bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 321: Metformin enhances the efficacy of enzalutamide treatment in castration-resistant prostate cancer
Authors: Simpson Kendall Elizabeth , Zhang ZhuangZhuang , Zhang Qiongsi , Liu Xiaoqi
Published: June 2023

Prostate cancer remains to be the most commonly diagnosed cancer among men in the United states with approximately 268,500 new cases a year. One of the first treatment options for men with prostate cancer is androgen deprivation therapy (ADT), however, recurrence and metastases after ADT remain an issue and are categorized as castration-resistant prostate cancer (CRPC). Enzalutamide, an FDA approved drug currently prescribed to patients with CRPC, inhibits androgen receptor (AR) nuclear translocation preventing the transcriptional activity of AR. This therapy typically only extends the survival of patients with CRPC and can lead to Enzalutamide-resistance over time accenting the importance of new therapy targets. Metformin, a commonly used FDA approved therapy for type 2 diabetes, has been recently investigated as potentially having anti-tumor effects in many cancer types. In this study, we use Enzalutamide and Metformin in combination to explore possible re-sensitization of Enzalutamide-resistant CRPC to Enzalutamide. Recently, we have tested the effects of this combination treatment on cell viability, drug synergy, and cell proliferation in Enzalutamide-resistant CRPC. After treatment, we see a decrease in cell proliferation and viability as well as a synergistic effect of both Enzalutamide and Metformin in Enzalutamide-resistant CRPC. Following these results, we sought to explore how this combination treatment effected key signaling markers for EMT, WNT signaling, and TGFβ signaling as well as key signaling markers for prostate cancer such as AR and prostate specific antigen (PSA). Since Metformin inhibits Complex I of oxidative phosphorylation, we also tested this combinational treatment using seahorse analysis to determine the mitochondrial respiration. After initial testing, we will continue to move forward in testing the effects of this combination treatment in Enzalutamide-resistant CRPC as well as further elucidate the underlying mechanisms contributing to this phenotype.

Citation Format: Kendall Elizabeth Simpson, ZhuangZhuang Zhang, Qiongsi Zhang, Xiaoqi Liu. Metformin enhances the efficacy of enzalutamide treatment in castration-resistant prostate 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 321.
Abstract 3264: Sotorasib, a selective G12C K-RAS inhibitor, modulates the tumor microenvironment to promote TNFα and IFNγ signaling and augment immunotherapy response
Authors: Ibrahim Mohammed L. , Foresman Dana , Zheng Hong , Haura Eric B. , Abate-Daga Daniel , Yu Xiaoqing , Beg Amer A.
Published: June 2023

KRAS-G12C mutation represents a highly prevalent mutation in lung and colon carcinoma with unfavorable clinical outcome. Recently, new compounds were developed that selectively bind to K-RAS with G12C mutation. While recent clinical results demonstrate activity of these compounds, their underlying mechanisms of action and impact on the antitumor immune response are not well understood. Here, we tested G12C-Kras inhibitor (Sotorasib/AMG-510) using multiple approaches. We found that Sotorasib significantly promoted the tumor regression and the expansion of tumor infiltrating lymphocytes (TILs) in both CT26-G12C heterotopic colon cancer, and KP-G12C autochthonous GEMM lung cancer mouse models, with more profound impact when combined with anti-PD1 ICB immunotherapy. In addition, it generated a “hot” tumor microenvironment with strikingly augmented T cell effector phenotype, endorsed by enrichment of MHC-II high macrophage and reduction of suppressive myeloid cells. Interestingly, Single cell RNA sequencing of CT26-G12C tumors shows an enrichment of tumor clusters with TNFα and IFNγ as top upregulated pathways in Sotorasib-treated tumors, generating a more immune active TME. These immune active tumor clusters were conserved or further expanded upon cotreatment with anti-PD1 ICI. Conversely, immune-deficient, or Sotorasib-resistant tumors exhibited a remarkable contraction of the immune active clusters, substantiating that the immune-modulatory impact of Sotorasib relies on tumor cells adaptation and their cross talk with the tumor infiltrated lymphocytes. In vitro, Single cell RNA sequencing of CT26-G12C cells also revealed that Sotorasib and TNFα/IFNγ cytokines are affecting different tumor clusters. Moreover, further in vitro testing demonstrated that Sotorasib dramatically enhances KRAS-G12C mutant cancer cell killing by PSCA-specific CART cells. These findings provide insight into the molecular basis of G12C-Kras inhibitor-mediated immune modulation in the TME and provide insight for combinatorial regimens with checkpoint inhibitor and adoptive T cell immunotherapy to achieve optimum anti-tumor activity.

Citation Format: Mohammed L. Ibrahim, Dana Foresman, Hong Zheng, Eric B. Haura, Daniel Abate-Daga, Xiaoqing Yu, Amer A. Beg. Sotorasib, a selective G12C K-RAS inhibitor, modulates the tumor microenvironment to promote TNFα and IFNγ signaling and augment immunotherapy response [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 3264.
Abstract 3338: Targeted serum metabolomics for noninvasive detection of colorectal neoplasia
Authors: Fitzgerald Liam W. , Orton Dennis J. , Kopciuk Karen A. , Vogel Hans J. , Hilsden Robert J. , Bathe Oliver F.
Published: June 2023

Background: Using a shotgun metabolomics approach, we have previously reported distinct features in the circulating metabolome of patients with colorectal cancer (CRC) and adenoma. A clinically useful metabolomic assay requires a targeted approach that detects CRC with high sensitivity, and which also detects pre-malignant precursor lesions. Our objective was to identify a metabolomic signature for CRC, adenomatous polyps (AP), and sessile serrated adenomas (SSA), using a targeted approach.

Methods: Serum from patients with all stages of CRC (N=86), AP (N=48), SSA (N=46), as well as age- and sex-matched disease-free controls (DFC; N=120) were analyzed using a targeted metabolomic assay (Biocrates MxP® Quant 500). The assay was performed on liquid chromatography-tandem mass spectrometry (LC-MS/MS), quantifying 630 metabolites across 26 biochemical classes. Orthogonal partial least squares-discriminant analysis (OPLS-DA) was used to identify metabolite patterns distinguishing each neoplastic condition relative to DFCs. A small validation cohort (N=78) was additionally analyzed to test our meta-biomarkers.

Results: In comparisons with DFCs, statistically significant OPLS-DA models were constructed for all three neoplastic conditions. The OPLS-DA models for CRC, AP, and SSA consisted of 39, 14, and 22 metabolites, respectively, with the most frequently represented biochemical classes being triglycerides, phosphatidylcholines, bile acids, and acylcarnitines. Based on 7-fold internal cross validation, receiver operating characteristic (ROC) analysis demonstrated an area under the curve (AUC) of 0.85, 0.89, and 0.82 for CRC, AP, and SSA, respectively. When applied to the independent validation cohort, the sensitivity and specificity of each model were: 87.0% and 87.0% for CRC; 53.3% and 93.3% for AP; and 78.6% and 57.1% for SSA.

Conclusion: Metabolomic meta-biomarkers for CRC and its precursors demonstrated excellent performance based on internal validation. Importantly, these lesions were each detected with superior sensitivity compared to readily available stool-tests. A large external validation study in progress will be used to confirm these findings.

Citation Format: Liam W. Fitzgerald, Dennis J. Orton, Karen A. Kopciuk, Hans J. Vogel, Robert J. Hilsden, Oliver F. Bathe. Targeted serum metabolomics for noninvasive detection of colorectal neoplasia [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 3338.
Abstract 3339: Circulating long non-coding RNA HULC as an early indicator of hepatocellular carcinoma irrespective of etiology
Authors: George Joseph , Tsuchishima Mutsumi , Tsutsumi Mikihiro
Published: June 2023

Background: Hepatocellular carcinoma (HCC) is the 6th most common cancer worldwide and is highly prevalent in Asian countries. Long noncoding RNAs (lncRNAs) are non-coding transcripts with a size of more than 200 bases in length. Several studies reported that lncRNAs are highly deregulated in HCC and could be used as biomarkers for early diagnosis and prognostic purposes. In the present study, we measured the circulating lncRNA HULC as an early diagnostic marker for HCC with different etiology.

Methods: Serum samples were collected from healthy controls, liver cirrhosis (LC), and HCC patients with different etiology and stored at -80°C until assay. Total RNA was extracted using Qiagen RNeasy kit and tested for RNA quantity and purity using NanoDrop spectrophotometer. The RNA extract that contains lncRNA HULC was transcribed into cDNA using a reverse transcription kit. Employing target specific primers for the lncRNA HULC, qRT-PCR was performed with the synthesized cDNA using Qiagen QuantiFast SYBR Green PCR kit on a Rotor-Gene qRT-PCR system.

Results: Quantification of lncRNA HULC using qRT-PCR in the serum samples demonstrated significant increase in LC cases and in all HCC cases irrespective of etiology compared to the normal healthy controls. The highest concentration was observed in serum samples from HCC patients with hepatitis B virus (HBV) infection, which was about 8 fold higher compared to healthy controls. There was no significant difference between the mean HULC lncRNA levels in HCC patients with different etiology such as alcoholic cirrhosis, LC due to other reasons, hepatitis B and hepatitis C virus (HCV) infections, and HCC developed from other unknown causes. However, serum HULC lncRNA levels in LC cases without HCC was significantly lower compared to all HCC samples.

Conclusions: The results of the present study demonstrated that there is a significant increase in HULC lncRNA levels in the serum samples of all HCC patients irrespective of etiology. Therefore, the circulating lncRNA HULC could be used as an early diagnostic biomarker for HCC before appearing other clinical symptoms.

Citation Format: Joseph George, Mutsumi Tsuchishima, Mikihiro Tsutsumi. Circulating long non-coding RNA HULC as an early indicator of hepatocellular carcinoma irrespective of etiology [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 3339.
Abstract 3344: Early detection biomarkers of LncRNA H19 genotypes for determining childhood leukemia susceptibility
Authors: Tsai Chia-Wen , Chang Wen-Shin , Chen Chao-Chun , Hsu Pei-Chen , Wang Yun-Chi , Pei Jen-Sheng , Bau Da-Tian
Published: June 2023

The purpose of our study was to investigate whether genetic variations in lncRNA H19 were associated with susceptibility to childhood leukemia. 266 childhood leukemia patients and 266 healthy controls were enrolled in Taiwan, and two single nucleotide polymorphisms (SNPs), rs2839698 and rs217727, in H19 were genotyped and analyzed. There was a significant difference in the genotypic distribution of rs2839698 between patients and healthy controls (p=0.0277). Compared to the wild-type CC genotype, the heterozygous variant CT and homozygous variant TT genotypes were associated with significantly increased risks of childhood leukemia with an adjusted odds ratio (OR) of 1.46 (95% confidence interval [CI], 1.08-2.14, p=0.0429) and 1.94 (95%CI, 1.15-3.31, p=0.0169), respectively (p for tread=0.0277). The difference in allelic frequencies between childhood leukemia patients and controls was also significant (T versus C,

adjusted OR = 1.53, 95%CI, 1.13-1.79, p=0.0077). There were no significant differences in the genotypic and allelic distributions of rs217727 between cases and controls. Our results indicated that H19 SNP rs2839698, but not rs217727, may serve as a novel susceptibility marker for childhood leukemia.

Citation Format: Chia-Wen Tsai, Wen-Shin Chang, Chao-Chun Chen, Pei-Chen Hsu, Yun-Chi Wang, Jen-Sheng Pei, Da-Tian Bau. Early detection biomarkers of LncRNA H19 genotypes for determining childhood leukemia susceptibility [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 3344.
Abstract 3589: The use of a novel orthotopic xenograft model of fusion negative rhabdomyosarcoma to study the effect of ASAP1 on tumor progression and metastasis
Authors: Hammoudeh Sarah M. , Ng Yeap , Yadav Mukesh P. , Weigert Roberto , Randazzo Paul A.
Published: June 2023

ASAP1 is a multidomain ADP-ribosylation factor (Arf) GTPase-activating protein (GAP) involved in the regulation of the actin cytoskeleton, focal adhesions dynamics and receptor tyrosine kinases trafficking. ASAP1 expression levels correlate with progression of solid tumors (e.g. breast, colorectal, pancreatic, ovarian, gastric, and prostate cancer). The first evidence for a role of ASAP1 in cancer progression was the discovery that the amplification of ASAP1 gene on chromosome 8 correlates with poor prognosis in uveal melanoma. A similar amplification of chromosome 8 has been reported in ~72% of fusion negative rhabdomyosarcoma (FN-RMS) tumors, suggesting a potential role for ASAP1 signaling in the progression of FN-RMS. To determine the function of ASAP1 in tumor growth, invasion and metastasis, we used a tongue orthotopic xenograft of FN-RMS which recapitulates physiologically relevant tumor progression and metastasis to lymph nodes and lungs, two of the common metastatic sites in patients. Using intravital microscopy, we observed that injected FN-RMS cells formed elongated structures and exhibited dynamic branching, which might represent local invasion. H&E stained of fixed tissue revealed invasion into the tongue and local metastasis. Metastases to the regional lymph nodes, lower mandible and lungs were also observed. Using this system, we are now able to determine effect of knocking down and knocking out ASAP1 on the progression of the xenografts, including invasion in real time and metastasis.

Citation Format: Sarah M. Hammoudeh, Yeap Ng, Mukesh P. Yadav, Roberto Weigert, Paul A. Randazzo. The use of a novel orthotopic xenograft model of fusion negative rhabdomyosarcoma to study the effect of ASAP1 on tumor progression 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 3589.
Abstract 3622: Characterizing the metastatic potential and clonal dynamics of invasive subpopulations in TNBC cells in response to chemotherapy
Authors: De Santiago Carolina , Brock Amy
Published: June 2023

Triple negative breast cancer (TNBC) is one of the deadliest forms of breast cancer and is characterized by its high rates of metastasis, significant intra-tumor heterogeneity and lack of hormonal treatments. Through utilization of a novel genetic cell tracking technology, ClonMapper, we identify and characterize the origins of invasive subpopulations.

Genetic barcodes are stably integrated into HCC1806 cells to generate starting parental populations of ~1000 unique barcoded cells. Invasive barcoded subpopulations were selected by migration through transwell assays for 24 hours and then collected for expansion. Clonal abundance sequencing across all replicates showed a >8-fold decrease in the number of unique barcodes. Similarity was quantified using the Bray-Curtis and Cosine similarity indexes. 5 out of 8 replicates shared the same lineages, where more than 50% of the replicates were composed of the same 9 unique barcodes thus indicating the pre-existence of invasive clones. Following cell subpopulation isolation, we observed that isolated subpopulations display faster growth rates than parental populations. A scratch assay verified that the 5 similar invasive subpopulations displayed elevated migration rates in this 2D migration format (invasive populations vs parental population, n= 6). Separate transwell assays also confirmed the 3-dimensional migration rates of isolated subpopulations.

Parental HCC1806 and invasive subpopulations were treated with doxorubicin, cisplatin, paclitaxel and 5-flourouracil (5-FU) at their respective LD50 for 48 hours then allowed to recover to confluency before assessment by scratch assay. We observed that paclitaxel significantly decreased the migration rate, but that both doxorubicin and 5-FU resistant cells had a significantly higher migration rate (chemoresistant populations vs parental population, n=5) indicating that these drugs select for more migratory populations. Future studies will elucidate whether invasive clones are pre-existing or de novo and determine the effect of chemotherapeutics on these populations.

Citation Format: Carolina De Santiago, Amy Brock. Characterizing the metastatic potential and clonal dynamics of invasive subpopulations in TNBC cells in response to chemotherapy. [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 3622.
Abstract 364: Multi-omics approaches revealed the involvement of immune cells in skeletal muscles of lung cancer cachexia mouse model
Authors: Park Se-Young , Kim Bomi , Lee Myunggyo , Lee Haeseung , Song Na-Young
Published: June 2023

Cancer-associated cachexia (CAC) is a multifactorial syndrome characterized by loss of body weight and skeletal muscle mass that cannot be fully recovered by nutritional support. It accounts for approximately 20 % of cancer-related deaths. However, there are no approved treatments for CAC yet. In order to better understand CAC, we wanted to apply the multi-omics approaches to simultaneously assess tumors and skeletal muscles in CAC mouse models. First, we established a lung cancer cachexia mouse model by orthotopic transplantation of KRAS-mutant lung cancer cells. Lung tumor-bearing mice developed cachexia at 7 weeks post injection, showing over 20 % of body weight loss and wasting of skeletal muscles and fats. The histological and immunoblotting analyses revealed degradation of gastrocnemius (GA) and tibialis anterior (TA) muscles in CAC group. Then, we performed the bulk RNA sequencing to assess transcriptomic changes in lung tumors and GA muscles of CAC mice. In lung tumors, genes associated to ATP metabolic process was upregulated. However, skeletal muscles of CAC mice showed upregulation of genes involved in protein degradation and immune responses, while genes related to energy metabolism were downregulated. Notably, the immunohistochemical staining revealed the increased numbers of infiltrating macrophages and CD8+ T cells in skeletal muscles of CAC mice. These data suggest the involvement of immune cells in lung cancer-associated cachexia. To further identify the distinct immune cell populations involved in CAC, the single cell RNA sequencing was conducted using CD45+ immune cells infiltrated into GA muscles of CAC mice. The enrichment of natural killer cells, dendritic cells and T cells was detected in GA of CAC mice. Moreover, we confirmed the roles of the infiltrating immune cells by using the neutralizing antibodies in CAC mice. Taken together, these data suggest the involvement of the immune cells in the skeletal muscle wasting of CAC mice. These multi-omics approaches will shed light on development of therapeutics for cancer-induced cachexia.

Citation Format: Se-Young Park, Bomi Kim, Myunggyo Lee, Haeseung Lee, Na-Young Song. Multi-omics approaches revealed the involvement of immune cells in skeletal muscles of lung cancer cachexia mouse model [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 364.
Abstract 3800: The disruption of miR-125b-SLC1A5 cascade defines the oncogenicity and differential immune profile in oral carcinoma
Authors: Liu Ying-Chieh , Liu So-Yu , Lin Yu-Cheng , Liu Chung-Ji , Chang Kuo-Wei , Lin Shu-Chun
Published: June 2023

Head and neck squamous carcinoma (HNSCC) including oral SCC (OSCC) is a worldwide malignancy that requires vigorous efforts for interception. The metabolic reprogramming sustains malignant tumor cells to overcome stressful microenvironments. As the glutamine is the one of the essential components which helps cells to erase reactive oxygen species and energy production, the increased uptake of glutamine is a common metabolic hallmark in cancers. Bioinformatic analysis of TCGA HNSCC datasets and RNAseq results in our cohort indicated that solute carrier family 1 member 5 (SLC1A5) is the most eminently dysregulated transporter member among the seven glutamate or neutral amino acid transporters in SLC1A family. This work identified that the knockdown of SLC1A5 expression decreased in vitro oncogenicity, xenografic tumorigenesis and glutamine uptake; increased oxidative stress, cell cycle arrest, and sensitivity to cisplatin and ferroptosis inducer in OSCC cells. By way of contrast, the upregulation of endogenous SLC1A5 expression mediated by CRISPR/dCas9 activation system drove the OSCC oncogenesis. The reporter assay and functional analysis demonstrated that suppressor miRNA miR-125b targeted and attenuated SLC1A5, while NEAT1 lncRNA sponged and ablated miR-125b-SLC1A5 axis. The analysis of TCGA and GEO datasets confirmed the concordant upregulation of NEAT1 and downregulation of miR-125b along with the SLC1A5 upregulation in tumors. Besides, SLC1A5 upregulation defined the worse HNSCC patient survival. In our tumor cohort, OSCC harboring higher SLC1A5 expression had lower CD4+ immune score, and higher scores in M0 and M1 macrophage. GSEA algorithm further deciphered that tumors having higher SLC1A5 expression manifested the repression in glycolysis, apoptosis and p53 signaling. This study concludes that the frequent disruption of NEAT1-miR-125b-SLC1A5 cascade modulates the pluripotent oncogenic activities, the treatment efficacy and the immune landscape in oral carcinoma.

Citation Format: Ying-Chieh Liu, So-Yu Liu, Yu-Cheng Lin, Chung-Ji Liu, Kuo-Wei Chang, Shu-Chun Lin. The disruption of miR-125b-SLC1A5 cascade defines the oncogenicity and differential immune profile in oral 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 3800.
Abstract 3810: MicroRNA-483-5p inhibits cell steatosis and fibrosis by targeting PPARa, TIMP2 and HCC cell proliferation by modulating notch signaling
Authors: Niture Suryakant , Gadi Sashi , Qi Qi , Gyamfi Maxwell , Rios-Colon Leslimar , Varghese Rency , Ressom Habtom , Kumar Deepak
Published: June 2023

MicroRNAs (miRNAs) are small non-coding RNA molecules that bind with 3’ untranslated regions (UTRs) of genes and regulate expression. Downregulation of miR-483-5p (miR-483), is associated with the progression of hepatocellular carcinoma (HCC). However, the molecular and mechanistic role of miR-483 in nonalcoholic fatty liver disease (NAFLD), alcoholic fatty liver diseases (AFLD), and HCC remain elusive. In the current study, we investigated the biological significance of miR-483-5p in NAFLD, AFLD, and HCC in vitro and in vivo. Our data show that the expression of miR-483 was downregulated in tumor samples from HCC patients. Overexpression of miR-483 in HCC cells dysregulated Notch signaling, inhibited cell proliferation/migration, induced apoptosis, and increased sensitivity towards sorafenib/regorafenib; anti-HCC drugs. Interestingly, miR-483 overexpression downregulated cell steatosis and fibrogenic signaling in HCC cells. Mechanistically, miR-483 targets peroxisome proliferator-activated receptor α (PPARα) and tissue inhibitor of metalloproteinases 2 (TIMP2) gene expression by binding to 3’-UTR and suppress cell steatosis and fibrosis. Down-regulation of miR-483 was observed in mice fed with a high-fat diet (HFD) or alcohol which lead to increased hepatic steatosis/fibrosis. Our data suggest that miR-483 inhibits cell steatosis and fibrogenic signaling, and functions as a tumor suppressor in HCC. Therefore, miR-483 may be used as a novel therapeutic target for NAFLD/AFLD/HCC management in patients with fatty live diseases/HCC.

Citation Format: Suryakant Niture, Sashi Gadi, Qi Qi, Maxwell Gyamfi, Leslimar Rios-Colon, Rency Varghese, Habtom Ressom, Deepak Kumar. MicroRNA-483-5p inhibits cell steatosis and fibrosis by targeting PPARa, TIMP2 and HCC cell proliferation by modulating notch 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 3810.
Abstract 3812: Circular RNAs from the PAX5 oncogene are overexpressed in B-cell malignancies and modulate cancer processes
Authors: Martin Danick M. , Hannay Brandon , Martin Alexis , Veilleux Vanessa , Finn Nicholas , Robichaud Gilles A.
Published: June 2023

Circular RNAs (circRNAs) represent a new class of gene products involved in many biological processes. Due to their ability to bind and modulate co-interacting proteins and microRNAs (miRs), circRNAs represent new signaling mediators in cell biology and cancer processes. Recently, we identified circRNAs from the PAX5 oncogene (circPAX5) in B-cells. Given that PAX5 gene products are well established as potent oncogenic regulators of B-cell cancer lesions, we hypothesized that circPAX5 may also be potentially involved in pathogenic processes of B-cell malignancies. In this study, we wanted to understand the role of circPAX5 in B-cell cancer processes and elucidate new potential targets for diagnostic and therapeutic avenues. Using a series of B-cell cancer models and clinical samples, we sequenced, mapped, and profiled circPAX5 expression profiles and found that circPAX5 products are indeed overexpressed in various cancer cell types where the predominant expressed isoform consisted of exons 2 to 5 (circPAX5_2-5) of the PAX5 locus. In addition, we isolated circPAX5 complexes from cancer cells and identified co-interacting small non-coding RNAs, notably microARNs (miRs) by deep sequencing. To further elucidate the functional role of circPAX5 and miR co-interactions, selected miRs from the list were then conditionally overexpressed in B-cells followed by cellular phenotypic assays. Altogether, our results characterize new signaling products and pathways in B-cells. Our data also suggests that circPAX5 products are involved in B-cell cancer processes and could potentially represent new cancer pathways targeted for strategic therapeutic and/or diagnostic strategies in B-cell cancer lesions

Citation Format: Danick M. Martin, Brandon Hannay, Alexis Martin, Vanessa Veilleux, Nicholas Finn, Gilles A. Robichaud. Circular RNAs from the PAX5 oncogene are overexpressed in B-cell malignancies and modulate cancer processes. [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 3812.
Abstract 3863: Aminoflavone confers activity against aromatase inhibitor-resistant breast cancer cells in part via miR125b2-3p restoration
Authors: Brantley Eileen J. , Mavingire Nicole , Khan Salma , Nagaraj Gayathri , Angeloni Shawnee , Wang Charles , Soto Ubaldo
Published: June 2023

Aromatase inhibitor therapy is the most crucial adjuvant treatment option for postmenopausal women with hormone receptor-positive (HR+) non-metastatic breast cancer. However, up to 30% of these patients will experience relapse due in part to aromatase inhibitor resistance. We recently identified miR125b2-3p as a putative tumor suppressor in HR+ breast cancer. We also found that small molecule Aminoflavone (AF) induces miR125b2-3p expression in HR+ breast cancer cells. In the current study, we hypothesized that AF confers anticancer actions in breast cancer cells resistant to aromatase inhibitors via miR125b2-3p reactivation. Using the Alamar Blue and colony forming assays, we discovered that AF demonstrated anticancer activity in estrogen-deprived resistant (EDR) cells designed to mimic aromatase inhibitor resistance. AntagomiR125b2-3p enhanced, while miR125b2-3p mimics diminished colony formation of EDR cells. AF-mediated suppression of colony formation in EDR cells was attenuated during co-treatment with antagomiR125b2-3p and was enhanced during co-treatment with miR125b2-3p mimics. Both miR125b2-3p mimics and AF suppressed EDR cell migration. AntagomiR125b2-3p counteracted AF-mediated suppression of cell migration. Our in silico profiling studies identified the alpha6-integrin, a mediator of breast cancer stem cell activity and metastasis, as one miR-125b2-3p target. Immunohistochemistry of patient tumor samples revealed that alpha 6-integrin protein expression intensified following relapse on the aromatase inhibitor anastrozole. MCF-7 cells transfected to overexpress the alpha 6-integrin were less responsive to anastrozole than those transfected with empty vector. These data suggest that AF restores the tumor suppressor function of miR125b2-3p to inhibit the alpha 6-integrin and confer activity against aromatase inhibitor-resistant breast cancer.

Citation Format: Eileen J. Brantley, Nicole Mavingire, Salma Khan, Gayathri Nagaraj, Shawnee Angeloni, Charles Wang, Ubaldo Soto. Aminoflavone confers activity against aromatase inhibitor-resistant breast cancer cells in part via miR125b2-3p restoration. [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 3863.
Abstract 488: New therapeutic strategies suppressing NAD+ and ATP production in cancer metabolism specific to 3D cultured ovarian cancer cells
Authors: Kudo Kei , Harrington Brittney S. , Ning Franklin , Kamdar Rahul , Shibuya Yusuke , Korrapati Soumya , Fergusson Maria , Kucera Calen , Massimo Fantini , Annunziata Christina M.
Published: June 2023

Ovarian cancer is the gynecological cancer with the highest mortality rate. As resistance to treatment is acquired and many patients relapse, new treatment strategies are needed. A model that strongly reflects the features of recurrent ovarian cancer in the peritoneal cavity is the "cancer stem cells (CSCs)," which are enriched when cells are cultured in three dimensions (3D) on low-attachment plates. The unique metabolic system of the "3D CSCs" is the subject of a novel therapeutic approach. Here, we show that ovarian cancer 3D CSCs have strong NAD+ production capacity, which is particularly dependent on nicotinamide phosphoribosyl transferase (NAMPT), known as the rate-limiting enzyme in the salvage pathway of NAD+ production. We found that NAMPT inhibition blocks NADPH and ATP production as well as NAD+, and subsequently suppresses the synthesis capacity of proteins, fat droplets, and cholesterol in the 3D ovarian cancer cells. In addition, we found that the combination of NAMPT inhibitor and the ALDH inhibitor disulfiram had a pronounced synergistic effect. This synergy inhibits oxidative phosphorylation and ALDH activity and promotes reactive oxygen species and apoptosis via caspase 3/7 cleavage, which leads to death of the 3D cultured cells. Our results demonstrate that high NAMPT expression in ovarian cancer is likely to be a strong poor prognostic factor and that the combination of a NAMPT inhibitor and disulfiram could be a new treatment option to treat or prevent recurrent ovarian cancer. Our novel findings focus on NAMPT addiction as a property of ovarian cancer 3D CSCs, and strongly suggest the combined inhibition of NAMPT and ALDH as a potential therapeutic strategy.

Citation Format: Kei Kudo, Brittney S. Harrington, Franklin Ning, Rahul Kamdar, Yusuke Shibuya, Soumya Korrapati, Maria Fergusson, Calen Kucera, Fantini Massimo, Christina M. Annunziata. New therapeutic strategies suppressing NAD+ and ATP production in cancer metabolism specific to 3D cultured ovarian cancer cells [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 488.
Abstract 5257: Activation of NRF2-mediated oxidative stress response pathway by curcumin analog FLLL12: Implication for chemoprevention
Authors: Lukmon Raji , Adeluola Adeoluwa A. , Amin A.R.M. Ruhul
Published: June 2023

Purpose: Carcinogenesis is a multi-step complex process initiated with irreversible genetic mutation due to endogenously derived or carcinogen-induced oxidative stress. Activation of the antioxidant pathways ameliorates oxidative stress, protects cells from carcinogenic insults, and plays a pivotal role in chemoprevention. We have previously reported that FLLL12 is a potent curcumin analog, possesses in vitro and in vivo anticancer activity, and has better pharmacokinetic profiles than curcumin. The purpose of the current study is to identify novel pathways activated by FLLL12 using RNASeq analysis.

Methods: MDA686, a head and neck cancer cell line, were treated with 2 µM FLLL12 for 24h. Total RNA was isolated and used for RNASeq analysis. 2-fold differentially expressed genes were used for Ingenuity Pathway Analysis to identify the most significantly affected pathways. Real-time qPCR and western blotting were used to confirm the expression of the genes associated with the most significantly affected pathways and their protein products respectively in normal, premalignant, and malignant cell lines.

Results: Whole transcriptome analysis using RNASeq identified 641 genes that were either upregulated or downregulated by 2-folds after FLLL12 treatment. Enrichment of this gene set with Ingenuity Pathway Analysis identified the ferroptosis signaling pathway, the tumor microenvironment pathway, and the NRF2-mediated oxidative response pathway as the top three most significantly affected pathways by FLLL12. We confirmed the activation of HMOX-1, NQO1, SLC7A11, and GCLC mRNA and proteins in HOK (normal), MSK-LEUK1 (premalignant), and MDA686TU (malignant) cells. Although these genes are common for the ferroptosis signaling pathway and the NRF2-mediated oxidative response pathway, treatment of cells with ferroptosis inhibitors, ferrostatin 1, and deferoxamine, had no effect on FLLL12-induced cell death suggesting that these genes are associated with oxidative stress response pathway.

Conclusions: Our results strongly suggest that FLLL12 activates the oxidative stress response pathway in normal, premalignant, and malignant head and neck cancer cell lines and has strong promise for chemoprevention.

Citation Format: Raji Lukmon, Adeoluwa A. Adeluola, A.R.M. Ruhul Amin. Activation of NRF2-mediated oxidative stress response pathway by curcumin analog FLLL12: Implication for chemoprevention. [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 5257.
Abstract 5264: Cucurbitacin D modulates the Nrf2 signaling pathway and protects against benzo[a]pyrene-induced liver injury
Authors: Malik Shabnam , Rodriguiez Anyssa , Sikander Mohammed , Zubieta Daniel , Halaweish Fathi T. , Chauhan Subhash C. , Jaggi Meena
Published: June 2023

Background: Accumulating studies have shown strong correlation of HCC and co-morbidity factors including smoking. Tobacco smoke contains benzo[a]pyrene, which is extremely carcinogenic and contributes to liver damage. Cucurbitacin, a triterpene, has a wide range of biological activities, including antioxidant, anti-inflammatory, and anti-cancer properties. However, their hepatoprotective effects remain poorly understood. In the current study, we examined the hepatoprotective activity of cucurbitacin D, a novel analog of cucurbitacin, against benzo[a]pyrene-induced liver injury in human HepG2 cells.

Method: To investigate the hepatoprotective effect of cucurbitacin D against benzo[a]pyrene-induced liver damage, proliferation, clonogenicity, migration, invasion, Western blotting, and qPCR analyses were performed. The DCFDA assay was performed to determine the level of intracellular reactive oxygen species (ROS) in liver cells.

Results: Functional assays showed that cucurbitacin D exhibited cytoprotective effects against dose-dependent growth inhibition by benzo[a]pyrene in human HepG2 cells. This protective effect was likely associated with antioxidant potential of cucurbitacin D, as evidenced by the attenuation of ROS observed by fluorimeter and fluorescence microscopy. Western blotting analysis demonstrated Cucurbitacin D targets Nrf-2 signaling pathway and associated effector proteins including HO-1 and LC3A in protecting liver cells against benzo[a]pyrene induce oxidative damage. Further studies are underway to understand the underlying molecular mechanism of action.

Conclusion: These findings demonstrate the hepatoprotective effects of cucurbitacin D against benzo[a]pyrene-induced liver damage, making it a promising ingredient for nutritional supplements.

Citation Format: Shabnam Malik, Anyssa Rodriguiez, Mohammed Sikander, Daniel Zubieta, Fathi T. Halaweish, Subhash C. Chauhan, Meena Jaggi. Cucurbitacin D modulates the Nrf2 signaling pathway and protects against benzo[a]pyrene-induced liver injury. [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 5264.
Abstract 5277: Activation of TRPM8 channel suppresses prostate cancer growth and progression
Authors: Asuthkar Swapna
Published: June 2023

Prostate cancer (PC) is one of the most prevalent male malignancies and a leading cause of cancer-related deaths in men. While androgen-deprivation therapy is successful in the early stages of PC, tumor cells eventually become resistant to its effects. With a median survival of 18 months, the shift to androgen-independent prostate cancer (AIPC) has a poor prognosis. Mechanisms causing this change have yet to be explained, however mounting evidence implicates the loss of transient receptor potential melastatin 8 (TRPM8) as a significant contributor. The prostate epithelium normally expresses the ionotropic receptor TRPM8. Our team has recently shown that testosterone-induced activation of TRPM8 enhances Ca2+ absorption and causes apoptosis. This prompted us to propose that increased TRPM8 activity on the plasma membrane is cytotoxic to PC cells and that TRPM8 internalization is a crucial step in the pathogenesis of PC. We examined the amounts of TRPM8 mRNA in benign tumor and metastatic PC patient datasets. TRPM8 mRNA is initially elevated in the early stages of PC but is increasingly lost during the progression to AIPC. In addition, male and female TRPM8−/− mice exhibited increased serum testosterone levels, heightened AR activity, and activation of cell cycle, invasion, and adhesion-related effectors. In both AR+ and AR− xenograft models, our research reveals that TRPM8 possesses potent antitumor properties. Given this, investigating the in vivo role of TRPM8 in PC has the potential to significantly enhance patient outcomes by preventing progression to the androgen-independent state.

Citation Format: Swapna Asuthkar. Activation of TRPM8 channel suppresses prostate cancer growth and progression. [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 5277.
Abstract 536: The role of CCN1 in mutant K-RAS addiction in pancreatic ductal adenocarcinoma
Authors: De Archana , Gupta Vijayalaxmi , Das Amlan , Ghosh Arnab , Haque Inamul , Lau Laster , Banerjee Sushanta K. , Banerjee Snigdha
Published: June 2023

Mutant K-RAS addiction of pancreatic ductal adenocarcinoma cells (PDAC) coordinates their transformation, proliferation, and survival. Therefore, initially, it had an impression that targeting mutant K-RAS could be an ideal strategy for treating PDAC. However, after decades of studies, no effective RAS inhibitors reached the clinic, indicating that RAS-oncoprotein is an undruggable target in PDAC. Thus, identifying a new target linked with K-RAS signaling was most needed. Using genetically engineered cell lines and K-RAS-driven genetically engineered mouse models (KC or KPC) with or without pancreas-specific CCN1 conditional knockout background, we identified a positive feedback circuit between CCN1 and mutant K-RAS expression. Moreover, blocking this feedback via conditional knockout of CCN1 in the pancreas significantly reduced invasive phenotypes, PDAC growth, and metastasis in the lung in KC and KPC mice. Mechanistically we found that the K-RAS-CCN1 feedback circuit is regulated by microRNA miR-145 via a feedforward mechanism. Collectively, these studies highlight the clinical potential of targeting CCN1 in PDAC. (This work is supported by VA Merit grants).

Citation Format: Archana De, Vijayalaxmi Gupta, Amlan Das, Arnab Ghosh, Inamul Haque, Laster Lau, Sushanta K. Banerjee, Snigdha Banerjee. The role of CCN1 in mutant K-RAS addiction in pancreatic ductal 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 536.
Abstract 5398: Optimization generator app (OpGen) for radiotherapy treatment planning using case-based reasoning
Authors: Reiazi Reza , Prajapati Surendra , Mohamed Abdallah Sherif , Fuller Clifton David , Salehpour Mohammad
Published: June 2023

Case-based reasoning (CBR) means adopting previous experiences (i.e, data from previous patients) to meet new demands (new patients). In this scenario, we developed an application named OpGen to generate optimization objective values for head and neck radiotherapy inverse treatment planning, using the previously treated patient with an acceptable outcome. OpGen will be helpful in radiotherapy treatment planning automation and efficiently reducing the required time to reach the target goals and improving the quality and safety of the treatment.OpGen has been developed in Python and has four main modules: 1- Analyzing radiotherapy treatment planning parameters of the previous patients in the database. We have developed (a) a powerful graphical user interface (GUI) that allows quantitative and qualitative analysis of the previous knowledge, including the type of frequently used objective parameter (Min Dose, Max Dose, Min DVH dose, etc.); (b) The range of values used for each specific objective type, and (c) Histogram of the number of patients vs. values used for a specific objective type. 2- Importing patients from treatment planning and retrieving a user-defined number of treatment planning information from the previously treated patients in the database corresponding to the given patient using the user-defined quantitative features (geometrical, distance or geo-distance) and retrieval algorithms (k-Nearest Neighbor and Random Forest). 3- Generating optimization objective values based on the following available options: (a) The most similar patient in the database; (b)- Average or median value over the number of patients retrieved from the database; and (c) Deep learning generated values using a model trained over 100 similar cases retrieved from the database. 4- Generate a portable database of the objective values and patient geometric information for any inter- or intra-institutional usage of the algorithm. OpGen is compatible with RayStation (RaySearch Laboratories™) treatment planning and can retrieve, analyze and generate treatment planning optimization parameters in less than 3 minutes. OpGen can potentially increase the quality and safety of radiotherapy treatment planning by improving the efficiency of the optimization process.

Citation Format: Reza Reiazi, Surendra Prajapati, Abdallah Sherif Mohamed, Clifton David Fuller, Mohammad Salehpour. Optimization generator app (OpGen) for radiotherapy treatment planning using case-based reasoning. [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 5398.
Abstract 547: Role of KDM4A in bladder cancer
Authors: Warren Bailey O. , Abdelfattah Ammar Y. , Ghoneum Alia , Salsbury Freddie , Said Neveen A.
Published: June 2023

Bladder cancer (BCa) is the most common malignancy of the urinary system in the United States. Most patients present with non-muscle invasive bladder cancer (NMIBC) that is treated with bladder preserving approaches. However, recurrence is high with a 50% chance to progression into muscle invasive bladder cancer (MIBC). Patients with MI disease are treated with aggressive surgery and/or systemic chemo-radiation, and more recently immune- and targeted therapies. Patients with BCa require life-long surveillance with expensive imaging, and repeated treatment courses, making BCa one of the most expensive malignancies throughout the patients’ lifespan. BCa therapies have been stagnant for almost four decades until 2016 when new immune and targeted therapies were approved for patients with locally advanced, recurrent, or metastatic disease, with an overall response of 15-29%. Thus, better understanding of the pathobiology of BCa is needed to identify actionable targets and develop novel therapeutics. In this respect, we have identified histone lysine demethylase 4A (KDM4A), also known as Jumanji D2A (JMJD2A) as a BCa onco-protein. We found that the expression of nuclear KDM4A significantly correlated with disease stage in patients’ tumors and in tobacco carcinogen-induced murine bladder tumors. KDM4A transcript expression significantly increased in bladder tumors compared to normal in TCGA data and three independent publicly available datasets. Knockdown of KDM4A in BCa cell lines significantly inhibited their proliferation and invasiveness. Interestingly, KDM4A transcripts exhibited significant inverse correlation with BCa tumor suppressor SPARC in five independent datasets. In addition, using phenotypic screening and computational modeling, we found that KDM4A is a direct target of verteporfin, a potent inducer of SPARC expression in BCa cell lines. Mechanistically, knockdown of KDM4A in BCa cell lines increased SPARC expression, whereas its overexpression inhibited basal and verteporfin-induced SPARC expression. Chromatin immune-precipitation (ChIP) assays indicated that verteporfin significantly inhibited the association of KDM4A with SPARC promoter. Together, our data underscore the oncogenic role of KDM4A in BCa and its potential as an actionable target.

Citation Format: Bailey O. Warren, Ammar Y. Abdelfattah, Alia Ghoneum, Freddie Salsbury, Neveen A. Said. Role of KDM4A in bladder 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 547.