bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of the Protoscolicidal Activity of Humulus lupulus Methanolic Extracts on Echinococcus granulosus Sensu Stricto
Authors: Albani Clara María , Iglesias Azucena , Albanese Adriana , Fuentes Giselle , Orallo Dalila , Maggi Matías , Elissondo María Celina
Published: April 2024

The larval stage of the parasite Echinococcus granulosus sensu lato (s.l) is responsible for cystic echinococcosis (CE), a long-term infection affecting humans and animals worldwide, and constitutes a serious public health concern. If left untreated, CE can cause serious damage to multiple organs, especially the liver and lungs. Regarding the treatment, in the last few years, the use of pharmacological treatment has increased, suggesting that in the future, drug therapy may replace surgery for uncomplicated cysts. However, the only available anthelmintic drug to treat this infection is the albendazole, which has an efficacy that does not exceed 50%. On the basis of the above-mentioned evidence, new and improved alternative treatments are urgently needed. The use of natural products and their active fractions and components holds great promise as a valuable resource for the development of novel and effective therapies. Hop (Humulus lupulus L.) is a bittering agent in the brewing industry for which the sedative, digestive, anti-inflammatory, and antimicrobial effects have been reported. The purpose of this study was to assess the in vitro efficacy of methanolic extracts from the leaves of hop varieties against E. granulosus sensu stricto (s.s) protoscoleces. Varieties Mapuche and Victoria caused a stronger protoscolicidal effect compared to the Bullion, Cascade, and Traful varieties (P<0.01), coinciding with their highest content of flavonoids, total polyphenols, and saponins. The viability of protoscoleces treated with the varieties Mapuche and Victoria decreased to approximately 50% at days 5 y 8, respectively, showing alterations such as soma contraction and impaired microtriches. After 18 days of treatment with both varieties, protoscoleces were completely altered both structurally and ultrastructurally. In conclusion, the methanolic extracts of the H. lupulus varieties Mapuche and Victoria demonstrated a marked in vitro effect against E. granulosus s.s. protoscoleces. The beer-making industry exclusively uses hop cones, leaving behind large amounts of hop leaves as an agricultural by-product that is not being utilized. On the basis of our study, we propose that hop leaves could also be used as a source of secondary metabolites with anthelmintic activity.
Co-encapsulation of β-D-Galactosidase and Ascorbic Acid in the Milk Protein-Based Microcapsules: Optimization and Characterization
Authors: Hosseinnia Mahmoud , Khaledabad Mohammad Alizadeh , Almasi Hadi
Published: April 2024

This research is aimed at preparing the β-galactosidase (βg) and vitamin C (VC) cocapsules stabilized by milk proteins. The effect of different independent parameters including core-coating ratio (10-100%), whey protein isolate (0 : 1), sodium caseinate (0 : 1), and ultrasound power (50-150 W) on physicochemical properties of microcapsules was investigated. The response surface methodology (RSM) defined the optimal conditions. Increasing the WPI values had different effects on the particle size and polydispersity index (PDI). The zeta potential values decreased by decreasing SC values. The βg had better encapsulation efficiency in comparison to VC. Increasing the core-coating ratio showed a negative effect on the enzyme activity. Among the test parameters, the core-coating ratio was effective on the viscosity of microcapsules. Two optimum conditions for co-encapsulation were determined as WPI, SC, core-coating ratio, and ultrasound power of 0, 1, 100%, and 79.4 W and 0.2, 0.8, 100%, and 75 W for microcapsules I and II, respectively. In the next step, the structural and morphological properties of the optimum samples were analyzed. The heterogeneous morphology of microcapsules was observed by SEM analysis. The formation of new interactions between wall materials, βg, and VC was confirmed by FT-IR analysis. XRD analysis revealed that the WPI-coated sample had a higher crystallinity index. Generally, the successful co-encapsulation of βg and VC exhibited the potential of the resultant microcapsules for the industrial production of VC fortified and lactose-free milk.
New Means and Challenges in the Targeting of BTK
Authors: Nawaratne Vindhya , Sondhi Anya K. , Abdel-Wahab Omar , Taylor Justin
Published: April 2024

Bruton's tyrosine kinase (BTK) is central to the survival of malignant and normal B lymphocytes and has been a crucial therapeutic target of several generations of kinase inhibitors and newly developed degraders. These new means for targeting BTK have added additional agents to the armamentarium for battling cancers dependent on B-cell receptor (BCR) signaling, including chronic lymphocytic leukemia and other non–Hodgkin lymphomas. However, the development of acquired resistance mutations to each of these classes of BTK inhibitors has led to new challenges in targeting BTK as well as novel insights into BCR signaling. The first-generation covalent BTK inhibitor ibrutinib is susceptible to mutations affecting the covalent binding site, cysteine 481 (C481). Newer noncovalent BTK inhibitors, such as pirtobrutinib, overcome C481 mutation–mediated resistance but are susceptible to other kinase domain mutations, particularly at residues Threonine 474 and Leucine 528. In addition, these novel BTK inhibitor resistance mutations have been shown biochemically and in patients to cause cross-resistance to some covalent BTK inhibitors. Importantly, newer generation covalent BTK inhibitors zanubrutinib and acalabrutinib are susceptible to the same mutations that confer resistance to noncovalent inhibitors. The BTK L528W mutation is of particular interest as it disrupts the kinase activity of BTK, rendering it kinase dead. This observation suggests that BTK may act independently of its kinase activity as a scaffold. Thus, the timely development of BTK degrading proteolysis targeting drugs has allowed for degradation, rather than just enzymatic inhibition, of BTK in B-cell lymphomas, and early clinical trials to evaluate BTK degraders are underway.
Abstract CT287: LINNOVATE: A Phase 1/2 study of safety/efficacy using lurbinectedin, combined with ipilimumab, and nivolumab for advanced soft tissue sarcomas (NCT05876715)
Authors: Gordon Erlinda M. , Omelchenko Nadezhda , Jeffrey Samantha , Kumar Vaishali , Chawla Neal S. , Seetharam Mahesh , Chua Victoria , Moradkhani Ania , Quon Doris , Wong Steven , Chawla Sant
Published: April 2024

Background: Sarcoma cells are most immunogenic earlier in the course of the disease. Therefore, immune checkpoint inhibitors such as ipilimumab (ipi) and nivolumab (nivo), that promote sustained T-cell activation by suppressing T regs would be most effective when given as first line therapy, together with a tumoricidal agent, such as lurbinectedin, whose plausible mechanism of action is not only to destroy the cancer cells and expose the tumor neoantigens for immune recognition, but also to destroy growth promoting factors in the tumor microenvironment. In the SAINT phase 2 study using ipi, nivo and trabectedin as first line therapy for advanced soft tissue sarcomas (STS; n=79), there were 6 complete responses, 14 partial responses, 49 stable disease, 25.3% best response rate, 87.3% disease control rate; median PFS, 6.7 months, median OS, 24.6 months (Gordon et al, Cancers vol. 15, 906, 2023). Lurbinectedin is a synthetic marine-derived agent and an analog of trabectedin. Therefore, we expect that lurbinectedin will be as effective as trabectedin when used together with ipi and nivo as first line therapy for advanced STS.

Methods: Objective 1: Assess MTD of lurbinectedin. Objective 2: Evaluate PFS, OS, incidence and severity of adverse events; Correlative: Correlate response with ctDNA using Signatera. This is an open label, dose-seeking phase 1/2 study. The phase 1 part employs standard “cohort of three” design with a DLT window of 6 weeks. Treatment schedule: Ipi 1 mg/kg IV q 12 weeks; Nivo 3 mg/kg IV q 2 weeks; escalating doses of lurbinectedin from 2.6 mg/m2 to 3.2 mg/m2 IV q 3 weeks. Phase 2 part follows completion of phase 1 part wherein additional 28-34 participants will receive lurbinectedin at the MTD with fixed doses of Ipi and Nivo. Participants may continue treatment until significant disease progression or unacceptable toxicity occurs. Eligible patients are 18 years of age or older, previously treated in phase 1 and previously untreated in phase 2 with confirmed diagnosis of advanced STS, adequate hematologic and organ function, and no history of autoimmune disorder. To date, 6 patients in phase 1 have been dosed.

Citation Format: Erlinda M. Gordon, Nadezhda Omelchenko, Samantha Jeffrey, Vaishali Kumar, Neal S. Chawla, Mahesh Seetharam, Victoria Chua, Ania Moradkhani, Doris Quon, Steven Wong, Sant Chawla. LINNOVATE: A Phase 1/2 study of safety/efficacy using lurbinectedin, combined with ipilimumab, and nivolumab for advanced soft tissue sarcomas (NCT05876715) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT287.
Abstract LB010: Identification of malignant lung indeterminate pulmonary nodules with Plasma WGS using exon read depth of short DNA fragments
Authors: Yip Shun H. , Swarup Neeti , Hyun Jason C. , Cheng Jordan C. , Ramamurthy Raghuraman , Wong Edmund , Choi Irene , Arora Akanksha , Aberle Denise , Wong David TW , Tang Cheuk Y.
Published: April 2024

Background: Plasma WGS for non-invasive cancer detection has been an active area of research, where fragmentomics studies have been gaining attention for its promising results. Previous studies showed that the quantity of short and long fragments could be used for the prognosis of malignant tumor. In this study, the efficacy of short fragment read depth on the reference genome for malignant nodule detection was evaluated.

Methods: Plasma samples from 40 patients with Indeterminate Pulmonary Nodules (IPN) (20 with Benign nodule, 20 Malignant) were collected for WGS. Low quality samples were filtered, and 31 patients (15 Benign, 16 Cancer) remained. Short fragments in each sample were extracted (30-99 bp) and exon read depth for each sample was obtained through human reference genome alignment. A proprietary data analytic machine learning pipeline using XGBoost was trained and tested using 10 fold cross validation.

Results: XGBoost model testing accuracy was 96.77% in differentiating Benign and Cancer samples using read depth information from only 3 exons. GPR35’s ENSE00003591008.1, ZNF594-DT’s ENSE00002678218.2 and IGHGP’s ENSE00001819760.2 were the 3 exons used in the model. Hierarchical clustering using top features with differential read depth (p<0.001) was able to perfectly cluster the samples into the Benign group and the Malignant group.

Conclusion: Read depth information of exon by short fragments from WGS plasma was able to differentiate patients with Lung benign nodules from patients with malignant nodules. This analysis demonstrates promises of developing diagnosis tools using exon read depths from plasma samples.

Citation Format: Shun H. Yip, Neeti Swarup, Jason C. Hyun, Jordan C. Cheng, Raghuraman Ramamurthy, Edmund Wong, Irene Choi, Akanksha Arora, Denise Aberle, David TW Wong, Cheuk Y. Tang. Identification of malignant lung indeterminate pulmonary nodules with Plasma WGS using exon read depth of short DNA fragments [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB010.
Abstract LB034: First-in-class compounds targeting the MIEN1 signaling pathway: A breakthrough approach for breast and prostate cancer therapy
Authors: Tripathi Amit Kumar , Vishwanatha Jamboor K.
Published: April 2024

We report the identification and complete characterization of first-in-class molecules targeting the MIEN1 signaling pathway in cancer. Migration and invasion enhancer 1 (MIEN1) overexpression characterizes several cancers and facilitates cancer cell migration and invasion. Leveraging conserved ITAM and prenylation motifs within MIEN1, we identified potent anti-cancer peptides. Among them, bioactive peptides LA3IK and RP-7 induced pronounced transcriptomic and protein expression changes at sub-IC50 concentrations. The peptides effectively inhibited genes and proteins driving cancer cell migration, invasion, and EMT pathways, concurrently suppressing EGF-induced NF-kB nuclear translocation in metastatic breast cancer cells. Specifically, peptides targeted the same signal transduction pathway initiated by MIEN1. Molecular docking and circular dichroism spectroscopy indicated the formation of MIEN1-peptide complexes. The third-positioned isoleucine in LA3IK and CVIL motif in RP-7 were crucial for inhibiting breast cancer cell migration. This is evident from the limited migration inhibition observed when MDA-MB-231 cells were treated with scrambled peptides LA3IK-SCR and RP-7-SCR. Additionally, LA3IK and RP-7 effectively suppressed tumor growth in an orthotopic breast cancer model. Notably, mice tolerated high peptide doses of up to 90 mg/Kg well, surpassing significantly lower doses of 5 mg/Kg intravenously (iv) and 30 mg/Kg intraperitoneally (ip) used in both in vivo pharmacokinetic studies and orthotopic mouse model assays. D-isomers of LA3IK and RP-7 showed enhanced anti-cancer activity compared to their L-isomers. D-LA3IK remained stable in mouse plasma for 24 h with 75% remaining, exhibiting superior pharmacokinetic properties over D/L-RP-7. In summary, our findings mark the first report of short peptides based on MIEN1 protein sequence capable of inhibiting cancer signaling pathways, effectively impeding cancer progression both in vitro and in vivo.

Citation Format: Amit Kumar Tripathi, Jamboor K. Vishwanatha. First-in-class compounds targeting the MIEN1 signaling pathway: A breakthrough approach for breast and prostate cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB034.
Abstract LB052: PTP1B inhibitors activate a novel hypoxia-induced inflammatory cell death pathway in breast cancer
Authors: Neel Benjamin G. , Bhardwaj Abhishek , Antonelli Maria , Ueberheide Beatrix
Published: April 2024

Dual PTP1B/TC-PTP (PTPN1/2) inhibitors are in clinical trials for multiple cancer types, primarily because they augment anti-tumor immunity, but they can also have cancer cell-autonomous effects. Hypoxic cancer cells resist many anti-neoplastic therapies and can seed recurrence. We found previously that PTP1B deficiency/inhibition promotes HER2+ breast cancer cell death in hypoxia by activating RNF213, an ~600kDa protein containing AAA-ATPase domains and two ubiquitin ligase domains (RING and RZ) that also is implicated in Moyamoya disease (MMD), lipotoxicity, and innate immunity. We now report that PTP1B and ABL1/2 reciprocally control RNF213 phosphorylation on tyrosine-1275. This phosphorylation promotes RNF213 oligomerization and RZ domain activation. The RZ domain ubiquitylates CYLD/SPATA2, and together with the LUBAC complex, induces their degradation. Decreased CYLD/SPATA2 causes NF-κB activation, which together with hypoxia-induced ER-stress triggers GDSMD-dependent pyroptosis. Mutagenesis experiments show that the RING domain negatively regulates the RZ domain. CYLD-deleted HER2+ cell-derived xenografts phencopy the effects of PTP1B deficiency, and reconstituting RNF213 knockout lines with RNF213 mutants shows that the RZ domain mediates PTP1B-dependent tumor cell death. Our results identify a novel, potentially targetable PTP1B/RNF213/CYCLD/SPATA pathway critical for controlling inflammatory cell death in hypoxic tumors that could be exploited to target hypoxic tumor cells, potentially turning “cold” tumors “hot”, thereby increasing the potential clinical utility of PTP1B inhibitors.

Citation Format: Benjamin G. Neel, Abhishek Bhardwaj, Maria Antonelli, Beatrix Ueberheide. PTP1B inhibitors activate a novel hypoxia-induced inflammatory cell death pathway in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB052.
Abstract LB055: IBI3001: A potentially first-in-class site-specifically conjugated B7-H3/EGFR bispecific ADC for multiple solid tumors
Authors: Guan Jian , Zhang Xiao , Wu Weiwei , Cao Lei , Liao Zhengguang , Zhu Tianyu , Liu Chentao , Zhou Shuaixiang , Lu Jia , Li Ninghuan , Luo Nana , Zhang Ying , Dian Kang , Chia Tiong Sun , Chen Bingliang , He Kaijie
Published: April 2024

B7-H3 and EGFR are both clinically validated TAAs in solid tumors. We previously demonstrated in IBI334 (B7-H3/EGFR bsAb) study that (1) B7-H3 and EGFR are co-expressed in multiple solid tumors; (2) B7-H3-aided EGFR signaling inhibition was significantly more potent than Zalutumumab (EGFR mAb) and Amivantamab (c-met/EGFR bsAb); and (3) IBI334 has favorable safety profile in cynomolgus monkey at 120 mg/kg/week. Building on this optimized bispecific antibody, we site-specifically conjugated IBI334 with Exatecan (drug-antibody-ratio = 4) using the clinically validated SyntecanE platform. The resulting bispecific ADC, IBI3001, has multiple mechanism of actions: (1) enhanced EGFR signaling blockage; (2) EGFR- and B7-H3-aided payload internalization and cytotoxicity; and (3) bystander killing effects of ADC. More importantly, the optimized B7-H3 arm not only enhances the EGFR signaling blockage but also reduces EGFR on-target toxicities. IBI3001 demonstrated potent in vitro cytotoxicity in all B7-H3LowEGFRLow, B7-H3HighEGFRLow, B7-H3LowEGFRHigh and B7-H3HighEGFRHigh cancer cells across multiple solid tumors, including lung cancer, colorectal cancer, pancreatic cancer, breast cancer, head and neck cancer, and stomach cancer. In addition, it had strong bystander effects, killing both EGFR+ B7-H3+ cancer cells and EGFR- B7-H3- cancer cells. All in vitro studies were further validated in vivo with robust tumor growth inhibition in NCI-H508 (colorectal cancer), JIMT-1 (breast cancer), BxPC3 (pancreatic cancer) and NCI-1975 (lung cancer) xenograft models. IBI3001 has a favorable PK profile in BALB/c mice with the half-life at 282 hours, and is well-tolerated in cynomolgus monkeys up to 90 mg/kg/week. In conclusion, IBI3001 is a novel bispecific ADC that demonstrated strong anti-tumor efficacy cross multiple solid tumors, and has excellent PK and safety profile.

Citation Format: Jian Guan, Xiao Zhang, Weiwei Wu, Lei Cao, Zhengguang Liao, Tianyu Zhu, Chentao Liu, Shuaixiang Zhou, Jia Lu, Ninghuan Li, Nana Luo, Ying Zhang, Kang Dian, Tiong Sun Chia, Bingliang Chen, Kaijie He. IBI3001: A potentially first-in-class site-specifically conjugated B7-H3/EGFR bispecific ADC for multiple solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB055.
Abstract LB140: Age difference in cervical cancer stigma among Caribbean population
Authors: Thomas-Purcell Kamilah , Song Gaole , Sealy Diadrey-Anne , Bailey Althea , Ashing Kimlin T.
Published: April 2024

Introduction Cervical cancer (CCA) is the second most common cancer and one of the leading causes of death among women in the Caribbean region. CCA stigma impedes women from seeking and obtaining relevant healthcare. This study aimed to evaluate age difference in cancer stigma among non-patient populations in Jamaica, Grenada, and Trinidad and Tobago, utilizing a culturally trans-created scale.

Methods A cross-sectional study involving 1207 participants was conducted. The survey comprised the adapted Cancer Stigma Scale, supplemented with questions on cervical cancer knowledge and beliefs. Participants' knowledge and attitudes towards the HPV vaccine were also assessed. Data was collected online from October 2022 to March 2023. Statistical analysis was performed using SAS.

Results This study included 33.4% of total participants aged 18-25 years, 23.4% aged 26-35 years, 21.4 aged 36-45 years, 10.7% aged 46-55 years, 7.5% aged 56-65 years, and 3.6% aged 65+ years. Chi-squared tests showed age is significantly related to respondents’ opinions on women with CCA are treated with less respect than usual by others (p<.0001), women with CCA must keep it a secret (p=0.03), women with CCA are rejected by family members (p=0.0007), women with CCA are rejected by intimate partners (p<.0001), and intimate partners blame women for having CCA (p=0.003). Additionally, age has significant associations with agreements on negative views of cervical cancer from the community (p<.0001) and family (p<.0001) may discourage women from seeking and obtaining screening and treatment and community members believe CCA is viewed as shameful (p=.0001). Notably, there are more younger respondents (18-25 years) who agreed that women with CCA are rejected by intimate partners (32.3%), and the fear of family reaction and rejection may discourage women from getting further diagnosis after an abnormal Pap smear (48.1%).

Discussion Age is an important factor that influences cervical cancer stigma even in Caribbean contexts. Findings of this study indicate younger people may have a more serious stigma, which calls for proper educational programs for cervical cancer screening and treatment.

Citation Format: Kamilah Thomas-Purcell, Gaole Song, Diadrey-Anne Sealy, Althea Bailey, Kimlin T. Ashing. Age difference in cervical cancer stigma among Caribbean population [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB140.
Abstract LB147: Fine-scale mapping of chromosome 9q22.33 identifies candidate causal variant in ovarian cancer
Authors: Li an , Xing Tongyu , Zhao Yanrui , Chen Kexin , Zheng Hong
Published: April 2024

Ovarian cancer is a complex polygenic disease in which genetic factors play a significant role in disease etiology. Genome-wide association study(GWAS) identified a novel variant on chromosome 9q22.33as a susceptibility locus of epithelial ovarian cancer (EOC) in Han Chinese population. However, the underline mechanism of this region was still unknown. In this study, we conducted a fine-mapping analysis of 130kb region including 1,039 variants in 200 health women. Ten variants were selected to evaluate the association with EOC risk in 1,099 EOC cases and 1,591 controls. We identified two variants were significantly associated with ovarian cancer risk (rs7027650, P=1.91 × 10−7; rs1889268, P=3.71 × 10−2). Expression quantitative trait locus (eQTL) analysis found thatrs7027650was significantly correlated with COL15A1 gene expression (P=0.009). Luciferase reporter gene assay confirmed that rs7027650 could interact with the promoter region of COL15A1 to reduce its activity. Electrophoretic Mobility ShiftAssay (EMSA) showed theallele-specific binding capacity of rs7027650. These findings revealed that rs7027650 may be a potential causal variant at 9q22.33 region and may regulate the expression level of COL15A1. This study provided a clue for the molecular mechanism of potential causal variantaffecting the risk of ovarian cancer.

Citation Format: an Li, Tongyu Xing, Yanrui Zhao, Kexin Chen, Hong Zheng. Fine-scale mapping of chromosome 9q22.33 identifies candidate causal variant in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB147.
Abstract LB167: Acetalax (Oxyphenisatin acetate, NSC 59687) and bisacodyl (dulcolax) cause oncosis in triple negative breast cancer by poisoning the ion exchange membrane protein TRPM4
Authors: Mizunuma Makito , Redon Christophe , Saha Liton , Takebe Naoko , Pommier Yves G.
Published: April 2024

Triple negative breast cancer (TNBC) is generally unresponsive to current therapies, and the development of new anticancer agents is needed. Oxyphenisatin acetate, called Acetalax, which was used as a laxative in the 1950’s, has recently been reported to have anticancer activity in several murine cancer models. In this study we demonstrate that Acetalax and its structural analog laxative analog bisacodyl (Dulcolax) exhibit potent antiproliferative activity in TNBC cell lines. We show that both cause oncosis, a non-apoptotic cell death characterized by cellular and nuclear swelling and cell membrane blebbing that leads to mitochondrial dysfunction and ATP depletion. Acetalax was further shown to enhance immune and inflammatory responses and inhibit protein synthesis. Mechanistically, we provide evidence that TRPM4 (Transient Receptor Potential Melastatin member 4) is poisoned by Acetalax and bisacodyl. By blocking TRPM4, a monovalent cation channel, Acetalax and Bisacodyl cause oncosis, and TNBC cells without endogenous TRPM4 expression as well as TRPM 4 knockout TNBC cells were found to be Acetalax- and Bisacodyl resistant. TRPM4 was also lost in cells with acquired Acetalax resistance. Moreover TRPM4 is rapidly degraded the ubiquitin-proteasome system upon acute exposure to Acetalax and bisacodyl. Together, these results demonstrate that TRPM4 is a previously unknown target of Acetalax and Bisacodyl and that TRPM4 expression in cancer cells is a predictor of Acetalax and Bisacodyl efficacy, and could be used for the clinical development of these drugs as anticancer agents.

Citation Format: Makito Mizunuma, Christophe Redon, Liton Saha, Naoko Takebe, Yves G. Pommier. Acetalax (Oxyphenisatin acetate, NSC 59687) and bisacodyl (dulcolax) cause oncosis in triple negative breast cancer by poisoning the ion exchange membrane protein TRPM4 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB167.
Abstract LB209: YBX1 is a negative regulator of post-transcriptional program associated to EMT, invasion and metastasis in cSCC
Authors: Sol Stefano , Boncimino Fabiana , Todorova Kristina , Mandinova Anna
Published: April 2024

Y-box binding protein 1 (YB-1), which is encoded by YBX1 gene, is a multifunctional cold-shock protein that binds both DNA and RNA to orchestrate transcription as well as RNA processing and translation. Recently, we demonstrated that YBX1 is a post-transcriptional effector required for the maintenance of epidermal homeostasis. Although it was well known that YBX1 plays prominent pro-oncogenic roles in malignant transformation and cell invasion in a wide variety of cancers, the role of YBX1 in cutaneous squamous cell carcinoma (cSCC) is poorly understood. Here, we report that YBX1 protein level is higher in cSCC tumor compared with non-neoplastic skin, but it goes down in metastatic states. According to this finding, YBX1-KD cSCC primary cell lines show an increase of cell migration and invasion in vitro assay and in a context that mimics the natural three-dimensional environment of tumor with organotypic skin equivalent cultures. Furthermore, we found that YBX1-KD promotes an epithelial-to-mesenchymal transition and leads to the detection of more metastases in vivo after tail vein injection of YBX1-KD cSCC cells when compared with the injection of control cells. YBX1 is predominantly found in the cytoplasm of SCC cells. Therefore, we focused on the role of YBX1 as a post-transcriptional regulatorand performed an eCLIP-seq (crosslinking immunoprecipitation sequencing) to identify its direct RNA targets. We found that YBX1 binds novel pro-invasive RNA regulons associated to cell-cell and cell-matrix adhesions. Through the interpolation of the analysis of RNA-seq and SILAC, we found that YBX1 KD up-modulates at post-transcriptional level several targets, involved into invasion and metastasis. Our study reveals that, in human cutaneous squamous cell carcinoma, loss of function of YBX1 promotes migration, invasiveness and metastasis, through the induction of an epithelial-to-mesenchymal transition regulated at post-transcriptional level.

Citation Format: Stefano Sol, Fabiana Boncimino, Kristina Todorova, Anna Mandinova. YBX1 is a negative regulator of post-transcriptional program associated to EMT, invasion and metastasis in cSCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB209.
Abstract LB225: Regulation of breast cancer stem cell identity by PLK1 and FOXC2
Authors: Maddela Joanna Joyce , Castaneda Maria , den Hollander Petra , Kuburich Nick , Mani Sendurai
Published: April 2024

One of the main drivers for the drastic reduction in survival rates caused by metastatic breast cancer is attributed to Epithelial-to-Mesenchymal Transition (EMT). During embryonic development and wound healing, EMT plays a vital role and during cancer pathogenesis, cancer cells reactivate this program to gain plasticity, migration, and invasion promoting metastasis. In addition, EMT also plays a crucial role in generating a subpopulation of highly resilient tumor cells known as cancer stem cells (CSCs). Moreover, following conventional treatment, residual tumor cells that remain after treatment tend to exhibit CSC properties. EMT-induced cancer stem cells exhibit heightened plasticity and stemness, significantly fueling the metastatic spread of cancer and enhancing the endurance and adaptability throughout the process of metastasis. Thus, understanding the molecular intricacies governing stem cell properties holds immense potential for developing targeted therapies to target metastasis and the development of resistance to treatments.

To further elucidate EMT-induced cancer stem cell generation, our lab has demonstrated that the Polo-like kinase 1 (PLK1), an important G2/M regulator, plays an important role in the generation of CSCs through Forkhead box protein C2 (FOXC2). Most importantly, we have shown that the FOXC2 transcription factor, known to be crucial in embryonic development, now has emerging roles in metastatic cancer progression. FOXC2 is positioned at the center of the complex network of signal transduction pathways and functions downstream of most EMT-inducing transcription factors. FOXC2 is necessary for the acquisition and maintenance of stem cell property. In addition, we found that phosphorylation of FOXC2 by PLK1 stabilizes, and inhibition of PLK1 causes a decrease in FOXC2 and stemness in cancer cells. This led us to hypothesize that EMT-induced CSC generation is driven by the PLK1-FOXC2 axis. By expressing phosphomimetic and non-phosphorylatable FOXC2 mutants in breast cancer cell lines, we show that PLK1 phosphorylation is necessary for gaining stemness and an increase in symmetric self-renewal type of stem cell division. Evidence will be presented as to how we are targeting the PLK1-FOXC2 axis to make tumors sensitive to standard-of-care treatments.

Citation Format: Joanna Joyce Maddela, Maria Castaneda, Petra den Hollander, Nick Kuburich, Sendurai Mani. Regulation of breast cancer stem cell identity by PLK1 and FOXC2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB225.
Abstract LB256: New mechanistic insights into cisplatin resistance in muscle-invasive bladder cancer
Authors: Feldman Lily Elizabeth R. , Mohapatra Saswat , Jones Robert , Tilton Charlene , Zuiverloon Tahlita , Costello James , Theodorescu Dan
Published: April 2024

The standard of care for eligible patients with muscle-invasive bladder cancer (MIBC) is cisplatin-based neoadjuvant chemotherapy followed by radical cystectomy. However, platinum-based treatments leave up to 70% of MIBC patients with residual disease, and the 5-year survival rate for these individuals is under 30%. Understanding cisplatin resistance mechanisms will improve MIBC treatment by developing more effective therapeutics and precision medicine strategies. We previously identified NPEPPS, an M1 aminopeptidase, as a novel and potentially targetable mediator of platinum drug response. Recently, we have established that NPEPPS controls platinum drug import through its interactions with volume regulated anion channels (VRACs), which have been shown to import cisplatin to cells directly. We have shown that NPEPPS interacts with key VRAC subunit SWELL1 at the cell membrane and that intact SWELL1 is required for NPEPPS-mediated platinum resistance. We further discovered that NPEPPS cannot bind SWELL1 when its enzymatic aminopeptidase function is disrupted genetically or pharmacologically. In vivo studies showed that catalytic-dead NPEPPS-mutant MIBC xenografts were sensitive to cisplatin treatment compared to NPEPPS-wildtype xenografts. Patient cohort data analysis supported the model that the expression of NPEPPS and its relative expression with VRAC proteins are associated with overall survival in treatment-resistant MIBC patients. These findings shed light on how bladder cancer cells resist chemotherapy by disrupting the transport of drugs into cells and suggest new ways to target NPEPPS to enhance the effectiveness of platinum-based therapeutics in treating bladder cancer.

Citation Format: Lily Elizabeth R. Feldman, Saswat Mohapatra, Robert Jones, Charlene Tilton, Tahlita Zuiverloon, James Costello, Dan Theodorescu. New mechanistic insights into cisplatin resistance in muscle-invasive bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB256.
Abstract LB267: Disrupting cytoplasmic proton motive force induces ferroptotic cell death in colorectal cancer: A novel anti-cancer mechanism
Authors: Gaikwad Shreyas R. , Eltokhy Mohamed A. , Moore Morgan , Kailath Keerthi , Srivastava Sanjay K.
Published: April 2024

Introduction: Chemotherapeutic resistance and tumor recurrence remain the leading causes of poor prognosis in colorectal cancer (CRC). Herein, we evaluated the anti-cancer effect of a novel repurposed drug HLN* in CRC.

Methods: The changes in membrane potential were studied using DiSC(3) fluorescence by confocal microscopy. Cytotoxic activity of compound was studied using Sulphorhodamine-B (SRB) assay. Changes in mitochondrial membrane potential were studied using JC-1 dye. ROS accumulation was analyzed using DCFDA fluorescence using FACS analysis. Ferrorange assay was used to check the ferrous ion influx within the cell. Western blot was used to identify mode of cell death enacted.

Results: Our results show that HLN treatment reduced the survival of human and murine CRC cells (CT26, HCT116, Colo 205 and MC38) with IC50 ranging 1-4 µM. HLN further inhibited proton motive force (PMF) by sequentially depolarizing and hyperpolarizing the cell membrane, as examined by cellular membrane potential using DiSC(3) dye by confocal microscopy. Disruption of the PMF leads to changes in various cellular signaling cascades including ROS production and mitochondrial membrane potential (MMP) changes. The MMP was significantly reduced by HLN treatment in CRC cells. Furthermore, we also observed an increase in ROS production by HLN. Pretreatment with N-acetyl cysteine (NAC), an antioxidant, blocked the effects of HLN. Interestingly, ferroptotic cell death markers GPX4, GPX1, SLC7A11/xCT, NRF2 and KEAP1 were significantly inhibited by HLN treatment in CRC cells as observed by western blotting. Ferroptotic induction was then confirmed using ferrorange assay wherein we observed a concentration dependent increase in ferrorange intensity indicating an increase in Fe+2. Overall, HLN dissipates ΔpH by modulating iron influx leading to ferroptotic cell death through ROS.

Conclusion: Taken together, our results demonstrate the anti-cancer activity of a novel repurposed drug HLN and its unique mechanism of action. Our results provide a promise for developing this drug as a potential treatment option for CRC, through a novel anti-cancer mechanism. * Due to Intellectual property issued, the name of the drug is not provided

Citation Format: Shreyas R. Gaikwad, Mohamed A. Eltokhy, Morgan Moore, Keerthi Kailath, Sanjay K. Srivastava. Disrupting cytoplasmic proton motive force induces ferroptotic cell death in colorectal cancer: A novel anti-cancer mechanism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB267.
Abstract LB322: ARC2-001, a CAF-targeting drug exhibits strong anticancer effect in in vivo setting
Authors: Yeo So-Young , Farh Mohamed , Sohn InSuk , Kim Seok-Hyung , Lee Keun-Woo
Published: April 2024

For more effective tumor growth suppression, a combinatorial treatment of CAF-targeting agent and other well-validated therapies such as chemotherapy, radiotherapy, and immune checkpoint therapy should be considered. Thus, we hypothesized that co-administration of the CAF-targeting drug, ARC2-001 (the in vitro-approved CAF-targeting candidate) and clinically practiced anticancer drugs could be useful strategy for treating under clinical settings. However, it is essentially important to evaluate this using well-established pre-clinical models. We thoroughly verified the anticancer effect of ARC-001 using two in vivo model systems. ( 2nd- in vivo systems for CAF targeted drugs Research). 1. (Subcutaneous model) Immune-deficient mice (NOD/SCID): in which human colon cancer cell line HCT116 was co-injected with human CAF (CAFs were obtained from stage IV CRC patients) followed by administration of ARC2-001 alone or in combination with 5-fluorouracil (5FU). 2. (Subcutaneous and Orthotopic model) Wild-type mice (C57BL/6): in which mouse colon cancer cells was co-injected with mouse colon CAFs (both cancer cells and CAFs were obtained from chemically-induced colorectal cancer model with rich desmoplastic stroma) followed by administration of ARC2-001 alone or in combination with aPD-1. We observed a magnificent inhibition of tumor growth and metastasis in the combined treatment group over singular treatment groups in both models. This denotes that ARC2-001 largely augments the anti-cancer effect of widely employed anticancer drugs, such as 5FU, and thus can be exploited as an adjuvant for enhanced colorectal cancer chemotherapy.Collectively, our results evidenced the necessity of considering CAF-targeting agents for a more regulated therapeutic strategy to treat desmoplastic cancers.

Citation Format: So-Young Yeo, Mohamed Farh, InSuk Sohn, Seok-Hyung Kim, Keun-Woo Lee. ARC2-001, a CAF-targeting drug exhibits strong anticancer effect in in vivo setting [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB322.
Abstract LB329: Interleukin6 as a Potential Prognostic Marker and Therapeutic Target in Oral Squamous Cell Carcinoma
Authors: Goda Hiroyuki , Adachi Tomoko , Nakashiro Koh-ichi , Kuribayashi Nobuyuki , Uchida Daisuke
Published: April 2024

This study investigates the role of interleukin (IL)-6 in oral squamous cell carcinoma (OSCC), evaluating its potential as a prognostic marker and therapeutic target. This study involved 95 untreated OSCC patients, with pre-surgical serum IL-6 levels measured by Electro chemiluminescence immunoassay (ECLIA). Patients were categorized into high (78 patients) and low (17 patients) IL-6 groups based on a cutoff value of 8 pg/mL.A strong correlation was observed between higher IL-6 levels and lower overall survival (P<0.0001) and disease-free survival (P=0.0283). Serum IL-6 levels also showed a strong positive correlation with certain blood test markers such as neutrophil and lymphocyte fractions and C-reactive protein (CRP), and a negative correlation with albumin levels. These findings suggest a link between systemic inflammation, malnutrition, and the Inflammation-based prognostic score (IBPS).Further analysis through microarray assessed the gene expression in primary tumor tissues, revealing 513 genes with more than a two-fold change in the high IL-6 group compared to controls. Gene Ontology analysis indicated increased metabolic activity in the high IL-6 group and suppressed inflammation and immune-related molecules in the low IL-6 group.Immunohistochemical staining for IL-6 in tumor tissues showed its presence mainly in the stroma, particularly in cancer-associated fibroblasts (CAFs) identified by vimentin and α-SMA positivity. Both OSCC cell lines and CAFs established from OSCC tissues demonstrated high IL-6 secretion, especially in cell lines derived from lymph node metastases. Furthermore, cell lines from primary, lymph node, and lung metastases exhibited high IL-6 secretion, with HRAS mutations also identified. Knockdown of HRAS using siRNA reduced IL-6 secretion.Finally, in vivo experiments using an anti-IL-6 receptor antibody showed that blocking IL-6 signaling reduced the proliferation of OSCC cell lines.The study concludes that IL-6 is a useful prognostic and therapeutic target in OSCC. The wide applicability of IL-6 testing and the clinical use of anti-human IL-6R antibody drugs, combined with aggressive nutritional intervention, suggest a potential paradigm shift in OSCC treatment.

Citation Format: Hiroyuki Goda, Tomoko Adachi, Koh-ichi Nakashiro, Nobuyuki Kuribayashi, Daisuke Uchida. Interleukin6 as a Potential Prognostic Marker and Therapeutic Target in Oral Squamous Cell Carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB329.
Abstract LB429: Discovery and identification of a novel cancer associated fibroblast target using Oxford Biotherapeutics (OBTs) OGAP discovery platform
Authors: Sarnecky Jeremy , Lewis Joseph , Allen Jason , Davies Daniel , Metzger Matthew , Thomas Ben , Rohlff Christian , Cairns Belinda
Published: April 2024

Cancer Associated Fibroblasts (CAFs) are a major component of the tumor stromal microenvironment and have been indicated in promoting tumor growth as well as inhibiting tumor suppression mechanisms. One of the challenges in this field has been a lack of molecules that target CAFs specifically and can interfere with the CAF function. Here we describe a novel target discovered using OBTs proprietary OGAP platform, that appears to be specific for CAFs in a variety of tumor tissues. Interestingly this target appears to be minimally expressed in normal tissue fibroblasts by in situ hybridization RNA analysis but is highly upregulated in solid tissue stroma fibroblasts of colon, breast and pancreatic cancer tissues. The OGAP Discovery Platform utilizes a mass spectrometry approach to identify membrane targets on the surface of tumor cells as well as tumor microenvironment associated cells. This suggests that this novel target is expressed on the membrane of the CAFs making it a unique target for drug intervention. The identification of such a novel target is both exciting and a wonderful validation of OBTs OGAP Discovery Platform.

Citation Format: Jeremy Sarnecky, Joseph Lewis, Jason Allen, Daniel Davies, Matthew Metzger, Ben Thomas, Christian Rohlff, Belinda Cairns. Discovery and identification of a novel cancer associated fibroblast target using Oxford Biotherapeutics (OBTs) OGAP discovery platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB429.
Abstract LB436: Using FCCS to monitor target occupancy in cells and patient biopsies
Authors: Becker Frank
Published: April 2024

Lack of target engagement is one of the major reasons for failures in clinical studies. We developed an assay platform to detect and quatify target occupancy values in patient and animal biopsies to guide the development and dosing during the drug development process.

FCCS (fluorescence cross correlation spectroscopy) is a powerful tool to characterize drug target interaction in physiological environments. Over the last decade this technology was used to understand the binding and kinetics of small molecules and antibodies to challenging target proteins in great details and accuracy. Over the last decade Intana Bioscience expanded the application of FCCS from affinity measurements to comprehensive interaction studies including kinetics, multi component complex formation, assessing stoichiometry and quantification of aggregation, PK studies and target engagement. This presentation will highlight examples of results obtained with FCCS in projects ranging from early stages in the drug discovery up to the support of clinical trials to quantify target occupancies in patient samples.

Citation Format: Frank Becker. Using FCCS to monitor target occupancy in cells and patient biopsies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB436.
Abstract SY07-01: p53-mediated stem cell competition: Insights into mechanisms of clonal dominance
Authors: Piddini Eugenia
Published: April 2024

TP53 Mutations confer clonal advantage in several tissues contributing to tumorigenesis. It has traditionally been proposed that heightened resistance to apoptosis/cell cycle arrest can account for clonal advantage. P53 mediated clonal dominance has also been observed in cultured embryonic stem (ES) cells, though it is not clear how this is established. Using live cell imaging we show that human ES cells (hESCs) mutant for TP53 induce death of hES-WT cells and outcompete them. Competition is mediated by apoptosis induction and requires direct cell contact, as in co-cultures of hES-P53KO and hES-WT cells where contact is prevented competition is not observed. To ask if hES-P53KO competition is mediated by mechanical interactions, we cultured cells on deformable membranes to measure their resistance to cell compaction. Interestingly, while hES-WT cells displayed apoptosis induction upon cell compaction, hES-P53KO cells were resistant to compaction-induced apoptosis. We suggest that differential sensitivity to compaction in crowded cultures/tissues can drive p53-induced competitive cell elimination. Identifying the mechanisms by which p53 confers resistance to cell compaction could lead to novel actionable targets to prevent the expansion of p53 mutant stem cells in cancerous and pre-cancerous lesions.

Citation Format: Eugenia Piddini. p53-mediated stem cell competition: Insights into mechanisms of clonal dominance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr SY07-01.