bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1479: FOXA2 induces invasive mucinous adenocarcinoma of the lung in the presence of mutant KRAS
Authors: Maeda Yutaka , Tomoshige Koichi , Stuart William D. , Fink-Baldauf Iris M. , Ito Masaoki , Tsuchiya Tomoshi , Nagayasu Takeshi , Yamatsuji Tomoki , Okada Morihito , Fukazawa Takuya , Guo Minzhe
Published: June 2023

Introduction: The role of the endoderm lineage transcription factor FOXA2 in autochthonous lung tumors has been elusive though the expression of FOXA2 has been reported in human lung cancers. Here, using GEMM (Genetically Engineered Mouse Models) and human lung cancer cell lines, we delineate the role of FOXA2 in lung cancers.

Experimental Procedures: GEMM that carry mutant KRAS or mutant EGFR along with FOXA2 (CCSP-rtTA; tetO-Foxa2; tetO-Kras.G12D or tetO-EGFR.L858R) were developed allowing for lung tumor induction by doxycycline administration. The growth of lung tumors affected by FOXA2 was assessed by microCT and pathologically. FOXA2 downstream genes were also evaluated in vivo by immunohistochemistry of mouse lung sections and in vitro by gain (ectopic lentivirus transduction) and loss (siRNA) of function approaches with human lung cancer cell lines using TaqMan gene expression, RNA-seq, ChIP-seq and CRISPRi analyses. FOXA2 protein expression in human lung cancer specimens was also assessed.

Data: FOXA2 suppressed the growth of EGFR-mutant lung tumors while promoting that of KRAS-mutant lung tumors in GEMM. Notably, half of the KRAS-mutant lung tumors presented invasive mucinous adenocarcinoma of the lung (IMA) with the expression of a master mucous transcription factor SPDEF and the two major IMA mucin genes MUC5AC and MUC5B. In vitro, FOXA2 induced the expression of SPDEF, MUC5AC and MUC5B by directly binding to their loci. H3K27ac was introduced at the enhancer region of MUC5AC (most highly expressed IMA gene) by FOXA2 and the expression of MUC5AC initiated by FOXA2 was enhanced by mutant KRAS. FOXA2 was highly expressed in human IMA compared to normal lung.

Conclusions: FOXA2 induces IMA in the presence of mutant KRAS by directly regulating the expression of mucous genes.

Citation Format: Yutaka Maeda, Koichi Tomoshige, William D. Stuart, Iris M. Fink-Baldauf, Masaoki Ito, Tomoshi Tsuchiya, Takeshi Nagayasu, Tomoki Yamatsuji, Morihito Okada, Takuya Fukazawa, Minzhe Guo. FOXA2 induces invasive mucinous adenocarcinoma of the lung in the presence of mutant KRAS [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 1479.
Abstract 1482: Critical role of OGT-mediated novel NF-κB o-glcNAcylation in pancreatic cancer
Authors: Motolani Aishat , Martin Matthew , Wang Benlian , Jiang Guanglong , Liu Yunlong , Lu Tao
Published: June 2023

Pancreatic ductal adenocarcinoma (PDAC) has one of the highest mortalities of all malignancies, with a mere 5-year survival of ~10%. However, the current first-line treatments have poor patient outcomes, highlighting the urgent need for innovative therapeutics. The nuclear factor κB (NF-κB) is a crucial transcription factor frequently activated constitutively in PDAC. It mediates the transcription of oncogenic and inflammatory genes that facilitate multiple PDAC phenotypes. Thus, a better understanding of the mechanistic underpinnings of NF-κB activation holds substantial promise in PDAC diagnosis and new therapeutics. The purpose of this study is to identify novel regulation of NF-κB, with the aim of providing new diagnostic and therapeutic strategies for PDAC. Here, we report protein O-GlcNAc transferase (OGT) - mediated NF-κB activation through novel serine O-GlcNAcylation of its p65 subunit. We show that overexpression of serine-to-alanine (S-A) mutant at the O-GlcNAcylation site impaired NF-κB nuclear translocation and transcriptional activity in PDAC cells. Moreover, these S-A p65 mutants downregulate NF-κB-target genes important to major cancer hallmarks and inhibit cellular proliferation, migration, and anchorage-independent growth of PDAC cells compared to WT-p65. Interestingly, this modification happens downstream of the inhibitor of NF-κB (IκBα) degradation. Collectively, we have identified OGT-mediated serine O-GlcNAcylation of NF-κB and determined its mechanistic and cellular function in driving PDAC phenotypes. Thus, our study holds great significance as it uncovers a brand-new strategy for PDAC diagnosis and effective therapeutics development.

Citation Format: Aishat Motolani, Matthew Martin, Benlian Wang, Guanglong Jiang, Yunlong Liu, Tao Lu. Critical role of OGT-mediated novel NF-κB o-glcNAcylation in pancreatic 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 1482.
Abstract 1544: MBK-103, a potent novel conjugation platform-based antibody-drug conjugate, changing therapeutic options in folate receptor alpha positive cancer patients
Authors: Viricel Warren , Conilh Louise , Leroy Edouard , Lafay Jean-Guillaume , Brune Frederique , Sadilkova Lenka Kyrych , Dumontet Charles
Published: June 2023

Folate receptor alpha (FOLR1) is a clinically validated target, that is overexpressed extracellularly in numerous solid malignancies with a high unmet medical need. Ovarian, non-small cell lung and breast adenocarcinomas are among indications with the highest frequency of FOLR1-positive patients with one out of two patients showing upregulated topoisomerase I expression. MBK-103, an antibody-drug conjugate targeting FOLR1, is based on Mablink’s novel proprietary hydrophilic drug-linker platform. It is comprised of: (i) an Fc-attenuated humanized IgG1 monoclonal antibody, that selectively binds to FOLR1; (ii) a polysarcosine hydrophobicity masking entity allowing for a high drug-to-antibody ratio (DAR) of 8, while improving the pharmacokinetics and tolerability of the drug; (iii) a proprietary dipeptide cleavable unit and iv) exatecan, a potent topoisomerase I inhibitor. MBK-103 showed excellent ex vivo stability in human plasma enabling a prolonged half-life, similar to that of the unconjugated monoclonal antibody. This led to an increased drug exposure, resulting in a potent antitumor efficacy observed in a collection of more than ten in vivo mouse tumor models with a diverse FOLR1 expression, already at doses as low as 1-3 mg/kg. In addition, the distinct mode of action of exatecan along with the unique characteristics of the proprietary drug-linker platform resulted in the in vivo antitumor potency obtained also in colorectal cancer models, in which tubulin-inhibiting agents exhibit only limited therapeutic effects. The compound was also very well tolerated in cynomolgus monkeys with HNSTD at 50 mg/kg and even when administered repeatedly. High conjugation yields along with 100% homogeneity are major drivers of the successful ongoing manufacturing process with planned IND submission in Q4/2023.

Citation Format: Warren Viricel, Louise Conilh, Edouard Leroy, Jean-Guillaume Lafay, Frederique Brune, Lenka Kyrych Sadilkova, Charles Dumontet. MBK-103, a potent novel conjugation platform-based antibody-drug conjugate, changing therapeutic options in folate receptor alpha positive cancer patients [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 1544.
Abstract 1664: Discovery of a novel and potent TACC3 protein-protein interaction inhibitor targeting highly aggressive cancers
Authors: Sahin Ozgur , Banoglu Erden , Lim Chaemin , Caliskan Burcu , Cruz Emmanuel , Stergiopoulos Sotirios , Vempati Sridhar
Published: June 2023

TACC3 is one of the most oncogenic members of the Transforming Acidic Coiled-Coil Containing (TACC) protein family highly overexpressed in multiple different cancers leading to worse prognosis. TACC3 is a key player in microtubule and centrosomal processes and is considered to have an important role in causing chromosomal instability leading to aggressive cancers. TACC3 also plays important role in DNA damage response mechanisms, hematopoiesis, epigenetics and immune functions by interacting with different protein partners. Inhibition of TACC3 by genomic approaches has shown significant reduction in growth of cancer cells in vitro and in vivo. Given its multiple roles in cancer, TACC3 is an ideal therapeutic target for highly aggressive cancers. Here, using phenotypic screening and rational drug design, we developed AO-252, a novel small molecule TACC3 protein-protein interaction (PPI) inhibitor. Binding assays and kinome screening suggested the specificity of AO-252 to TACC3. AO-252 targets the interactions of TACC3 with its known protein partners e.g., Clathrin/KIFC1, BARD1 and MBD2/HAT complexes regulating mitosis, DNA damage response and epigenetic functions. Gene expression profiling in multiple cell lines further confirmed the various TACC3 pathways affected by AO-252. AO-252 showed low nanomolar potency in >200 cell line panel covering various cancer indications and spared the normal cells. AO-252 showed excellent oral pharmacokinetics and demonstrated strong tumor growth inhibition or regression in multiple xenograft models without any safety concerns. Currently, AO-252 is prepared for IND submission for Q1 2023 and will enter clinic in the first half of 2023 for the treatment of advanced solid tumors including triple-negative breast cancer (TNBC), high-grade serous ovarian carcinoma (HGSOC), and endometrial cancer.

Citation Format: Ozgur Sahin, Erden Banoglu, Chaemin Lim, Burcu Caliskan, Emmanuel Cruz, Sotirios Stergiopoulos, Sridhar Vempati. Discovery of a novel and potent TACC3 protein-protein interaction inhibitor targeting highly aggressive cancers [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 1664.
Abstract 1665: Imipramine enhances the efficacy of lomustine by regulating WNT-β-catenin signaling in glioblastoma
Authors: Venkata Prabhakar Pitta , Rao Arhan , Timilsina Santosh , Subbarayalu Panneerdoss , Brenner Andrew , Sareddy Gangadhara Reddy , Rao Manjeet , Kelly William , Vadlamudi Ratna
Published: June 2023

Background: Glioblastoma (GBM) is one of the most aggressive and difficult-to-treat brain cancers. Surgical excision followed by concurrent temozolomide (TMZ), and radiation therapy are the current standard of care for GBM. Lomustine, another alkylating regimen, has also been approved for the treatment of patients with recurrent GBM in addition to TMZ. However, the primary GBM frequently recur as therapy-resistant relapses resulting in shorter survival of patients. To date, no systemic therapies have shown proven survival benefit in recurrent GBM. In this study, we examined whether FDA approved anti-depressant drug Imipramine serve as the potent option for treating patients with recurrent GBM and to improve efficacy of approved chemotherapy.

Methods: We investigated the anticancer effects of Imipramine using both established and patient-derived GBM cell lines. We tested the effect of Imipramine and Lomustine combination therapy using cell viability, colony formation, migration and apoptosis assays. Mechanistic studies were conducted using RNA-seq, western blotting, RT-PCR and reporter assays. In vivo efficacy of combination therapy was evaluated using orthotopic mouse models.

Results: Our results showed that antidepressant Imipramine blocked the growth of recurrent GBM. Furthermore, Imipramine enhanced the response of Lomustine in therapy resistant GBM cells as a combination therapy and significantly reduced the cell survival and migration, and increased the apoptosis in both established and patient derived GBM cells. Further, our results demonstrated that Imipramine inhibits growth of self-renewing cancer stem cells, which contribute to drug resistance and relapse of GBM. Mechanistic studies revealed that the Imipramine treatment abrogated the WNT-β-catenin signaling, which was confirmed by TOPFLASH reporter assay.

Conclusion: Collectively, the results from our study suggest that Imipramine alone and in combination with Lomustine can be used for treating recurrent GBM. Based on these results, a clinical trial is currently ongoing, where we are testing the efficacy of Imipramine for treating patients with recurrent GBM.

Citation Format: Prabhakar Pitta Venkata, Arhan Rao, Santosh Timilsina, Panneerdoss Subbarayalu, Andrew Brenner, Gangadhara Reddy Sareddy, Manjeet Rao, William Kelly, Ratna Vadlamudi. Imipramine enhances the efficacy of lomustine by regulating WNT-β-catenin signaling in glioblastoma [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 1665.
Abstract 1699: Targeting mercapturic acid pathway transporter in chemoradiotherapy resistant ovarian cancer
Authors: Singhal Sharad S. , Yue Er , Kulkarni Prakash , Horne David , Awasthi Sanjay , Wang Edward , Salgia Ravi
Published: June 2023

Ovarian cancer is the most common and the most lethal malignancies arising in the female reproductive system. Morbidity and mortality associated with ovarian cancer will continue to impact the female population. Although cisplatin centered chemotherapy is the first line anticancer agent for human ovarian cancer, chemoresistance and adverse side effects remain major hurdles to successful treatment. The effective targets for cancer therapy must be desirably expressed differentially in particular cancers as compared with normal cells. Also, there should be an essential dependence of cancer cells on the target compared to non malignant cells. Ideally, the target should be understood in the context of existing biochemical and signaling frameworks known to play a direct role in a particular carcinogenesis or in regulating the response to therapy. In the context of striking chemo radiotherapy resistance of ovarian cancer and the fundamental role of RLIP as an important mercapturic acid pathway transporter that is essential for survival and therapy resistance in cancers, we investigated the role of RLIP in regulating the critical signaling proteins involved in relaying the inputs from multiple upstream survival pathways and mechanisms contributing to chemo radiotherapy resistance in ovarian cancer. Compounds that inhibit, deplete, or downregulate RLIP will function as wide spectrum agents to treat ovarian cancer, independent of common signaling pathway mutations. Here, we demonstrate that RLIP expression is significantly greater in ovarian cancer cells than in non malignant cells, and RLIP is an essential requirement for cancer formation, and its depletion or inhibition will prevent ovarian carcinogenesis as well as metastases. Taken together, the results of our studies provided insights into a novel anticancer effect of RLIP depletion/inhibition on ovarian cancer and might open new therapeutic avenues for the treatment of ovarian cancer. Supported in part by the Department of Defense grant W81XWH 22 1 0331. Funding from the Beckman Research Institute of City of Hope is also acknowledged.

Citation Format: Sharad S. Singhal, Er Yue, Prakash Kulkarni, David Horne, Sanjay Awasthi, Edward Wang, Ravi Salgia. Targeting mercapturic acid pathway transporter in chemoradiotherapy resistant 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 1699.
Abstract 2028: Using bioinformatic analysis of proteomic data to probe novel drug mechanism of action
Authors: Chen Weining , Woods Nicholas , Qiao Fangfang , Law Henry Chun Hin , Jagadesan Sankarasubramanian , Guda Chittibabu , Bergan Raymond C.
Published: June 2023

Introduction: Metastasis is a major cause of death for patients with solid tumors. Our group has discovered a selective inhibitor of cell motility, KBU2046, that blocks the activation of Raf1 by inhibiting the phosphorylation of ser338 on its activation motif (Nature Communications 2018). To better understand the effect of KBU2046 on human prostate cancer (PCa) cells, we undertook a proteomic analysis. Experimental Procedures: After treatment of PC3 cells with KBU2046 or vehicle (N=3/group), protein expression of whole cell lysate was quantified by Liquid Chromatography/Tandem Mass Spectrometry analysis (LC-MS/MS). In separate experiments (N=4), membrane fractions were isolated, and LC-MS/MS was performed after Tandem Mass Tag (TMT) labeling. Resultant data were analyzed by several complementary bioinformatics tools, including: Gene Set Enrichment Analysis (GSEA), Gene Ontology (GO) term analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, Ingenuity Pathway Analysis (IPA), and protein-protein interaction (PPI) network analysis to characterize the proteins that were significantly differentially expressed upon KBU2046 treatment.

Results: Independent bioinformatic analyses of differentially expressed proteins affected by KBU2046 treatment identify significantly enriched processes related to cell motility, inclusive of effects on structural machinery. This constitutes orthogonal confirmation of KBU2046’s known effects on cell motility. Of importance, major effects on cellular energy-generating processes were also identified. This is consistent with prior studies that found changes in motility induce obligatorye changes in cellular energy requirements. Our focused analysis of membrane fractions identified changes in membranous compartments, providing a relevant positive control for bioinformatic analysis, which we confirmed by Western blot. Analytics also pointed to a major role for adhesion proteins in response to KBU2046, consistent with our prior identification of primary effects on cell migration.

Conclusions: These findings provide orthogonal mechanistically relevant support for a novel acting agent. They highlight the ability of proteomics and bioinformatic tools to efficiently probe the mechanism of action and global impact on PCa in response to treatment.

Citation Format: Weining Chen, Nicholas Woods, Fangfang Qiao, Henry Chun Hin Law, Sankarasubramanian Jagadesan, Chittibabu Guda, Raymond C. Bergan. Using bioinformatic analysis of proteomic data to probe novel drug mechanism of action [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 2028.
Abstract 2089: Vanishing deviance problem in high-dimensional penalized Cox regression
Authors: Yao Sijie , Li Tingyi , Cao Biwei , Wang Xuefeng
Published: June 2023

Penalized regression models, such as LASSO and elastic net, have been well-established for high-dimensional variable sections. For the analysis of censored survival outcomes, the penalized Cox regression model has been widely used. The parameter tuning in the penalized regression is often implemented based on the cross-validation (CV) via the “one standard error rule” (1se rule), resulting in a more parsimonious model. However, in a high-dimensional Cox regression model, a common observation is that the CV plot for tuning parameters against partial-likelihood deviance is often not U-shaped but L-shaped. In this case, the model selection will fail with either the “1se” or the“min” rule. We observed that this phenomenon is quite universal in the setting of high-dimensional prognostic biomarker selection, in which the standard CV steps often result in no or very few features included in the final model. We refer to this phenomenon as the “vanishing deviance” (VD) problem. In this study, we investigated this problem in more depth and demonstrated the issue and potential solution using real data from pan-cancer analysis of prognostic lncRNA biomarkers. To discover the cause behind the VD problem, we conducted an extensive set of simulations by varying sample sizes, feature dimensions, censoring rates, number of true biomarkers, and significance levels of true signals.Our results indicated that the main cause of the VD problem can be related to the limited sample size in the cancer omics dataset and a large number of true (or highly associated) biomarkers or signals underlying. To this end, we propose computational solutions that can alleviate the problem and we implemented them in the R package called CoxHive. We verified the performance of our solution based on both simulation studies and the analyses of the pan-cancer gene expression data. This software tool will greatly facilitate biomarker selection (including genomic and imaging biomarkers) in the high-dimensional setting.

Citation Format: Sijie Yao, Tingyi Li, Biwei Cao, Xuefeng Wang. Vanishing deviance problem in high-dimensional penalized Cox regression [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 2089.
Abstract 2100: Genomic profiles and immune cell infiltration landscapes for lymph node metastasis in T1 lung adenocarcinoma
Authors: Wu Fang , Pan Yue , Hu Chunhong , Chen Chen , Liu Wenliang , Fan Songqing , Shu Long , Zhao Lishu , Fu Yucheng , Zhang Sujuan , Liu Junqi , Zeng Yue , Peng Yurong , Zang Hongjing , Deng Chao , Qiu Zhenhua , Ma Fang , Yu Fenglei , Liu Xianling , Liu Lijuan , Yang Lingling , Shao Yang
Published: June 2023

Background: Comprehensive genomic and tumor immune microenvironment (TIME) analysis of T1 lung adenocarcinoma (LAD) may provide new insights into the biology of lymph node metastasis (LNM), thereby guiding surgical procedures and perioperative treatment.

Methods: A total of 212 cases of T1 LAD were included, of which 174 cases underwent whole exome sequencing to assess gene mutation and tumor mutation burden (TMB). Tumor mutation score (TMS) was constructed from LNM identified by LASSO regression. Immunohistochemical staining was performed to assess PD-L1 levels, tumor-infiltrating lymphocytes (TILs) and tumor-associated macrophages (TAMs) densities. Validation and supplementation were performed by RNA sequencing data from the TCGA database.

Results: A total of 25 genes were associated with the risk of LNM, of which STK11, NOTCH2, RHBDF2, EGFR, and CYP2D6 could be validated by RNA sequencing. The model predicted LNM (AUC = 0.904) and 2-year recurrence and metastasis (AUC = 0.700). Besides, TILs were negatively correlated with LNM of T1 LAD, while TAMs were positively (p<0.05). Moreover, the TMS was correlated with the TIME, particularly TAMs(r=0.331, p<0.001).

Conclusions: Tumor gene mutation promotes lymph node metastasis by changing the TIME, especially TAMs. Predictive models can identify high-risk patients and provide evidence for future studies of perioperative management.

Citation Format: Fang Wu, Yue Pan, Chunhong Hu, Chen Chen, Wenliang Liu, Songqing Fan, Long Shu, Lishu Zhao, Yucheng Fu, Sujuan Zhang, Junqi Liu, Yue Zeng, Yurong Peng, Hongjing Zang, Chao Deng, Zhenhua Qiu, Fang Ma, Fenglei Yu, Xianling Liu, Lijuan Liu, Lingling Yang, Yang Shao. Genomic profiles and immune cell infiltration landscapes for lymph node metastasis in T1 lung adenocarcinoma [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 2100.
Abstract 2101: Cancer-associated fibroblast activation predicts progression, metastasis, and prognosis of cutaneous squamous cell carcinoma
Authors: Knuutila Jaakko S. , Riihilä Pilvi , Nissinen Liisa , Kallionpää Roosa , Pellinen Teijo , Kähäri Veli-Matti
Published: June 2023

Purpose: Cutaneous squamous cell carcinoma (cSCC) is the most common metastatic skin cancer and the metastatic disease is associated with poor prognosis. At present, no established biomarkers are available for predicting the metastasis risk and prognosis of patients with cSCC. Cancer-associated fibroblasts (CAFs) promote the progression of cancer, but their role in cSCC is largely unknown. Here, we have examined the potential of CAF markers in the assessment of metastasis risk and prognosis of primary cSCC.

Experimental Design: We utilized multiplexed fluorescence immunohistochemistry for profiling of CAF landscape in metastatic and non-metastatic primary human cSCCs, the actual metastases, and in premalignant and benign epidermal lesions. Quantitative high-resolution image analysis was performed with two separate panels of antibodies for CAF markers and the results were correlated with clinical and histopathological parameters including disease-specific mortality.

Results: Increased stromal expression of fibroblast activation protein (FAP), α-smooth muscle actin, and secreted protein acidic and rich in cysteine (SPARC) were associated with progression of premalignant epidermal lesions to invasive cSCC. Furthermore, elevation of FAP and platelet-derived growth factor receptor-β (PDGFRβ) expression was associated with the risk of metastasis of primary cSCCs. High expression of PDGFRβ and periostin correlated with poor prognosis. Furthermore, a multi-marker defined CAF subset, PDGFRα-/PDGFRβ+/FAP+, was associated with invasion and metastasis and independently predicted poor disease-specific survival.

Conclusions: These results show, that fibroblast activation evolves during the progression of cSCC. High PDGFRβ expression alone and multimarker combination PDGFRα-/PDGFRβ+/FAP+ are potential biomarkers for the progression of primary cSCC to metastatic stage, and for poor prognosis.

Citation Format: Jaakko S. Knuutila, Pilvi Riihilä, Liisa Nissinen, Roosa Kallionpää, Teijo Pellinen, Veli-Matti Kähäri. Cancer-associated fibroblast activation predicts progression, metastasis, and prognosis of cutaneous squamous cell carcinoma [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 2101.
Abstract 2215: Periostin plays a crucial role in the recurrence of head and neck squamous cell carcinoma
Authors: Park Jung Je , Seo Ji Hyun , Won Seongjun , Ryu Somi , Baek Seung-Kuk , Koh Yoon Woo
Published: June 2023

Periostin (POSTN) is associated with many human tumors, such as colon, lung, breast, ovarian, prostate, and head and neck cancers. In this study, we focused on POSTN in Head and neck squamous cell carcinoma(HNSCC) because little is known about its possible role in carcinogenesis. This study aims to investigate the role of POSTN in the recurrence and prognosis of tongue cancer. A total of 97 patients diagnosed with tongue cancer in Gyeongsang National University Hospital between Jan 2012 and Dec 2017 were included. A retrospective review of medical records was performed, and the immunohistochemical staining with the POSTN antibody was done. Statistical analysis between cancer progression and POSTN expression was done. Referring to the clinical results that POSTN was related to the recurrence of tongue cancer, the effects of POSTN on HNSCC cell lines were investigated. Overexpression and depletion of POSTN were established in the HNSCC cell line. The results of POSTN in the HNSCC cell line were evaluated by colony-forming, migration, invasion, wound healing, and flow cytometry in vitro and in vivo. We also investigated the effects of POSTN overexpression and depletion on invasion, migration, and cell apoptosis. High expression of POSTN in HNSCC was related to cancer recurrence. In multivariate analyses, High expression of POSTN was a statistically significant risk factor for tongue cancer recurrence (HR, 16.8; p=0.040). Also, POSTN inhibits colony-forming, migration, invasion, and wound healing. This study showed that POSTN in HNSCC is a positive correlation with recurrence. And we also confirmed that POSTN promoted tumor progression in vitro and in vivo. POSTN plays an essential role in the growth and metastasis of tongue cancer and may be positioned as a therapeutic target for head and neck cancer.

Citation Format: Jung Je Park, Ji Hyun Seo, Seongjun Won, Somi Ryu, Seung-Kuk Baek, Yoon Woo Koh. Periostin plays a crucial role in the recurrence of head and neck squamous cell carcinoma [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 2215.
Abstract 2288: Longitudinal measurement of HPV copy number in cell-free DNA predicts progression-free survival in HPV-positive oropharyngeal cancer patients
Authors: Adamowicz Martyna Joanna , Carey Lara M. , Robert Christelle , Esnal-Zufiaurre Arantza , Warlow Sophie J. , Wescott Robert , Cuschieri Kate , Conn Brendan , Hay Ashley , Nixon Iain J. , Aitman Timothy J.
Published: June 2023

Oropharyngeal squamous cell carcinoma (OPSCC) is increasing in global prevalence and is divided into two types dependent on association with human papillomavirus (HPV), with a more favorable prognosis in HPV+ve tumors. Assay of HPV copy number in plasma cell-free DNA (cfDNA) provides a minimally invasive method for detecting and monitoring tumor-derived HPV, with potential for enhancing clinical care. We have evaluated the utility of cfDNA droplet digital PCR (ddPCR) as a method for characterization and longitudinal monitoring of patients with OPSCC in a prospectively recruited cohort of 163 HPV+ve OPSCC patients. ddPCR assay of cfDNA for five HPV types showed overall 96.4% concordance with both p16 immunohistochemistry (IHC) and broad spectrum PCR analysis of solid tumor tissue. Longitudinal sampling in 105 HPV+ve patients, with median follow-up of 29 months, strongly predicted patient outcomes. Progression-free survival, stratified respectively by the presence or absence of detectable HPV cfDNA at a median of 13 weeks post-treatment, was 50% and 88% (p=0.001, hazard ratio 10.0 (95% CI 2.1-47.1)). In three patients sequential HPV measurement indicated cancer recurrence before the presence of clinical symptoms or detection on imaging. In two patients, greater reliance on sequential HPV measurement would have avoided surgical intervention which ultimately did not confirm disease presence. The high concordance of pre-treatment plasma cfDNA-HPV analysis with p16 IHC and HPV-PCR of solid tissue, together with the predictive value and utility of sequentially measured post-treatment cfDNA-HPV copy number, provide a compelling case for the routine use of cfDNA-HPV ddPCR in diagnostic classification and longitudinal monitoring of OPSCC.

Citation Format: Martyna Joanna Adamowicz, Lara M. Carey, Christelle Robert, Arantza Esnal-Zufiaurre, Sophie J. Warlow, Robert Wescott, Kate Cuschieri, Brendan Conn, Ashley Hay, Iain J. Nixon, Timothy J. Aitman. Longitudinal measurement of HPV copy number in cell-free DNA predicts progression-free survival in HPV-positive oropharyngeal cancer patients [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 2288.
Abstract 2856: Age-induced chronic accumulation of cortisol drives chemotherapy resistance in lung cancer
Authors: Raizada Devesh , Drapela Stanislav , Tejero Joanne , Sarigul Nadir , Ilter Didem , Gomes Ana
Published: June 2023

Lung cancer accounts for the largest number of cancer-associated deaths in the United States. While great strides have been made due to the introduction of immunotherapies and targeted therapies against oncogenic drivers, chemotherapies remain the standard of care for the majority of lung cancer patients. However, many patients do not respond to these treatments or relapse following an initial response. We postulate that part of the problem is the lack of consideration in preclinical research and clinical trials of the main driver of lung tumorigenesis, the aging process. Here, we show that the organismal reprogramming that occurs with aging makes lung tumors in an aged host more conducive to withstand chemotherapies. We traced this to an age-induced chronic accumulation of the stress hormone cortisol in circulation. Mechanistically our data show that chronic accumulation of cortisol drives the upregulation of metallothioneins—small, highly conserved, cysteine-rich metal-binding proteins—through activation of the glucocorticoid receptor in lung cancer cells, which in turn rewire these cells to be refractory to chemotherapies. Together, our work demonstrates a role for age-induced cortisol levels in promoting resistance to standard of care chemotherapies in lung cancer and offers a rationale for using therapeutic agents that block the glucocorticoid receptor to sensitize the average and most vulnerable lung cancer patients, the elderly, to chemotherapy.

Citation Format: Devesh Raizada, Stanislav Drapela, Joanne Tejero, Nadir Sarigul, Didem Ilter, Ana Gomes. Age-induced chronic accumulation of cortisol drives chemotherapy resistance in lung 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 2856.
Abstract 2923: JAB-X1800: A potent immunostimulatory antibody-drug conjugate (iADC) targeting CD73
Authors: Ye Yang , Yang Guiqun , Wang Hongling , Sun Xin , Wang Yanping , Yan Fashun , Wang Lei , Long Wei
Published: June 2023

Background: STING emerges as a key immune sensor and activated STING pathway plays a key role in innate and adaptive immune response. STING agonist has shown promising potency to stimulate anti-tumor immunity. Considering immune-related toxicity due to systemic administration, as well as difficulty in intra-tumor administration, targeted delivery of STING agonist by iADC is warranted. We developed CD73-STING iADC JAB-X1800, which uses anti-CD73 mAb for targeted delivery of payload JAB-27670, a highly potent non-cyclic dinucleotide STING agonist.

Method: Binding assay and HEK293-STING-luciferase reporter assay were performed to test affinity of JAB-27670. Tumor cells with CD73 expression and THP-1 cells were co-cultured in the presences of iADC JAB-X1800 to assess the IFN-β production. The cell killing activity of iADC JAB-X1800 was evaluated when cancer cells co-cultured with human PBMCs. Furthermore, human and mouse plasma stability of iADC JAB-X1800 was assessed. Finally, the anti-tumor effects of iADC JAB-X1800 alone or in combination with anti-PD-1 were evaluated in animal models.

Results: JAB-27670 is a highly potent STING agonist with sub-nanomolar EC50 in binding and luciferase reporter assays. In CD73-expressing cancer cells and THP-1 co-culture experiments, iADC JAB-X1800 induced IFN-β production with significant improvement compared with the free payload. JAB-X1800 also induced potent killing in vitro. JAB-X1800 is stable in human and mouse plasma. A single dose of JAB-X1800 induced tumor regression in MDA-MB-231 xenograft model and hCD73-MC38 syngeneic model. JAB-X1800 conferred immunologic memory when tumor free mice were re-challenged. JAB-X1800 showed synergistic anti-tumor activity with anti-PD-1 in hCD73-MC38 syngeneic model.

Conclusions: The CD73-targeted STING agonist iADC JAB-X1800 demonstrated high stability in plasma, robust activity, and good tolerability in animal models, supporting clinical development of JAB-X1800 alone and in combination with anti-PD-1 for patients with CD73-expressing tumors.

Citation Format: Yang Ye, Guiqun Yang, Hongling Wang, Xin Sun, Yanping Wang, Fashun Yan, Lei Wang, Wei Long. JAB-X1800: A potent immunostimulatory antibody-drug conjugate (iADC) targeting CD73 [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 2923.
Abstract 3093: Synthesis and bioevaluation of LAT-1 efflux agonists for the treatment of pancreatic cancers
Authors: Moots Holly , Ikhlef Houssine , Sekhar Vandana , Phanstiel Otto
Published: June 2023

The purpose of this report is to describe the discovery and development of the world’s first L-type amino acid transporter 1 (LAT-1) efflux agonist. LAT-1 is used for the import of amino acid substrates (leucine, phenylalanine, and methionine) and LAT-1 expression has been showed to be increased in pancreatic ductal adenocarcinoma (PDAC) cells. LAT-1 is a promising target for the treatment of rapidly growing cancers, such as PDAC, in part, due to its key role in methionine import. Methionine is required for various cellular processes, including the generation of S-adenosylmethionine (SAM) and the maintenance of intracellular polyamine pools, which are essential for PDAC growth. Indeed, the biosynthesis of the higher polyamines (spermidine and spermine) requires the transfer of an aminopropyl group from decarboxylated S-adenosylmethionine (dc-SAM), illustrating the molecular connection between methionine and polyamine pools.

LAT-1 inhibition, such as with the known LAT-1 inhibitor JPH203, has shown efficacy in suppressing tumor growth in pancreatic cancer cell lines. While the early pre-clinical data is promising, competitive inhibitors of LAT-1 may have limited potency in the presence of high extracellular levels of LAT-1 substrates.

In contrast, LAT-1 efflux agonists are insensitive to extracellular LAT-1 substrates. These novel compounds reverse the flow of substrates through LAT-1, effectively depleting cells of key intracellular amino acids, rather than blocking the import of extracellular substrates like methionine.

The chiral piperazine (1), our lead efflux agonist, was discovered by a high throughput screen and was shown to deplete intracellular methionine, leucine and phenylalanine pools by LC-MS as well as decrease the synthesis of the higher polyamines (spermidine and spermine), a process which is reliant upon methionine as a precursor. A structure activity relationship (SAR) analysis of the four possible diastereomers of 1 was performed. The performance of these diastereomers and other derivatives were evaluated by LC-MS, HPLC, and Western blot. These molecules were shown to increase 3H-leucine efflux, decrease intracellular polyamine, SAM, and S-adenosylhomocysteine (SAH) levels, decrease phosphatidylethanolamine levels and induce the autophagic marker LC3-II. In summary, this novel method of methionine restriction allows one to alter intracellular methionine levels in a dose-dependent manner and provides a novel way to control pancreatic cancer cell growth.

Citation Format: Holly Moots, Houssine Ikhlef, Vandana Sekhar, Otto Phanstiel. Synthesis and bioevaluation of LAT-1 efflux agonists for the treatment of pancreatic cancers [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 3093.
Abstract 3111: The Sygnature CHARMD platform - combinatorial high-throughput assembly and review of molecular degraders
Authors: Hjerpe Roland , Birch Louise , Brien Alex , Cusin Lola , Duffy Lorna , Gourdet Benoit , Lawrence Rachel , MacFaul Philip , Mortlock Hannah , Neal Hannah , Pearce Christopher , Pitti Erica , Robles Gonzalo , Rodrigues Edith , Scott Catriona , Singh Rajesh , Thomson Stuart , Zhang Yujia , Jordan Allan
Published: June 2023

Recent interest the ability to degrade oncogenic proteins has grown exponentially, and the advent of Protein-Targeting Chimeras (PROTACs) has allowed many previously undruggable targets to be specifically targeted. However, the successful development of these bioavailable heterobifunctional degraders relies on the identification of a combination of ligase recruiter, linker and exit vector that allow formation of a competent ternary complex and target degradation. To this end, conventional single-compound synthesis is time-consuming and poses challenges for selection of degrader components with predictive SAR and ADME properties of the assembled bifunctional compound.

Here, we demonstrate a high-throughput integrated platform for generation and assessment of bifunctional degraders, incorporating combinatorial chemistry using both commercial and bespoke linkers, cell-based activity assays, and assessment of in vitro DMPK properties (lipophilicity, metabolic stability and experimental polar surface area). Alongside this, we have in place computational methods for ternary complex modelling, linker design and PROTAC prioritisation.

Altogether, our approach facilitates rapid screening and identification of bifunctional lead compounds for challenging oncology targets with the potential to deliver significant patient benefit. Our platform approach has been exemplified through application to oncology targets of significant current interests and our results in this area will be highlighted.

Citation Format: Roland Hjerpe, Louise Birch, Alex Brien, Lola Cusin, Lorna Duffy, Benoit Gourdet, Rachel Lawrence, Philip MacFaul, Hannah Mortlock, Hannah Neal, Christopher Pearce, Erica Pitti, Gonzalo Robles, Edith Rodrigues, Catriona Scott, Rajesh Singh, Stuart Thomson, Yujia Zhang, Allan Jordan. The Sygnature CHARMD platform - combinatorial high-throughput assembly and review of molecular degraders [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 3111.
Abstract 3161: Novel gene signature for predicting aggressive type of bladder cancer and the associated therapeutic strategies
Authors: Sheu Jim Jinn-Chyuan , Chang Chung , Lin Bo-Chen , Hwang Tritium
Published: June 2023

We have previously developed a unique HD-MAC (high-dimensional analysis of molecular alterations in cancer) platform that enables us to define independent tumor biomarkers from a group of clinically-significant genes. Using this platform, 26 genes were identified as independent prognosis biomarkers whose overexpression determine shorter overall survivals in patients with bladder cancer. Through clinical association study (including tumor stage, lymph node invasion, angiogenesis, and metastasis), a gene signature containing 6 biomarkers was established which can be validated by two independent cohorts. Gene knock down effects on cancer cell survival further revealed PPT2 and PRPF19 as potential targets for target-based therapy. Correlation study between genetic dependencies and drug sensitivities across cell lines suggested several potent lead compounds which are likely to function through similar mechanisms of gene effect actions. Our study therefore provides newly defined prognostic biomarkers as well as the associated therapeutic strategies for possible clinical applications in patients with bladder cancer.

Citation Format: Jim Jinn-Chyuan Sheu, Chung Chang, Bo-Chen Lin, Tritium Hwang. Novel gene signature for predicting aggressive type of bladder cancer and the associated therapeutic strategies [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 3161.
Abstract 3162: Genome-scale metabolic modeling suggests novel molecular targets in ER-positive breast cancer
Authors: Mudge Zachary B. , Kovacs Kristof , Kemp Melissa L. , Badve Sunil S. , Gokmen-Polar Yesim
Published: June 2023

High expression of the epithelial splicing regulatory protein 1 (ESRP1) is correlated with poor prognosis in estrogen receptor positive (ER+) breast cancer and acquired resistance to tamoxifen/endocrine therapies. Our previous study revealed a novel functionality of ESRP1 in metabolic dysregulation by affecting lipid metabolism and oxidoreductase processes. To further understand how metabolic programming is affected by ESRP1 expression and identify novel therapeutic targets, we developed systems biology models to explore ER+ breast cancer metabolism. Flux balance analysis (FBA) is a computational approach to predict steady-state fluxes through metabolic reactions by posing cellular “objectives” such as maximization of biomass, ATP, or minimization of reactive oxygen species that reflect selective pressures posed by the tumor microenvironment or therapeutic insults. Our custom FBA pipeline leverages transcriptomic data, mutation information, and thermodynamic constraints to generate personalized genome-scale metabolic models. In this study, 100 ESRP1-high and 100 ESRP1-low tumors from the TCGA Breast Cancer Invasive Carcinoma dataset were simulated and compared; in a secondary analysis, we simulated metabolic rerouting of tamoxifen-resistant LCC2 cells and fulvestrant-resistant LCC9 cells upon ESRP1 knockdown. The intersection of statistically significant (p < 0.05) reactions that were altered in both the TCGA and cell line contexts suggested that changes in eicosanoid metabolism and androgen and estrogen synthesis pathways are highly involved in ESRP1's role in breast cancer prognosis and resistance to endocrine therapies. Eicosanoid-utilizing enzymes are consistently altered between the ESRP1-low and high conditions; Hematopoietic prostaglandin D synthase (HPGDS), prostaglandin I2 synthase (PTDIS), and thromboxane A synthase 1 (TBXAS1) are druggable. Furthermore, reactions associated with cytosolic and lysosomal 4-hydroxy-17beta-estradiol production, catalyzed by the enzymes myeloperoxidase, lactoperoxidase, and peroxiredoxin 6, also emerged as consistently altered across the groups. The findings from this methodology have suggested novel metabolic targets to overcome tamoxifen/endocrine therapy resistance in ER+ breast cancer. Experimental validation of these targets is currently underway.

Citation Format: Zachary B. Mudge, Kristof Kovacs, Melissa L. Kemp, Sunil S. Badve, Yesim Gokmen-Polar. Genome-scale metabolic modeling suggests novel molecular targets in ER-positive breast 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 3162.