a single-cell transcriptomics approach. Our analysis has identified shared populations of malignant epithelial, stromal, and immune cells between lesions, allowing for a direct comparison of programming. Here, we have observed that the dominant populations of PDA cells in pancreas vs. liver tumors engage in distinct metabolic programs that suggest different actionable vulnerabilities.Beyond these, we also observed fibroblasts and myeloid sub-populations that are exclusive to liver tumors. These data further support our hypothesis that alternative mechanisms of metabolic, immune, and stromal interactions take place in the metastatic tumor microenvironment. To define these crosstalk networks, we employed the interactome tool CellChat, which has uncovered unique signaling interactions between the CAF and cancer cell populations in pancreatic or liver tumors. Overall, these data have the potential to lead to new avenues to directly target cancer cells or engage an effective immune response that can be designed to treat metastatic pancreatic cancer- an urgent clinical need.

Citation Format: Rima Singh, Christopher Halbrook. Single-cell transcriptomics reveals unique stromal and metabolic heterogeneity in liver metastatic pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1592.
Abstract 1597: CD144+cancer-associated fibroblasts drive the malignancy of pancreatic cancer via stimulating inflammatory paracrine
Authors: Sun Xugang , Yu Jun , Zhao Tiansuo , Hao Jihui
Published: March 2024

Purpose: Cancer-associated fibroblasts (CAFs) are highly heterogeneous in pancreatic ductal adenocarcinoma (PDAC). We previously reported that CAFs can acquire an endothelial phenotype under stress. Thus, we hypothesized that there are CAFs highly expressing endothelial markers and substantiated a novel subtype of CAFs promoting tumor progression.

Experimental design: We performed single-cell RNA sequencing to identify the existence of CD144+CAFs. The prospective and retrospective analysis were combined to figure out the correlation between CD144+CAFs proportion and clinical outcome. The proliferation and invasion assay of cell lines, organoids and pancreatic cancer xenograft model were used to detect the effect of CD144+CAFs on tumor progression. Cytokine array assay, RNA-sequencing, IP-mass spectrum, Ch-IP and luciferase analysis were conducted to elucidate the underlying mechanism. siRNA delivery nanosystem was designed and administrated to precisely target CD144+CAFs in vivo.

Results: CD144+CAFs were present in tumor microenvironment of PDAC and breast cancer and patients with a higher CD144+CAFs proportion have worse prognosis in PDAC. CD144+CAFs can promote the metastasis of PDAC through enhancing epithelial-mesenchymal transition. CD144-β-catenin-STAT3 signaling axis was activated and downstream inflammatory secreted cytokines were transcriptionally upregulated to maintain the functions and phenotype of CD144+CAFs. A novel CAF-targeting siRNA delivery nanosystem, loading FAPα and siCDH5, was designed and administrated to precisely target CD144+CAFs, which substantially inhibited the protumoral roles in vivo.

Conclusion: CD144+CAFs can promote the growth and metastasis of PDAC by activating CD144-β-catenin-STAT3 signaling axis and stimulating inflammatory secretion. CAF-targeting siRNA delivery nanosystem can inhibit tumor progression by precisely targeting CD144+CAFs.

Citation Format: Xugang Sun, Jun Yu, Tiansuo Zhao, Jihui Hao. CD144+cancer-associated fibroblasts drive the malignancy of pancreatic cancer via stimulating inflammatory paracrine [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 1597.
Abstract 1635: Inhibition of FANCA Suppresses Breast Cancer Development by Regulating Cell Cycle Progression
Authors: Luo Liang , Liu Wenjun , Yuan Fenghua , Li Fang , Palovcak Anna , Calkins Daniel , Briegel Karoline Briegel , Li Yan , Mason Christian , Liu Zhao-Jun , Daunert Sylvia , Zhang Yanbin
Published: March 2024

FANCA is one of the 22 known Fanconi anemia (FA) pathway genes that are involved in interstrand crosslink (ICL) repair. We have also shown that and FANCA plays a role in double strand break repair by promoting strand annealing and exchange. In this report, we describe that FANCA expression in breast cancer is significantly elevated within TCGA databases and in patient tumors-derived sections on both transcription and translation levels and this elevation is associated with poor outcomes of the breast cancer patients. Importantly, we have found that overexpressing FANCA in breast cancer cell line MDA-MB-231 promotes tumor growth both in vitro and in vivo. We have also demonstrated that FANCA inactivation by knocking out or knocking down inhibits growth of MDA-MB-231 in vitro and in vivo by causing cell cycle arrest at G1 phase resulting from hypophosphorylation of Rb protein. Our RNA-seq data has indicated that E2F targets are also downregulated in FANCA knockout (KO) cells compared to the wildtype (WT) MDA-MB-231 cells, which is consistent with the G1/S cell cycle arrest and Rb hypophosphorylation. Intriguingly, heterozygous FANCA KO in a breast cancer mouse model MMTV-PyMT model has shown reduction of breast tumor formation and volume. Our goal is to elucidate the role of FANCA in breast cancer development and to evaluate the potential of FANCA as a target for breast cancer treatment.

Citation Format: Liang Luo, Wenjun Liu, Fenghua Yuan, Fang Li, Anna Palovcak, Daniel Calkins, Karoline Briegel Briegel, Yan Li, Christian Mason, Zhao-Jun Liu, Sylvia Daunert, Yanbin Zhang. Inhibition of FANCA Suppresses Breast Cancer Development by Regulating Cell Cycle Progression [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 1635.
Abstract 1693: Mitochondrial-associated DNAJA3 variant and its role in NASH-driven hepatocellular carcinoma
Authors: Shiode Yuto , Chang Ching-Wen , Wang Xin Wei
Published: March 2024

Nonalcoholic fatty liver disease (NAFLD) is increasingly recognized as a major risk factor for hepatocellular carcinoma (HCC). Despite the known progression from NAFLD to nonalcoholic steatohepatitis (NASH) and eventually HCC, the genetic underpinnings driving this transition remain underexplored. A comprehensive genomics assessment was conducted, encompassing a genome-wide study of variants linked to body fat distribution in 344,369 individuals. Correlating these, significant variants associated with NASH and HCC were discerned in a cohort of 1,009 participants from the NCI-UMD study. Candidate variants underwent further eQTL analysis. Our investigation unveiled the rs3747579-TT variant emerged as significantly associated with NASH-related HCC, serving as an eQTL for mitochondrial DNAJA3. HCC patients harboring this variant presented with diminished DNAJA3 expression, correlating with an unfavorable prognosis. Exploration into the 3D genome architecture has identified potential chromatin topology sites that may influence the regulation of DNAJA3 by rs3747579. Specifically, enhancer loops exhibit allele-specific interactions with rs3747579 in the regulation of DNAJA3. Notably, the rs3747579-CC variant was found to interact with RBFOX2. Beyond its known RNA splicing functions, RBFOX2 might have implications in the chromatin landscape, influencing chromatin dynamics and gene transcription. RBFOX2 knockdown experiments showcased significantly reduced DNAJA3 expression, and luciferase assays highlighted increased activity for the rs3747579-CC allele versus its TT counterpart. In summary, our findings spotlight the rs3747579-TT variant as a potential oncogenic factor in DNAJA3, elucidating its role in the genesis of NASH and progression to HCC. This understanding could pave the way for novel therapeutic interventions in HCC stemming from NAFLD/NASH.

Citation Format: Yuto Shiode, Ching-Wen Chang, Xin Wei Wang. Mitochondrial-associated DNAJA3 variant and its role in NASH-driven hepatocellular carcinoma [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 1693.
Abstract 1696: ACTL6A deficiency drives cell death through KLF4-involved chromatin accessibility changes in colorectal cancer
Authors: Yang Hye-Ju , Kim Eun-Ju , Song Sang-Hyun , Kim Tae-You
Published: March 2024

ACTL6A is a subunit of the SWI/SNF and INO80 chromatin remodeling complexes and plays roles in neural development and cancer cell proliferation. Additionally, it is known to be upregulated in various types of cancer and is associated with a decrease in patient survival rates. However, the mechanism of how chromatin remodelers including ACTL6A cause cell death has yet to be defined. Therefore, we would like to investigate how the depletion of ACTL6A promotes cell growth reduction by mediating changes in the epigenetic landscape in colorectal cancer cells. We engineered ACTL6A knockout by CRISPR-Cas9. We performed Assay for transposase-accessible chromatin using sequencing (ATAC-seq), RNA sequencing (RNA-seq), and Chromatin Immunoprecipitation sequencing (ChIP-seq) for SMARCB1 and INO80, as representative members of their respective families, and KLF4. Moreover, HCT116 cells stably transduced with doxycycline-inducible shGFP or shACTL6A were implanted into the flank of nude mice. One week after tumor implantation, mice were randomly allocated to 4 groups (n=13-15 per group) and doxycycline treated group was administered with doxycycline (1mg/ml) and 5% sucrose in the drinking water. Tumor volume was measured with calipers. Upon depletion of ACTL6A, the regions of increased chromatin accessibility were correlated with the upregulated genes. After the loss of ACTL6A, there was a significant increase in KLF4 expression and genomic binding largely increased, coinciding with an increase in newly accessible regions containing KLF4 motifs. Interestingly, there is a strong positive correlation between changes in chromatin accessibility and overall changes in the binding of KLF4, SWI/SNF, and INO80 complexes. We find that changes in chromatin accessibility are co-regulated by KLF4, the SWI/SNF and the INO80 complex, while SWI/SNF and INO80 complex have a low correlation in binding between the SWI/SNF and INO80 complexes at newly accessible regions, indicating no physical interactions between these two chromatin complexes. Overall, we demonstrated that the redistribution of ACTL6A-deficient two chromatin remodeling complexes and KLF4 is associated with changes in chromatin accessibility, which are also linked to upregulated genes that promote the p53 pathway. These findings revealed the role of KLF4 along with the SWI/SNF and INO80 complexes, in regulating the chromatin landscape and gene expression in ACTL6A-depleted colorectal cancer cells.

Citation Format: Hye-Ju Yang, Eun-Ju Kim, Sang-Hyun Song, Tae-You Kim. ACTL6A deficiency drives cell death through KLF4-involved chromatin accessibility changes in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1696.
Abstract 1732: Linker histone h1 inhibits topoisomerase IIa in vitro and in vivo
Authors: Lasater Samantha , Hamdan Hiba , Ho Po-Yi , Wu Ting , Cao Kaixiang , Yang Ruiqiao , Zhang Yunzhe , Fan Yuhong
Published: March 2024

H1 linker histones play a crucial role in organizing chromatin into higher-order structures and regulating gene expression, contributing significantly to various cellular processes. Topoisomerase IIa (TopoIIa) is essential for multiple cellular functions via regulating DNA topology. Here, we report a direct interaction between H1 linker histones and TopoIIa through both N- and C-terminal domains of H1 variants. H1 suppresses the decatenation enzymatic activity of TopoIIa in vitro in a dose-dependent manner. Overexpressing specific H1 variants in HeLa cells results in a marked decrease in cell proliferation and a dramatic increase in the occurrence of cells displaying abnormal mitotic chromosomes. Additionally, we find that the inhibitory effect of the specific H1 variant on TopoIIa activity occurs prior to the formation of DNA-TopoIIa complexes. These results reveal a novel mechanistic link between histone H1 and Topo IIa in the regulation of chromatin function.

Citation Format: Samantha Lasater, Hiba Hamdan, Po-Yi Ho, Ting Wu, Kaixiang Cao, Ruiqiao Yang, Yunzhe Zhang, Yuhong Fan. Linker histone h1 inhibits topoisomerase IIa in vitro and in vivo [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 1732.
Abstract 1770: Right oncogene, wrong tumor - CBL mutations in pediatric CNS and solid tumors
Authors: Brown Lauren M. , Mayoh Chelsea , Mateos Pablo Acera , De Weck Antoine , Salomon Robert , Sadras Teresa , Manoharan Neevika , Wong Marie , Cowley Mark J. , Ekert Paul G.
Published: March 2024

The molecular analysis of individual patient tumors by personalized medicine programs like the Zero Childhood Cancer Program (ZERO) enables the detection of potentially targetable lesions, unlocking new therapeutic opportunities for patients. Somatic mutations in the E3-ubiquitin ligase, CBL, are known to activate receptor tyrosine kinases (RTKs) in cancer and have been exclusively characterized in hematological malignancies. This includes acute myeloid leukemia, where CBL mutations lead to FLT3 activation. Using molecular data from ZERO we have identified known and novel CBL variants in novel tumor contexts. Our findings raise the possibility that CBL mutation may be a marker of RTK activation in a range of paediatric cancer types and may represent a group of patients who could benefit from tyrosine kinase inhibitor (TKI) therapy.

We analyzed whole genome and RNA sequencing data from the ZERO cohort and identified 26 somatic CBL variants in 22 individual patients, the majority of which were in CNS tumors (14 patients). In 8 patients with CNS tumors, we identified missense and splicing variants, which were either novel or not previously seen in this tumor type, that are predicted to have a functional impact on CBL E3 ligase activity due to their location in the linker region or RING finger domain of CBL. Interestingly, most of these patients did not fall into a pre-defined DNA methylation-based subtype classification of CNS tumors and had no significant upregulation of RTK genes.

In addition to the novel variants, we identified an established oncogenic CBL deletion variant, CBL exon 8/9 deletion (CBL ex8/9Δ), in novel tumor types - neuroblastoma and a germ cell tumor. Functionally, we show that overexpression of CBL ex8/9Δ enhances cell proliferation and maintains EGFR signaling by blocking CBL mediated degradation of phosphorylated EGFR in neuroblastoma cells.

In this study, we demonstrate that CBL is mutated in a range of pediatric cancer types beyond hematological malignancies and may represent a marker of RTK activation and a new target for high-risk patients with limited therapeutic options.

Citation Format: Lauren M. Brown, Chelsea Mayoh, Pablo Acera Mateos, Antoine De Weck, Robert Salomon, Teresa Sadras, Neevika Manoharan, Marie Wong, Mark J. Cowley, Paul G. Ekert. Right oncogene, wrong tumor - CBL mutations in pediatric CNS and solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1770.
Abstract 1787: The β3-adrenergic receptor as novel target of metabolic phenotype in Ewing sarcoma
Authors: Banella Cristina , Carrozzo Francesco , Zocca Laura , Pasha Amada , Mattei Gianluca , Mola Marina , Ballerini Lara , Lorito Nicla , Tondo Annalisa , Favre Claudio , Calvani Maura
Published: March 2024

Purpose of study: Metabolic targeted therapies may represent an innovative strategy in Ewing sarcoma (ES).

Introduction: Ewing sarcoma (ES) is highly aggressive round cell mesenchymal neoplasm, most often occurring in children and young adults. Metabolic peculiarities of ES cells are indeed involved in tumor growth, survival and resistance to therapy. We have further investigated apoptotic role of 𝜷;;3-adrenergic receptor (𝜷;;3-AR) SR59230A (SR) antagonist for its ability to reduce cells viability due increased oxidative stress, now we underline the metabolic role of SR.

Materials & Methods: The metabolic profile after SR administration in A673 ES cell model was studied using the Seahorse XF Analyzer. Viability was evaluated with MTS. Changes in gene expression was evaluated with RNA sequencing. Western blot analysis and Immunofluorescence was performed for redox signature and metabolic alteration. Ros production and cell death was assessed by flow cytometry.

Results: Metabolic analysis reveals glucose and fatty acid dependency in A673 cells using the Fuel Flex Test of Seahorse XFe Analyzer. Quantification of transcript abundance using Salmon in A673 after SR treatment showed decrease in response to reactive oxygen species, increase of Thioredoxin Interacting Protein (TXNIP) inhibits the antioxidative function of thioredoxin, moreover reduced glucose uptake, consequently, decrease NADPH/NADP resulting in the accumulation of ROS coincident with a decrease of GLUT1, SOD2, TrDX an increase of LC3IIA protein expression. SR enhances A673 pyruvate dependency, and decrease glyco and mito ATP production rate, more over observed under glucose deprivation. Furthermore, apoptotic effect was significantly amplified when cells were under nutrient starvation, revealing a 56% apoptotic rate after 24 hours of SR treatment.

Conclusion: SR antagonist of 𝜷;;3-AR and combination of SR with BUF could be an innovative therapeutic option to raise awareness ES.

Citation Format: Cristina Banella, Francesco Carrozzo, Laura Zocca, Amada Pasha, Gianluca Mattei, Marina Mola, Lara Ballerini, Nicla Lorito, Annalisa Tondo, Claudio Favre, Maura Calvani. The β3-adrenergic receptor as novel target of metabolic phenotype in Ewing sarcoma [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 1787.
Abstract 1808: Discovery of highly selective novel MTA-cooperative PRMT5 inhibitors for the treatment of cancers
Authors: Wu Wen-Lian , Hu Taishan , Deng Zhilin , Li Honghai , Shen Quanrong , Zhang Lei , Ma Xiaochu , Sun Peihua , Cheng Cindy , Liu Fang , Chen Xin , Hua Ye , Huang Bryan
Published: March 2024

Protein arginine methyltransferase 5 (PRMT5), a type II PRMT catalyzing the formation of symmetric dimethylation of arginine residues on histone and non-histone proteins, regulates many biological pathways in mammalian cells, including cell growth and differentiation. Methylthioadenosine phosphorylase (MTAP) is required for the methionine salvage pathway, deletion of MTAP leads to the accumulation of inhibitory PRMT5 cofactor methylthioadenosine (MTA). MTAP gene is adjacent to and frequently co-deleted with CDKN2A gene, the most commonly deleted tumor suppressor gene in human cancers. The increase in MTA significantly reduced PRMT5 activity in MTAP-deficient cancer cells, rendering them more vulnerable to PRMT5 inhibition than normal cells. Selective Inhibitors of PRMT5-MTA complex (MTA-cooperative) are supposed to exhibit an increased therapeutic index compared to first generation PRMT5 inhibitors for the treatment of MTAP-deleted (MTAP-del) cancer patients. Guided by computation-aided drug design (CADD), binding mode analysis of known PRMT5 inhibitors has led to the design and synthesis of a novel series of MTA-cooperative PRMT5 inhibitors. Many compounds within this series were found to inhibit PRMT5/MEP50 complex with single digit nM of IC50 in enzymatic assay. Cell based activity of these compounds was assessed by measuring the symmetric demethylarginine (SDMA) and 10-day cell proliferation assays. Several compounds exhibited low double-digit nM of cellular potency in HCT116 MTAP-del cells, the inhibition of SDMA and anti-proliferation activities in HCT116 MTAP-del cells over HCT116 MTAP-WT cells have excellent selectivity (80 - 400 fold). Further optimization of in vitro and in vivo pharmacokinetic properties has yielded candidate compounds suitable for evaluation of pharmacodynamic effect in the LU99 MTAP-del NSCLC xenograft model. Tumor growth inhibition (TGI) was observed in a dose-dependent manner, accompanied by a reduction of PRMT5-mediated SDMA levels in both the tumor and bone marrow. In summary, we have identified compounds with novel scaffolds that are potent and highly selective MTA-cooperative PRMT5 inhibitors. Further development of the second-generation PRMT5 inhibitors for treatment of cancers is also planned.

Citation Format: Wen-Lian Wu, Taishan Hu, Zhilin Deng, Honghai Li, Quanrong Shen, Lei Zhang, Xiaochu Ma, Peihua Sun, Cindy Cheng, Fang Liu, Xin Chen, Ye Hua, Bryan Huang. Discovery of highly selective novel MTA-cooperative PRMT5 inhibitors for the treatment of cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1808.
Abstract 1810: Development of new phthalic hydrazide scaffold for potent TNKS inhibitors
Authors: Ryu Hwani , Seo Hye-Ran , Lee Ju Han , Lee Kyu Myung , Ahn Jiyeon
Published: March 2024

Tankyrase 1 and 2 (TNKS1/2) catalyze post-translational modifications of target proteins through poly-ADP-ribosylation. Inhibition of TNKS stabilizes AXIN and induces β-catenin degradation in adenomatous polyposis coli (APC)-mutated colorectal cancer (CRC) cells. Therefore, TNKS emerges as a therapeutic target for APC mutant CRC, and there is a critical need for drug development against it. Tankyrase 1 and 2 (TNKS1/2) catalyze the post-translational modification of target proteins through poly-ADP-ribosylation. Their inhibition stabilizes AXIN and induces β-catenin degradation. Therefore, TNKS is a therapeutic target for APC mutant colorectal cancer (CRC), and there is a need for drug development against it. However, reported TNKS inhibitors have serious limitations in that they cause intestinal toxicity. To address these issues and enhance efficacy, we synthesized 60 compounds based on the tetrahydrophthalazine scaffold using protein structure modeling-Al technology. Using an in vitro TNKS1 and TNKS2 enzyme assay and derivatizing Hit compounds, we confirmed that TI-61987 significantly inhibits TNKS enzyme activity at low nanomolar levels. TI-61987 stabilized AXIN2, reduced active β-catenin, and downregulated β-catenin target genes in COLO320DM, APC-mutated CRC cells. Additionally, TI-61987 exhibited excellent anticancer effects not only in COLO320DM cells but also in in vivo xenograft mouse models. Our findings demonstrate that TI-61987 is a therapeutic candidate targeting TNKS for the treatment of APC-mutated CRCs, and this anticancer drug may represent an effective therapy regimen for APC-mutated CRCs.

Citation Format: Hwani Ryu, Hye-Ran Seo, Ju Han Lee, Kyu Myung Lee, Jiyeon Ahn. Development of new phthalic hydrazide scaffold for potent TNKS inhibitors [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 1810.
Abstract 1811: Efficient identification of new small molecules targeting succinate dehydrogenase in non-small cell lung cancer
Authors: Silva Luis , Skiados Nicholas , Murugavel Nikitha , Luna Nastassja , Cover Karen , Gupta Manish K. , Contreras Stephanie C. , O'Brien Terrence E. , Zhang Wen Cai
Published: March 2024

Efficient lung cancer treatment remains challenging due to the lack of therapeutic targets. The succinate dehydrogenase (SDH) enzyme plays a critical metabolic role as an intermediate between the citric acid cycle and the electron transport chain associated with the cancer disease state. Although several existing compounds have been applied to target metabolic diseases in vitro, modulating SDH for lung cancer treatment is still elusive. By integrating the AtomNet® technology for compound identification with mitochondria- and cell-based enzyme activity assays, we identified four new small molecules from 96 predicted candidates. Cellular thermal shift assay confirmed that these small molecules bind directly to SDH subunits in lung cancer cells. Mechanistically, treatment with these small molecules increased cellular and mitochondrial reactive oxygen species, accumulated succinate, and DNA damage-induced apoptosis in lung cancer cell lines. Functionally, these small molecules reduced the growth, migration, and formation of 3D organoids in vitro in lung cancer cell lines, both in the short and long term. Our study sheds light on the function of SDH in metabolic dysfunction related to cancer and highlights the potential of modulating SDH as a viable therapeutic strategy for targeting lung cancer and other cancer types. LS, NS, and WCZ contributed equally to this work.

Citation Format: Luis Silva, Nicholas Skiados, Nikitha Murugavel, Nastassja Luna, Karen Cover, Manish K. Gupta, Stephanie C. Contreras, Terrence E. O'Brien, Wen Cai Zhang. Efficient identification of new small molecules targeting succinate dehydrogenase in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1811.
Abstract 1872: A next generation treatment for Nectin-4 positive cancers - Preclinical characterization of LY4052031, an anti-Nectin-4 antibody, conjugated to a novel camptothecin payload
Authors: Sagar Divya , Srinivasan Mohan , Lindquist Kevin , Guo Qianxu , Wong Wailin , Lebron Maria B. , Sattler Rose Marie , Zhou Jieyu , Helms Whitney , Boyles Jeffrey , Verdino Petra , Zhao Xuan , Mak Yvonne , Park Joshua , Holmgaard Rikke , Driscoll Kyla , Duramad Omar , Bedard Kristin
Published: March 2024

Nectin-4 is an immunoglobulin-like antigen overexpressed in a variety of cancers including bladder, breast, NSCLC, and gastric. Normal tissues including skin, salivary glands, bladder and esophagus express only low to medium levels of Nectin-4 making the protein an attractive target for cancer therapies. The approval of enfortumab vedotin-ejfv, an anti-Nectin-4 based antibody-drug conjugate (ADC) with a microtubule inhibitor payload, either as a single agent or in combination with pembrolizumab has been a significant advancement in the treatment of metastatic urothelial cancer. We have developed a novel anti-Nectin-4 ADC with a novel topoisomerase I (TOPO 1) inhibitor drug payload. Topoisomerase inhibitor payload-based ADCs offer potential advantages compared to microtubule payload-based ADCs including a different toxicity profile and the differentiated susceptibility to resistance mechanisms, such as P-gp mediated drug efflux. LY4052031 is a fully human monoclonal anti-Nectin-4 antibody conjugated to a novel TOPO 1 inhibitor by a cleavable peptide linker at a homogeneous drug antibody ratio (DAR) of 8:1. In vitro, both LY4052031 and its unconjugated parental antibody specifically bind to Nectin-4 expressing cell lines and are quickly and efficiently internalized. LY4052031 demonstrated potent cytotoxicity on tumor cells with high and low Nectin-4 expression. Additionally, bystander activity exerted by LY4052031 on antigen negative cells co-cultured with antigen positive cells was greater than that observed with the clinical benchmark enfortumab vedotin-ejfv. In vivo studies in xenografts expressing low to high levels of Nectin-4 showed significant tumor growth inhibition following treatment with LY4052031 as monotherapy. Overall, LY4052031 exhibits specificity, selectivity, potency, and effectiveness as a next generation therapy to treat Nectin-4 positive tumors. An IND submission is planned for 2024.

Citation Format: Divya Sagar, Mohan Srinivasan, Kevin Lindquist, Qianxu Guo, Wailin Wong, Maria B. Lebron, Rose Marie Sattler, Jieyu Zhou, Whitney Helms, Jeffrey Boyles, Petra Verdino, Xuan Zhao, Yvonne Mak, Joshua Park, Rikke Holmgaard, Kyla Driscoll, Omar Duramad, Kristin Bedard. A next generation treatment for Nectin-4 positive cancers - Preclinical characterization of LY4052031, an anti-Nectin-4 antibody, conjugated to a novel camptothecin payload [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 1872.
Abstract 1874: Common vulnerabilities of stem cells along the Barrett's-dysplasia-adenocarcinoma sequence
Authors: Xian Wa , McKeon Frank , Crum Christopher , Ajani Jaffer , Davies Peter , Vincent Matthew , Bachovchin William , Lai Hung-Sen , Pan Shuang , Su Amber , Hoffman Ashley , Montes Raul Caballero , Khorrami Melina , Khorrami Melika , Yamamoto Yusuke
Published: March 2024

Barrett's esophagus (BE) is an early precursor of esophageal adenocarcinoma and therefore a relevant target of preemptive therapeutics. Using robust stem cell cloning technologies, we have cloned stem cells from patient-matched biopsies of BE, low-grade dysplasia (LGD), high-grade dysplasia (HGD), and esophageal adenocarcinoma (EAC) from multiple patients. Whole exome sequencing of these stem cells reveals the mutational maturation along the sequence of BE, LGD, HGD, and EAC at unprecedented resolution of both copy number and single nucleotide variation. In vitro differentiation and xenografting of these clones recapitulates their original histology and shows that only the EAC stem cells yield robust tumors in immunodeficient mice. Leveraging the intrinsic immortality of these stem cells, we have performed high-throughput screens for small molecules that selectively kill BE stem cells without harming patient-matched normal esophageal stem cells. These screening efforts evolved into synthetic lethal strategies that identified drug combinations that selectively eliminate, in vitro, BE stem cells from 20 of 23 cases. Remarkably, this same synthetic lethal drug combination also eliminated in vitro stem cells of LGD, HGD, and EAC that were not components of the original screens. We have used synthetic chemistry to generate a series of novel molecules based on the identified drug combination that significantly improved the in vitro efficacy to sub-nanomolar levels. These novel molecules directed to the stem cell component of BE, LGD, HGD, and EAC also show potent effects against tumors generated from EAC stem cells in immunodeficient mice. We anticipate that these synthetically lethal drug combinations will have potential utility against precursor lesions to preempt EAC as well as against EAC itself.

Citation Format: Wa Xian, Frank McKeon, Christopher Crum, Jaffer Ajani, Peter Davies, Matthew Vincent, William Bachovchin, Hung-Sen Lai, Shuang Pan, Amber Su, Ashley Hoffman, Raul Caballero Montes, Melina Khorrami, Melika Khorrami, Yusuke Yamamoto. Common vulnerabilities of stem cells along the Barrett's-dysplasia-adenocarcinoma sequence [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 1874.
Abstract 1919: Activity and affinity tuning next-generation immunotoxins for targeted therapy
Authors: Liang Huazhu (Peter) , Beilhartz Greg L. , Bo Cao Shi , Melnyk Roman A.
Published: March 2024

Classic cancer-associated receptors, such as EGFR and Her2, are often overexpressed on malignant cells relative to healthy cells. Numerous molecules have been developed to leverage this differential expression with the goal of developing specific and safe targeted therapies, including antibody-drug-conjugates and immunotoxins. The general schematic of these molecules is to fuse toxic payloads -either small molecules or proteins - with a targeting moiety that binds to the cancer-specific receptor. Despite the general success of this approach, even low levels of these receptors on healthy tissues result in on-target/off-tumor toxicity thus limiting the dose of therapeutic molecules available to tumors. Targeted therapies with greater on-tumor efficacy and reduced toxicity are urgently needed in order to further advance therapeutic outcomes. To achieve this, we set out to exploit the unique features of bacterial toxins as highly modular protein delivery systems to design next-generation targeted therapies. We demonstrate the capacity of the immunotoxin platform to deliver various therapeutic proteins, with distinct activities from inhibiting protein synthesis to degrading oncogenic signaling pathways, such as the RAS/MAPK pathway. Further, we show how modifying the affinity, valency, and specificity of the receptor-binding moieties, as well as tuning the enzymatic activity of the toxic enzyme payloads interact to affect the on-target efficacy and off-target toxicity of the targeted molecules. Altogether, these data demonstrate the potential of immunotoxins as highly flexible and tunable platforms for the development of a new class of anti-cancer therapeutics.

Citation Format: Huazhu (Peter) Liang, Greg L. Beilhartz, Shi Bo Cao, Roman A. Melnyk. Activity and affinity tuning next-generation immunotoxins for targeted therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1919.
Abstract 1961: Discovery of a highly potent and brain penetrant fourth generation EGFR tyrosine kinase inhibitor
Authors: kumar sanjeev , wang junmei , li wenqian , pan weitao , wen zhiming , yu Kuo-Long , Zhao Genshi , yin tinggui
Published: March 2024

Third generation EGFR tyrosine kinase inhibitors (TKIs) such as Osimertinib have emerged as first-line standard of care therapy to treat non-small cell lung cancer (NSCLC) patients with activating EGFR mutations (del19, L858R or T790M). However, within 12 to 18 months following treatment, 15-20% of the patients develop C797S mutation, which drives resistance to these drugs; with the progression of disease brain metastasis is observed in 40-70% of the EGFRm+ NSCLC patients. Here, we report a highly potent and brain penetrant fourth generation EGFR-TKI ACE-34237307, designed to overcome C797S mutation (del19/C797S and L858R/C797S), and common single point mutations (del19 and L858R). ACE-34237307 is also designed to potently inhibit the more prevalent non-canonical EGFR mutations such as G719S and L861Q. ACE-34237307 showed robust anti-proliferative activity against EGFR double mutations in Ba/F3 (EGFR-del19/C797S and EGFR-L858R/C797S) cell lines with IC50 values of 0.1 and 0.23 nM, respectively. In phospho-EGFR assay, ACE-34237307 demonstrates over 79-fold selectivity against A431 (EGFR-WT) cell line when compared to PC-9 (EGFR-del 19) cell line. ACE-34237307 is highly brain penetrant with B/P ratio of 2.3 in rat and 1.2 in mouse. It has an excellent pharmacokinetic profile in rat, mouse and dog with good bioavailability, a flat PK curve and reduced Cmax, which will translate to a better safety profile. The in vivo anti-tumor efficacy was evaluated in Osimertinib resistant double mutant Ba/F3-EGFR-del19/C797S and L858R/C797S as well as del19 and L858R single mutant xenograft models. It has been shown that ACE-34237307 at doses of ≥ 0.6 mg/kg induces complete tumor regression or eradication in all tumor models tested. In brain tumor models with Ba/F3 (EGFR-del19/C797S) and PC-9 (EGFR-del19), ACE-34237307 showed complete inhibition or eradication of brain tumors and drastically extended animal survival at ≥ 0.6 mg/kg. In summary, ACE-34237307 is a potent fourth generation EGFR-TKI with high WT selectivity, excellent brain penetration, highly favorable DMPK profile, outstanding in vivo efficacy in subcutaneous and brain tumor models and low projected human dose amenable to QD dosing.

Citation Format: sanjeev kumar, junmei wang, wenqian li, weitao pan, zhiming wen, Kuo-Long yu, Genshi Zhao, tinggui yin. Discovery of a highly potent and brain penetrant fourth generation EGFR tyrosine kinase inhibitor [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 1961.
Abstract 1963: BB102, a highly selective and potent reversible-covalent FGFR4 inhibitor effective in multiple xenograft tumor models
Authors: Wang Qi , Duan Gongping , Hua Wei , Wei Xianglong , Li Min , Wang Junheng , Zhang Xingmin
Published: March 2024

Introduction: Fibroblast growth factor receptor 4 (FGFR4) belongs to the FGFR family which interact with fibroblast growth factors (FGFs) to regulate cell proliferation, differentiation and apoptosis. Due to unique structural domain, FGFR4 is specifically bound by FGF19 with high affinity in the presence of co-receptor β-Klotho. Hyperactivated FGF19/FGFR4 signaling, induced by FGF19/FGFR4 overexpression and FGFR4 mutation, was reported to be associated with the growth, metastasis and invasion of various cancer. Thus, FGFR4 is considered as a novel target to treat cancer with hyperactivated FGF19/FGFR4 signaling. Here we report BB102 as a highly selective and potent reversible-covalent FGFR4 inhibitor.

Methods: BB102 was developed through structure-based drug design, and optimized by SAR analysis and medicinal chemistry iteration. Biochemical and cell assays were applied in evaluation of BB102 inhibitory activity. Preclinical studies including pharmacokinetic (PK) and toxicity were performed in rats and dogs. Xenograft tumor models in mice were conducted to demonstrate the in vivo efficacy of BB102.

Results: As a reversible-covalent FGFR4 inhibitor, about half compound of BB102 dissociated from FGFR4 kinase within 24 h, which was conducive to overcome the fast re-synthesis rate of FGFR4 (< 2 h) in hepatocellular carcinoma cells. BB102 only inhibited kinase activity of FGFR4WT, FGFR4N535K and FGFR4V550L at nanomolar levels in 207 kinase panel screening. BB102 completely blocked the phosphorylation of the downstream protein ERK at 16.7 nM. BB102 exhibited strong inhibitory effects on the cell proliferation in vitro and tumor growth in mouse models inoculated with the high expression of both FGF19 and FGFR4, or FGFR4 mutated tumor cells. PK profile of BB102 showed good oral bioavailability (F>70%) with a nearly linear dose-dependent exposure. The compound had no effects on the respiratory and central nervous systems in rats, cardiovascular system in Beagle dogs or hERG liability in vitro. In comprehensive safety evaluation, the Severely Toxic Dose in 10% (STD10) in rats was 300 mg/kg/day and the Highest Non-Severely Toxic Dose (HNSTD) in dogs were 100 mg/kg/day.

Conclusion: BB102 is a highly selective and potent reversible-covalent FGFR4 inhibitor with promising efficacy in high expression of both FGF19 and FGFR4, and/or FGFR4 mutated tumor models and good safety in preclinical studies. The phase I study of BB102 is in progress.

Citation Format: Qi Wang, Gongping Duan, Wei Hua, Xianglong Wei, Min Li, Junheng Wang, Xingmin Zhang. BB102, a highly selective and potent reversible-covalent FGFR4 inhibitor effective in multiple xenograft tumor models [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 1963.