bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2455: Investigating cancer stem cell plasticity in ovarian cancer
Authors: Metcalfe Tara X. , Zhang Shu , Hassel Christiane , O'Hagan Heather M. , Nephew Kenneth P.
Published: April 2023

Most women diagnosed with late-stage high grade serous ovarian cancer (HGSOC) develop recurrent, platinum-resistant tumors. Ovarian cancer stem cells (OCSCs) are hypothesized to contribute to the emergence of these resistant tumors. CSCs have been postulated to reside in a plastic state, which can allow for the conversion of non-CSC to CSC. This process of dedifferentiation continues during tumor development and chemotherapeutic agents like platinum can exaggerate CSC plasticity. We have previously demonstrated that acute platinum treatment enriched for OCSCs. However, whether platinum transforms non-OCSCs into OCSCs to contribute to this subpopulation of cells remains unclear, and the underlying mechanism remains incompletely understood. To examine OCSC plasticity, aldehyde dehydrogenase (ALDH; functional marker) and fluorescence activated cell sorting were used to isolate OCSCs (ALDH+) and non-OCSCs (ALDH-) from HGSOC cell lines, OVCAR5 and OVCAR3. To determine the stability of the non-OCSC phenotype, ALDH- cells were cultured for 3 and 5 days and ALDH activity was measured using flow cytometry. At both timepoints examined, ALDH- cells remained ALDH-, with approximately less than 1% being ALDH+ (p<0.05). To determine if platinum can induce conversion of non-OCSC to OCSC, ALDH- cells were treated with cisplatin (12µM for 16h), and the percent of ALDH+ cells was measured using flow cytometry. Treatment of ALDH- cells with cisplatin resulted in conversion of approximately 3-4% of ALDH- cells into ALDH+ cells (p<0.05). Furthermore, increased expression (p<0.05) of stemness genes BMI1, NANOG, OCT4, and SOX2, analyzed using qRT-PCR, was observed in converted OCSCs compared to parental ALDH+ and whole cell populations treated with cisplatin (12µM for 16h), suggesting that platinum induced the observed differences in the stemness phenotype. With the goal of targeting key genes and pathways to inhibit platinum-induced OCSC conversion, we are carrying out RNA-sequencing analysis of the converted OCSCs and investigating the genes and pathways driving the platinum-induced conversion on non-OCSCs to OCSCs. Understanding the mechanisms involved in OCSC plasticity is critical to developing targeted therapies to block the persistence of OCSCs and ultimately reduce mortality in patients.

Citation Format: Tara X. Metcalfe, Shu Zhang, Christiane Hassel, Heather M. O'Hagan, Kenneth P. Nephew. Investigating cancer stem cell plasticity in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2455.
Abstract 2456: Transcriptional signatures of stem cell phenotypes in normal and clonal hematopoiesis
Authors: Soldatov Ruslan , Schiroli Giulia , Kharchenko Peter , Scadden David
Published: April 2023

Clonal hematopoiesis (CH) is an age-related clonal expansion of hematopoietic stem cells (HSCs). Factors that shape the progression of CH are only beginning to be characterized. HSCs are a heterogeneous population of cells with distinct clonal outputs over time, which are hypothesized to be largely scripted in epigenetic and transcriptional cell state. Molecular properties of HSC states that mediate clonal behaviors of two major driver mutations in CH - DNMT3A and TET2 - represent potential therapeutical targets. Recent studies have identified the landscape of human CH mutant cells compared to non-mutated ones. However, the notion of a specific molecular target identified in CH cells associated with selective advantage or predisposition for subsequent transformation requires a formal validation by longitudinal studies. To uncover molecular underpinnings of CH-driven clonal phenotypes in human HSCs in vivo, we combined clonal analysis using heritable expressed DNA barcodes with single-cell transcriptomics using transplant in immunodeficient mice. We developed a statistical approach to associate transcriptional differences of HSCs with the clone phenotypes, such as size and HSC density, across more than a hundred of detected clones. It revealed robust transcriptional signatures underlying clone size and HSC density, which are largely complementary to transcriptional programs activated in TET2 and DNMT3A mutant HSCs. Of note, the clone size and density of HSCs in the clone tend to be uncoupled and associated with distinct transcriptional processes.

Citation Format: Ruslan Soldatov, Giulia Schiroli, Peter Kharchenko, David Scadden. Transcriptional signatures of stem cell phenotypes in normal and clonal hematopoiesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2456.
Abstract 2470: ARG1 is potential prognostic biomarker for development of distant metastatic endometrial cancer in mice with Pten deficiency
Authors: Jeong Jae-Wook , Tran Dinh , Rozen Valery , Kim Tae Hoon , Yoo Jung-Yoon
Published: April 2023

Endometrial cancer is the most common gynecologic malignancy. The most common histological type is endometrioid endometrial cancer (EEC). While the majority of patients present with early-stage and low-grade EEC and have an excellent prognosis, a subset has metastatic disease at presentation, or develops distant recurrence after initial treatment of the primary. However, the lack of prognostic biomarkers for metastatic EEC is a critical barrier. We have developed a genetically engineered mouse model for metastatic EEC that implicates coexistent Pten and Mig-6 mutations in EEC. Pten mutation in the uterus is not sufficient for distant metastasis, but mice with concurrent ablation of Mig-6 and Pten develop distant metastasis. Arginase 1 (ARG1) regulates the last step of the urea cycle, and an increase in ARG1 has been correlated as a poor prognostic factor in a variety of cancers including ovarian carcinoma, colorectal cancer, and neuroblastoma. In the present study, ARG1 expression was evaluated with immunohistochemistry in endometrial hyperplasia and cancer of mice with Pten mutation as well as Pten and Mig-6 double mutations. Our immunostaining analysis revealed that the expression of ARG1 in early stage of EEC from mice deficient in Mig-6 and Pten mutations significantly increased compared to Pten mutation in the uterus. High levels of ARG1 are associated with distant metastatic endometrial cancer. The results demonstrate that a high level of ARG1 is associated with poor prognosis in association with EEC of mouse. ARG1 has the potential to identify patients that require additional treatment at an early stage of endometrial cancer.

Citation Format: Jae-Wook Jeong, Dinh Tran, Valery Rozen, Tae Hoon Kim, Jung-Yoon Yoo. ARG1 is potential prognostic biomarker for development of distant metastatic endometrial cancer in mice with Pten deficiency [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2470.
Abstract 2579: The tumor landscape is molded by quantitative and qualitative signaling differences between different oncogenic KRAS mutations
Authors: Roux Ozgun Le , Counter Christopher M.
Published: April 2023

Despite over 50 possible oncogenic RAS mutations, cancers tend to have a bias towards a specific subset, often unique to each cancer type. As RAS mutation occur early, if not being the first mutations, these mutation patterns ostensibly reflect the selection of specific RAS mutations in each tissue to initiate tumorigenesis. Different RAS mutations can affect the level of active oncprotein and/or the type of pathways activated thereof. To attempt to determine the contribution of these two forms of signaling on the process of RAS mutation patterns in cancer, we generated four conditional murine Kras alleles containing one of two distinct oncogenic mutations, G12D or Q61R, encoded by native rare or common codons to induce low or high protein expression, respectively. These four alleles were globally activated throughout the mouse, revealing different tissue susceptibilities to the oncogenic Kras alleles. We observed that hematolymphopoietic lesions were preferentially initiated by the level of active RAS, squamous tumor were preferentially induced by one mutation over the other, while pulmonary carcinomas are susceptible to both activity level and the mutation type of the Kras allele. To further explore the signaling responses to the oncogenic Kras allele, we performed RNAseq analysis at the moment of tumor initiation, revealing that each allele induced a unique cellular response, ranging from transcriptional features of a plastic state to high oncogenic signaling and a stress response. We thus suggest that distinct RAS mutations impart a unique quantitative or qualitative signaling response that are selected in a tissue-specific fashion to initiate tumorigenesis, which has implications for early detection, interception, and prevention.

Citation Format: Ozgun Le Roux, Christopher M. Counter. The tumor landscape is molded by quantitative and qualitative signaling differences between different oncogenic KRAS mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2579.
Abstract 261: Whole genome sequencing provides a more comprehensive genetic characterization of lung cancer compared to a targeted panel sequencing
Authors: Park Ji-Hyung , Kim Young Tae , Connolly-Strong Erin , Ju Young Seok
Published: April 2023

Non-small cell lung cancer (NSCLC) remains the leading cause of cancer death in the United States despite a decline in mortality. Adenocarcinoma of the lung usually evolves from the mucosal glands and represents about 40% of all lung cancers. A significant proportion of lung adenocarcinoma patients present alterations in oncogenic driver genes. Next-Generation Sequencing (NGS) allows the sequencing of several genomic regions in a single test. There are multiple NGS approaches including Targeted Panel Sequencing (TPS), Whole Exome Sequencing (WES), and Whole Genome Sequencing (WGS). Methods: The detection of actionable genomic mutations in 60 lung adenocarcinoma surgical specimens was evaluated through two NGS approaches TPS and WGS. TPS was performed using the Seoul National University Hospital (SNUH) TPS consisting of 75 total genes, with a total coverage of 1.92Mb (exon 0.69Mb, intron 1.23Mb). WGS was performed using Illumina’s NovaSeq 6000 platform and data were processed with BWA and variant callers including Mutect2 and Delly. Functional mutations were determined by Ensembl VEP, while putative driver mutations were identified via the Genome Insight pipeline (San Diego). Results: A total of 120 driver mutations were detected by TPS. WGS identified 114 out of the 120 driver mutations (95% match). An additional 9 driver mutations were detected by WGS, along with thousands of additional non-driver mutations. (Table 1) Conclusion: An NGS-based approach allows for the identification of actionable molecular changes which leads to more informed clinical decision making. These results validate that Genome Insight’s WGS workflow and pipeline are comparable to TPS in detecting key driver mutations. Furthermore, this study demonstrated that WGS provides a more comprehensive genetic characterization of lung cancer, which may impact the therapeutic options and patients’ prognosis.

Table 1. Driver mutations # of mutations detected by TPS # of mutations detected by WGS Detected by TPS EGFR 20 19 KRAS 9 9 TP53 30 30 Fusion oncogenes 17 17 PIK3 genes 1 1 BRAF 2 2 Secondary drivers detected by TPS 41 36 Sub-total 120 114 Missed by TPS Other drivers not detected by TPS 0 9 Total 120 123

Citation Format: Ji-Hyung Park, Young Tae Kim, Erin Connolly-Strong, Young Seok Ju. Whole genome sequencing provides a more comprehensive genetic characterization of lung cancer compared to a targeted panel sequencing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 261.
Abstract 2870: Inhibition of DNA methylation and histone deacetylation synergistically reprograms M2-polarized macrophages and inhibits tumor growth by upregulating miR-7083-5p
Authors: Murugan Poongkavithai Vadevoo Sri , Rangaswamy Gunassekaran Gowri , Lee Byungheon
Published: April 2023

Reactivation of epigenetically suppressed miRs, in tumors, have become increasingly relevant in clinical practice. But less epigenetic studies have been performed on tumor associated M2 macrophages that plays a key role in the functional regulation of epithelial cancer development. In this study, we used 5-Aza-2’5’Aza-deoxycytidine (Aza) or decitabine (5-Aza) plus Trichostatin A (TSA) as epigenetic drugs to study the M2 macrophage modulation in the tumor microenvironment. Epigenetic therapy, not only modulated the M2 macrophages to a tumoricidal phenotype, but also strengthened the tumor immune surveillance and eventually attenuated tumor growth. In vitro studies on M2 macrophages, showed that these epigenetic modifiers, namely 5Aza+TSA, fine-tuned the M2 macrophages into a ‘no more tumor friendly’ phenotype. Using comparative miR profiling, we identified nearly 79 miRs that were upregulated in M2 macrophages after epigenetic drug treatment. Of all miRs, miR-7083-5p showed 32 fold upregulation in M2 after epigenetic therapy and csf2ra, a GM-CSF receptor that helps in IL3 and GM-CSF signaling to M2 polarization, and CD43,a leukocyte marker, were one two of the important genes targeted by this miR-7083-5p. Nevertheless, re-expression of miR-7083-5p in M2 Macrophages in vitro, modulated M2 Macrophage to an anti-tumor phenotype and 4T1 tumor tissues showed inversed correlation between the expression of miR-7083-5p and its target,csf2ra and CD43,implying the pathological relevance of targeting. Considering the vital role of M2-TAM in tumor growth and metastasis, epigenetic drug mediated reactivation of miRs in M2-TAM could act as a promising therapeutic approach in cancer treatment.

Citation Format: Poongkavithai Vadevoo Sri Murugan, Gunassekaran Gowri Rangaswamy, Byungheon Lee. Inhibition of DNA methylation and histone deacetylation synergistically reprograms M2-polarized macrophages and inhibits tumor growth by upregulating miR-7083-5p [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2870.
Abstract 2939: ATOR-4066, a Neo-X-Prime bispecific antibody targeting CD40 and CEA, activates myeloid cells in primary human tumors in vitro and induces anti-tumor immunity in vivo
Authors: Hägerbrand Karin , Sundstedt Anette , Levin Mattias , de Coaña Yago Pico , Varas Laura , Ljung Lill , Nilsson Anneli , Celander Mona , Jimenez David Gomez , Andersson Hampus , Lindstedt Malin , Ellmark Peter
Published: April 2023

Alligator’s Neo-X-Prime platform aims to enable antigen presenting cells to efficiently enhance priming of tumor neoantigen-specific T cells. We have demonstrated that binding of a CD40 x TAA bispecific antibody (bsAb) to CD40 on dendritic cells (DCs) and a tumor-associated antigen (TAA) on tumor exosomes or tumor debris leads to activation of the DC, uptake of the tumor material, cross-presentation of a tumor-derived antigen and priming of tumor antigen-specific T cells. This has the potential to result in an increased quantity and/or quality of the tumor-targeting T cell pool and enhanced anti-tumor activity. Herein, we present ATOR-4066, a Neo-X-Prime bsAb targeting CD40 and CEA (CEACAM5), a TAA highly expressed in several cancers. Using in vitro models, we have demonstrated CEA-dependent activation of CD40-expressing cells, and an ability to mediate co-localization of CEA-expressing tumor debris and CD40 expressing antigen presenting cells. In addition, ATOR-4066 was shown to activate CD40-expressing cells in the presence of primary human tumor cells from CEA+ colorectal and gastric cancer patients. Furthermore, in vitro treatment of primary colorectal and gastric tumor-derived cell cultures and tumoroids with ATOR-4066 induced activation of tumor-infiltrating immune cells. In vivo studies using human CD40 transgenic mice bearing CEA-transfected MC38 tumors showed superior anti-tumor effects of ATOR-4066 compared to a CD40 mAb, demonstrating the ability of ATOR-4066 to efficiently induce a tumor-targeting immune response. Overall, these data show the ability of ATOR-4066 to remodel the immune microenvironment and activate tumor-infiltrating immune cells in primary human tumors expressing CEA, demonstrating the promise of this new candidate drug for further clinical development.

Citation Format: Karin Hägerbrand, Anette Sundstedt, Mattias Levin, Yago Pico de Coaña, Laura Varas, Lill Ljung, Anneli Nilsson, Mona Celander, David Gomez Jimenez, Hampus Andersson, Malin Lindstedt, Peter Ellmark. ATOR-4066, a Neo-X-Prime bispecific antibody targeting CD40 and CEA, activates myeloid cells in primary human tumors in vitro and induces anti-tumor immunity in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2939.
Abstract 3149: Prediction of metabolic drug targets for cancer patients with poor prognosis by using genome-scale metabolic models and survival analyses
Authors: Lee Sang Mi , Lee GaRyoung , Kim Hyun Uk
Published: April 2023

Cancer metabolism has been extensively explored to identify biomarkers and novel drug targets for effective cancer treatment. In particular, it is expected that metabolism-derived data somehow reflects unique features associated with a cancer patient’s prognosis. Here, we introduce a computational framework that predicts metabolic drug targets by considering metabolites associated with poor prognosis. For inputs, the computational framework requires genome-scale metabolic models (GEMs) and survival data that represent each of cancer patients with diverse prognoses. GEM is a computational model that allows predicting fluxes of entire metabolic reactions in a patient-specific manner. By using this computational framework, a risk score, which reflects the prognosis of cancer patients, was first established on the basis of metabolites significantly associated with survival. Finally, metabolic reaction targets were predicted that could reduce the risk score (i.e., improving the prognosis) upon their knockdown in silico. In this study, this framework was applied to bladder cancer patients as a proof-of-concept demonstration, and predicted 13 metabolic reaction targets. These metabolic reaction targets were expected to be also effective to bladder cancer patients with poor prognosis. The computational framework developed in this study is distinct from previous drug targeting methods because it also considers unique metabolic features associated with cancer patients with poor prognosis in order to identify drug targets that are effective to both low-risk and high-risk groups of cancer patients.

Citation Format: Sang Mi Lee, GaRyoung Lee, Hyun Uk Kim. Prediction of metabolic drug targets for cancer patients with poor prognosis by using genome-scale metabolic models and survival analyses [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3149.
Abstract 3170: Predicting the signals of adverse drug reactions on the basis of medical data
Authors: Jeon Junhyeok , Hong Eujin , Kim Hyun Uk
Published: April 2023

Multiple drugs are often prescribed for the better treatment of a disease. While the use of multiple drugs is to improve the overall therapeutic efficacy, it can also lead to unintended adverse drug reactions (ADRs) from drug-drug interactions (DDIs). More than 30% of ADRs have been reported to be caused by DDIs. Because of the importance of DDIs and ADRs, many computational models have been developed to predict DDIs and/or ADRs. However, these computational models mostly make predictions on the basis of molecular features of drugs without considering clinical factors, which consequently does not allow predictions in a person-specific manner. To address this problem, we present a machine learning model that predicts the signals of ADRs potentially caused by DDIs on the basis of medical data. Medical data used for model development come from MIMIC-IV, and include age, ethnicity, gender, patients’ disease information (i.e., ICD-10) and information on drugs administered (e.g., molecular structure and dose). The medical data were subjected to extensive preprocessing based on a domain knowledge, and multiple machine learning methods were examined to predict the ADR signals using the preprocessed dataset. ADR signals are captured by classifying whether each of the selected laboratory measurements (e.g., hematocrit, creatinine, and hemoglobin) is normal. The best-performing model showed the average AUROC of more than 0.8. The machine learning model developed in this study can be useful for examining potential harms when considering multiple drugs, including anticancer drugs.

Citation Format: Junhyeok Jeon, Eujin Hong, Hyun Uk Kim. Predicting the signals of adverse drug reactions on the basis of medical data [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3170.
Abstract 3202: NanoBiT technologies for CAR-T characterization and potency testing
Authors: Gilden Julia K. , Stecha Pete , Paguio Aileen , Hartnett Jim , Binkowski Brock , Lazar Dan , Fan Frank , Cong Mei
Published: April 2023

Despite significant clinical advances in CAR-T cell therapies for cancer, the ability to precisely and reproducibly analyze the potency of these drug products remains a challenge. As new CAR-T products and production methodologies are developed, it is becoming increasingly important to be able to rigorously evaluate the relative potencies of each batch. We have developed two MOA-based bioluminescent assays that overcome challenges in quantitative assessment of T cell therapies. Both assays rely on split NanoBiT luciferase technology. In the HiBiT Target Cell Killing Assay, incubation of CAR-T cells with target cells stably expressing HiBiT results in lysis of the target cells and release of HiBiT proteins. These HiBiT proteins bind with high affinity to LgBiT in the detection reagent and form functional NanoBiT luciferase, resulting in production of light in proportion to target cell lysis. Lumit Immunoassays are a simple and homogeneous method for measuring cytokine production from engineered T cells. Monoclonal antibodies are covalently labeled with LgBiT and a low-affinity tag, SmBiT, which have negligible association in solution. When the antibodies are brought together by a common analyte, such as a cytokine, SmBiT and LgBiT can compliment to form NanoBiT luciferase and produce light in proportion to the concentration of analyte in the sample. Lumit Immunoassays can be run directly on cells, removing media transfer, dilution, and wash steps that can be common sources of error. These luminescent assays are homogenous, fast, highly sensitive, and have robust assay windows. They represent a new set of tools for development and characterization of T cell-based immunotherapies.

Citation Format: Julia K. Gilden, Pete Stecha, Aileen Paguio, Jim Hartnett, Brock Binkowski, Dan Lazar, Frank Fan, Mei Cong. NanoBiT technologies for CAR-T characterization and potency testing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3202.
Abstract 328: Acetylation regulates the oncogenic functions of PRMT5
Authors: Vo Tam Thuy Lu , Do Yen Thi , Pham Thi Tuyet Mai , Shin So-Jin , Kim Jin Young , Lee Seungmee , Ha Eunyoung , Seo Ji Hae
Published: April 2023

Protein arginine methyltransferase 5 (PRMT5) is an enzyme responsible for symmetric arginine methylation, which governs multiple crucial cellular events. PRMT5 expression and activity are dysregulated in many malignancies. PRMT5 dynamically shuttles between the cytosol and nucleus, and different subcellular localizations of PRMT5 imposes distinct cellular functions. However, the biological significance and underlying regulatory mechanism of PRMT5 translocation remain elusive. Herein, we found that PRMT5 acetylation/deacetylation status is a major determinant of its translocation and oncogenic function. PRMT5 acetylation not only drives its cytoplasmic localization but also enhances its methyltransferase activity, promoting cancer cell proliferation. On the other hand, PRMT5 deacetylation translocates PRMT5 into nucleus, to protect cells under the stress condition. Our current findings demonstrate that the acetylation/deacetylation-dependent subcellular localization of PRMT5 is a key regulator of its oncogenic properties.

Citation Format: Tam Thuy Lu Vo, Yen Thi Do, Thi Tuyet Mai Pham, So-Jin Shin, Jin Young Kim, Seungmee Lee, Eunyoung Ha, Ji Hae Seo. Acetylation regulates the oncogenic functions of PRMT5 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 328.
Abstract 3293: CADM1 is a serum marker for small-cell lung cancer associated with poor prognosis and metastasis
Authors: Ito Takeshi , Funaki Toko , Iwanari Hiroko , Tanaka Goh , Nagase Takahide , Hamakubo Takao , Murakami Yoshinori
Published: April 2023

Small-cell lung cancer (SCLC) is aggressive neuroendocrine cancer that accounts for approximately 15% of lung cancer. Although chemotherapy is effective for SCLC, resistant tumors develop rapidly; therefore, early detection in the initial screening and after chemotherapy is one of the critical issues in controlling SCLC progression. For this purpose, neuron-specific enolase (NSE) and pro-gastrin-releasing peptide (ProGRP) are used as tumor markers for SCLC but are not sensitive enough, especially in the early stages. Previously we found that Cell adhesion molecule 1 (CADM1) is highly expressed in SCLC, and an SCLC-specific splicing variant, CADM1v8/9, is selectively cleaved by ADAM17 protease to release the extracellular fragments. Here, we report that soluble CADM1v8/9 fragments (sCADM1v8/9) can be a serum marker of SCLC associated with poor prognosis and metastasis. Immunohistochemical analysis showed that the expression of CADM1 was observed in 83% of the resected SCLC cases and strongly correlated with that of INSM1, a marker of neuroendocrine differentiation. The high expression of CADM1 was associated with poor prognosis and nodal involvement, which was supported by experimental data showing that CADM1 promoted subcutaneous tumor growth and lymph node metastasis of SCLC cells in nude mice. Moreover, sCADM1v8/9 distinguished 47% of SCLC, including a significant portion of patients with limited disease SCLC, independent of and partly overlapped with the cases detected by ProGRP or NSE. Notably, a high amount of sCADM1v8/9 was observed in SCLC cases with liver metastasis or pleural effusion. Furthermore, serum levels of sCADM1v8/9 well reflected the change in tumor burden before and after chemotherapy. These findings indicate that detecting sCADM1v8/9 in sera would provide clinical benefits in diagnosis, follow-up, and metastasis prediction of SCLC cases.

Citation Format: Takeshi Ito, Toko Funaki, Hiroko Iwanari, Goh Tanaka, Takahide Nagase, Takao Hamakubo, Yoshinori Murakami. CADM1 is a serum marker for small-cell lung cancer associated with poor prognosis and metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3293.
Abstract 330: EP300 loss drives tumorigenesis in bladder cancer via activation of IL-6/JAK/STAT3 signaling
Authors: Gao Sizhi P. , Rodrigues James A. , Sabel Amanda R. , Luo Jiaqian , Ziyu Chen , Tang Xinran , Mascareno Eduardo A. , Rustgi Naryan , Al-Ahmadie Hikmat , Kim Kwanghee , Pietzak Eugene J. , Iyer Gopakumar V. , Solit David B.
Published: April 2023

Bladder cancer (BLCA) is the sixth most common cancer in the U.S with 83,730 new cases, accounting for 17,200 deaths in the year 2021 and 212,536 deaths worldwide. EP300, a histone acetyltransferase gene, is mutated in ~17% of BLCA and in other cancer types such as endometrial cancer. The mechanisms by which EP300 mutations contribute to tumorigenicity of BLCA is not currently understood. To determine the phenotypic effects of loss-of-function EP300 mutations, we generated isogenic BLCA cells with EP300 knocked out (KO) using CRISPR/CAS9. Conspicuous changes in phenotypes consistent with more aggressive forms of cancer were observed in EP300-null BLCA clones, including luminal-basal histotype plasticity, anchorage-independent growth, enhanced in vitro and in vivo cell proliferation and enhanced invasion in Boyden chamber assays. To identify mechanisms whereby EP300 KO promotes increased oncogenicity, we performed phospho Tandem Tag Mass Spectrometry (TMT MS) using lysates from parental and EP300 KO cells. A notable finding was significantly increased expression of phosphorylated STAT3 (pSTAT3), which we subsequently confirmed by western blot. siRNA knockdown and selective inhibitor experiments indicated JAK1 as the upstream activator of pSTAT3 in BLCA cells. Studies of conditioned media (CM) from the EP300 KO cells using ELISA and neutralizing antibodies demonstrated that secreted IL-6 ligand and soluble IL-6R in the CM drove STAT3 activation through the IL-6 family common signaling subunit, gp130, i.e. trans-signaling. BLCA RT112 cells express an oncogenic FGFR3-TACC3 fusion and are sensitive to the FDA-approved FGFR kinase inhibitor erdafitinib. pSTAT3 expression was not dependent on FGFR3 signaling in EP300 KO RT112 cells, and EP300 KO was sufficient to confer erdafitinib resistance in a BLCA context. In sum, our results uncover that EP300 loss enhances IL-6/JAK/STAT3 signaling which promotes BLCA tumorigenesis and FGFR inhibitor resistance. Our findings elucidate a role for cytokine induced sterile inflammation in EP300-mediated tumorigenesis in bladder cancers and suggest that targeting the IL-6/JAK/STAT3 axis may represent a novel therapy strategy to overcome FGFR3 inhibitor resistance.

Citation Format: Sizhi P. Gao, James A. Rodrigues, Amanda R. Sabel, Jiaqian Luo, Chen Ziyu, Xinran Tang, Eduardo A. Mascareno, Naryan Rustgi, Hikmat Al-Ahmadie, Kwanghee Kim, Eugene J. Pietzak, Gopakumar V. Iyer, David B. Solit. EP300 loss drives tumorigenesis in bladder cancer via activation of IL-6/JAK/STAT3 signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 330.
Abstract 3317: Expression of glucose transporter-1 in follicular lymphoma affected tumor-infiltrating immunocytes and was related to progression of disease within 24 months
Authors: Ma Jianli , Deng Yuwei , Zhao Shu , Yang Ming , Yu Yutian , Zhang Qingyuan , Yu Jinming
Published: April 2023

Objective Follicular lymphoma (FL) occurring progression within 24 months (POD24) after initial immunochemotherapy has poor prognosis. GLUT1 affects glycolysis within tumor microenvironment (TME) and promotes tumor progression. However, its specific mediated mechanism remains unclear in FL.

Methods Baseline GLUT1 expression, infiltrations of M2 macrophage, and CD8+ T-cells were assessed by immunohistochemistry in FL with POD24 and long-term remission respectively. The spatial features of TME were assessed by MIBI-TOF and proteomics. Predictive immunophenotypes for POD24 occurrence was analyzed by random forest algorithm. The lactate production and the induction of M2 macrophages were detected when GLUT1 was transfected or knocked down in DOHH2. The activation of PI3K/Akt/mTOR signaling in DOHH2 and WSU-FSCCL cells co-cultured with induced inhibitory immunocytes was tracked by western blotting.

Results The FL with POD24 exhibited higher baseline GLUT1 expression and increased infiltration of various inhibitory immunocytes. Spatial signatures of 69 immunophenotypes could predict POD24 occurrence. The activation of PI3K/Akt/mTOR signaling pathway was not significant in both groups. The supernatant of DOHH2-GLUT1 cells which had more lactate content could induce more M2-type macrophages than that of DOHH2/siRNA GLUT1 cells. When co-cultured with exhausted CD8+ T cells, M2-type macrophages and Tregs, compared with WSU-FSCCL cells, DOHH2 cells with high GLUT1 expression induced more M2-type macrophages and was triggered activation of PI3K/Akt/mTOR signaling pathway.

Conclusion Tumor cells overexpressing GLUT1 could domesticate immunocytes to form an immunosuppressive TME, which promotes occurrence of POD24 and gradually activates PI3K/Akt/mTOR pathway of tumor cells in FL.

Clinical correlation between GLUT1 expression and tumor-infiltrating macrophages and CD8+ T cells GLUT1 expression High n(%) Low n(%) P value CD206 expression High 16(43.2) 5(18.5) 0.037 Low 21(56.8) 22(81.5) CD8 expression High 27(73.0) 12(44.4) 0.021 Low 10(27.0) 15(55.6)

Citation Format: Jianli Ma, Yuwei Deng, Shu Zhao, Ming Yang, Yutian Yu, Qingyuan Zhang, Jinming Yu. Expression of glucose transporter-1 in follicular lymphoma affected tumor-infiltrating immunocytes and was related to progression of disease within 24 months [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3317.
Abstract 3392: Antitumor activity of a potent MNK inhibitor NSP-1047 in acute myeloid leukemia
Authors: Li Bing , Huang Qionglin , Zhang Ke , Zhang Jianming
Published: April 2023

MAPK-interacting kinases (MNKs) phosphorylate the eukaryotic initiation factor 4E (eIF4E) and promote the synthesis of tumorigenic proteins. MAPK/MNK/eIF4E pathway is activated in many kinds of haematological and solid tumors. Here, we report the discovery of a novel polycyclic compound NSP-1047, which potently inhibits MNK1 and MNK2, and has sub-nanomolar cellular potency inhibiting phosphorylation of eIF4E. NSP-1047 potently inhibits the expression of multiple immune checkpoint proteins, including PD-1, Tim-3, and Lag-3, and it also inhibits the secretion of several inflammatory cytokines, such as IL-6, IL-8 and IL-10. NSP-1047 displays excellent ADME and pharmacokinetic properties, and demonstrates potent in vivo antitumor activity in acute myeloid leukemia animal models with tumor regression.

Citation Format: Bing Li, Qionglin Huang, Ke Zhang, Jianming Zhang. Antitumor activity of a potent MNK inhibitor NSP-1047 in acute myeloid leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3392.
Abstract 340: Investigating the role of HMGA2 and AR crosstalk in prostate cancer cell proliferation
Authors: Awolowo Mojisoluwa Simisoluwa , Campbell Taaliah , Hawsawi Ohuod , Odero-Marah Valerie
Published: April 2023

Prostate cancer (PCa) is the most common non-skin cancer in men, and second leading cause of death. PCa occurs when cells within the prostate grow uncontrollably, creating small tumors. Generally, the growth/proliferation of the prostate is regulated by the male hormone, androgen, that binds androgen receptor (AR), a nuclear receptor that functions as a transcription factor and regulates the development and proliferation of the prostate as well as PCa. Interestingly, AR has become one of the leading pathways targets to slow down prostate cancer progression. In fact, enzalutamide a second-generation antiandrogen drug works to inhibit AR translocation into the nucleus. High Mobility Group AT-Hook (HMGA2) is a transcription factor protein that alters the structure of DNA by binding to the promoter region to regulate the transcription of genes. The expression of HMGA2 is associated with cancer proliferation and metastasis and is higher in PCa patients compared to men without cancer. Additionally, this gene has two isoforms HMGA2 full-length/wild-type and truncated (missing the carboxyl terminus). Preliminary studies shown that AR is localized in the nucleus of HMGA2 wild-type but not truncated overexpressing cells. Our hypothesis is that HMGA2 wild-type promotes AR translocation to the nucleus which leads to increased cell proliferation in PCa cells. We utilized HMGA2-wild-type and HMGA2-truncated overexpressing LNCaP cells and C4-2B enzalutamide resistant cells to knockdown either AR or HMGA2 with siRNA followed by western blot analysis, cell proliferation assay, and immunofluorescence. Our results show that HMGA2 siRNA led to decreased expression of wild-type (25 kDa long form) and truncated isoform (17 kDa) protein expression, and cell proliferation when compared to control siRNA. However, AR knockdown decreased AR expression in both cell lines but only decreased cell proliferation in HMGA2-wild-type and C4-2B enzalutamide-resistant cells. This suggests that HMGA2 wild-type works with AR to promote cell proliferation, whereas HMGA2-truncated promotes cell proliferation by an AR-independent mechanism. Therefore, we need to examine how current PCa treatments such as enzalutamide that target AR work in patients expressing differing HMGA2 isoforms.

Grant/Other Support: RISE: 5R25GM0058904, NIH/NIMHD 2U54MD007590; 5U54MD013376-8281; NSF S-STEM 2030608.

Citation Format: Mojisoluwa Simisoluwa Awolowo, Taaliah Campbell, Ohuod Hawsawi, Valerie Odero-Marah. Investigating the role of HMGA2 and AR crosstalk in prostate cancer cell proliferation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 340.
Abstract 3444: Role of AKR1C3 in converting reservoirs of DHEA-S into potent androgens that drive prostate cancer cell growth
Authors: Detlefsen Andrea J. , Duan Ling , Mesaros Clementina , Penning Trevor M.
Published: April 2023

Castration resistant prostate cancer (CRPC) treatment aims to stop a resurgence of androgen receptor (AR) signaling with a class of therapeutics known as androgen receptor signaling inhibitors (ARSI). Through inhibition of CYP17A1, Abiraterone (Abi) prevents an upstream step in androgen biosynthesis, while Enzalutamide (ENZ) acts as an AR antagonist. Despite the early success of ARSI, a subset of patients develop resistance and succumb to the disease. One major mode of resistance is thought to occur when steroidogenic enzymes that are upregulated in CRPC convert weaker circulating precursors to testosterone (T) and dihydrotestosterone (DHT) to restore AR signaling. In this study, we identify a mechanism driving intratumoral steroidogenesis that involves AKR1C3 (type 5 17β-hydroxysteroid dehydrogenase type 5). AKR1C3 is a major steroidogenic enzyme in the prostate that is involved in all androgen biosynthetic pathways to T and DHT, upregulated in CRPC patients, and is a component of Abi and ENZ drug resistance. Previously with Taplin et al. we observed in patients on Leuprolide that T was reduced to 5 ng/dL, and the addition of Abi lowered DHEA-S by 90% (from 230,000 ng/dL to 22,000 ng/dL). However, the DHEA-S that remained was 4,000x greater than circulating T, making it a reservoir for intratumoral steroidogenesis post Abi treatment. Here, we demonstrate that post-Abi concentrations of DHEA-S, and castrate levels of DHEA-S (post-leuprolide but pre-Abi treatment), which also corresponds to DHEA-S levels patients on ENZ are 1) converted to T in an AKR1C3-dependent manner in prostate cancer (PCa) cells, and 2) sufficient to stimulate AKR1C3-dependent cell growth. Our observations are consistent across both primary and metastatic PCa cell lines, CWR22PC and DuCaP, respectively. Androgen measurements were made using our highly sensitive and specific stable isotope dilution liquid chromatography tandem mass spectrometry (SID-LC-MS/MS) method. Additionally, we demonstrate AKR1C3-dependence using genetic and pharmacological approaches with an AKR1C3 stable knockdown for each cell line and two AKR1C3 competitive inhibitors. Taken together, we have identified a mechanism of resistance to ARSIs common to both primary and metastatic cell lines that operates through AKR1C3’s intratumoral conversion of DHEA-S to potent androgens that drive prostate cancer growth.

Citation Format: Andrea J. Detlefsen, Ling Duan, Clementina Mesaros, Trevor M. Penning. Role of AKR1C3 in converting reservoirs of DHEA-S into potent androgens that drive prostate cancer cell growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3444.
Abstract 3465: HDAC8 regulates immune escape in melanoma through increased transcription of inhibitory cytokines that increase accumulation of myeloid-derived suppressor cells (MDSCs)
Authors: Zhang Chao , Emmons Michael , Bennett Richard , Licht Jonathan , Smalley Keiran
Published: April 2023

Immune checkpoint blockade leads to improved and durable responses in 30-40% of melanoma, yet nearly 1/3 of patients relapse from complete remission within 3 years. Recent studies from our group and others demonstrated that upfront use of targeted therapy leads to decreased immune infiltrate and confers resistance to immunotherapy. We have previously showed that HDAC8 reprograming cell transcriptional states leads to reduced recognition by immune system. However, it’s unclear how HDAC8 activity modulates the tumor immune microenvironment (TIME). In this study, we used integrated single cell RNA-seq, ATAC-seq and a novel BRAF-V600E; PTEN+/- melanoma genetically engineered mouse model (GEMM) to explore “cold” TIME modulated by HDAC8. Analysis of human melanoma cells overexpressing HDAC8 by ATAC-seq, ChIP-Seq and RNA-Seq showed that HDAC8 expression significantly increases chromatin accessibility, H3K27 promoter region acetylation and gene expression of IL-11. Increased IL-11 protein expression was confirmed by cytokine array in isogenic SM1 mouse melanoma cells expressing HDAC8.Overexpression of HDAC8 in the GEMM accelerated melanoma development and was associated with worse overall survival rate. Analysis of HDAC8 and control tumors by scRNA-seq and immunohistochemistry (IHC) showed that HDAC8 overexpressing tumors dramatically decreased CD4 and CD8 T cell infiltration and increased MDSCs and IL-11 expression. We next explored whether IL-11 drives the expansion and polarization of MDSCs and subsequent reduction in T cell infiltration and activation. A MDSCs polarization assay demonstrated that addition of IL-11 led to increased expansion and polarization of MDSCs from mouse bone marrow myeloid cells. A MDSCs suppression assay confirmed IL-11 induced MDSCs accumulation significantly reduced T cell proliferation and activation. We show for the first time that HDAC8 expression regulates immune escape by activating IL-11 expression and driving MDSCs expansion. Going forward, inhibition of HDAC8 activity is intriguing as a novel therapeutic strategy to reverse tumor derived immune suppression and to ultimately improve patient responses to immunotherapy.

Citation Format: Chao Zhang, Michael Emmons, Richard Bennett, Jonathan Licht, Keiran Smalley. HDAC8 regulates immune escape in melanoma through increased transcription of inhibitory cytokines that increase accumulation of myeloid-derived suppressor cells (MDSCs) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3465.
Abstract 3466: Microbial neoantigen vectors for cancer immunotherapy
Authors: Redenti Andrew A. , Danino Tal , Arpaia Nicholas
Published: April 2023

Cancers employ a variety of immunosuppressive and evasive mechanisms to avoid detection by the immune system and prevent tumor neoantigen-mediated elimination. In recent years, tumor-specific neoantigen vaccines have shown significant promise—promoting tumor regression and increasing survival benefit in certain malignancies by therapeutically directing the immune system to target previously protected tumor specific antigens. However, despite encouraging preclinical data, objective and durable responses have remained elusive—highlighting a requirement for improved therapeutic platforms that permit delivery and targeting of diverse sets of tumor antigens to enhance the efficacy of cancer vaccines. Through synthetic biology, we have engineered living microbial vectors harboring constructs encoding synthetic arrays of tumor-specific neoantigens that specifically colonize tumors following intravenous injection and deliver high levels of neoantigen cargo—both extracellularly (within the tumor microenvironment) and intracellularly (to the cytosol of tumor-resident phagocytes). These engineered vectors robustly activate innate immune cascades and effectively prime tumor neoantigen-directed T cell responses to promote tumor regression and survival in advanced murine solid tumor and lung metastasis models. Extensive remodeling of the tumor immune microenvironment accompanies therapeutic efficacy, with increased activation of tumor neoantigen-specific CD4+ and CD8+ T cells, broadened stimulation of tumor-infiltrating B and T cells, and reduced numbers of intratumoral myeloid cells exhibiting immunosuppressive phenotypes. This work provides a foundational platform for the development of a new class of next-generation tumor vaccines, leveraging the advantages of living medicines to deliver diverse arrays of tumor-specific neoantigens within the optimal immunostimulatory context to induce potent anti-tumor immunity.

Citation Format: Andrew A. Redenti, Tal Danino, Nicholas Arpaia. Microbial neoantigen vectors for cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3466.