bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4482: Development of small molecule inhibitors of lipocalin 2 for the treatment of aggressive cancers
Authors: Flores-Colón Marienid , Peña-Vázquez Cristal A. , Hernández-O'Farril Eliud , Vázquez-Quiñones Luis E. , Vivas-Mejía Pablo E.
Published: March 2024

Lipocalin-2 (LCN2), a secreted protein pivotal in iron homeostasis, immune responses, siderophore transport, and epithelial cell differentiation. LCN2 is highly abundant in aggressive forms of many cancers including breast, pancreatic, ovarian, and esophageal cancer. Elevated LCN2 levels in aggressive tumors correlate with heightened cancer cell motility, proliferation, angiogenesis, invasion, and metastasis. After the protein folds it makes a barrel structure which encloses a three-calyx pocket that can bind and transport molecules. A first generation of small molecule inhibitors (SMIs) targeting LCN2 demonstrated efficacy in attenuating the proliferative capacity of Inflammatory breast cancer (IBC) cells. IBC stands as a rare and highly aggressive variant, accounting for 1 to 5% of all BC cases, with 20-30% presenting as de novo metastatic disease. In the previous studies the concentration of LCN2 inhibitors to reduce cell proliferation in 50% was around 10 µM. In pursuit of better LCN2 SMIs, we conducted in-silico analysis using a library of 369,000 ligands. Through the High-Performance Computer Facility at UPR Río Piedras, we employed AutoDock Vina for virtual screening, targeting the LCN2-calyx with Lys134 as the central coordinate. Docking protocols, predicting binding affinities, identified 8,162 out of 327,638 ligands that have been docked already with values below -9.6 kcal/mol for further consideration. The next phase involves subjecting these potential inhibitors to the SwissADME tool for a comprehensive evaluation of absorption, distribution, metabolism, and excretion parameters, as well as pharmacokinetics, drug-likeness, and medicinal chemistry considerations. The parameters to be evaluated are the Lipinski rules, which states that an orally active drug may have a maximum of one violation and the pann-assay interference compounds (PAINS) alerts tool, a source of false positives for biologically active compounds. The top candidates, exhibit binding affinities within the range of -11.5 to -12.2. After SwissADME filtering, we will re-rank the ligand list, identifying 30 commercially available candidates for subsequent cell viability assays in SUM149 (TNBC) and SUM190 (HER2+) cell lines. This multifaceted approach integrates computational methodologies with rigorous experimental validation, paving the way for potential therapeutic interventions in IBC treatment.

Citation Format: Marienid Flores-Colón, Cristal A. Peña-Vázquez, Eliud Hernández-O'Farril, Luis E. Vázquez-Quiñones, Pablo E. Vivas-Mejía. Development of small molecule inhibitors of lipocalin 2 for the treatment of aggressive cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4482.
Abstract 4501: First reported PIM kinase degraders: Design, profiling & optimization
Authors: Legros Celine , Ratcliffe Paul , Porkolab Vanessa , Modugno Michele , Mirguet Olivier
Published: March 2024

The PIM serine/threonine kinases (PIM1/PIM2/PIM3) are downstream effectors of ABL, JAK2 and Flt-3 oncogenes and are required for tumorigenesis. Overexpression has been reported in hematological and solid tumors, myeloma, lymphoma, leukemia and adenocarcinoma. The first generation of PIM inhibitors to make it to the clinic, including SGI-1776, AZD1208 and PIM447, are known to be pan-PIM inhibitors, while the aim of this project was to identify novel & selective PIM3 inhibitors. To this end, several Hit-finding approaches were employed to rapidly generate novel chemical matter, which were then assessed for their activity against PIM3 using a robust ADP-Glo™ assay. In parallel to this Hit-findings activity, we used AZD1208 scaffold as a starting point to generate PIM3 bifunctional protein degraders and designed a specific screening cascade in order to profile such modalities. The PIM degrader library, synthetized using Automated Robotics Lab, was characterized using ADP-Glo assays. It was then screened on our binding and activity kinase platforms, KINOMEscan® and KinaseProfiler™, and then fully profiled including ternary complex formation with biophysics and in vitro Safety Pharmacology Profiling Panels. One example is the AZD1208 derivate degrader, i.e. EC108154-1, which showed nanomolar range affinity for PIM3 and the E3-ligase CRBN-DDB1, and its affinity range was also confirmed with the ternary complex in SPR and MST-TRIC. This comprehensive screening cascade ensures an in-depth characterization and optimization of protein degraders, with the possibility of monitoring the ternary complex formation with biophysics methods. It is a good illustration of how this combination of tests can be instrumental in selecting the best degraders to progress to the next step of drug discovery.

Citation Format: Celine Legros, Paul Ratcliffe, Vanessa Porkolab, Michele Modugno, Olivier Mirguet. First reported PIM kinase degraders: Design, profiling & optimization [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4501.
Abstract 4525: Discovery of a highly potent and selective PARP1 inhibitor with superior PK and safety profiles
Authors: Wei Yi , Li Wenming , Li Zehui , Li Manhua , Kang Kai , Wen Zhiming , Yin Tinggui , Yu Kuo-Long , Zhao Genshi , Yuan Hongbin
Published: March 2024

PARP inhibitors have been an effective treatment option for cancers deficient in homologous recombination repair, such as ovarian, breast, prostate, and pancreatic cancers, particularly those harboring BRCA mutations. However, their severe overlapping hematological and gastrointestinal toxicities have limited their clinical use, especially in the combination therapies with other anti-cancer agents. More recently, studies have shown that inhibiting PARP1 mainly drives anti-tumor activity while inhibiting PARP2 contributes to the hematological toxicity such as anemia due to its essential role in the survival of hematopoietic stem and progenitor cells. Thus, there is an urgent need for developing a selective PARP1 inhibitor. In this study, we report the discovery and characterization of a unique PARP1 inhibitor, ACE-86225106. This molecule is highly selective at inhibiting PARP1 over PARP2 as demonstrated in biochemical (72x) and DNA-trapping (131x) assays. It demonstrates potent anti-proliferative activities against BRCA mutant cancer cell lines (IC50 = 1-9 nM) but not against BRCA wild type cancer cells (IC50 >30,000 nM). As compared to the BRCA mutant cancer cells, this molecule is much less active at inhibiting the differentiation of CD34+ hematopoietic stem cells (IC50 ≈ 450-1,400 nM), i.e.: erythroid, myeloid and megakaryocyte. The results of these studies show that ACE-86225106 is a highly potent and selective PARP1 inhibitor. This molecule has a unique PK profile characterized by its long half-life and low volume of distribution. As such, it demonstrates robust efficacy at low dose (1 mg/kg) with a less frequent dosing schedule (once a week). Importantly, there was no significant body weight loss observed in the treatment group even at much higher doses (10 mg/kg), but there was a significant body weight gain. The GLP toxicology studies have confirmed its superior safety profile as the maximum recommended safe starting first-in-human dose was >5-fold higher than the predicted human efficacious dose. Taken together, ACE-86225106 is a highly potent, selective PARP1 inhibitor with excellent PK and safety profile and expected to have a better safety profile, deeper and more durable clinical responses, and broader utility in the clinic as a preferred combination partner with other anti-cancer therapies. A phase 1 clinical trial to evaluate its safety, PK, and clinical utility is to begin in early 2024.

Citation Format: Yi Wei, Wenming Li, Zehui Li, Manhua Li, Kai Kang, Zhiming Wen, Tinggui Yin, Kuo-Long Yu, Genshi Zhao, Hongbin Yuan. Discovery of a highly potent and selective PARP1 inhibitor with superior PK and safety profiles [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4525.
Abstract 4557: Overexpressed CPAP promotes cancer stemness and enhances chemo-resistance in HCC
Authors: Hung Liang-Yi
Published: March 2024

Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and is highly resistant to chemotherapy and targeted therapy. It was reported that HCC cancer stem cells (CSCs) are involved in chemo-drug resistance by regulating multidrug resistance (MDR) gene expression, such as ABC transporters. Our previous reports indicated that CPAP, a centrosomal protein, can promote HCC progression by up-regulating NF-κB and STAT3 transcriptional activities. Here, we demonstrated that overexpressed CPAP can increase cancer stemness features and CSCs-derived sorafenib resistance in HCC by up-regulating the transcriptional activities of NF-κB and β-catenin. Importantly, CPAP stably expressed HCC spheroids are significantly resistance to sorafenib in vitro and in vivo. Our results suggested that CD24 and ABCG2 are two potential downstream effectors in CPAP overexpressed HCC. CPAP can transcriptionally up-regulate CD24 and ABCG2 expression via Wnt/β-catenin and NF-κB signaling pathways, respectively; high CPAP/ABCG2/CD24 expression levels are negatively correlated with disease-specific survival rates in HCC patients. Combined treatment with sorafenib and Wnt/β-catenin pathway inhibitor PRI-724 significantly inhibited CPAP-overexpressing Hep3B spheroids-derived tumor growth in a xenograft mouse model. Our study highlights the oncogenic role of CPAP in promoting CSCs-induced sorafenib resistance in HCC and provides insights into potential therapeutic strategies.

Citation Format: Liang-Yi Hung. Overexpressed CPAP promotes cancer stemness and enhances chemo-resistance in HCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4557.
Abstract 4579: DNMT1 determines the sensitivity of Class I HDAC inhibitor Psammaplin A to induce apoptosis and DNA damage in hematological malignant cells
Authors: Bao Yu , Liu Dan , Wu Zi , He Yumei , Gong Ping , Waxman Samuel , Jing Yongkui
Published: March 2024

Histone deacetylases (HDACs) and DNA methyltransferases (DNMTs) are overexpressed in cancer cells associated with dysregulated gene expression and tumor progression. DNMT1 partnered with Class I HDAC as complex to inhibit gene transcription. Several pan-HDAC inhibitors have been approved for T-cell lymphoma with evident side effects. More selective HDAC inhibitors need to be developed. Psammaplin A (PsA), a symmetrical bromotyrosine-derived disulfide dimer isolated from the Psammaplysilla sponge, is a selective Class I HDAC inhibitor. We tested its cell growth inhibition and apoptosis induction in 12 hematological malignant cell lines with 8 lines sensitive (more than 50% growth inhibition at 1 μM) and 4 lines insensitive (less than 20% growth inhibition at 1 μM). PsA induces acetylation of H3, but not α-tubulin, supporting the Class I HDAC inhibition selection. In the sensitive cell lines, PsA induces apoptosis with down-regulation of c-Flip and upregulation of NOXA as well as DNA damage detected by γ-H2A.X. RNA sequence analysis results confirm the regulation of PsA on NOXA, c-Flip and p21 gene expression. The four insensitive cell lines express high levels of DNMT1 with resistance to apoptosis induction. Silencing DNMT1 with siRNA sensitizes PsA-induced apoptosis with induction of NOXA and γ-H2A.X. Azacytidine, the DNMT1 degrader, in combination with PsA induce synergistic cell growth inhibition and apoptosis in vitro. PsA (20 mg/kg) plus Azacytidine (2.5 mg/kg) synergistically inhibit tumor growth in insensitive mantle cell lymphoma Jeko-1 xenografts, without detected toxic signs in organ pathological analysis. We find a new mechanism of DNMT1 controlling the sensitivity of Class I HDAC inhibitor PsA and provide a rationale of PsA with azacytidine for combination therapy.

Citation Format: Yu Bao, Dan Liu, Zi Wu, Yumei He, Ping Gong, Samuel Waxman, Yongkui Jing. DNMT1 determines the sensitivity of Class I HDAC inhibitor Psammaplin A to induce apoptosis and DNA damage in hematological malignant cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4579.
Abstract 4585: Discovery of ETS-006, a highly potent YAP/TEADs PPI inhibitor with broad anti-tumor activity as a single agent
Authors: Lu Jiajun , Gao Ming , Du Wenpei , Fan Xiaofei , Li Jinping , Wang Mengya , Feng Lijian , Li Yingjie , Yao Jiaqi , Lu Jiting , Cui Wenqi , Zheng Qiangang , Zhu Jidong
Published: March 2024

The transcription co-activators YAP, pairing with the TEAD family of transcription factors, serve as essential effectors of the conserved Hippo signaling pathway. Hippo kinase cascade controls YAP nuclear entry and binding with TEADs by modulating its phosphorylation and stability. Overexpression and activation of YAP/TAZ are frequently observed in a variety of cancers. It has been well established that YAP aberration contributes to cancer development, metastasis, immunosurveillance escape and drug resistance. Disruption of YAP/TEADs PPI is an effective approach to inhibit YAP transcriptional activity. We previously have reported the identification of YAP/TEADs PPI hit compound through the fragment-based screen and advanced the hit to lead compound ETS-003. Here we reported the preclinical candidate of YAP/TEADs PPI inhibitor ETS-006 with much more potent anti-tumor activity and favorable safety profile. Compared to TEAD palmitoylation inhibitor, ETS-006 could completely disrupt the interaction of YAP/TEADs, resulting in a more profound downregulation of YAP target gene expression. ETS-006 demonstrated more robust anti-tumor activity than the competitor’s YAP/TEADs PPI inhibitor in MPM CDX model with favorable ADME properties. Notably, ETS-006 showed promising efficacy across multiple solid tumor types, including head and neck squamous cell carcinoma, osteosarcoma, and triple-negative breast cancer with urgent unmet clinical needs. In conclusion, ETS-006 is a highly potent and orally available YAP/TEADs PPI inhibitor with broad anti-tumor activity as a single agent.

Citation Format: Jiajun Lu, Ming Gao, Wenpei Du, Xiaofei Fan, Jinping Li, Mengya Wang, Lijian Feng, Yingjie Li, Jiaqi Yao, Jiting Lu, Wenqi Cui, Qiangang Zheng, Jidong Zhu. Discovery of ETS-006, a highly potent YAP/TEADs PPI inhibitor with broad anti-tumor activity as a single agent [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4585.
Abstract 4633: Differentiation driver gene HOXD4 as a potential prognostic indicator and therapeutic target in anaplastic thyroid cancer
Authors: Kopec Kaci , Jarboe Tara , DeSouza Nicole R. , Islam Humayun K. , Geliebter Jan , Tiwari Raj K.
Published: March 2024

Anaplastic thyroid cancer (ATC) is always diagnosed as stage IV with a 5 year survival rate of less than 3% due to its aggressive metastatic phenotype. ATC lacks differentiation and loses its thyroid-like functions making it a difficult cancer to treat due to its lack of targetable genetic lesions and unresponsiveness to conventional treatments. Therefore, we investigated genes that had aberrant expression in ATC in relation to differentiation. Using NCBI Gene expression omnibus (GSE33630, GSE85457), which provides publicly available transcriptomic datasets, and bioinformatics software, GEO2R, we found that Homeobox D4 (HOXD4) has 12.5FC higher expression in ATC vs. normal tissue (p=0.000552) and 4.36FC higher in ATC vs. papillary thyroid cancer (PTC) in human patients (p=6.69e-08). HOXD4 is a transcription factor that plays a crucial role in cell differentiation and has been associated with a worse prognosis in cancers such as ovarian serous carcinoma. As loss of differentiation is essential to ATC’s refractory nature and therapeutic resistance, we are exploring modulation of differentiation by all-trans retinoic acid (ATRA). ATRA was used to induce differentiation in ATC cell line T238 therefore abrogating the aggressive phenotype of this cancer. T238 treated with ATRA had a 26% decrease in proliferation at 24 hours, 46% at 48 hours, and 87% at 72 hours measured using a trypan blue exclusion assay. T238 treated with ATRA also had a 44% decrease in migratory capacity at 24 hours and 24% at 48 hours measured by a cell scratch wound assay. Immortalized normal thyroid cell line NTHY-ori-3-1 did not have a significant decrease in either proliferation or migratory capacity when treated with ATRA. T238 cells treated with ATRA for 24 hours had a 30% decrease in HOXD4 protein levels. Lastly, immunohistochemistry staining of HOXD4 in human patient tissue samples confirmed the over-expression of HOXD4 in ATC (19.5% staining) vs. normal tissue (2.5%) and PTC (7.5%). Overall, we hypothesize that the aberrant expression of HOXD4 in ATC plays a crucial role in the aggressive metastatic phenotype and therefore differentiation agents such as ATRA can target this biomarker to modulate the ATC phenotype. Therefore, HOXD4 can be used as a potential prognostic indicator for early detection of ATC. Future studies will involve CRISPRi based knockdown of HOXD4 in vitro and validation of these results in animal in vivo models.

Citation Format: Kaci Kopec, Tara Jarboe, Nicole R. DeSouza, Humayun K. Islam, Jan Geliebter, Raj K. Tiwari. Differentiation driver gene HOXD4 as a potential prognostic indicator and therapeutic target in anaplastic thyroid cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4633.
Abstract 4636: Genome-wide drug anchor screens identify CAAP1 and AKAP17A as regulators of PRMT5 inhibitor sensitivity
Authors: Yoda Satoshi , Tonini Matthew R. , Nicolson Hilary E. , Poleretzky Samuel , Fenoglio Silvia , Lombardo Steven , Grove Lauren , Meier Samuel R. , Choi Ashley , Yu Yi , Cottrell Kevin M. , Maxwell John P. , Teng  , Andersen Jannik N. , Briggs Kimberly J.
Published: March 2024

MTA-cooperative PRMT5 inhibitors are an emerging treatment option for patients with one of the 10-15% of all human cancers harboring MTAP homozygous deletion. To identify potential regulators of sensitivity to PRMT5 inhibitors, we performed genome-wide CRISPR knockout screens in the presence and absence of an MTA-cooperative PRMT5 inhibitor. Knockout of CAAP1 and AKAP17A were among the strongest sensitizing hits across multiple MTAP-deleted cancer cell lines representing different histologies. Strikingly, the CAAP1 gene co-localizes with MTAP and CDKN2A on chromosome 9p21. Co-deletion of CAAP1 is reported in 20 percent of MTAP-deleted cancers in the TCGA PanCancer Atlas. CAAP1 or AKAP17A knockout in MTAP-deleted cancer cell lines sensitized the cells to PRMT5 inhibitors including the clinical stage MTA-cooperative inhibitors, TNG908 and TNG462, and the non-MTA-cooperative inhibitor, GSK3326595. Moreover, we discovered that CAAP1 and AKAP17A protein levels are interdependent, as knockout of either gene caused decreased protein levels for the other. Consistent with this finding, CAAP1 reconstitution in CAAP1-deleted cell lines led to increased AKAP17A levels. Endogenous CAAP1 and AKAP17A protein levels are positively correlated across a panel of cancer cell lines and MTAP-deleted patient-derived xenograft models. Consistent with a previous report (Ni et al., 2023), exogenous CAAP1 and AKAP17A co-immunoprecipitation studies suggest that the proteins form a protein complex. AKAP17A and CAAP1 are not well-characterized proteins, but PRMT5 inhibitors induce global alternative splicing events (ASEs) in cancer cells, and based on preliminary studies a possible function for the CAAP1/AKAP17A complex could be to mitigate ASEs induced by PRMT5 inhibition. Collectively, these data indicate that CAAP1 and AKAP17A exist interdependently and mediate sensitivity to PRMT5 inhibitors. The colocalization and 20 percent incidence of CAAP1 deletion in the setting of MTAP deletion may suggest that such patients will have improved responses to PRMT5-targeted therapy. S. Yoda and M. R. Tonini contributed equally.

Citation Format: Satoshi Yoda, Matthew R. Tonini, Hilary E. Nicolson, Samuel Poleretzky, Silvia Fenoglio, Steven Lombardo, Lauren Grove, Samuel R. Meier, Ashley Choi, Yi Yu, Kevin M. Cottrell, John P. Maxwell, Teng, Jannik N. Andersen, Kimberly J. Briggs. Genome-wide drug anchor screens identify CAAP1 and AKAP17A as regulators of PRMT5 inhibitor sensitivity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4636.
Abstract 471: Attenuation of circulating miR-17-5p, miR-20b-5p, and miR-106a-5p in low-risk prostate cancer patients following proton therapy
Authors: Velasquez Johnny , Bryant Curtis , Mendenhall Nancy , Ricks-Santi Luisel
Published: March 2024

Introduction: Liquid biopsies offer a novel approach to diagnose, predict cancer outcomes, and monitor treatment response by analyzing tumor-derived substances. Circulating cell-free microRNAs (miRNAs) show clinical promise as biomarkers within prostate cancer (PCa) and have been implicated with PCa prognosis and outcomes. However, their potential for monitoring and assessing radiotherapy (RT) treatment response remains understudied. Understanding miRNA expression changes during RT is crucial, as they impact RT efficacy by directly affecting genes and proteins within RT-response pathways.

Aim: This study aimed to profile the expression of three oncogenes, miR-17-5p, miR-20b-5p, and miR-106a-5p and to determine their association with PCa risk groups in patients receiving proton therapy (PT).

Patients and Methods: Plasma was obtained from the University of Florida Health Proton Therapy Institute Outcomes Tracking Protocol biobank. Plasma was collected from men diagnosed with localized PCa without metastatic disease at three time points: prior to PT (PRE-PT), at 4 weeks following the start of PT (4WKSPD1), and at 8 weeks following the start of PT (8WKSPD1). The pilot study consisted of 38 patients, with 23 categorized as low-risk and 15 categorized as high-risk for PCa. MiRNA expression levels were evaluated using reverse transcription-polymerase chain reaction and quantified using the Livak method.

Results: The study revealed a downregulation of miR-17-5p in response to PT, with median gene expression levels decreasing from 0.73 at PRE-PT to 0.587 at 8WKSPD1. Low-risk PCa patients also showed distinct miRNA expression changes with all three miRNAs (miR-17-5p, miR-20b-5p, and miR-106a-5p) downregulated from 0.953, 1.037, and 1.010 at PRE-PT to 0.398, 0.216, and 0.298 at 8WKSPD1, respectively. Statistical analysis using a Kruskal-Wallis test followed by a Dunn's post-hoc test identified significant differences (p=0.035) specifically between PRE-PT and 8WKSPD1 for miR-17-5p.

Conclusion: The study revealed that PT led to a time-dependent reduction in miRNA levels, indicating a higher responsiveness in low-risk compared to high-risk PCa patients. This differential response highlights miR-17-5p, miR-20b-5p, and miR-106a-5p as potential PT response biomarkers, with clinical implications for personalizing PCa treatment. These results underscore the necessity for further validation of these miRNAs to maximize their clinical utility.

Citation Format: Johnny Velasquez, Curtis Bryant, Nancy Mendenhall, Luisel Ricks-Santi. Attenuation of circulating miR-17-5p, miR-20b-5p, and miR-106a-5p in low-risk prostate cancer patients following proton therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 471.
Abstract 4715: Azathioprine and Z36 induced cell death in human colorectal cancer HCT-116 and HCT-15 cells
Authors: Ali Yousuf Tahir , Fatima Mahwish , Ahmed Muhammad Areeb , Siddiqi Hasan Salman , Hussain Azhar , Rehman Mati Ur
Published: March 2024

Rationale and Objectives: Cancer accounts for a huge global health burden and is the third highest cause of death across the globe. Cancer cells are known to have higher basal reactive oxygen species (ROS) levels, owing to their higher metabolism and proliferation. Higher basal ROS in cancer cells made them susceptible to exogenous ROS stimuli that lead to cell death. Therefore, in the present study, we have analyzed the effects of Azathioprine, a glutathione (GSH) inhibitor and an intracellular ROS producer. The effect of Azathioprine was analyzed alone or in combination with a BCL-2 family protein inhibitor Z36, on human colorectal cancer cell lines HCT-116 and HCT-15.

Methods: The effects of the drugs have been investigated using various methods including cell viability assay, DNA fragmentation assay, and Annexin V-FITC/PI apoptosis assay. In addition, morphological changes evident in cell death were detected by Giemsa staining. Combination dosage for drug synergistic effects was computed through the use of CompuSyn software. Western blot was performed to detect changes in the expression of cell death-related proteins following drug treatments.

Results and conclusion: Azathioprine at a dose of 5 µM has been demonstrated to induce cell death in both HCT116 and HCT15 cells in a concentration and time-dependent manner. On the contrary, Z36, a novel BCLXL inhibitor, has demonstrated similar cell viability results at an even lower dose of 2.5 µM. When both drugs were combined at lower doses, no significant effects were observed on the cell viability of either cell line, demonstrating that the combined treatment has no synergistic or additive effects. This suggests that both drugs do not augment each other and might induce cell death via upregulation of ROS using a similar mechanism.

Citation Format: Yousuf Tahir Ali, Mahwish Fatima, Muhammad Areeb Ahmed, Hasan Salman Siddiqi, Azhar Hussain, Mati Ur Rehman. Azathioprine and Z36 induced cell death in human colorectal cancer HCT-116 and HCT-15 cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4715.
Abstract 4719: Modulation of HR-HPV viral protein E7 by SHetA2 in cervical cancer
Authors: Garland Justin , Hussain Showket , Benbrook Doris M.
Published: March 2024

Introduction: The purpose of this study is to determine the effect of the chemotherapeutic sulfur heteroarotinoid A2 (SHetA2) on high-risk human papillomavirus (HR-HPV) proteins and the modulation of cell survival and proliferation in cervical cancer. Cervical cancer is caused by persistent HR-HPV infection. HR-HPV tumorigenesis is driven by its early 6 and 7 (E6, E7) oncoproteins, which increase cell proliferation and survival. Cellular pathways regulated by E6 and E7 are counter-regulated by the investigational new drug SHetA2, which is currently in phase 1 clinical trial for advanced and recurrent cancers. SHetA2 disrupts complexes of 70-kDa heat shock protein (HSP70) family members, hsc70, GRP78, and mortalin, with their client proteins. We hypothesized that SHetA2 reduces E6 and E7 levels and their binding to HSP70 chaperones in association with the reduction of cervical cancer cells and tumor growth.

Methods: SHetA2 effects on specific genes and proteins in HR-HPV-positive cervical cancer cell lines and xenograft tumors were assessed by western blot, quantitative polymerase chain reaction, immunofluorescence ligation assays, and coimmunoprecipitation assays.

Results: SHetA2 significantly reduced E6 and E7 mRNA, and protein levels of E7 and the E7-regulated p16, but not E6 in cervical cancer cells and xenograft tumors. MG132 inhibition of proteasome function attenuated the E7 reduction. HSP70/hsc70 proteins bound E7 and SHetA2 disrupted these complexes.

Conclusions/Implications: SHetA2 reduces E7 proteins in cervical cancer cells through a mechanism that involves proteasomal degradation in association with disruption of HSP70/E7 complexes, p16 reduction, and decreased cell and tumor growth. This is the first demonstration of HSP70/E7 complexes. SHetA2-induced release of E7 from HSP70s could be causing increased susceptibility of E7 to proteasomal degradation. This data supports the development of SHetA2 and HSP70 inhibitors for cervical cancer.

Citation Format: Justin Garland, Showket Hussain, Doris M. Benbrook. Modulation of HR-HPV viral protein E7 by SHetA2 in cervical cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4719.
Abstract 4746: PIM kinase inhibition synergizes with a MERTK inhibitor to treat osimertinib resistant EGFR-mutant non-small cell lung cancer
Authors: Yan Dan , Wang Xiaodong , Frye Stephen V. , Earp H. Shelton , DeRyckere Deborah , Graham Douglas K.
Published: March 2024

There is an unmet need for effective treatment in the setting of acquired osimertinib resistance in EGFR-mutant non-small cell lung cancer (NSCLC) patients and MERTK tyrosine kinase is a potential therapeutic target in this context. Treatment with the MERTK inhibitor MRX-2843 was sufficient to abrogate downstream PI3K-AKT and MAPK-ERK signaling, cell expansion, and colony formation in osimertinib-resistant EGFR-mutant NSCLC cell lines, suggesting dependence on MERTK signaling for tumor cell proliferation/survival. Further, two structurally distinct PIM kinase inhibitors, SGI-1776 and PIM447, provided synergistic inhibition of cell expansion and colony formation in osimertinib-resistant cell line cultures when combined with MRX-2843 treatment. Mechanistically, treatment with MRX-2843 in combination with either PIM kinase inhibitor decreased downstream PI3K-AKT and MAPK-ERK signaling more effectively than single agents. Furthermore, knockdown of PIM kinases (PIM1, 2, 3) using nine sets of siRNAs led to various changes in expression of TAM receptor family members (TYRO3, AXL, MERTK), indicating a role for PIM kinases in regulation of TAM kinase expression. These studies suggest a novel treatment strategy for osimertinib-resistant EGFR-mutant NSCLC using a first-in-class MERTK kinase inhibitor that is currently under evaluation in multiple Phase I clinical trials.

Citation Format: Dan Yan, Xiaodong Wang, Stephen V. Frye, H. Shelton Earp, Deborah DeRyckere, Douglas K. Graham. PIM kinase inhibition synergizes with a MERTK inhibitor to treat osimertinib resistant EGFR-mutant non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4746.
Abstract 4748: Aronia berry overcomes gemcitabine resistance by regulating the MYD88/NF-kB/P-glycoprotein pathway in pancreatic ductal adenocarcinoma
Authors: Li Yuan , Xu Caiming , Han Haiyong , Goel Ajay
Published: March 2024

Background: Pancreatic ductal adenocarcinoma (PDAC) is lethal, which is partly due to the limited efficacy of Gemcitabine (Gem)-based chemotherapy, resulting from the development of acquired resistance to this regimen. Aronia berry extract (ABE) is a natural source of phenolic compounds known for its anti-cancer properties. Previous research has highlighted the ability of ABE to help overcome chemoresistance to conventional chemotherapies in various cancers. In this study, we interrogated the ability of ABE to overcome Gem-resistance (Gem-R) in PDAC and identify specific pathway(s) responsible for this anti-cancer efficacy. Methods: We performed a series of in-vitro experiments in Gem-R cells to evaluate the synergistic effect of combined treatment with Gem and ABE. Furthermore, we conducted a genomewide transcriptomic analysis to identify the critical regulatory pathways associated with chemoresistance and potential therapeutic targets of ABE in Gem-R PDAC cells. The cell culture findings were validated in patient-derived 3D organoids. Results: The combined treatment with ABE and Gem in Gem-R cells exhibited significant synergistic anticancer effects on cell viability, proliferation, migration, and invasion. The transcriptomic analysis revealed that the NF-κB signaling pathway was associated with Gem-R (p<0.05), with a significant upregulation of MYD88. Additionally, MYD88 was significantly associated with overall survival in PDAC patients in the TCGA cohort (HR=1.58, P=0.045). The MYD88/NF-κB signaling is associated with chemoresistance as it can upregulate the efflux transporters like P-glycoprotein (P-gp). Our results confirmed that combined treatment inhibited the NF-κB signaling pathway by suppressing MYD88 and downregulating P-gp expression to overcome Gem-R. Finally, the combined treatment demonstrated superior anti-cancer activity in 3D organoids by reducing both their number and size (P<0.05). Conclusion: We provide novel insights into the potential of ABE in overcoming Gem-R in PDAC cells by targeting the MYD88/NF-κb/P-gp axis, which offers a safe and inexpensive approach for improving therapeutic outcomes in this fatal malignancy.

Citation Format: Yuan Li, Caiming Xu, Haiyong Han, Ajay Goel. Aronia berry overcomes gemcitabine resistance by regulating the MYD88/NF-kB/P-glycoprotein pathway in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4748.
Abstract 4765: Tumor, serum and plasma metabolome in patients with high-grade bladder cancer
Authors: Tsamouri Maria-Malvina , Karthikeyan Akshaya , Lucchesi Christopher A. , Durbin-Johnson Blythe P. , Dall'Era Marc A. , Ghosh Paramita M.
Published: March 2024

Introduction: Bladder cancer (BlCa) ranks as the sixth most prevalent cancer in the world. A number of urine based BlCa detection tests have recently been approved for clinical use, including the NMP22 tests and the bladder tumor antigen (BTA) tests for both detection and monitoring of patients with known bladder cancer. The FDA have approved them both for surveillance in BlCa only in conjunction with cystoscopy. Unfortunately, the sensitivity and specificity of these tests are poor, and cannot distinguish between aggressive and non-aggressive tumors. This project aims to identify alternate markers that can overcome these drawbacks.

Methods: We characterized the global metabolome of 47 bladder tumors retrieved by radical cystectomy (RC) or TURBT, including 14 specimens with matching plasma and serum samples. All specimens were subjected to untargeted metabolomic approaches with gas chromatography- time-of-flight mass spectrometry (GC-TOF MS) to profile primary metabolites, hydrophilic interaction liquid chromatography MS (HILIC-QTOF MS) to profile biogenic amines, and liquid chromatography charged surface hybrid MS (LC-CSH-QTOF MS) to characterize complex lipids. Differentially expressed metabolites were analyzed with respect to grade, stage, metastases, and the use of neoadjuvant chemotherapy. For selected metabolites, area under the ROC curve (AUC) is calculated as % sensitivity vs. 100% specificity. Expression of highly discriminatory metabolites were then validated in the plasma and serum samples to determine which forum would provide greater specificity.

Results: We first assembled a discovery cohort of 33 patients who did not have matching blood specimen for primary metabolites, biogenic amines and complex lipids. Comparison of high vs low grade tumors yielded two lipids, a sphingomyelin (SM (d38) and a phosphatidylcholine PC(p-38), that were significantly different (adjusted p-value<0.05, log2fold change>2.5). These results were validated in the 14-sample cohort with matching plasma and serum of which the latter showed significant correlation between tumor and plasma (Spearman R=0.872, adj p=0.03).

Conclusions: Our results indicate that certain sphingomyelins and phosphatidylcholines may be useful as plasma markers of high grade BlCa. Further analyses are required to evaluate whether these markers are more discriminatory compared to existing urinary markers.

Citation Format: Maria-Malvina Tsamouri, Akshaya Karthikeyan, Christopher A. Lucchesi, Blythe P. Durbin-Johnson, Marc A. Dall'Era, Paramita M. Ghosh. Tumor, serum and plasma metabolome in patients with high-grade bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4765.
Abstract 4790: Hospitalization as an opportunity to improve lung cancer screening in high-risk patients
Authors: Nielsen Ellen M. , Zhang Jingwen , Marsden Justin , Bays Chloe , Moran William P. , Mauldin Patrick D. , Lenert Leslie A. , Toll Benjamin A. , Schreiner Andrew D. , Heincelman Marc
Published: March 2024

Background: Lung cancer screening with annual low-dose computed tomography (LDCT) in high-risk patients with exposure to smoking reduces lung cancer-related mortality, yet the screening rate of eligible adults is low. As hospitalization is a critical moment to engage patients in their overall health, it may be an opportunity to improve rates of lung cancer screening. Prior to implementing a hospital-based lung cancer screening referral program, this study assesses the association between hospitalization and completion of lung cancer screening.

Methods: A retrospective cohort study of evaluated completion of at least one LDCT from 2014-2021 using electronic health record data using hospitalization as the primary exposure. Patients aged 55-80 who received care from a university-based internal medicine clinic and reported cigarette use were included. Univariate analysis and logistic regression models evaluated the association of hospitalization and completion of LDCT. The secondary outcome was completion of any CT of the chest.

Results: Of the 1,935 current smokers identified, 47% had at least one hospitalization, and 21% completed a LDCT during the study period. While a higher proportion of patients with a hospitalization had a LDCT (24%) compared to patients without a hospitalization (18%, p<0.001), there was no association between hospitalization and completion of a LDCT after adjusting for potentially confounding covariates (95%CI 0.680 - 1.149). Of cohort patients, 38% completed any CT of the chest and hospitalization was associated with increased odds of receiving chest CT imaging in the adjusted model (OR 1.72; 95%CI 1.37 - 2.17).

Conclusions: In a cohort of patients at risk for lung cancer, only 1 in 5 completed lung cancer screening with LDCT and hospitalization events were not associated with LDCT completion. A hospitalist driven lung cancer screening program has the potential to increase the suboptimal rates of lung cancer screening in high-risk patients.

Citation Format: Ellen M. Nielsen, Jingwen Zhang, Justin Marsden, Chloe Bays, William P. Moran, Patrick D. Mauldin, Leslie A. Lenert, Benjamin A. Toll, Andrew D. Schreiner, Marc Heincelman. Hospitalization as an opportunity to improve lung cancer screening in high-risk patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4790.
Abstract 4840: Creating a culturally sensitive manual to increase clinical trial participation among African Americans and African Nationals
Authors: Aristizabal Carolina , Ibarra Eduardo , Baezconde-Garbanati Lourdes A.
Published: March 2024

Background: Racial and ethnic minorities are immensely underrepresented in clinical trial studies in the United States. Black/African Americans, Latinos/as, Asians, American Indians, and Pacific Islanders make up roughly 30% of all clinical trials (CT) participation across all race groups. Being inclusive of all populations in CT ensures that everyone has access to modern medicines and treatments, but also, that these latest treatments work and react well to all people.

Methods: We developed a manual on clinical trials with a target focus to African Americans and African Nationals. We hosted two focus groups with Blacks/African Americans in which we found they are more comfortable getting cancer related information through trusted community services, such as coming from churches. We have connected with over 80 Pastors who agreed to collaborate in clinical trials education.

Results: Through outreach and engagement to African American churches in Los Angeles County, we have distributed a copy of this clinical trials educational manual to 90 churches to be disseminated during their services. Our office of Community Outreach and Engagement (COE) regularly attend community health and resource events and provide hard copies of the clinical trials manual at tables/booths designated for our participation. We have attended 50 community events and have disseminated about 1,900 copies of the CT manual so far this year 2023. This manual is also available for free download on our cancer center’s website, in which over 100 toolkits have been downloaded within the last year.

Conclusions: By creating a culturally sensitive and adapted educational manual and partnering with trusted services in the community such as faith-based organizations, we can begin to break down barriers to clinical trials participation among all race and ethnic minority groups.

Citation Format: Carolina Aristizabal, Eduardo Ibarra, Lourdes A. Baezconde-Garbanati. Creating a culturally sensitive manual to increase clinical trial participation among African Americans and African Nationals [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4840.
Abstract 485: Cancer killing effect of ultrasound-responsive mechanophore nanoparticles
Authors: Wang Jian , Zhao Shensheng , Yi Junxi , Sun Yunyan , Agrawal Megha , Li King , Moore Jeffrey S. , Oelze Michael , Chen Yun-Sheng
Published: March 2024

Radiotherapy, a mainstay in cancer treatment, often inadvertently damages healthy tissues while targeting malignancies with free radicals. As an alternative, Photodynamic Therapy (PDT) employs non-ionizing photons to generate reactive oxygen species (ROS) in tumors, offering precise targeting with minimal long-term effects. However, its efficacy is constrained to superficial tumors due to the limited penetration of light. Sonodynamic Therapy (SDT), an evolution of PDT, uses ultrasound to enhance tissue penetration while maintaining its non-invasive nature. Initial SDT studies suggest that ultrasound can activate PDT's photosensitizers to produce ROS, offering a potential pathway for tumor eradication. Nevertheless, the principal mechanisms of ROS generation in SDT, namely pyrolysis and sonoluminescence, raise concerns about potential tissue injury. In our research, we introduce an innovative approach employing mechanophores into nanoparticles. Mechanophores, molecular units within polymers sensitive to mechanical stress, undergo transformations under such conditions, including the release of small molecules or free radicals. We have developed injectable nano-sized mechanophore nanoparticles, designed to respond to high-frequency ultrasound (>100 kHz) more efficiently and with reduced energy requirements. This study focuses on nano-engineering the echogenicity of these mechanophore nanoparticles to enhance ROS generation. When agitated with focused ultrasound (FUS), these nanoparticles successfully induced ROS, leading to significant cell death, a result not observed with FUS alone in the absence of nanoparticles. In vivo experiments using mouse orthotopic cancer models demonstrated these nanoparticles' ability to inhibit tumor growth effectively. Under biocompatible ultrasound conditions, the nanoparticles cause effective cancer cell eradication with minimal impact on surrounding healthy tissues. Our findings not only introduce a novel approach to cancer treatment but also pave the way for broader applications of mechanophore nanoparticles in biomedicine. This study underscores the potential of ultrasound-activatable mechanophore nanoparticles as a versatile, effective, and safe option for cancer therapy.

Citation Format: Jian Wang, Shensheng Zhao, Junxi Yi, Yunyan Sun, Megha Agrawal, King Li, Jeffrey S. Moore, Michael Oelze, Yun-Sheng Chen. Cancer killing effect of ultrasound-responsive mechanophore nanoparticles [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 485.