bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5691: Preclinical evaluation of TFX05-01, a selective AKR1C3- targeted prodrug for solid tumor
Authors: Ding Charles Z , Cai Zhe , Sha Wei
Published: June 2022

Objective: Aldo-keto reductase 1C3 (AKR1C3), is a member of the aldo-keto reductase superfamily, catalyzing the NADP(H)-dependent stereospecific reduction of various aldehydes and ketones. AKR1C3 is overexpressed in a variety of cancer cells, including hepatocellular carcinoma (HCC), castration-resistant prostate cancer (CRPC), and leukemia cells. This study is to investigate the in vitro and in vivo antitumor activity of TFX05-01, a selective AKR1C3-activated anticancer prodrug, in preclinical models of AKR1C3 overexpressing cells. The studies provide solid foundation for nomination of TFX05-01 for clinical trials.

Method: Enzymatic activation of TFX05-01 was investigated in vitro using recombinant human AKR1C3. Cellular anti-proliferative activity was evaluated with a range of AKR1C3 overexpressed or low expressed cell lines. The in vivo antitumor activity was evaluated in various CDX mouse models with high expression of AKR1C3, including HepG2 and castration-resistant prostate VCaP, as well as PDX leukemia models

Result: TFX05-01 is a prodrug which selectively releases a DNA alkylating agent (TFX05-01A) upon exposure to AKR1C3. TFX05-01 inhibited cell proliferation in several AKR1C3 overexpressed HCC cell lines with an IC50 ≤ 100 nM. In contrast, TFX05-01 showed much lower anti-proliferative activity in low or no AKR1C3 expressed cells with an IC50 value greater than 1 μM. The cytotoxic potency of TFX05-01 in cancer cells correlates with the level of AKR1C3 expression, and the cytotoxic activity of TFX05-01 was inhibited when used in combination with a specific AKR1C3 inhibitor (IC50: 0.7 nM without inhibitor vs 188 nM with inhibitor in SNU-878 cells). TFX05-01 showed antitumor efficacy in AKR1C3 highly expressed HepG2 subcutaneously implanted xenograft model (TGI = 97% @2 mpk, QW, 3 doses IV). Concomitant PK studies showed that the concentration of active metabolites (TFX05-01A) in tumor was significantly higher (15~50 times) than that in plasma, which correlates with the selective release of the active DNA alkylating agent upon exposure to AKR1C3 in tumor tissues.

Conclusion: TFX05-01 is a novel selective AKR1C3-activated prodrug, which is different from traditional non-selective alkylating agents for cancer therapy. Preclinical studies have shown profound in vivo efficacy of TFX05-01 in AKR1C3 overexpressed mouse tumor xenograft models without marked toxicity. TFX05-01 represents a novel treatment option for AKR1C3 expressing solid cancers. Preclinical GLP safety evaluations were promising and TFX05-01 was nominated for clinical trials.

Citation Format: Charles Z Ding, Zhe Cai, Wei Sha. Preclinical evaluation of TFX05-01, a selective AKR1C3- targeted prodrug for solid tumor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5691.
Abstract 5701: HDAC inhibition decreases YAP1 expression but parodoxically activates TEAD transcription by the AP-1 pathway in breast cancer
Authors: Lin Ting-Yi , Tseng Yi-Ru , Wang Naikuan , Chao Ta-Chung , Liu Chun-Yu , Tseng Ling-Ming , Lai Jiun-I
Published: June 2022

The Hippo pathway is an evolutionally conserved pathway that is crucial in numerous aspects of cell biology, including contact inhibition, cell density, and others. Hippo pathway is well reported to be dysregulated in multiple cancer types and plays important oncogenic roles in human cancers. In mammalian cells, YAP is regulated by a phosphorylated cascade mediated by the kinases MST1/2 and LATS1/2, known as the canonical Hippo pathway. These upstream kinases phosphorylate YAP/TAZ and leads to its cytosolic degradation. Perturbation of the Hippo pathway leads to YAP intranuclear translocation and downstream activation of target genes, inducing proliferative signals and thus initiating oncogenesis. Novel mechanism linking the hippo pathway to various oncogenic mechanisms are rapidly discovered and reported, further establishing the significance of this pathway and its role in cancer biology. Breast cancer is the top leading cause of cancer in women and poses a significant morbidity and economic burden on society. Hormone positive (defined as positivity of the estrogen receptor ER and/or the progesterone receptor PR) breast cancer composes 70% of breast cancer. Endocrine therapy, chemotherapy, and more recently, CDK4/6 inhibitors and PI3K inhibitors are currently approved strategies towards breast cancer. Histone deacetylase (HDAC) inhibitors have been FDA approved for several hematological cancers, and recently have shown inconsistent results in clinical trials for breast cancer. In our study, we demonstrate and characterize a link between HDAC inhibition and the Hippo pathway. We discover that HDAC inhibitors suppress YAP expression both transcriptionally and translationally. Paradoxically, we demonstrate that although YAP is downregulated, TEAD mediated transcription of downstream Hippo targets is activated. Mechanistically, HDAC inhibition leads to upregulation of the AP-1 transcriptional machinery that cooperates with TEAD to facilitate transcription. By RNA-seq, chromatin studies and CRISPR knockout studies, we confirm that HDAC inhibition have the ability to activate downstream of Hippo pathway, suggesting a possible mechanism for breast cancer resistance towards HDAC inhibitor. Co-targeting HDAC inhibition and AP-1 results in enhanced efficacy of HDAC inhibitors and may be a promising approach for development towards breast cancer.

Citation Format: Ting-Yi Lin, Yi-Ru Tseng, Naikuan Wang, Ta-Chung Chao, Chun-Yu Liu, Ling-Ming Tseng, Jiun-I Lai. HDAC inhibition decreases YAP1 expression but parodoxically activates TEAD transcription by the AP-1 pathway in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5701.
Abstract 5709: Transcriptional circuitry of NKX2-1 and SOX1 defines a previously unrecognized lineage subtype of small cell lung cancer
Authors: kong ranran , Patel Ayushi S. , Sato Takashi , Yoo Seungyeul , Sinha Abhilasha , Tian Yang , Jiang Feng , Powell Charles A. , Snyder Eric , Jiang Jiantao , Li Shaomin , Watanabe Hideo
Published: June 2022

Introduction: The current molecular classification of small cell lung cancer (SCLC) based on expression of four lineage transcription factors, SCLC-A (ASCL1), SCLC-N (NEUROD1), SCLC-P (POU2F3), and SCLC-Y (YAP1) still leaves its major subtype SCLC-A as a large heterogeneous group, necessitating more precise characterization of lineage subclasses.

Experimental procedure: To refine the current SCLC classification and to identify specific lineage features of the SCLC subtypes, we performed unsupervised hierarchical clustering of H3K27ac profiles on transcriptional regulators from 25 SCLC cell lines and determined the epigenomic features for each cluster. Functional significance of the transcriptional lineage regulators for the identified cluster was evaluated by cell growth, apoptosis and xenograft using CRISPR-Cas9-mediated deletion. The specific cistromic profiles by ChIP-seq and its functional transcriptional partners using co-immunoprecipitation followed by mass spectrometry were determined to reveal their functional output in the identified subtype. Rb1fl/flTrp53fl/fl and Rb1fl/flTrp53fl/flNkx2-1fl/fl genetic engineered mouse model were generated to explore the function of Nkx2-1 in tumor initiation and differentiation. H3K27ac profiles were further analyzed to reveal 6 human SCLC specimen and 20 mice tumors epigenomic landscapes.

Summary: We identified previously uncharacterized epigenomic sub-clusters of the major SCLC-A subtype, named SCLC-A1 and SCLC-A2. SCLC-A1 was characterized by the presence of a super-enhancer at the NKX2-1 locus, which was observed in human SCLC specimens and a murine SCLC model. We found NKX2-1, a dual lung and neural lineage factor, is uniquely relevant in SCLC-A1, where it maintains neural lineage rather than pulmonary epithelial identity. We further found maintenance of this neural identity in SCLC-A1 is mediated by collaborative transcriptional activity with another neuronal transcriptional factor SOX1. ?

Conclusions: We comprehensively describe an additional epigenomic heterogeneity of the major SCLC-A subtype, and define SCLC-A1 subtype by the core regulatory circuitry representing NKX2-1 and SOX1 super-enhancers and their functional collaborations to maintain a neuronal linage state.

Citation Format: ranran kong, Ayushi S. Patel, Takashi Sato, Seungyeul Yoo, Abhilasha Sinha, Yang Tian, Feng Jiang, Charles A. Powell, Eric Snyder, Jiantao Jiang, Shaomin Li, Hideo Watanabe. Transcriptional circuitry of NKX2-1 and SOX1 defines a previously unrecognized lineage subtype of small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5709.
Abstract 5712: Dietary polyphenols as BRD7 and 9 inhibitors for cancer interception
Authors: Kapoor Sabeeta , Dashwood Roderick H.
Published: June 2022

Introduction Epigenetic mechanisms play an important role in the initiation and advancement of colorectal cancer (CRC) and other malignancies due, in part, to deregulated bromodomain (BRD) functions1,2. We examined compounds from natural sources as BRD7/9 inhibitors, seeking lead candidates outside of the widely studied bromodomain and extraterminal (BET) family3.

Methods As reported4,5, docking in silico involved STRAP and AutoDock Vina, ligand-protein interactions employed PDBePISA and LPC/CSU, and BROMOscan®-Bromodomain arrays were used to screen for BRD7/9 interacting compounds. BRD7/9 inhibitors also were assessed by isothermal titration calorimetry (ITC) and by anticancer activities in human colon cancer cells.

Results Docking in silico identified several dietary polyphenols as potential BRD7/9 inhibitors, with binding energies ≤ -8.0 kcal/mol. BROMOscan® and ITC experiments prioritized aspalathin, orientin, epigallocatechin-3-gallate, quercetin and equol for further investigation. Treatment with selected polyphenols mimicked BRD7/9 siRNA-mediated knockdown in reducing colon cancer cell viability and inhibiting colony formation, leading to DNA damage and apoptosis induction. Normal colonic epithelial cells were unaffected by the treatments, signifying cancer-specific targeting. These findings suggest that dietary polyphenols can recognize and target BRD7/9 proteins for potential cancer interception.

Acknowledgements Research supported in part by NCI grant CA122959, the John S. Dunn Foundation, and a Chancellor's Research Initiative.1)Jung G et al., Nat Rev Gastroenterol Hepatol 2020;17:111-130. 2)Fujisawa T, Filippakopoulos P. Nat Rev Mol Cell Biol 2017;18:246-262.3)Damiani E et al., J Cancer Prev 2020;25:189-203. 4)Rajendran P et al., Cancer Res 2019;79:918-927. 5)Kapoor S et al., Cancers (Basel) 2021;13:1438.

Citation Format: Sabeeta Kapoor, Roderick H. Dashwood. Dietary polyphenols as BRD7 and 9 inhibitors for cancer interception [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5712.
Abstract 5723: JDP2 interacts with p53 and enhances p53 transactivation and stability
Authors: Yang Wei-Hsiung , Price Kasey , Cardoso Leticia , Yang William , Wang Chiung-Min , Yang Richard
Published: June 2022

USAAP-1 protein mainly consists of proteins belonging to c-Jun, c-Fos, activating transcription factor (ATF), and Jun dimerization protein (JDP) family. In addition to AP-1 site, JDP2 also binds to cAMP responsive element (CRE) site in numerous cis-elements of the target genes. JDP2 has been shown to be involved in cancer development and cell-cycle regulation, suppression of adipocyte differentiation, and controlling bone homeostasis and antibacterial immunity. ATF3 and JDP2 share high similarity in C-terminal domains with identities of 65%. Previous studies have demonstrated that ATF3 can bind to p53 and regulate p53 transcriptional activity and stability. Though couple of studies have shown that JDP2 inhibits expression of p53 and p53 represses the transcriptional activity of JDP2 promoter; however, whether JDP2 directly interacts and regulates p53 transactivation remains largely unknown. Herein, we demonstrate for the first time that JDP2 can directly interact with p53. First, we found that both ATF3 and JDP2 can bind to p53 and the C-terminus of JDP2 is required for p53 binding. Secondly, in p53-null H1299 cells, JDP2 dose-dependently enhances p53 transactivation using p53RE-Luc (14X). Moreover, we demonstrated that JDP2 dose-dependently decreases p53 protein level in consistence with the previous reports. Finally, JDP2 increases p53 stability in CHX chase experiments. Taken together, our results demonstrate that JDP2 directly binds to p53 and enhances p53 transactivation and stability, suggesting that JDP2 is a novel regulator of p53.

Citation Format: Wei-Hsiung Yang, Kasey Price, Leticia Cardoso, William Yang, Chiung-Min Wang, Richard Yang. JDP2 interacts with p53 and enhances p53 transactivation and stability [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5723.
Abstract 5796: Targeting poorly differentiated and anaplastic thyroid cancer microenvironment via TNF-α/TGF-β/LOX signaling to improve drug delivery and treatment efficacy
Authors: LAHA DIPRANJAN
Published: June 2022

A major limitation of current systemic solid cancer therapies is the failure to effectively deliver drugs to cancer cells. Solid tumors have high cell density and poor lymphatic drainage, leading to higher interstitial fluid pressure (IFP). We previously showed that the tumor microenvironment of poorly-differentiated (PDTC) and anaplastic thyroid cancer (ATC) can be selectively and effectively treated with nanomedicine carrying recombinant human Tumor Necrosis Factor α (TNFα)(CYT6091), resulting in vascular leakage, decreased IFP, and increased intratumoral concentration of paclitaxel in vivo. The purpose of this study is to investigate the mechanisms involved in a TNFα-induced reduction of IFP. We investigated the effects of TNF-α on cytokines and extracellular matrix (ECM) proteins in tumor microenvironment. Our data showed that TNFα treatment downregulated transforming growth factor-beta (TGF-β) and Lysyl oxidase (LOX) and upregulated several ECM proteins. However, we observed LOX upregulation when we treated thyroid cancer with siTNF-α-receptor 1 or 2 followed by TNF-α, suggesting the involvement of TNF-α-receptors in treatment resistance. In addition to the role of LOX in stiffening the ECM, causing a barrier to drug delivery, we showed overexpression of LOX in aggressive thyroid cancer. We observed increased vascular leakage in siLOX-treated thyroid cancer xenografts, mimicking the effect of CYT-6091. Furthermore, we confirmed that the LOX expression in thyroid cancer is positively associated with transforming growth factor receptor 1 (TGFBR1). In TCGA thyroid cancer database the mRNA expression of TGFBR1 was positively correlated with those of ECM regulatory genes (ITGB2, ITGA2 and COL1A1) which were downregulated with TNF-α treated thyroid cancer cells. To assess the role of TGF-β on vascular permeability, we found that the TGF-β inhibitor (SB-431542) induced in vitro vascular leakage, while the recombinant TGF-β reversed TNF-α-induced in vitro vascular leakage in HUVEC cells. SB-431542 and TGF-β neutralizing antibodies independently and markedly improved treatment efficacy of paclitaxel in in vitro thyroid cancer 3D tumor spheroids. Moreover, TGF-β1 inhibition downregulates the protein levels of Smad3 molecules in thyroid cancer cells. TCGA database studies of thyroid cancer patient samples showed that SMAD3 positively correlated with TGFBR1 and LOX, suggesting the involvement of SMAD3 in TGF-β/LOX signaling in the TME.

Citation Format: DIPRANJAN LAHA. Targeting poorly differentiated and anaplastic thyroid cancer microenvironment via TNF-α/TGF-β/LOX signaling to improve drug delivery and treatment efficacy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5796.
Abstract 5807: MDM2 mitochondrial translocation mediates metabolic reprogramming towards OXPHOS in TKI-resistant oncogene-addicted cancer
Authors: Eu Jie Qing , Kong Li Ren , Hirpara Jayshree , Goh Boon Cher , Wong Andrea LA , Pervaiz Shazib
Published: June 2022

There is mounting evidence of oxidative phosphorylation (OXPHOS) dependency in cancers resistant to tyrosine kinase inhibitors (TKI), but the cause of this metabolic switch remains elusive. Mitochondria-bound MDM2 (mtMDM2), the fraction of MDM2 oncoprotein which is actively imported into the mitochondria, has been found to dysregulate electron transport chain (ETC) complex I function and OXPHOS. AKT is known to be a regulator of MDM2 protein stability. Given that AKT phosphorylation is commonly detected in oncogene-addicted tumours, we investigated the role of mtMDM2 and AKT in promoting TKI resistance through reprogrammed cellular metabolism. EGFR+ NSCLC [HCC827 and HCC827-GR (gefitinib-resistant)] and BRAF+ melanoma [A375 and A375-VR (vemurafenib-resistant)] cells were established via repetitive pulsed strategy. Translocation of MDM2 was examined through subcellular fractionation and Western Blotting. To investigate the role of MDM2 in mediating OXPHOS, genetic knockdown of MDM2 was performed via siRNA followed by measurement of oxygen consumption rate (Seahorse Analyzer); mRNA expression of mtDNA-encoded ETC subunits by real-time PCR (qPCR). ChIP-qPCR analysis was performed to examine the binding affinity of MDM2 and TFAM (mitochondrial transcription factor A) to mitochondrial DNA (mtDNA). Mitochondrial translocation of MDM2 was predominant in TKI-sensitive cells; MDM2 silencing upregulated OXPHOS and induced mRNA expression of mtDNA-encoded ETC complex I subunits, suggesting that mtMDM2 inhibits OXPHOS. ChIP analysis revealed competitive binding of MDM2 and TFAM at the LSP region of mtDNA. Conversely, MDM2 was primarily localized the cytoplasm in TKI-resistant cells and was associated with increased OXPHOS in TKI-resistant cells. Mechanistically, both pAKT and pMDM2 (Ser166/Ser186) were higher in TKI-resistant cells. Collectively, we describe a novel observation that OXPHOS upregulation underpinning TKI resistance in cancer cells could be mediated by AKT upregulation and MDM2 localisation.

Citation Format: Jie Qing Eu, Li Ren Kong, Jayshree Hirpara, Boon Cher Goh, Andrea LA Wong, Shazib Pervaiz. MDM2 mitochondrial translocation mediates metabolic reprogramming towards OXPHOS in TKI-resistant oncogene-addicted cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5807.
Abstract 5819: Tumor nutrient conditions confer drug resistance in pancreatic cancer
Authors: Sheehan Colin , Martin James , Croley Grace , McKeever Joshua , Thomas-Toth Anika , Jonker Patrick , LaBelle James , Koh Andrew , Muir Alexander
Published: June 2022

Drug resistance is a major obstacle to the treatment of pancreatic ductal adenocarcinoma (PDAC). Current standard of care includes combination chemotherapies; however, the majority of patients fail to respond to these treatments. Despite significant advances in our understanding of PDAC biology, it remains unclear what biological processes drives the drug resistance observed in most PDAC cases. Contributing to our limited understanding of this problem, in vitro models of PDAC fail to recapitulate in vivo drug resistance, suggesting the tumor microenvironment is a critical regulator.A defining feature of PDAC tumors is an abnormal nutrient environment, which may carry important biological cues driving many of the undesirable cell behaviors related to disease progression. Using a novel medium formulation that mimics the nutrient conditions observed within PDAC tumors (TIFM), we find that PDAC cells cultured in TIFM become highly resistant to common chemotherapeutic agents, better recapitulating the in vivo drug resistance phenotype than standard cultures. Several of these agents engage with their intended target in TIFM cultured cells, discouraging the idea that drug resistance is primarily mediated by drug efflux. However, TIFM cultured cells display reduced apoptotic priming, enabling their survival to drug exposure. Drug resistance enabled by TIFM nutrients is not immediately reversible with standard culture and can be inherited over multiple cell generations. Consistent with this, we find that TIFM nutrient conditions results in significant alterations to the epigenetic landscape of pancreatic cancer cells, rewiring expression of genes that may function as critical regulators of cell survival in PDAC. Ultimately, the work from this project will help elucidate how poor nutrient perfusion within solid tumors regulates cell survival processes and may lead towards better treatment options for PDAC patients beyond combination chemotherapy.

Citation Format: Colin Sheehan, James Martin, Grace Croley, Joshua McKeever, Anika Thomas-Toth, Patrick Jonker, James LaBelle, Andrew Koh, Alexander Muir. Tumor nutrient conditions confer drug resistance in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5819.
Abstract 5832: Utilizing small noncoding RNAs as biomarkers for selinexor treatment in gastric cancer
Authors: Sexton Rachel E. , Aboukameel Amro , Li Yiwei , Khan Husain Y. , Uddin Md Hafiz , Kashyap Trishyan , Landesman Yosef , Tesfaye Anteneh , Azmi Asfar S.
Published: June 2022

Gastric cancer is a deadly disease with a low 5-year survival rate of 32%. This disease is characterized by high rates of chemo resistance and late initial detection. We have previously reported that inhibition of nuclear exporter protein Exportin 1 (XPO1/CRM1) with the small molecule inhibitor selinexor (KPT-330) is a viable therapeutic approach in gastric cancer. Inhibition of XPO1 blocks proliferation and suppresses tumor growth through nuclear retention of tumor suppressor proteins such as TP53. Besides nuclear retention, we also demonstrated that XPO1 inhibition leads to a perturbation of cancer specific microRNAs including microRNA-7974 downregulation (miR-7974) and microRNA-129-1-3p upregulation (miR-129-1-3p) within a small RNA sequencing study. In this study we evaluated the role of these microRNAs in gastric cancer perpetuation in respect to their naive functions in five distinct subtypes of gastric cancer. Our results show over-expression of miR-7974 and loss of miR-129-1-3p promotes gastric cancer growth and progression, based on specific subtype of disease. We have further found that the differential expression of these microRNAs leads to gastric cancer cellular changes including cell cycle alterations, upregulation of autophagy processes, chemoresistance and alterations in the cellular framework leading to more invasive phenotypes. Validation with normal gastric epithelial cells revealed these microRNAs are relevant as they are perturbed in gastric cancer as well as the proteins in which they modulate. Finally, we have validated treatment of gastric cancer cells with the XPO1 inhibitor selinexor altered the expression of miR-7974 and miR-129-1-3p differently, based on subtype, as we have previously observed in small RNA sequencing screen, leading to a reversal of the pro-cancerous phenotypes previously observed. In vivo investigation is currently ongoing to identify whether we can capture either miR-7974 or miR-129-1-3p within the serum as further evidence of microRNA perturbation as a predictive biomarker for selinexor efficacy.

Citation Format: Rachel E. Sexton, Amro Aboukameel, Yiwei Li, Husain Y. Khan, Md Hafiz Uddin, Trishyan Kashyap, Yosef Landesman, Anteneh Tesfaye, Asfar S. Azmi. Utilizing small noncoding RNAs as biomarkers for selinexor treatment in gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5832.
Abstract 5834: Examining the potential oncogenic function of septins and their interaction with Chmp1A tumor suppressor in pancreatic cancer
Authors: Cai Mei , Amor Joseph , Park Maiyon
Published: June 2022

Introduction: Chmp1A functions as a tumor suppressor by the activation of ATM and p53 in pancreatic cancer. The nuclear localization signal (NLS) of Chmp1A is required for cell growth inhibition and activation of ATM and p53. Proteomic analysis has identified Septins (a group of GTP-binding proteins) to be associated with the NLS-deleted Chmp1A. Since NLS-deleted Chmp1A promotes cancer cell growth, we hypothesized that Septins may function as oncogenes and that Chmp1A functions as a tumor suppressor partly by inhibiting the oncogenic action of Septin proteins.

Objective: To examine whether Septin proteins exhibit oncogenic activity and to investigate whether Chmp1A inhibits the oncogenic function of Septins.

Methodology: Using normal pancreatic and cancer cells, we compared Septin transcripts using PCR assays, Septin protein expression using Western blot, and Septin cellular expression using immunocytochemistry. We performed immunohistochemical analysis to examine whether Septin expression is increased in pancreatic cancer tissues compared to corresponding normal tissues. Silencing technology was used to test whether Septin expression is regulated by Chmp1A.

Results: Our preliminary data indicates that the transcript and protein of Septins are increased in various pancreatic cancer cell lines compared to normal. Septin protein expression was increased or altered in human pancreatic ductal adenocarcinoma tissues compared to corresponding normal tissues. Additionally, our data implies that Chmp1A negatively regulates Septin expression, since Septin transcripts and proteins are increased when Chmp1A protein is silenced.

Summary and future direction: Our data suggests that Septin functions as an oncogene in pancreatic cancer cells and that Chmp1A functions as a tumor suppressor partly by inhibiting the oncogenic action of Septin proteins. For the completion of the project in the future, we plan to confirm the function of Septins by investigating the effect of Septin overexpression on pancreatic cancer cell proliferation. In addition, we will further investigate the interaction between Septins and Chmp1A by examining Septin expression and localization in pancreatic cancer cells that overexpress Chmp1A.

Citation Format: Mei Cai, Joseph Amor, Maiyon Park. Examining the potential oncogenic function of septins and their interaction with Chmp1A tumor suppressor in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5834.
Abstract 5838: The landscape of EGFR exon 20 insertion mutations in Chinese patients with non-small cell lung cancer
Authors: xu shun , Fu Zhichao
Published: June 2022

Background: The epidermal growth factor receptor (EGFR) is one of the most commonly altered genes in non-small cell lung cancer(NSCLC). Of these, insertion mutation in 20 exon (Ex20ins) of the EGFR occurs less frequently. In comparison with the common EGFR mutations (19Del/L858R), the NSCLC patients with Ex20ins have a worse prognosis. Recently, the FDA approved targeted oral therapeutic agent for NSCLC patients with EX20ins mutation, which has markedly improved their survival. Here we carry out a complete analysis of the frequency and types of EGFR 20 Exon insertion mutations in the large-scale NSCLC cohort of the Chinese population.

Methods: From January 2020 to October 2021, a total of 4,091 patients were confirmed as NSCLC were included in the study. The Next-generation sequencing was utilized for detecting genetic mutations.

Results: Among the 4,091 NSCLC patients, 2,411 (58.9%) experienced EGFR mutations. In which we identified 90 patients(2.2%) with Ex20ins. All of the Ex20ins were localized between amino acids S767-V774, and after C-helix of the EGFR kinase domain. Different from the exon 19 deletion, we found that Ex20ins exhibits various types of mutations. A total of 34 Ex20ins types were detected, with the most common A767_V769dupASV and S768_D770dupSVD mutations, accounting for 26% and 22.2% respectively. In addition, H773dupH (6.67%), H773_V774dupHV(5.56%), H773_V774insAH (2.22%), D770_N771insG (2.22%), and N771_P772insG(2.22%) were also observed in the NSCLC patients.

Conclusions: We described the landscape of EGFR Ex20ins in Chinese NSCLC patients cohort. Based on the large sample size, the frequency of the Ex20ins mutation in Chinese NSCLC patients was 2.2%, and the types of EGFR Ex20ins mutation were highly heterogeneous.

Citation Format: shun xu, Zhichao Fu. The landscape of EGFR exon 20 insertion mutations in Chinese patients with non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5838.
Abstract 5841: Discovery of a novel epigenetic regulatory axis that mediates sorafenib's antineoplastic activity
Authors: Wei Tianzi , Wang Hao , Lin Risheng , Li Yueming , Lu Yi , Zhang Jian
Published: June 2022

As a multi-kinase inhibitor, sorafenib has been widely used to treat advanced hepatocellular carcinoma patients since 2007. Sorafenib, however, is associated with severe drug adverse effects and drug resistance. Therefore, the strategies to improve the antineoplastic effects of sorafenib are urgently needed. In this study, we observed that metallothionein-1G (MT1G), was significantly activated by sorafenib and mediated sorafenib’s antineoplastic activity both in vitro and in vivo. Our study also proved that sorafenib effectively inhibited the activity of several critical enzymes required for maintaining methylation level in hepatoma cells to activate MT1G. In addition, the ATAC-seq results demonstrated that the promotor region of MT1G become more accessible upon the treatment of sorafenib, which may make a few key transcription factors easier to bind to the promoter region of MT1G to initiate the transcription. Finally, next-generation sequencing (NGS) results showed that the expression of two tumor associated genes: KLF4 and CA9, were significantly altered by MT1G overexpression. KLF4 was required to bind to the promotor region of CA9 and then inhibit the expression of CA9, a hypoxia-induced gene involved in modulating tumor microenvironment. The decrement of CA9 expression was related to hepatoma cells suppression. Taken together, we deduce that MT1G-KLF4-CA9 axis is a potential novel pathway to suppress hepatoma cell progression regulated by sorafenib. The regulation of MT1G-KLF4-CA9 is plausible to synergize with sorafenib to improve the therapeutic effects. Supported by NSFC projects 81773146, 81972766, 81972420, 82173336; Shenzhen Science and Technology Commission Project JCYJ20180302174235893; Sanming Project of Medicine in Shenzhen SZSM202003009; Guangdong Basic and Applied Basic Research Foundation 2021A1515012161; and Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research (2017B030301018)

Citation Format: Tianzi Wei, Hao Wang, Risheng Lin, Yueming Li, Yi Lu, Jian Zhang. Discovery of a novel epigenetic regulatory axis that mediates sorafenib's antineoplastic activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5841.
Abstract 5845: The fusion gene landscape in Chinese patients with non-small cell lung cancer
Authors: lin qiuju , Fu Zhichao
Published: June 2022

Background: Gene fusion is one of the common types of pathogenic mutation in non-small cell lung cancer(NSCLC). Numerous fusion genes have been identified as important biomarkers and therapeutic targets in NSCLC. The identification and analysis of fusion genes will provide important insights into the mechanisms of NSCLC development and devise novel therapeutic strategies. Here, our primary objective was to describe the landscape of fusion in Chinese patients with NSCLC to explore the new treatment option in NSCLC.

Methods: A total of 1192 pathologically confirmed NSCLC samples were collected. The fusion genes were detected using next-generation sequence(NGS).

Results: A total of 168 fusion events (14.1%) were identified in 1,192 patients with NSCLC. The most common type of fusion was ALK (64.9%), RET (25%), ROS (8.93%), NTRK1 (3.57%), and FGFR3 (3.57%). In these cases, 109 patients exhibited ALK fusion, including EML4/ALK (97.2%), KIF5B/ALK (0.92%), KLC1/ALK (0.92%), and STRN/ALK (0.92%). A total of 42 patients had RET fusions, of which 83.3%, 11.9%, 2.38%, and 2.38% with KIF5B, CCDC6, GOPC, and TET1 respectively. Notably, RET gene fusions with GOPC (GOPC/RET1), TET1 (TET1/RET1) were first reported in NSCLC and the two novel fusions retained the complete kinase domain of RET. We also found three types of ROS fusions in 15 patients, CD74/ROS1(66.7%) and EZR/ROS1(20%) were the common fusions. Additionally, a new fusion LRIG3/ROS1 which retained the complete kinase domain was identified for the first time in NSCLC patients.

Conclusions: We conducted a comprehensive review of fusion genes in Chinese patients with NSCLC. With the exception of common gene fusions, we found three cases of rare fusion mutations, which help to understand the potential pathogenic mechanisms of NSCLC and translate into therapeutic applications.

Citation Format: qiuju lin, Zhichao Fu. The fusion gene landscape in Chinese patients with non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5845.
Abstract 5884: Comprehensive analyzing the expression of IDO1 and TDO2 in bladder cancer
Authors: Pham Quoc Thang , Nguyen Thanh Tu , Nguyen Thao Quyen , Luu Duc Tung , Pham Thi Nhu Diem , Bui Thi Thanh Tam , Duong Thanh Tu , Phan Dang Anh Thu , Ngo Quoc Dat
Published: June 2022

Background: Several lines of evidence have been focused on the roles of tryptophan metabolism genes like indoleamine 2,3-dioxygenase 1 (IDO1) and tryptophan 2,3-dioxygenase (TDO2) in bladder cancer progression. However, the alternative way of targeting in cancer immunotherapy has been not yet investigated in bladder cancer. Method: We explored and validated the clinicopathological significance and the correlation with immune infiltrates of IDO1 and TDO2 expression in bladder cancer using Bladder urothelial carcinoma (BLCA) and other Gene Expression Omnibus (GEO) datasets (GSE13570, GSE31684, GSE48277) downloaded from https://xenabrowser.net/ and https://www.ncbi.nlm.nih.gov/geo/. The correlation between IDO1, TDO2 expression and the immune infiltrates as well as PD-L1 gene expression in BLCA dataset was explored with TIMER2.0, http://timer.comp-genomics.org/. Immunohistochemistry was performed to validate the relationship between TDO2 and PD-L1 expression in bladder cancer. Results: We figured out the IDO1 and TDO2 expressions were significantly upregulated in basal type compared with other molecular subtypes in BLCA. P53 mutations were associated with upregulation of IDO1 and TDO2 expression (p<0.01) whereas FGFR3 mutation was associated with downregulation of IDO1 and TDO2 expression (p<0.001). The overexpression of IDO1, TDO2 overexpression was significantly associated with advanced disease in BC. IDO1 and TDO2 expression were significantly correlated with purity and immune cell infiltrates, including T cells CD8+, T cells CD4+, B cells, neutrophils, and macrophages as wells as myeloid dendritic cells. Interestingly, TDO2 were positively correlated with cancer-associated fibroblasts (CAFs) (rho=0.5, p=1.14e-24). PD-L1 expression was also correlated with IDO1 (rho=0.64, p=4.42e-49) and TDO2 expression (rho=0.42, p=5.01e-19). Immunohistochemistry revealed the correlation between TDO2 and PD-L1 expression (p=0.011). Conclusion: Our results pointed out that IDO1 and TDO2 could play an essential role in regulating the tumor microenvironment as well as immune tolerance in bladder cancer. Suggesting that IDO1, TDO2 might be a promising novel immunotherapy target for bladder cancer patients.

Citation Format: Quoc Thang Pham, Thanh Tu Nguyen, Thao Quyen Nguyen, Duc Tung Luu, Thi Nhu Diem Pham, Thi Thanh Tam Bui, Thanh Tu Duong, Dang Anh Thu Phan, Quoc Dat Ngo. Comprehensive analyzing the expression of IDO1 and TDO2 in bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5884.
Abstract 589: Discovery of NTX-001, a potent Cbl-b inhibitor with antitumor activity in syngeneic models
Authors: Csibi-Levin Fred , Leit Silvana , Laughton David L. , Baker Tom , Jacques Suzanne L. , Browning Beth , Toms Angela V. , Garside Samantha , D'Archivio Simon , Kopycka Katarzyna , Zhu Xiaohua , Jordan Allan M. , Thomson Stuart , Loh Christine , Tummino Peter , Ciccone David N.
Published: June 2022

Introduction: Casitas B-lineage lymphoma b (Cbl-b), a RING finger E3 ligase and a member of a highly conserved family of Cbl proteins, catalyzes the ubiquitination of substrate proteins to regulate multiple signaling events in a variety of cell types, including immune cells. In T cells, Cbl-b negatively regulates adaptive immune system signaling by establishing the threshold for the activation of antigen receptors. Additionally, Cbl-b regulates the function of other immune cell types, including NK cells, dendritic cells (DC) and monocytes. Cbl-b deficient T cells no longer require a costimulatory signal to be fully activated. Cbl-b KO mice spontaneously reject tumors via an enhanced immune response. Taken together, these findings point to Cbl-b inhibitors as having the potential to be highly efficacious immuno-oncology agents.

Experimental Procedures: A structure-based drug design approach was used to identify potent inhibitors of Cbl-b. Biochemical and cellular binding assays, a cellular NFAT-luciferase reporter assay, as well as primary in vitro human and mouse T cell activation assays measuring IL-2 production were used to profile inhibitor compounds. In vivo activity of Cb-b inhibitors was evaluated using an anti-CD3 mouse model and a CT-26 syngeneic mouse model

Results: Cbl-b inhibitor NTX-001 potently binds to Cbl-b, preventing Cbl-b phosphorylation and binding to E2. In cells, NTX-001 treatment results in enhanced phosphorylation of ZAP-70, a protein proximal to the TCR and reported to be a direct substrate of Cbl-b. Additionally, inhibition of Cbl-b potently enhances cytokine secretion from primary human and mouse T cells. In vivo, an increase in cytokines and T cell activation markers was observed after a single dose of NTX-001. Repeated dosing of NTX-001 showed dose-dependent anti-tumor activity in the colorectal CT-26 syngeneic model.

Conclusions: NTX-001 is a potent Cbl-b inhibitor that demonstrated strong T cell activation and anti-tumor activity in a syngeneic tumor model. These data support Cbl-b inhibitors as a promising therapeutic opportunity for cancer treatment.

Citation Format: Fred Csibi-Levin, Silvana Leit, David L. Laughton, Tom Baker, Suzanne L. Jacques, Beth Browning, Angela V. Toms, Samantha Garside, Simon D'Archivio, Katarzyna Kopycka, Xiaohua Zhu, Allan M. Jordan, Stuart Thomson, Christine Loh, Peter Tummino, David N. Ciccone. Discovery of NTX-001, a potent Cbl-b inhibitor with antitumor activity in syngeneic models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 589.
Abstract 5920: Multi-cancer early detection with a spectroscopic liquid biopsy platform
Authors: Cameron James M. , Sala Alexandra , Antoniou Georgios , Brennan Paul M. , Conn Justin J. A. , Connal Siobhan , Palmer David S. , Smith Benjamin R. , Baker Matthew J.
Published: June 2022

The development of a robust multi-cancer early detection (MCED) test would be transformational within the cancer diagnostics field, as earlier detection is vital to inhibit disease progression, improve patient prognosis and reduce mortality rates. In this large-scale spectroscopic study, we present the Dxcover® Cancer Liquid Biopsy, the world’s first infrared spectroscopy-based blood test for the early detection of multiple cancers. The full cohort (n=2094) was comprised of patients with a confirmed cancer diagnosis (n=1543), symptomatic non-cancer patients (n=460) and healthy volunteers (n=91). For the cancer versus non-cancer classification, receiver operating characteristic analysis produced a mean curve with an area under the curve (AUC) value of 0.86. When adjusted for greater sensitivity, our model achieved 90% sensitivity with 61% specificity, and yielded a sensitivity of 56% with 91% specificity when tailored for higher specificity. We also tuned for maximized sensitivity or specificity, whilst the other parameter was fixed above a minimum value of 45% for the cross-validation result, based upon performance levels of currently available clinically benchmarked tests. This resulted in a 94% sensitivity where specificity was 47%, and a 94% specificity with 48% sensitivity. The overall detection rates were 93% for stage I, 84% for stage II, 92% for stage III and 95% for stage IV for the sensitivity-tuned model. The specificity-tuned model detected 90% of non-cancer patients accurately, resulting in few false negatives (54/551). For the binary classifiers of individual cancer types against symptomatic control patients, promising AUC values were reported for all cancers: brain (0.90), breast (0.74), colorectal (0.91), kidney (0.91), lung (0.90), ovarian (0.85), pancreatic (0.81) and prostate (0.85). Importantly, the detection rates did not vary significantly according to cancer stage, suggesting the technology is very effective at detecting early-stage tumors, which is a necessity for MCED tests.

Citation Format: James M. Cameron, Alexandra Sala, Georgios Antoniou, Paul M. Brennan, Justin J. A. Conn, Siobhan Connal, David S. Palmer, Benjamin R. Smith, Matthew J. Baker. Multi-cancer early detection with a spectroscopic liquid biopsy platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5920.
Abstract 5949: Aspirins metabolites inhibit human colon cancer cell growth: Potential role in colorectal cancer prevention
Authors: Tummala Poojitha , Sankaranarayanan Ranjini , Dachineni Rakesh , Bhat Jayarama Gunaje
Published: June 2022

Aspirin’s role in colorectal cancer (CRC) prevention is evidential from several epidemiological data. Numerous theories and targets have been proposed, a consensus has not been reached regarding its mechanism of action. One of the unexplored area in aspirin’s CRC prevention is the role of Dihydroxy benzoic acids (DHBAs), which are generated from aspirin’s metabolism by gut microflora and cytochrome P450 (CYP450). Out of the screened DHBAs, 2,3-DHBA and 2,5-DHBA showed concentration dependent inhibition of CDK1, CDK2, CDK4 and CDK6 enzyme activity in-vitro. We demonstrated that 2,3-DHBA and 2,5-DHBA inhibits CDK-1 enzyme activity beginning at 500 µM, while CDK2 and CDK4 activity was inhibited only at higher concentrations (>750 µM). 2,3-DHBA inhibited CDK6 enzyme activity from 250 µM, while 2,5-DHBA inhibited its activity >750 µM. Using colony formation assays, we further demonstrated the role of 2,3-DHBA and 2,5-DHBA in cancer cell cycle inhibition in 3 different cancer cell lines. Out of the 2 DHBAs, 2,5-DHBA was highly effective in inhibiting clonal formation in HCT-116 and HT-29 CRC cell lines (250-500 µM), and in the MDA-MB-231 breast cancer cell line (~100 µM); whereas both aspirin and salicylic acid failed to inhibit all four CDKs and colony formation. Based on the present results, it is suggested that 2,3-DHBA and 2,5-DHBA may contribute to the chemopreventive properties of aspirin, possibly through the inhibition of CDKs. The present data and the proposed mechanisms should open new areas for future investigations.

Citation Format: Poojitha Tummala, Ranjini Sankaranarayanan, Rakesh Dachineni, Jayarama Gunaje Bhat. Aspirins metabolites inhibit human colon cancer cell growth: Potential role in colorectal cancer prevention [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5949.
Abstract 5975: Dual-channel near-infrared fluorescence imaging for simultaneous identification of lung cancer and intersegmental plane
Authors: Quan Yu Hua , Bao Kai , Kim Kyungsu , Wang Haoran , Yokomizo Shinya , Park G. Kate , Choi Byeong Hyeon , Rho Jiyun , Kim Chungyeul , Choi Hak Soo , Kim Hyun Koo
Published: June 2022

Intraoperative delineation of the tumor with minimal negative margins is the holy grail of lung cancer surgery. Segmentectomy is a minimal resection surgery required for elderly patients or small sized early-stage lung cancer patients, in which accurate detection of tumor margins with simultaneous identification of the lung intersegment plane is the key to success. However, non-small cell lung cancer is difficult to detect intraoperatively, and the identification of intersegmental plane is challenging during minimal resection surgery. Here, we report dual-channel image-guided lung cancer surgery using renal clearable and physiochemically stable targeted fluorophores to visualize the tumor margins and intersegmental plane separately with different colors but simultaneously: cRGD-ZW800-PEG (800 nm channel) allows for tumor-specific targeting and ZW700-1C (700 nm channel) for discrimination of segmental planes. Armed with the dual-channel imaging, image-guided surgery enables complete tumor resection with minimal negative margins that can reduce the recurrence rate and increase the survival rate of lung cancer patients.

Citation Format: Yu Hua Quan, Kai Bao, Kyungsu Kim, Haoran Wang, Shinya Yokomizo, G. Kate Park, Byeong Hyeon Choi, Jiyun Rho, Chungyeul Kim, Hak Soo Choi, Hyun Koo Kim. Dual-channel near-infrared fluorescence imaging for simultaneous identification of lung cancer and intersegmental plane [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5975.