bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4457: Skeletal muscle-infiltrating CD8 T cells exacerbate muscle atrophy in a syngeneic lung cancer cachexia mouse model
Authors: Park Se-Young , Kim Bomi , Lee Myunggyo , Lee Haeseung , Song Na-Young
Published: March 2024

Cancer-associated cachexia (CAC) is a devastating syndrome of irreversible body weight loss and muscle wasting. CAC has been reported to often lead to poor outcomes of immune checkpoint inhibitors (ICIs), emerging as a standard treatment for various types of cancer. Previously, we have demonstrated that KRAS-mutant lung tumor tissues display an immunosuppressive microenvironment with decreased CD8 T cells and increased regulatory T cells (Tregs). In this orthotopic lung cancer mouse model, treatment with an anti-PD-L1 antibody, a representative ICI, reduced tumor burden, whereas accelerated loss of body weight and skeletal muscle mass. We thus assumed that CD8 T cells could facilitate skeletal muscle wasting in lung tumor-bearing mice, a key feature of CAC. First, we established a syngeneic CAC mouse model through orthotopic transplantation of murine lung cancer cells. At 7 weeks post-injection, these mice developed CAC, displaying over 20 % body weight loss, almost total loss of epididymal fats, and profound skeletal muscle wasting. In line with our assumption, CAC mice showed an increased number of CD8 T cells but a reduced number of Tregs in skeletal muscles, compared to tumor-free control mice. We then investigated whether skeletal muscle-infiltrating CD8 T cells directly induce muscle atrophy indeed. In CAC mice, CD8 neutralization led to restoration of muscle mass and downregulation of muscle atrophy markers, but was not able to significantly affect lung tumor burden. Moreover, direct co-culture with splenic CD8 T cells exacerbated C2C12 myotube atrophy in a conditioned medium from lung cancer cells, further supporting the involvement of CD8 T cells in CAC. To elucidate its underlying mechanism, RNA sequencing analysis was conducted in both skeletal muscles and lung tumors of CAC mice. Based on the differently expressed gene analysis, Cxcl13 was upregulated in skeletal muscles of CAC mice, while downregulated in lung tumors. It is well documented that CXCL13 can mediate recruitment of CD8 T cells. In this regard, CXCL13 might be a plausible regulator in CD8 T cell-induced skeletal muscle atrophy, which requires further investigation to fully understand the molecular mechanism.

Citation Format: Se-Young Park, Bomi Kim, Myunggyo Lee, Haeseung Lee, Na-Young Song. Skeletal muscle-infiltrating CD8 T cells exacerbate muscle atrophy in a syngeneic lung cancer cachexia mouse model [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 4457.
Abstract 4461: Unveiling the links between methionine metabolism and epigenomic reprogramming in upper aerodigestive squamous cell carcinoma
Authors: Nam Chehyun
Published: March 2024

Upper aerodigestive squamous cell carcinoma (UASCC) presents as a prevalent and aggressive malignancy, posing challenges in terms of effective therapeutic interventions. In this study, we explored the intricacies of amino acid metabolism within UASCC, revealing an unexpected observation that distinguishes UASCC among all human cancers, having the highest methionine levels, driven by the overexpression of its transporter LAT1. Notably, LAT1 exhibits peak expression levels in UASCC, regulated at the transcriptional level by UASCC-specific promoters and enhancers, co-regulated by SCC master regulators TP63/KLF5/SREBF1.Surprisingly, our unbiased bioinformatic screen identifies EZH2 as a pivotal downstream target of the LAT1-methionine pathway, establishing a direct link between methionine metabolism and epigenomic reprogramming. This cascade emerges as crucial for the survival and proliferation of UASCC patient-derived tumor organoids. Furthermore, we find that LAT1 expression closely correlates with cellular sensitivity to inhibition of the LAT1-methionine-EZH2 axis.Significantly, the newly discovered LAT1-methionine-EZH2 cascade emerges as a promising target for intervention, with effective outcomes achievable through both pharmacological approaches and dietary interventions in vivo. In summary, this research unveils a novel mechanistic crosstalk connecting epigenomic reprogramming with methionine metabolism, revealing its biological significance in UASCC. Moreover, it identifies a unique tumor-specific vulnerability, providing avenues for exploitation through both pharmacological and dietary strategies

Citation Format: Chehyun Nam. Unveiling the links between methionine metabolism and epigenomic reprogramming in upper aerodigestive squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4461.
Abstract 4476: Novel auristatins with improved tolerability and unique bystander activity profile
Authors: Moquist Philip N. , Eng-Duncan Nicole M. , Bovee Tim , Snead Katie , Wright Jenn , Smith Kaleb , Blevins Teri , Blackburn Brittney , Ulrich Michelle , Magunda Forgivemore , Simmons Jessica , Senter Peter D. , Doronina Svetlana O.
Published: March 2024

Auristatins are a class of clinically validated payloads used extensively in antibody-drug conjugate (ADC) technology. There are currently 4 FDA approved ADCs utilizing the vedotin drug linker to treat an array of solid tumors and lymphomas and several more in clinical trials. Auristatin payloads have several desirable properties, including strong anticancer cytotoxicity, bystander activity, and induction of immunologic cell death. While auristatin-based ADCs have been remarkably successful, there remain opportunities to further understand and improve payload properties. Bystander activity is one such property that can greatly impact the balance between efficacy and tolerability. Herein, we present a medicinal chemistry investigation into a series of auristatins with similar tubulin binding and biochemical potency that differ in lipophilicity and ability to cross the cell membrane. This design enabled us to focus on and characterize the impact of bystander activity while maintaining all other parameters. Ultimately, this effort resulted in ADCs with similar on-target in vitro cytotoxicity but differed in the extent of off-target bystander activity. Evaluation of in vivo efficacy, in vivo bystander activity, and rodent tolerability afforded a lead drug linker that was well-tolerated as an ADC and exhibited strong antitumor growth delay.

Citation Format: Philip N. Moquist, Nicole M. Eng-Duncan, Tim Bovee, Katie Snead, Jenn Wright, Kaleb Smith, Teri Blevins, Brittney Blackburn, Michelle Ulrich, Forgivemore Magunda, Jessica Simmons, Peter D. Senter, Svetlana O. Doronina. Novel auristatins with improved tolerability and unique bystander activity profile [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 4476.
Abstract 4509: Discovery of potent and highly efficacious STAT3 PROTAC degraders capable of achieving long-lasting tumor regression
Authors: Acharyya Ranjan K. , Bai Longchuan , Zhao Haibin , Wu Dimin , Hoda Metwally , Donna McEachern , Bo Wen , Sun Duxin , Wang Shaomeng
Published: March 2024

STAT3 (signal transducer and activator of transcription 3) is a transcription factor and a promising therapeutic targets for cancer and other human diseases. We have previously reported the discovery of SD-36 and SD-91 as potent, selective and highly efficacious STAT3 degraders. In the present study, we report the discovery of highly potent, selective and efficacious new STAT3 degraders. These STAT3 degraders were designed using our high-affinity STAT3 ligands and high-affinity VHL-1 ligands, with UM-STAT3-3100 being the best. In direct comparison, UM-STAT3-3100 is >10-times more potent than SD-36 and SD-91 in inducing STAT3 degradation in cells and are highly selective over other STAT members in vitro and in vivo. A single intravenous administration of UM-STAT3-3100 resulted in complete and long-lasting depletion of STAT3 protein for >48 hrs in tumor and native tissues. Weekly administration of UM-STAT3-3100 at 10 mg/kg achieved complete and long-lasting tumor regression in animal models of human cancer without any signs of toxicity. UM-STAT3-3100 is a promising compound for extensive evaluation for the treatment of human cancers and other human diseases in which STAT3 plays a key role.

Citation Format: Ranjan K. Acharyya, Longchuan Bai, Haibin Zhao, Dimin Wu, Metwally Hoda, McEachern Donna, Wen Bo, Duxin Sun, Shaomeng Wang. Discovery of potent and highly efficacious STAT3 PROTAC degraders capable of achieving long-lasting tumor regression [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 4509.
Abstract 4540: SY-7021, a novel DNA-PK inhibitor, exhibits significant anti-tumor activity in vitro and in vivo
Authors: Zhang Kai , Liu Zhihua , Gao Yuhao , Lu Chang , Cheng Shikang , Liu Xijie , Luo Hong , Sun Yinghui
Published: March 2024

Cancer therapies such as ionizing radiation or topoisomerase II inhibitors (for example, doxorubicin) induce DNA double-strand breaks (DSBs), which can subsequently be repaired by homologous recombination or non-homologous end joining (NHEJ). DNA-dependent protein kinase (DNA-PK), a nuclear serine/threonine protein kinase complex, is a pivotal component of the NHEJ process that maintain genome integrity. The critical role of DNA-PK in DNA damage response (DDR) and the dysregulated DNA-PK expression in cancer make it an intriguing therapeutic target for cancer, especially when combined with DSBs agents. Here, we identified a highly potent and selective DNA-PK inhibitor, SY-7021, with an IC50 value of 0.242 nM on DNA-PK kinase and high selectivity on ATM and ATR (greater than 400-fold) in biochemical assays. In a reporter assay for NHEJ repair, SY-7021 was able to dose-dependently reduce cellular NHEJ efficiency. Functionally, SY-7021 inhibited cell proliferation in multiple cancer cell lines alone or combined with doxorubicin. Basically, combination of SY-7021 and doxorubicin induced significant G2/M phase arrest and cell apoptosis, as well as enhanced the phosphorylation on Ser139 of γH2AX, Tyrosine 68 of CHK2 and Serine15 of p53 in MDA-MB-468 cells. In a subcutaneous NCI-H1703 xenograft tumor model, SY-7021 administered orally twice daily achieved dose-dependent tumor growth inhibition in vivo, with a tumor growth inhibition (TGI) of 105.6% at 60 mg/kg and no significant weight loss was observed. In addition, SY-7021 demonstrates good PK properties and acceptable safety profiles in in vivo studies. Collectively, SY-7021, a potent and selective DNA-PK inhibitor, shows significant inhibition on tumor growth in in vitro and in vivo studies, providing a rationale treatment for multiple tumors in monotherapy or in combination with other agents.

Citation Format: Kai Zhang, Zhihua Liu, Yuhao Gao, Chang Lu, Shikang Cheng, Xijie Liu, Hong Luo, Yinghui Sun. SY-7021, a novel DNA-PK inhibitor, exhibits significant anti-tumor activity in vitro and in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4540.
Abstract 456: A first comprehensive analysis of transcribed ultra conserved regions uncovers important regulatory functions of novel non-coding transcripts in gliomas
Authors: Gibert Myron K. , Abounader Roger
Published: March 2024

Transcribed Ultra-Conserved Regions (TUCRs) represent a severely understudied class of putative non-coding RNAs (ncRNAs) that are 100% conserved across multiple species. We performed the first-ever analysis of TUCRs in glioblastoma (GBM) and low-grade gliomas (LGG). We leveraged large human datasets to identify the genomic locations, chromatin accessibility, transcription, differential expression, correlation with survival, and predicted functions of all 481 TUCRs, and identified TUCRs that are relevant to glioma biology. Of these, we investigated the expression, function, and mechanism of action of the most highly upregulated intergenic TUCR, uc.110, identifying it as a new oncogene. Uc.110 was highly overexpressed in GBM and LGG, where it promoted malignancy and tumor growth. Uc.110 activated the WNT pathway by upregulating the expression of membrane frizzled-related protein, by sponging the tumor suppressor miR-544. This pioneering study shows important roles for TUCRs in gliomas and provides an extensive database and novel methods for future TUCR research.

Citation Format: Myron K. Gibert, Roger Abounader. A first comprehensive analysis of transcribed ultra conserved regions uncovers important regulatory functions of novel non-coding transcripts in gliomas [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 456.
Abstract 4568: Rational targeting of BCL-xL and/or mTOR enhance the efficacy of KRASG12D inhibitor in pancreatic cancer
Authors: Khan Sajid , Cao Lin , Vudhata Vignesh , Yang Yang , Zhang Peiyi , Zheng Guangrong , Trevino Jose G. , Zhou Daohong
Published: March 2024

Background: Pancreatic cancer (PaCa) is a highly aggressive malignancy with a very low five-year survival rate secondary to late diagnosis, limited treatment options and resistance to existing therapies. While the emergence of mutant-selective KRASG12D (G12D) inhibitors brings promise for treating PaCa patients carrying these mutations, their effectiveness as single agents is expected to be limited as evident from the clinical efficacy of KRASG12C inhibitors.

Experimental Design: First, multiple G12D-mutated PaCa cell lines were treated with a G12D inhibitor (MRTX1133) and subjected to immunoblot analysis for the expression of KRAS effectors and BCL-2 family proteins. Next, a proteolysis-targeting chimera (PROTAC)-based platelet-sparing BCL-xL degrader (DT2216) and/or an mTOR inhibitor (everolimus) were evaluated in combination with MRTX1133 for viability, clonogenicity and apoptosis. Subsequently, mechanistic investigations were carried out using immunoblotting, senescence-associated β-galactosidase staining and qPCR. Finally, therapeutic efficacy and safety of the combinations were evaluated in cell-derived and patient-derived xenograft models.

Results: MRTX1133 treatment creates apoptotic vulnerabilities by inducing aberrations in BCL-2 family proteins, with elevated BIM levels and decreased NOXA levels, in G12D-mutated PaCa cells. Furthermore, DT2216 and everolimus were shown to release BIM and stabilize NOXA, respectively, and their combination with MRTX1133 synergistically killed G12D-mutated PaCa cells in vitro and caused significant tumor growth inhibition or regression compared to single agents in vivo. Additionally, MRTX1133 treatment led to induction of senescence in a fraction of PaCa cells. These senescent cells were found to be dependent on BCL-xL and MCL-1 for their survival and were effectively killed with a combination of DT2216 and everolimus through BCL-xL degradation and NOXA-mediated MCL-1 inhibition, respectively.

Conclusions: Our findings suggest that the single agent efficacy of MRTX1133 is limited due to apoptosis inhibition and complimentary senescence induction, and therefore its combination with DT2216 and/or everolimus is synergistic, which can potentially overcome resistance, by enhanced apoptosis and clearance of senescent cells.

Citation Format: Sajid Khan, Lin Cao, Vignesh Vudhata, Yang Yang, Peiyi Zhang, Guangrong Zheng, Jose G. Trevino, Daohong Zhou. Rational targeting of BCL-xL and/or mTOR enhance the efficacy of KRASG12D inhibitor in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4568.
Abstract 4581: Identifying class 1 HDACs as a vulnerability for aggressive uveal melanoma
Authors: Yenisehirli Gulum , Zuniga Ashley , Rodriguez Sara , Adis Emily V. , Quintana Gabriela , Borges Sebastian , Lopez Sofia , Lopez Kassandra , Braun Steffanie , Volmar Claude H. , Harbour J. W. , Kurtenbach Stefan
Published: March 2024

Uveal melanoma (UM) is the most common primary intraocular cancer in adults, with a strong propensity to metastasize, primarily to the liver. Metastatic UM is notoriously unresponsive to existing therapies, rendering it almost uniformly lethal. Virtually all UM tumors have activating mutations in the Gq signaling pathway, predominantly in GNAQ/11, and around half of tumors have additional loss-of-function mutations in the tumor suppressor BAP1. Mutations in the epigenetic modulator BAP1 are the most significant clinical marker of metastatic risk. The limited efficacy of current treatment options necessitates an urgent exploration of novel therapeutic approaches to improve patient prognosis. Thus, we conducted a large epigenetic compound library screen with the currently most well characterized, focused epigenetic library available, which consisted of 960 potent, cell permeable, medically active, epigenetic-directed small molecule modulators (TargetMol, L1200). We screened BAP1-mutant as well as BAP1-wildtype uveal melanoma cell lines and followed up with comprehensive dose-response and synergy tests. Our lead compounds were highly efficient in reducing viability of UM cells. We find that inhibitors with high specificity for class 1 HDAC inhibition are especially cytotoxic to UM cells. Together, our results reveal a possible mechanism of interfering with UM progression and identify lead compounds to be tested in clinical trials.

Citation Format: Gulum Yenisehirli, Ashley Zuniga, Sara Rodriguez, Emily V. Adis, Gabriela Quintana, Sebastian Borges, Sofia Lopez, Kassandra Lopez, Steffanie Braun, Claude H. Volmar, J. W. Harbour, Stefan Kurtenbach. Identifying class 1 HDACs as a vulnerability for aggressive uveal melanoma [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 4581.
Abstract 4590: Antitumor effects of histone deacetylase inhibitors on breast cancer cell viability and mammosphere formation
Authors: Tehrani Golnaz Asaadi , Kubick Becca , Datta Meenal
Published: March 2024

Histone deacetylases (HDACs) are overexpressed in various types of primary human cancer and have become attractive targets for cancer therapy. This study aimed to evaluate the cytotoxic properties of three HDAC inhibitors (suberoylanilide hydroxamic acid (SAHA, or Vorinostat), Valproic Acid (VPA) and CAY10603) and compare them to Paclitaxel (PTX), a chemotherapy agent used to treat Triple Negative Breast Cancers (TNBCs). In the present study, drug response in the MDA-MB-231, MDA-MB-468, and MCF-10A cell lines was evaluated by cell viability assays. Apoptosis and cell cycle distribution were analyzed by flow cytometry. Mammospheres generated from MDA-MB-231 and MDA-MB-468 breast cancer cells were also subjected to drug treatment; their response was assessed under both normoxic and hypoxic conditions.

We found that SAHA and CAY10603 inhibited breast cancer cell viability without significantly affecting MCF-10A cells. SAHA, CAY10603, and VPA all arrested a significant percentage of MDA-MB-231 cells at G2/M phase after 24h compared to controls. TNBC cells experienced an increase in both early and late apoptosis following treatment with the most effective concentrations of SAHA, CAY10603, and VPA (1.8µM, 2µM, and 0.4mM, respectively). SAHA proved to be the most effective drug for induction of apoptosis compared to the control, followed by CAY10603 (8.26% and 6%, 3.67%). In both TNBCs and mammospheres under hypoxic conditions, SAHA and CAY10603 down regulated HIF-1α expression significantly, but PTX did not. In contrast, VPA and PTX down regulated CTNNB1 gene expression.

This study indicates that epi-drugs exert an anti-tumor effect on TNBCs, suggesting a potential role for HDAC inhibitors as novel therapeutics for breast cancer patients.

Citation Format: Golnaz Asaadi Tehrani, Becca Kubick, Meenal Datta. Antitumor effects of histone deacetylase inhibitors on breast cancer cell viability and mammosphere formation [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 4590.
Abstract 4593: A synergy between MDM2 and AURKB pathways in driving cell cycle and cell survival in lung cancer cells
Authors: Natarajan Umamaheswari , Rathinavelu Appu
Published: March 2024

Lung cancer is the leading cause of cancer-related deaths worldwide and non-small cell lung cancer (NSCLC) is the most prevalent type among lung cancers, contributing to the high mortality seen with this deadly disease. Despite aggressive treatments, the 5-year survival rate for lung cancer is only 23.7% in the United States. MDM2 and AURKB are two cancer-driving genes that are highly expressed in a broad range of cancer types including lung cancers. Furthermore, MDM2 overexpression can positively impact AURKB expression and contribute to oncogenesis and cancer progression. Therefore, blocking MDM2 with specific inhibitors has become a promising strategy for the restoration of p53 function that could be used for the treatment of different types of cancers including lung cancers. In this regard, both MDM2 and AURKB inhibitors have shown promising antitumor activity against NSCLC. Therefore, the main objective of this study was to understand the molecular mechanisms linking MDM2 and ARKB using RG7388, CM272, BMS-582949 (p38 inhibitor), and Ralimetinib (p38 inhibitor) in A549 and H460 lung cancer cells. We analyzed the individual or combination treatment effects of these drugs on epigenetic modifications, cell cycle arrest, and cell death mechanisms, using the in vitro experimental model. When RG7388 was used individually, or in combination with BMS-582949 or Ralimetinib, a significant increase in the cytotoxic effect was observed in A549 and H460 cells. Induction of cell cycle arrest by RG7388 in cancer cells was evidenced by elevated p21 expression levels. In addition, RG7388 and combination treatments in NSCLC cells showed significant upregulation of p53, p27, Bax, cleaved PARP, phospho-FOXO3a and down-regulation in DNMT3a, DNMT1, FOXM1, AURKB, CDC25, and CDK4 levels. It appears that during treatments with a combination of MDM2 with DNMT or p38 inhibitor, two pathways might be blocked simultaneously, which could lead to increased cancer cell death. RG7388 appears to induce cell death in a p21-dependent manner and CM272, BMS-582949, and Ralimetinib seem to induce cell death via a p21-independent manner by inhibiting a downstream pathway. However, further studies are needed to fully understand the intracellular link between MDM2 and AURKB that is disrupted by the anti-cancer agents used in our study. (This project was supported by the PFRDG Grant of Nova Southeastern University and by the Generous support from the Royal Dames of Cancer Research Ft. Lauderdale, FL)

Citation Format: Umamaheswari Natarajan, Appu Rathinavelu. A synergy between MDM2 and AURKB pathways in driving cell cycle and cell survival in lung cancer 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 4593.
Abstract 4597: RNA m1A methyltransferase TRMT61A induces MAPK/ERK signaling and is a therapeutic target in colorectal cancer
Authors: Zhang Xiaoting , Qin Na , Ji Fenfen , Su Hao , Chen Hongyan , Shang Haiyun , Chen Huarong
Published: March 2024

The widespread presence of N1-methyladenosine (m1A) modifications controls RNA metabolism and is pivotal to fundamental biological processes. However, the understanding of RNA m1A involved in cancer is still minimal. Here we reported the oncogenic role and the therapeutic targeting of RNA m1A methyltransferase TRMT61A in colorectal cancer (CRC). We observed consistent elevation of TRMT61A expression and RNA m1A levels in primary CRC tissues, which was significantly associated with poor patient survival in two independent cohorts (both P<.001). CRISPR/Cas9 screenings revealed that TRMT61A is the most essential gene among m1A regulators. TRMT61A facilitates the growth and metastasis of CRC cells by increasing the stability of mRNA of crucial targets, such as ONECUT2, which leads to the activation of MAPK/ERK signaling in an m1A-dependent manner. Notably, depletion of ONECUT2 or inhibition of MAPK/ERK could abrogate the tumor-promoting role of TRMT61A, implying that TRMT61A induces ONECUT2-MAPK/ERK axis to facilitate CRC. Inhibiting TRMT61A with nanoparticle-encapsulated siTRMT61A or our recently identified small molecule compound PGG shows promising anti-CRC effects in vitro and in vivo. Together, our study reveals the promoting role of TRMT61A-induced m1A modification in MAPK/ERK signaling and highlights the potential of targeting TRMT61A as a promising therapeutic approach in CRC treatment.

Citation Format: Xiaoting Zhang, Na Qin, Fenfen Ji, Hao Su, Hongyan Chen, Haiyun Shang, Huarong Chen. RNA m1A methyltransferase TRMT61A induces MAPK/ERK signaling and is a therapeutic target in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4597.
Abstract 4651: A macrocyclic kinase inhibitor overcomes the compound mutations that cause the resistance to all approved EGFR inhibitors in EGFR-positive lung cancer
Authors: Katayama Ryohei , Uchibori Ken , Nishio Makoto , Fujita Naoya
Published: March 2024

EGFR mutant cancers account for 20-30 % of non-small cell lung cancers and show marked sensitivity to EGFR tyrosine kinase inhibitors (TKIs). However, the tumor almost inevitably relapses due to acquired resistance. To date, several EGFR-TKIs have been approved, and the third-generation EGFR-TKI, osimertinib, has been widely used both as first-line therapy and for patients with the EGFR-T790M mutation after 1st or 2nd generation EGFR-TKI therapy. However, the EGFR-T790M+C797S compound mutation confers resistance to all approved EGFR-TKIs. In our previous study, we experimentally demonstrated that brigatinib with anti-EGFR antibody was effective against osimertinib-resistant EGFR-C797S and EGFR-T790M+C797S mutants, and is currently undergoing clinical studies. However, the tumor would relapse because of the acquisition of additional resistance mutations. Here, we first demonstrated the binding mode of brigatinib to the EGFR-T790M/C797S mutant by crystal structure analysis and predicted brigatinib-resistant mutations through a cell-based assay, including N-ethyl-N-nitrosourea (ENU) mutagenesis. We found that clinically reported L718 and G796 compound mutations appeared, consistent with their proximity to the binding site of brigatinib, and brigatinib-resistant quadruple mutants, such as EGFR-activating mutation/T790M/C797S/L718M, were resistant to all clinically available EGFR-TKIs. BI-4020, a fourth-generation macrocyclic EGFR inhibitor plus anti-EGFR antibody, overcomes the quadruple and major EGFR-activating mutants in vitro and in vivo, but not the minor mutants such as L747P. Microsecond molecular dynamics simulations revealed the binding mode and affinity between BI-4020 and the EGFR mutants. This study identified potential therapeutic strategies using new-generation macrocyclic EGFR inhibitor to overcome the emerging resistance mutants.

Citation Format: Ryohei Katayama, Ken Uchibori, Makoto Nishio, Naoya Fujita. A macrocyclic kinase inhibitor overcomes the compound mutations that cause the resistance to all approved EGFR inhibitors in EGFR-positive 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 4651.
Abstract 4674: Reserpine analog modulates transport function of ABCG2
Authors: Ohnuma Shinobu , Kokubo Shoji , Sugisawa Norihiko , Murakami Megumi , Hatsuzawa Yuuri , Kobayashi Minoru , Kajiwara Taiki , Suzuki Hideyuki , Karasawa Hideaki , Watanabe Kazuhiro , Doi Takayuki , Ambudkar Suresh V. , Kamei Takashi , Unno Michiaki
Published: March 2024

Backgrounds: Reserpine is an agent that is used for the treatment of hypertension. ABCG2 also known as breast cancer resistance protein (BCRP) is a member of the ABC transporters associated with multidrug resistance (MDR). Our high-throughput screening assay has revealed that reserpine analog inhibits transport function of ABCG2.

Aim: To investigate the effect of reserpine analog on ABCG2 function.

Results: Flow cytometry assay showed that reserpine analog blocked the efflux of Pheophorbide a (PhA: fluorescent substrate of ABCG2) from ABCG2-expressing HCT-116/BCRP colon cancer cells in a concentration-dependent manner. Furthermore, cell proliferation assay (MTS assay) revealed that 5 and 10 μM reserpine analog sensitized HCT-116/BCRP cells to SN38 with IC50 value 0.61 + 0.12 μM and 0.22 + 0.036 μM, respectively. Finally, reserpine analog did not stimulate ABCG2-mediated ATP hydrolysis, but rather inhibited it (concentration required for 50% inhibition = 2.7 μM), suggesting that reserpine analog might prevent conformational changes required for ATP hydrolysis.

Conclusion: Reserpine analog could be developed as a potent modulator to overcome ABCG2-mediated MDR in cancer cells.

Citation Format: Shinobu Ohnuma, Shoji Kokubo, Norihiko Sugisawa, Megumi Murakami, Yuuri Hatsuzawa, Minoru Kobayashi, Taiki Kajiwara, Hideyuki Suzuki, Hideaki Karasawa, Kazuhiro Watanabe, Takayuki Doi, Suresh V. Ambudkar, Takashi Kamei, Michiaki Unno. Reserpine analog modulates transport function of ABCG2 [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 4674.
Abstract 470: AR-induced miRNA124-3p.2 suppresses the metastasis-promoting factor CCL2
Authors: Aoyama Shuhei , Izumi Kouji , Mizokami Atsushi
Published: March 2024

Objective: The standard treatment for advanced prostate cancer is androgen deprivation therapy that blocks androgen receptor (AR) signaling, but this therapy becomes ineffective within a few years, resulting in castration-resistant prostate cancer. We have previously shown that blockade of AR signaling induces CCL2 and activates cancer cells through an autocrine mechanism, but why AR blockade induces CCL2 secretion has not been elucidated. Micro RNAs (miRs) are endogenous non-coding RNAs that regulate the translation of various proteins. In this study, we investigated whether miRs are involved in CCL2 induction.

Subjects and Methods: We used miRNA PCR Plate Array and Target Scan (https://www.targetscan.org) to search for miRs that are controlled by AR and bind to and repress the CCL2 gene. We knocked down the AR of human prostate cancer cell lines LNCaP and C4-2B with siRNA (siAR) and examined the expression of target miRs and CCL2. siAR-transfected LNCaP and C4-2B were transfected with target miR mimic and CCL2 expression was examined. siAR-transfected LNCaP, C4-2B, and C4-2B were transfected with target miR mimic and CCL2 expression was examined. siAR-transfected LNCaP and C4-2B were transfected with target miR mimic and CCL2 expression was examined. The target miR mimic was introduced into them, and the changes in migration ability were evaluated.

Results: miR-124-3p.2 was extracted as a candidate from miRNA PCR Plate Array and Target Scan. miR-124-3p.2 expression was upregulated in both LNCaP and C4-2B when DHT was added and downregulated when CCL2 was added. In siAR LNCaP and C4-2B, the expression of CCL2 was decreased when miR-124-3p.2 mimic was introduced. Furthermore, in siAR-transfected LNCaP, C4-2B, the introduction of miR-124-3p.2 mimic suppressed the migration.

Conclusion: AR suppresses CCL2 via miR124-3p.2. miR124-3p.2-mediated regulation of CCL2 may be a new therapeutic target for advanced prostate cancer.

Citation Format: Shuhei Aoyama, Kouji Izumi, Atsushi Mizokami. AR-induced miRNA124-3p.2 suppresses the metastasis-promoting factor CCL2 [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 470.
Abstract 4724: Optimizing 500 pooled PRISM cell lines for a 10-day assay to elucidate viability effects and mechanism of action for slower-acting therapeutics
Authors: Ronan Melissa , Ip Blanche C. , Nguyen Ellen , Widocki Ursula , Sangpo Tenzin , Kalathungal Alvin , Diaz-Gallegos Nashielli , Fazio Anthony , Golabi Aydin , Naim Maha , George Ashish Bino , Liu YuhJong , Kocak Mustafa , Puram Rishi , Rees Matthew G. , Roth Jennifer
Published: March 2024

PRISM (Profiling Relative Inhibition Simultaneously in Mixtures), using DNA-barcoded cell lines cultured as ~25 cell line pools, has been an effective drug discovery screening platform to rapidly test large numbers of potential therapeutics on over 900 different cancer cell lines. However, the conventional 5-day assay may be too short to offer the resolution needed to identify the full cellular response for slower-acting therapeutics such as those directed against epigenetic pathways. Here, we describe development of the PRISM 10-day extended assay with ~500 adherent solid tumor cancer cell lines to overcome this challenge. We found that the ranges of cell line growth rates within the standard PRISM pools were too large and not amenable for a long-term assay beyond 6 days—after 10 days of growth, slower cell lines dropped below the limit of barcode detection as faster cell lines overtook the pools. We therefore regenerated pools to reduce the range of growth rates within a pool. We show that this re-pooling, coupled with seeding density optimization, enables recovery of > 98% of cell lines at day 10 in vehicle-treated wells, with acceptable data variability. The average estimated number of cell doublings across our cell lines, 7, is significantly greater than the number of doublings in the PRISM standard 5-day assay (~3 doublings). Importantly, we observe greater drug effects on day 10 versus day 6 when cells are treated with slow-acting DNA methyltransferase inhibitors such as decitabine; this also results in increased sensitivity to identify cellular biomarkers underlying differential response. In summary, we have successfully engineered a PRISM 10-day assay in ~500 adherent cell lines, where the extended duration can be beneficial in elucidating the effects and mechanism of action for epigenetic drug candidates.

Citation Format: Melissa Ronan, Blanche C. Ip, Ellen Nguyen, Ursula Widocki, Tenzin Sangpo, Alvin Kalathungal, Nashielli Diaz-Gallegos, Anthony Fazio, Aydin Golabi, Maha Naim, Ashish Bino George, YuhJong Liu, Mustafa Kocak, Rishi Puram, Matthew G. Rees, Jennifer Roth. Optimizing 500 pooled PRISM cell lines for a 10-day assay to elucidate viability effects and mechanism of action for slower-acting therapeutics [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 4724.
Abstract 479: Enhanced payload delivery for acute myeloid leukemia treatment through CD37-targeting DNA nanobots
Authors: Vantangoli Nicholas , Halley Patrick D. , Spitzner Jeffrey R. , Byrd John C. , Larkin Karilyn T. , Castro Carlos E. , Lucas Christopher R.
Published: March 2024

Acute Myeloid Leukemia (AML) treatment faces considerable challenges, in particular the delivery of therapeutics with sufficient potency and specificity. Traditional Antibody-Drug Conjugates (ADCs) offer a strategic approach to cancer therapy by combining the targeting capabilities of monoclonal antibodies with the cell-killing effect of cytotoxic drugs. However, the limitations in drug-to-antibody ratio (DAR) and the challenges in controlling the release kinetics have often hindered their therapeutic window suggesting novel approaches are necessary. Our approach integrates DNA nanobot technology to facilitate targeted delivery of anthracycline agents directly to CD37-expressing leukemic cells. CD37, a tetraspanin superfamily antigen, has shown promise as a target for AML due to its selective expression on immune cells and favorable internalization properties. By harnessing DNA origami assembly techniques, we developed a customizable platform capable of accommodating a significantly higher payload of chemotherapeutic agents than traditional ADCs. This increased payload potential, paired with the precision targeting of CD37, presents a potent therapeutic strategy with the possibility of reduced systemic toxicity. Here we present preclinical evidence demonstrating targeted delivery of our DNA nanobot drug delivery device and anthracycline payload to CD37+ AML target cells in vitro. In addition, we show targeted efficacy in vitro and in vivo of our anthracycline-loaded DNA nanobot-enabled delivery system, which significantly outperformed free anthracycline in AML target cells. Collectively, our findings suggest an improvement in the therapeutic index of anthracycline and antimitotic agents as well as the ability to combine multiple drug pathways in a single delivery device. The implications of this work extend beyond AML, offering a versatile platform that could revolutionize ADC chemistry and targeted drug delivery. We propose that our DNA nanobot delivery system signifies a pivotal advancement in the field of targeted cancer therapy, with the potential to overcome longstanding barriers in the treatment of AML and potentially other malignancies.

Citation Format: Nicholas Vantangoli, Patrick D. Halley, Jeffrey R. Spitzner, John C. Byrd, Karilyn T. Larkin, Carlos E. Castro, Christopher R. Lucas. Enhanced payload delivery for acute myeloid leukemia treatment through CD37-targeting DNA nanobots [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 479.
Abstract 4802: Accuracy of the GALAD serologic model in the diagnosis of hepatocellular carcinoma in Ghanaian patients
Authors: Nartey Yvonne Ayerki , Yang Ju Dong , Zemla Tyler , Ayawin Joshua , Asibey Shadrack Osei , El-Kassas Mohamed , Bampoh Sally Afua , Duah Amoako , Agyei-Nkansah Adwoa , Awuku Yaw Asante , Afihene Mary Yeboah , Yamada Hiroyuki , Yin Jun , Plymoth Amelie , Roberts Lewis Rowland
Published: March 2024

Background: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related mortality worldwide. In sub-Saharan Africa, HCC is the second most common cancer in men and the sixth in women. African patients present with HCC at a young age and at advanced stage of disease. Improved surveillance can increase early HCC detection. The GALAD score has good sensitivity in the diagnosis of HCC in Asia, Europe and North America, however, it has not been validated in an African cohort. The aim of this study was to assess the performance of the GALAD score in the diagnosis of HCC in Ghanaian patients.

Methods: Clinical data from patients with cirrhosis (n=93) or HCC (n=78) from out-patient hepatology clinics at 3 teaching hospitals in Ghana were abstracted, and serum samples analyzed. A logistic regression model predicting HCC status based on GALAD score was constructed to obtain the Receiver Operating Characteristics (ROC) curve for GALAD. The area under the curve (AUC) with 95% confidence interval was calculated and used to assess the performance of the GALAD score in surveillance for HCC.

Results: The median AFP (5010.2 vs. 3.4), L3% (28.3 vs. 0.3) and DCP (507.6 vs. 0.2) levels were all significantly higher among HCC cases than the controls (p<0.0001). The median GALAD score was also higher among HCC patients (8.0 vs. -4.1, p<0.0001). The AUC of the GALAD score in the diagnosis of HCC was 0.86 (95% CI 0.79 - 0.92). At a cut-off value of -0.37, the GALAD score had a sensitivity of 0.81 and a specificity of 0.86. In a re-fit of the logistic regression model, the most significant factors in the GALAD model in our cohort were AFP-L3% and log(DCP). The AUC of the new GALAD model after model selection was 0.89.

Conclusion: The GALAD score can be used to detect HCC in Ghana and has the potential to improve HCC surveillance in low-resource settings in sub-Saharan Africa.

Citation Format: Yvonne Ayerki Nartey, Ju Dong Yang, Tyler Zemla, Joshua Ayawin, Shadrack Osei Asibey, Mohamed El-Kassas, Sally Afua Bampoh, Amoako Duah, Adwoa Agyei-Nkansah, Yaw Asante Awuku, Mary Yeboah Afihene, Hiroyuki Yamada, Jun Yin, Amelie Plymoth, Lewis Rowland Roberts. Accuracy of the GALAD serologic model in the diagnosis of hepatocellular carcinoma in Ghanaian 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 4802.
Abstract 4844: Epidemiology of bone and soft-tissue sarcomas: A two-decade analysis of the surveillance, epidemiology, and end results program
Authors: El Beaino Marc , Baum Rachel , Connors Katherine M. , Masrouha Karim , Lozano-Calderon Santiago A. , Lin Patrick P.
Published: March 2024

In the United States, ~15000 cases of bone and soft-tissue sarcomas are diagnosed annually, representing roughly 1% of all incident cancers. While several studies have analyzed independent predictors of outcomes for these sarcomas, small samples and confounding bias have limited their ability to establish reliable conclusions. The present analysis provides a comprehensive epidemiological description of patients with any of 16 distinct bone and soft-tissue sarcomas by querying the Surveillance, Epidemiology, and End Results database between 2000 and 2020. Incidence, prevalence, incidence-based mortality, survival rates, and case-fatality ratios were computed. Using patient age, sex, race, and ethnicity, as well as tumor size, grade, and stage as covariates, multivariable survival Cox proportional hazards regression analyses were performed to model hazard ratios (HRs) and identify independent predictors of worse prognosis. Only patients with myosarcoma and fibrous histiocytoma witnessed a decrease in the prevalence of their disease over time (both, p<0.05). The incidence of chondrosarcoma and synovial sarcoma increased in White and Asian patients but decreased in Black individuals (both, p<0.001), while their case-fatality rates remained constant and increased, respectively, regardless of race and ethnicity (all, p<0.05). Case-fatality rates only declined significantly for White and Non-Hispanic patients with solitary fibrous tumor (both, p<0.05). Aside from alveolar soft-part sarcoma and clear cell sarcoma, a 10-year increase in age of patients with bone and soft-tissue sarcomas resulted in 18% to 61% higher likelihood of death (all, p<0.05). For most malignancies, having a tumor size greater than 5 cm yielded a 37% to 189% higher likelihood of death. While incidence trends suggest improvements in diagnostic techniques and early tumor detection, survival and case-fatality trends indicate potential for advancement in treatment for most bone and soft-tissue sarcomas.

Citation Format: Marc El Beaino, Rachel Baum, Katherine M. Connors, Karim Masrouha, Santiago A. Lozano-Calderon, Patrick P. Lin. Epidemiology of bone and soft-tissue sarcomas: A two-decade analysis of the surveillance, epidemiology, and end results program [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 4844.
Abstract 4900: Spatial Atlas of Human Anatomy (SAHA): A subcellular, multiscale spatial odyssey of immune and gastrointestinal tissues and tumors
Authors: Park Jiwoon , Gregorio Roberto , Hissong Erika , Patel Sanjay , Robinson Brian , Socciarelli Fabio , Metzger Evenlyn , Liang Yan , Reeves Jason , Beechem Joseph , Elemento Olivier , Alonso Alicia , Houlihan Shauna , Schwartz Robert , Mason Christopher E.
Published: March 2024

The Spatial Atlas of Human Anatomy (SAHA) is a foundational effort to map 250 million cells and transcriptomes and proteomes of 30 non-diseased organs from healthy adults at two spatial scales: whole transcriptome of histological features (50 µm to 2 mm), and 1,000-plex RNA and 64-plex protein panels at spatial subcellular resolution (50 nm across 1 cm2). The project aims to establish and validate best practices in experimental design, sample processing, data analysis, and data standards for high-content spatial analysis across multiple human organs at whole transcriptome and proteome levels. The profiled samples will capture variability across genders and ancestries. All results, including raw and processed data, will be made available to the scientific community through the SAHA data portal and AtoMxTM Spatial Informatics Platform.

Here we present the part of Phase I data collected specifically around immune (bone marrow and lymph node) and gastrointestinal tissues (ileum, appendix, colon, and liver), and compare these data to colon and liver cancer samples. On serial sections, we perform spatial whole transcriptome data (with GeoMx® Digital Spatial Profiler) and 1,000-plex RNA profiles and 64-plex protein profiles collected by the CosMx™ Spatial Molecular Imager (SMI) matched to the exact shape of functional histological organ features from H&E and immunofluorescence stainings. Our data, which spans ~1.5 million cells collected across ~1,100 fields of view, enables the highest-ever subcellular resolution maps of cell types, lineage states, metabolic capacity, cellular neighborhoods, subcellular movements of organelles, and spatially resolved (and novel) ligand-receptor interactions across normal and cancer tissues. Through comparing these data to several cancer samples collected across multiple spatial platforms, we show how spatial organ atlasing at multiple scales can uncover unique insights into organ development, health, and cancer. We also show how the SAHA data can serve as a benchmark reference for spatial precision medicine.

Citation Format: Jiwoon Park, Roberto Gregorio, Erika Hissong, Sanjay Patel, Brian Robinson, Fabio Socciarelli, Evenlyn Metzger, Yan Liang, Jason Reeves, Joseph Beechem, Olivier Elemento, Alicia Alonso, Shauna Houlihan, Robert Schwartz, Christopher E. Mason. Spatial Atlas of Human Anatomy (SAHA): A subcellular, multiscale spatial odyssey of immune and gastrointestinal tissues and tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4900.