bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1188: IL-17/IL-17RA signaling in the pancreatic epithelium upregulates B7-H4 to promote tumorigenesis
Authors: Pando Susana Castro , Li Le , Howel Rian Morgan , Mascaro Marilina , Le Roux Olivereen , Romanin David , Riquelme Erick , Zhang Yu , Kolls Jay , Moghaddam Seyed Javad , McAllister Florencia
Published: June 2023

Pancreatic ductal adenocarcinoma (PDAC) is expected to overtake other cancers as the second leading cause of death from cancer. PDAC is distinguished by the early formation of a highly immunosuppressive fibro-cellular tumor microenvironment (TME). Our group has demonstrated that IL-17 is necessary for the development and spread of pancreatic cancer, but we have not yet identified the cellular compartment responsible for IL-17-mediated tumor promotion. We propose that early pancreatic carcinogenesis is mediated by modification of an immunosuppressive program and is dependent on pancreatic epithelial IL-17/IL-17RA signaling. We created mice with Kras activated and IL-17RA selectively deleted from the pancreatic compartment (KC; IL-17RAfl/fl). In order to specifically remove IL-17RA from hematopoietic cells, we implanted IL-17RA deficient (IL17-RA−/−) bone marrow cells into KC animals. While deletion of IL-17RA in the hematological compartment had no effect on the progression of pancreatic carcinogenesis, deletion of IL-17RA from the epithelial compartment delayed the development of premalignant lesions, indicating that the pancreatic epithelium is the compartment required for IL-17/IL-17RA signaling during tumorigenesis. Deletion of IL-17RA from the oncogenic epithelium resulted in increased cytotoxic CD8+T cells infiltration in the TME. To learn more about the transcriptional alterations brought on by the lack of IL-17RA in the pancreatic compartment we performed single cell sequencing of pancreas tissue of KC and KC; IL-17RAfl/fl. We found that the absence of IL-17RA in the pancreatic compartment had an impact on epithelial cells transcriptional profiles, with modulation of several immune pathways including regulation of B7-H4, a negative regulator of T cells through Eomes activation. We have generated pancreatic cancer transgenic animals that lack B7-H4 and found that KC; B7-H4−/− mice have delayed tumorigenesis and decreased immunosuppression. In sum, we describe a novel IL-17-mediated regulation of B7-H4 and unravel its function in pancreatic tumorigenesis.

Citation Format: Susana Castro Pando, Le Li, Rian Morgan Howel, Marilina Mascaro, Olivereen Le Roux, David Romanin, Erick Riquelme, Yu Zhang, Jay Kolls, Seyed Javad Moghaddam, Florencia McAllister. IL-17/IL-17RA signaling in the pancreatic epithelium upregulates B7-H4 to promote tumorigenesis [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 1188.
Abstract 1301: Evolution of heterogenous BRAFi/MEKi+CDK4/6i resistance mechanisms and targeting site-specific resistant populations in metastatic cutaneous melanoma
Authors: Wilson Haley P. , Purwin Timothy J. , Capparelli Claudia , Cheng Phil F. , Levesque Mitch P. , Dummer Reinhard , Teh Jessica , Aplin Andrew E.
Published: June 2023

Intrapatient heterogeneity among different metastatic sites contributes to targeted therapy resistance. Determining the proteomic profile of metastatic melanoma lesions that develop within the same patient could help elucidate adaptive mechanisms that drive tumor progression during therapy. Tumors that persist through treatment and colonize distant organs adapt to and rely on organ-specific microenvironments. To assess how protein expression is altered in cutaneous melanoma derived from different metastatic sites in a single patient, we utilized patient-derived BRAFi/MEKi and BRAFi/MEKi+CDK4/6i-treated samples from the LOGIC2 clinical trial. Eight cell lines were established from patient biopsies at chronological time points throughout treatment and subsequent post-mortem resection of the bone, ovary, and brain metastatic sites. While the initial metastatic tumor in the breast was BRAFV600E mutant, NRASQ61 mutations were acquired after treatment with CDK4/6i in the bone and ovary tumors. RPPA was performed on six of the cell lines to characterize site-specific differences in protein expression. We detected upregulation of ErbB3 in the brain metastatic tumor. Since neuregulin 1, a ligand for ErbB3, is abundant in brain tissue, we tested its ability to promote resistance to BRAFi/MEKi in melanoma brain metastases. Neuregulin 1 rescued the growth of brain metastasis cells and restored phospho-AKT and phospho-ERK1/2 levels in the presence of BRAFi/MEKi. Therefore, the ErbB3-Neuregulin axis may be targeted, for example by using ErbB3 inhibitors to improve response to MAPK inhibitors in melanoma brain metastases. Further studies aim to investigate the evolution of metastatic cutaneous melanoma longitudinally in response to targeted therapies. Site-specific signaling mechanisms will be functionally examined as possible avenues for metastasis-specific targeted therapies to improve patient outcomes.

Citation Format: Haley P. Wilson, Timothy J. Purwin, Claudia Capparelli, Phil F. Cheng, Mitch P. Levesque, Reinhard Dummer, Jessica Teh, Andrew E. Aplin. Evolution of heterogenous BRAFi/MEKi+CDK4/6i resistance mechanisms and targeting site-specific resistant populations in metastatic cutaneous melanoma [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 1301.
Abstract 1375: Paranuclear retention of c-KIT by Merkel cell polyomavirus large T-antigen suppresses autophagy through interaction with Beclin 1 essential for Merkel cell carcinoma cell survival
Authors: Shi Hao , Yang Yajie , Larsson Catharina , Lui Weng-Onn
Published: June 2023

Merkel cell carcinoma (MCC) is a very aggressive skin cancer most commonly caused by Merkel cell polyomavirus (MCPyV). While the strong expression of the human proto-oncogene c-KIT in MCC is well established, the functional relevance of this expression is unknown. Here, we demonstrate that paranuclear presence of c-KIT suppresses autophagy which is essential for maintenance of MCC cells. Specifically, after its phosphorylation, c-KIT traffics by clathrin-dependent endocytosis to late endosomes where it is sequestered in the paranuclear compartment by the MCPyV-encoded large transforming antigen (LT) via its Vam6p binding site, which also blocks c-KIT's lysosomal degradation. In this paranuclear compartment, c-KIT interacts directly with both the BH3 and the evolutionarily conserved domain (ECD) of Beclin 1, decreasing its binding to VPS34 and increasing its binding to BCL2, resulting in suppression of autophagy. Indeed, silencing of c-KIT induces autophagy and apoptosis in MCPyV-positive MCC cell lines. It is noteworthy that the positive feedback loop of LT and c-KIT also applies in the reverse direction, as LT is degraded via autophagy. This notion explains why an autophagy-inducing peptide efficiently induces apoptosis-independent cell death and inhibits tumor growth of MCC xenografts in mice. In conclusion, we identified a novel mechanism of MCPyV LT to suppress autophagy through sequestering and stabilizing c-KIT in the paranuclear compartment, which is mandatory to maintain the oncogenic phenotype of MCC cells. Autophagy-inducing agents therefore represent a therapeutic strategy for patients with advanced MCPyV-associated MCC.

Citation Format: Hao Shi, Yajie Yang, Catharina Larsson, Weng-Onn Lui. Paranuclear retention of c-KIT by Merkel cell polyomavirus large T-antigen suppresses autophagy through interaction with Beclin 1 essential for Merkel cell carcinoma cell survival [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 1375.
Abstract 1433: Targeting the mitotic kinase NEK2 potentiates CDK4/6 inhibitor efficacy in breast cancer
Authors: Bobbitt Jessica R. , Cuellar-Vite Leslie , Bryson Benjamin , Weber-Bonk Kristen , Keri Ruth
Published: June 2023

Mitotic kinases are essential for cell cycle progression, and several mitotic kinase inhibitors have been developed to target the uncontrolled proliferation of cancer cells. However, despite their efficacy in preclinical models, mitotic kinase inhibitors are generally considered poor therapeutic targets due to their off target effects in other rapidly proliferating tissues and subsequent toxicity. However, preliminary data in our lab shows that relative to non-transformed cells, triple negative breast cancer (TNBC) cells are selectively sensitive to inhibition of the mitotic kinase NEK2 compared to other mitotic kinase inhibitors. Therefore, we hypothesize that NEK2 is a targetable vulnerability of TNBC. To begin to understand the role of NEK2 in TNBC, we performed RNA-sequencing following NEK2 silencing and found increased expression of genes important at the G1/S cell cycle transition. This transition is largely controlled by the RB-E2F signaling axis, where CDK4 and CDK6 are key players. CDK4/6 inhibitors have been FDA approved for the treatment of a subset of breast cancers. We investigated the impact of silencing NEK2 and treating with CDK4/6 inhibitors and found that loss of NEK2 sensitizes TNBC cells to CDK4/6 inhibition. Mechanistically, this combination causes a significant increase in genomic instability phenotypes. We tested the combination of NEK2 inhibition and CDK4/6 inhibition in vivo, and found that the combination significantly decreases mouse tumor volume without inducing toxicity. Mechanistically, the combination altered mitotic spindle genes in vivo, indicating an increase in genomic instability. Together, these data suggest that that NEK2 is a viable therapeutic target that could be used in combination with FDA approved CDK4/6 inhibitors for the treatment of aggressive breast cancers such as TNBC.

Citation Format: Jessica R. Bobbitt, Leslie Cuellar-Vite, Benjamin Bryson, Kristen Weber-Bonk, Ruth Keri. Targeting the mitotic kinase NEK2 potentiates CDK4/6 inhibitor efficacy in 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 1433.
Abstract 1713: TRMT6-mediated N1-methyladenosine methylation promotes tumorigenesis in colorectal cancer
Authors: SUI SILEI , XU CAIMING , Qing Ying , Baba Hideo , Goel Ajay
Published: June 2023

Background: N1-methyladenosine (m1A) within tRNAs, an essential and highly conserved RNA modification, has recently emerged as a significant epigenetic modification that plays a pivotal role in tumor progression in some cancers. However, the role of m1A methylation in colorectal cancer (CRC) remains largely unexplored. TRMT6 is the most critical methyltransferase that catalyzes N1-methyladenine modification within tRNA, and early evidence suggests that this TRMT6-mediated m1A methylation is also required for the self-renewal of cancer stem cells (CSCs). Herein, we systematically interrogated the role of TRMT6 in mediating stemness and pathogenesis of CRC.

Methods: We first analyzed the expression levels of all m1A regulators in paired tumor and adjacent-normal samples from TCGA and GEO databases. Subsequently, real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting (WB) assays were performed to validate the dysregulation of TRMT6 in CRC tissues and cell lines. m1A dot-blot assays were undertaken to detect methylation levels in CRC tissues and RKO and SW620 CRC cell lines. Cell viability, colony formation, spheroid formation, wound healing, and invasion assays were conducted to investigate the biological functions of TRMT6 in CRC.

Results: Higher m1A methylation levels were detected in CRC than in normal tissues. By analyzing the expression of ten m1A regulators in public datasets, and subsequent qRT-PCR validation in a CRC clinical cohort, we identified that high TRMT6 expression was significantly associated with advanced tumor stages (III&IV vs. I&II, p<0.05) and poor prognosis (5-year relapse-free survival rates in high vs. low expression groups: 50.9% vs. 65.1%, p<0.05). TRMT6 silencing in CRC cell lines inhibited cell growth, colony formation, and metastatic efficiency (p<0.01-0.001). Furthermore, we discovered that the TRMT6 could enhance stem cell-like properties required for CSC self-renewal. Transcriptomic profiling identified that the epidermal growth factor receptor (EGFR) pathway is one of the critical signaling regulators of TRMT6 in CRC. The TRMT6 knockdown resulted in significant inhibition of p-EGFR and p-ERK, but not p-AKT in CRC cell lines (all p<0.05), which suggested that m1A modification exerts its oncogenic effects in CRC by activating EGFR downstream pathway.

Conclusion: Our study elucidates for the first time the oncogenic effects of TRMT6 on catalyzing m1A modification through the EGFR/ERK pathway, which could offer a promising therapeutic strategy in CRC.

Citation Format: SILEI SUI, CAIMING XU, Ying Qing, Hideo Baba, Ajay Goel. TRMT6-mediated N1-methyladenosine methylation promotes tumorigenesis in colorectal 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 1713.
Abstract 1867: Pre-clinical characterization of anti-CEACAM1 antibody
Authors: Dong Jinyu , Zhang Yu , Wang Baiyang , Pu Tingting , Shao Liegang , Wang Binbin , Wang Dong , Ni Jie , Li Sunan , Dong Xin
Published: June 2023

Targeting checkpoint molecules cytotoxic T lymphocyte antigen-4 (CTLA-4) or programmed death receptor-1 (PD-1) expressed on immune cells has shown promising results in the treatment of cancer patients. Nevertheless only a small portion of population benefited from current ICI treatments, so there should be other mechanisms for immune evasion potentially. The carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1) is an activation-induced inhibitory molecule on T and NK cells. In a wide range of cancers, it’s also expressed on tumor cells and other immune cells including macrophages and neutrophils. Homophilic interaction between CEACAM1 expressed on tumor cells and immune cells dampens NK and T cell function, mediates tumor migration, and promotes tumor angiogenesis, which implies that this homophilic interaction is an important mechanism for tumor progression. Here, we describe the identification and characterization of a novel humanized anti-human CEACAM1 therapeutic antibody developed via Neologics’ novel target validation and antibody screening Tier-A platform. This anti-CEACAM1 antibody specifically binds to recombinant human CEACAM1 with nM level affinity and significantly enhances T cell activities and NK killing on CEACAM1 expressing tumor cells, inhibits tumor cell migration in vitro as well. In humanized mouse models, this antibody evidently reduces tumor growth alone or combined with other immune checkpoint blockers. Taken together, we have demonstrated this anti-CEACAM1 antibody specifically enhance NK cell activity, modulate T cell immune activation and inhibit tumor migration supporting further clinical investigation for various indications.

Citation Format: Jinyu Dong, Yu Zhang, Baiyang Wang, Tingting Pu, Liegang Shao, Binbin Wang, Dong Wang, Jie Ni, Sunan Li, Xin Dong. Pre-clinical characterization of anti-CEACAM1 antibody [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 1867.
Abstract 2111: Identification of molecular markers of pseudomyxoma peritonei using integrative analysis of bulk and single cell RNA-seq data
Authors: Park Seong-Hwan , Go Hanyong , Kim Jae-Yoon , Kim Seon-Young , Lee Jong Lyul , Ha Ye Jin , Kim Seon-Kyu
Published: June 2023

Backgrounds: Psuedomyxoma peritonei (PMP) is a rare colon disease, whose symptoms and treatments have been numerously investigated, however, there are insufficient known diagnostic or prognostic markers. Here, we aim to identify molecular markers exhibiting distinct characteristics of PMP.

Methods: We analyzed RNA-seq data to identify gene signatures associated with PMP development using 19 fresh frozen tissue samples including 12 PMPs (discovery cohort). For validation, we applied those signatures into an independent cohort containing 34 formalin-fixed paraffin embedded (FFPE) samples including 25 PMPs (validation cohort). We also verified protein expression levels in the signature using immunohistochemical staining (IHC) in other independent cohort in PMP. Additionally, a transcriptomic profiling using single-cell RNA-seq was carried out to confirm the cell type-specific gene expression patterns in a separated patient group with five PMPs.

Results: In the discovery cohort of fresh-frozen tissues, unsupervised hierarchical clustering analysis exhibited a distinct gene expression pattern clearly dichotomizing normal and PMP samples. Genes involved in cell movement or immune response were highly expressed in PMPs. Interestingly, we found that PMP disease had a molecular heterogeneity showing epithelial as well as mesenchymal characteristics, in which both CDH1 and CDH2 were significantly elaborated in PMPs. We also estimated protein expression levels of PMP-associated key markers using IHC, in which a significant positive correlation between transcript and protein expressions in CDH1, whereas we could not observe such a significance but a clear tendency toward highly expression of CDH2 in PMPs. However, in a single-cell RNA-seq analysis, we discovered distinct eight cell type-specific clusters including an epithelial cell group, in which CDH1 was highly expressed, and a mesenchymal cell group, in which CDH2 showed a significant higher expression pattern, suggesting that PMP show dual molecular characteristics of epithelial as well as mesenchymal.

Conclusion: We explore genetic characteristics and suggest key markers of PMP development. Our findings would be practically utilized for clinical practice in PMP diagnosis or prognosis.

Citation Format: Seong-Hwan Park, Hanyong Go, Jae-Yoon Kim, Seon-Young Kim, Jong Lyul Lee, Ye Jin Ha, Seon-Kyu Kim. Identification of molecular markers of pseudomyxoma peritonei using integrative analysis of bulk and single cell RNA-seq data [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2111.
Abstract 2184: Short Peptides derived from MIEN1 and their analogs exhibit anti-cancer activity in breast and prostate cancer cells
Authors: Tripathi AMIT Kumar , Vishwanatha Jamboor K.
Published: June 2023

Migration and invasion enhancer 1 (MIEN1) is overexpressed in various cancers and plays an important role in cancer cell migration and invasion. Conserved regions of ITAM and prenylation motif in MIEN1 were used as a template to identify anti-cancer peptides. RNA-seq, qPCR analysis and western blots showed changes in the transcriptome and protein expression after peptide treatment. The two bioactive peptides inhibited genes and proteins responsible for cancer cell migration and invasion in both MDA-MB-231 breast cancer and DU-145 prostate cancer cells. The mechanism of the action of the peptides involves the inhibition of key pathways like Epithelial-Mesenchymal transition and Epidermal Growth Factor-mediated NF-κB pathway to exert their anti-cancer activity. Interestingly, the peptides targeted the same signal transduction pathways followed by parental MIEN1 to show its cancer properties. Thus, the two peptides acted as dominant negative effectors of MIEN1 activity. Additionally, Peptide-treatments induced apoptosis in mice groups bearing tumors derived from MDA-MB-231 cells as evidenced by increased levels of cleaved caspase-3 and PARP proteins in western blots. Interestingly, the MIEN1 mRNA and protein expression was lowered in the in vivo breast cancer tumor models which remained unchanged in MDA-MB-231 breast cancer cells indicating an improved therapeutic activity in living system. The peptides did not cause any toxicity in the mice group which received peptides only, at three times the dose used during in vivo assays.

Citation Format: AMIT Kumar Tripathi, Jamboor K. Vishwanatha. Short Peptides derived from MIEN1 and their analogs exhibit anti-cancer activity in breast and prostate 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 2184.
Abstract 289: B7-H3 mediated metabolic reprogramming promotes small cell lung cancer progression
Authors: Fatima Mahek , Khan Parvez , Rehman Asad Ur , Khan Md Arafat , Maurya Shailendra Kumar , Ahmad Aatiya , Zaidi Mohd Ali , Gautam Shailendra , Lele Subodh , Batra Surinder Kumar , Nasser Mohd Wasim
Published: June 2023

Lung cancer is the leading cause of cancer-related deaths worldwide. Small cell lung cancer (SCLC) is an aggressive subtype of lung cancer. The overall survival rate of SCLC is dismal owing to early metastasis, chemoresistance, higher rate of recurrence, and lack of available treatment options. Although some immunotherapeutic drugs have been approved for SCLC, they are effective only in a small fraction of the patient population. This necessitates the quest to find promising vulnerabilities against SCLC. Bioinformatic analysis on SCLC data set in the backdrop of various immune checkpoint regulators revealed B7-H3 as a promising target. The bioinformatics data was recapitulated in SCLC cell lines and human SCLC tissues. To delineate the effects of B7-H3 targeting, we performed CRISPR-Cas9 mediated B7-H3 knockout in SCLC cell lines. B7-H3 knockout in SCLC cells showed a decrease in colony formation, migration, and wound healing properties. Our results suggest that deletion of B7-H3 decrease the functional activities and activation of oncogenic signaling pathways, such as Erk, Akt, and Stat3. Further, we assessed if B7-H3 regulates SCLC metabolism. Preliminary data showed that B7-H3 knockout in SCLC cells inhibits glucose uptake, extracellular acidification rate (ECAR), and oxygen consumption rate (OCR), suggesting its role in SCLC metabolic reprogramming. Altogether, our data indicate that B7-H3 plays a crucial role in SCLC pathogenesis and could serve as a potential therapeutic target for SCLC.

Citation Format: Mahek Fatima, Parvez Khan, Asad Ur Rehman, Md Arafat Khan, Shailendra Kumar Maurya, Aatiya Ahmad, Mohd Ali Zaidi, Shailendra Gautam, Subodh Lele, Surinder Kumar Batra, Mohd Wasim Nasser. B7-H3 mediated metabolic reprogramming promotes small cell lung cancer 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 289.
Abstract 309: ATM phosphorylation of NRF2 promotes DNA damage repair
Authors: Schneider Cheyenne A. , Lin Tin-Yu , Falzone Aimee , Caldwell Samantha , Johnson Jared L. , DeNicola Gina M.
Published: June 2023

Nuclear factor erythroid 2-related factor 2 (NRF2) is a transcription factor that responds to cellular and oxidative stress by promoting the transcription of antioxidant response element-containing genes. In an unstressed cellular environment, NRF2 is bound by Kelch-like ECH-associated protein 1 (KEAP1) and tagged for proteasomal degradation. In many cancers, the interaction between NRF2 and KEAP1 is interrupted due to mutations in either protein. Previous work has suggested that NRF2 can promote DNA damage repair, and our preliminary data demonstrates that NRF2 is phosphorylated at Serine-40 by the DNA damage response kinase, Ataxia Telangiectasia Mutated (ATM) in response to DNA damage. To create a system to better study the relationship between NRF2 Serine-40 phosphorylation and DNA damage, we modified the mouse Nfe2l2 locus to generate a mouse model in which Nrf2 Serine-40 is mutated to Alanine on a C57BL/6 background. Utilizing this model, we administered 7.5 Gy of total body irradiation and found that Nrf2S40A/S40A mice had significantly shortened survival compared to Nrf2WT mice. Immunohistochemical analysis of radiation-sensitive tissues including bone marrow and spleen revealed a prolonged persistence of the DNA damage marker γH2AX following irradiation in the Nrf2S40A/S40A group compared to Nrf2WT. We also observed a significantly higher level of cell death in the Nrf2S40A/S40A splenic tissue compared with Nrf2WT as indicated by the marker of apoptotic cell death, Cleaved Caspase-3. Our findings indicate that phosphorylation of Nrf2 at Serine-40 in response to DNA damage promotes DNA damage repair. Our ongoing work is focused on examining these phenotypes in cellular systems to determine the specific mechanisms by which NRF2 promotes DNA damage repair. Future work will include incorporating the systems generated thus far into non-small cell lung cancer (NSCLC) and liver cancer mouse models to further investigate the impact of this phosphorylation site on tumorigenesis and radio-resistance.

Citation Format: Cheyenne A. Schneider, Tin-Yu Lin, Aimee Falzone, Samantha Caldwell, Jared L. Johnson, Gina M. DeNicola. ATM phosphorylation of NRF2 promotes DNA damage repair [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 309.
Abstract 3818: Isoandrographolide derivative, (3,19)-4-Bromobenzylidene-isoandrographolide induces apoptosis and causes loss of mitochondrial membrane potential in ER positive Breast cancer
Authors: Karanam Balasubramanyam , Tamang Nitesh , Inabathina Yashwanth , Mavileti Sai Kiran , Sripada Lakshminath , Nanduri Srinivas , Golakoti Nageswara Rao
Published: June 2023

Natural product, Andrographolide.is diterpene lactone extracted from Andrographis paniculata. In traditional form of medicine, it is being used in India, China and Malaysia for treating various diseases. While in the last two decades there are many reports on andrographolides and its derivatives with various biological activities such as anti-cancer and anti-inflammatory. We synthesized compounds along with isoandrographolide were characterized and tested for their anticancer activity against 60 cancer cell line at NCI. Most compounds were found to be active against leukemia and breast cancer with (3,19)-4-Bromobenzylidene-isoandrographolide having best growth inhibition of 76.90% and 87.54% against leukemia (MOLT-4) and breast cancer (BT-549) respectively. Upon further testing compound (3,19)-4-Bromobenzylidene-isoandrographolide was found to be more activate than the tamoxifen against MCF-7. Compound (3,19)-4-Bromobenzylidene-isoandrographolide arrested cell cycle at G1-Phase of MCF-7 and was observed to be inducing apoptosis and loss of mitochondrial membrane potential. We discuss our studies on crystal structure, cell cycle progression assay, apoptotic studies on ER-positive breast cancer MCF-7.

Citation Format: Balasubramanyam Karanam, Nitesh Tamang, Yashwanth Inabathina, Sai Kiran Mavileti, Lakshminath Sripada, Srinivas Nanduri, Nageswara Rao Golakoti. Isoandrographolide derivative, (3,19)-4-Bromobenzylidene-isoandrographolide induces apoptosis and causes loss of mitochondrial membrane potential 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 3818.
Abstract 3861: c-MYC mediated resistance to trametinib plus hydroxychloroquine in pancreatic cancer is overcome by CDK4/6 and lysosomal inhibition
Authors: Silvis Mark , Silva Dilru , Riley Rohweder , Schuman Sophia , Gudipaty Swapna , Truong Amanda , Yap Jeffery , Affolter Kajsa , McMahon Martin , Kinsey Conan
Published: June 2023

Pharmacological inhibition of KRAS>RAF>MEK>ERK signaling has provided no clinical benefit to patients with pancreatic ductal adenocarcinoma (PDA). Interestingly, combined inhibition of MEK1/2 (with trametinib, T) plus autophagy (with chloroquine, CQ, or hydroxychloroquine, HCQ) demonstrated striking anti-tumor effects in preclinical models and in a patient (Patient 1). However, not all patients respond to the T/HCQ regimen, and Patient 1 eventually developed resistant disease. Here we report that primary or acquired resistance is associated with focal DNA copy number gains encompassing c-MYC. Furthermore, ectopic expression of c-MYC in PDA cell lines rendered them T/HCQ resistant. Interestingly, a CDK4/6 inhibitor, palbociclib (P), also induced autophagy and overrode c-MYC-mediated T/HCQ resistance, such that P/HCQ promoted regression of T/HCQ resistant PDA tumors with elevated c-MYC expression. Finally, P/HCQ treatment of Patient 1 resulted in a biochemical disease response. These data suggest that elevated c-MYC expression is both a marker and a mediator of T/HCQ resistance, which may be overcome by the use of palbociclib plus hydroxychloroquine.

Citation Format: Mark Silvis, Dilru Silva, Rohweder Riley, Sophia Schuman, Swapna Gudipaty, Amanda Truong, Jeffery Yap, Kajsa Affolter, Martin McMahon, Conan Kinsey. c-MYC mediated resistance to trametinib plus hydroxychloroquine in pancreatic cancer is overcome by CDK4/6 and lysosomal inhibition. [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 3861.
Abstract 5368: AI-enabled prediction of lung cancer specific hot spot gene alterations from histology images
Authors: Shafi Gowhar , Shivamurthy Shiva , Ulle Anand , Chinglemba Chongtham Cha , Haldar Sumit , Moubeen Fauzul , Uttarwar Mohan
Published: June 2023

Background: The clinical intervention relies on molecular testing of primary biomarkers. We developed an AI enabled platform to predict NCCN guidelines recommended hot-spot genomic alterations from the image analysis using H&E slide.

Methods: AI engine was trained using clinically observed hot spot genomic alterations present in lung cancer patients as a ground truth based on histological FFPE whole-slide images. The H & E stained histological images of the tumor having matching pathogenic mutations were downloaded from the TCGA database for the training and testing purpose. The program extracted histopathological features, followed by pattern mapping to predict genomics status based on the results of NGS testing from 480 samples. ResNet AI algorithm was trained to predict lung specific hot spot genomic alterations. An H & E stained slide was imaged with a 40x objective lens to obtain a brightfield image as an input data. The input image was segmented into 65 x 103 tiles, each of 64 µm2 in size for an individual pathological class. 80% of the data set was used for training the engine while validation and blinded study was respectively performed on remaining 10% data.

Results: Using single blinded clinical samples. AI model analyzed whole slides. Table 1 summarize parameters for clinically significant variants. Higher sensitivity and specificity was observed for variants encoding mutated cell surface proteins compared to variants encoding intracellular mutant targets when compared on similar training and testing parameters.

Conclusions: AI-enabled prediction of hotspot genomic alterations resulted comparable to NGS with shortest TAT, cost and resources. It predicted actionable gene alterations with high positive and negative prediction values.

Table 1. Gene Sample (n = 480) Evaluation parameters for tested samples Training Testing Sensitivity (%) Specificity (%) PPV (%) NPV (%) F1-score (%) Accuracy (%) KRAS 248 62 100 94 94 100 97 97 ALK 16 4 100 100 100 100 100 100 EGFR 72 18 88 100 100 90 94 94 MET 16 4 50 100 100 66 66 75 ROS 16 4 100 100 100 100 100 100 STK11 88 22 82 73 75 80 78 77 RET 24 6 50 100 100 60 50 67 BRAF 14 2 100 100 100 100 100 100 NTRK 92 24 78 56 64 71 70 67 ERBB2 12 4 50 50 50 50 50 50

Citation Format: Gowhar Shafi, Shiva Shivamurthy, Anand Ulle, Chongtham Cha Chinglemba, Sumit Haldar, Fauzul Moubeen, Mohan Uttarwar. AI-enabled prediction of lung cancer specific hot spot gene alterations from histology images. [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 5368.
Abstract 5520: Research Ready graphic-style story to support future research participation among adolescents
Authors: Bates Lauren , Gertz Susan , Hershberger Susan , Kovacic Melinda Butsch ,   We Engage 4 Health Community-Academic Partnership
Published: June 2023

Historically, marginalized adults are more likely to be mistrustful of health research worrying both about its invasiveness and their own safety. As a result, many decline to participate. Adolescents have a much lower perception of risk compared to adults and are also susceptible to peer pressure and socialized trust or mistrust from their parents and family members, which can influence risk perception. As encouraging equitable participation in research becomes an important tool in the fight against cancer disparities, We Engage 4 Health created a graphic-style story “Research Ready” to help community members learn the 3 P’s: the purpose of cancer research, how they are kept safe while participating, and why people from diverse backgrounds are needed to participate. An accompanying Research Review Activity was also included to further support participants’ decision-making regarding research participation. Two groups of diverse adolescents were asked to read the story aloud together in small groups and then discuss an active research study targeting adolescents using the review activity with a scientist to consider whether the first the teen story character (Vito) and ultimately themselves, should consider participation in the study and to identify what information was needed from study staff to better be able to make a decision based on their values and accurate information provided by study staff. Group 1 included 45 adolescents that were 68% African American and 10% European American. Group 2 included 73 high schoolers that were 78% Asian, 15% European American, and 5% African American. Post-discussion surveys indicated that only 9% and 3% in group 1 and group 2 respectively had previously participated in health research. However, after the story discussion, 83% and 89% indicated they thought it would be safe to participate in research and 76% and 89% indicated a willingness to participate in research in the future. Finally, 60% collectively enjoyed the story’s graphic-style format. These data suggest Research Ready to be potentially useful in promoting future research participation and may be especially useful in engaging ethnic/racial groups commonly underrepresented in cancer research.

Citation Format: Lauren Bates, Susan Gertz, Susan Hershberger, Melinda Butsch Kovacic, We Engage 4 Health Community-Academic Partnership. Research Ready graphic-style story to support future research participation among adolescents. [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 5520.
Abstract 6032: Exploiting metabolic vulnerabilities of ovarian cancer with upregulated succinate dehydrogenase
Authors: Wang Lin , Bulicz Magdalena , Mucha Patryk , Kinter Michael , Pranay Atul , Griffin Timothy M. , Miller Benjamin F. , Bieniasz Magdalena
Published: June 2023

Ovarian cancer remains the deadliest of all gynecologic malignancies nationwide due to limited therapeutic options and chemotherapy resistance. The response to chemotherapy is substantially influenced by the high metabolic flexibility of ovarian cancer cells, which allows the cells to reprogram their metabolism and survive the treatment. Better understanding mechanisms regulating ovarian tumor metabolism could be exploited to develop new improved therapies. Our work demonstrated that the upregulation of mitochondrial enzyme succinate dehydrogenase (SDHA) is particularly prevalent (~20% of patients) in ovarian carcinoma and contributes to high energy tumor metabolism and cell survival. We observed that SDHA overexpressing ovarian cancer cells showed improved ability to survive and generate colonies in anchorage-independent conditions, which is an important feature of ovarian tumor cells surviving and spreading in peritoneal fluid (ascites). Proteomics data revealed that the SDHA overexpressing tumor cells showed reduced levels of proteins promoting anaerobic glycolysis (LDHA and HIF1α), and increased levels of mitochondrial pyruvate carrier (MPC) proteins. In addition, our tracer-based metabolomics data revealed that the elevated SDHA stimulates glutaminolysis by increasing mitochondrial glutamine supply fueling the TCA cycle with essential metabolites. We also demonstrated that the overexpression of SDHA is associated with a significant increase of OXPHOS and ATP production rate. Further, we reasoned that the SDHA overexpressing tumor cells could be particularly vulnerable to drugs disrupting glucose and/or glutamine metabolism. We performed a drug screening and identified an anti-metabolic compound shikonin known to disrupt glucose and amino acid metabolism. In in vitro studies, shikonin exhibited a profound anti-tumor efficacy and selectivity towards SDHA overexpressing tumor cells superior to that observed with chemotherapy. Importantly, our in vivo study validated the in vitro observations, and showed that shikonin is highly effective in suppressing SDHA overexpressing patient-derived tumor growth. In summary, the unique metabolic state of ovarian cancer associated with SDHA amplification could be successfully targetable offering a potential new treatment strategy for ovarian cancer patients.

Citation Format: Lin Wang, Magdalena Bulicz, Patryk Mucha, Michael Kinter, Atul Pranay, Timothy M. Griffin, Benjamin F. Miller, Magdalena Bieniasz. Exploiting metabolic vulnerabilities of ovarian cancer with upregulated succinate dehydrogenase [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 6032.
Abstract 6284: SUV420H1 as a novel target in HPV-negative head and neck squamous cell carcinoma
Authors: Moshiri Arfa , Cheng Hui , Kim Sohyoung , Saloura Vassiliki
Published: June 2023

Human Papilloma Virus (HPV)-negative head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer type in the world and its outcomes are mostly unchanged for the past few decades, especially for HPV-negative HNSCC. The TCGA recently revealed a plethora of genetic alterations in chromatin modifiers in HPV-negative HNSCC, including protein lysine methyltransferases which methylate lysine residues on histone tails and regulate gene expression. One of these enzymes, SUV420H1, is known to trimethylate H4K20 and is recurrently amplified in approximately 6% of HPV-negative HNSCC tumors (TCGA). It has also been found to be significantly overexpressed in multiple HNSCC cell lines compared with normal keratinocytes. This study aims to investigate whether SUV420H1 has oncogenic activity and could be a novel therapeutic target in HPV-negative HNSCC. To evaluate the effect of SUV420H1 depletion on HPV-negative HNSCC cell lines, siRNA-mediated knockdown was performed in four SUV420H1 overexpressing HPV-negative HNSCC cell lines, and MTT assays showed significantly reduced cell viability by approximately 80-90% after 12 days of siRNA treatment. Colony formation assays, cell cycle analysis and apoptosis assays are ongoing, and these assays are planned in SUV420H1 CRISPR KO cell lines. Gene Set Enrichment Analysis in 434 HPV-negative HNSCC tumors (TCGA) showed significant enrichment of immune signatures, EMT, cell cycle and apoptosis pathways. Ongoing experiments aim to investigate the effects of SUV420H1 knockdown on the global histone methylome, and to identify direct downstream targets of SUV420H1 through combined genome-wide mapping of SUV420H1 using CUT&RUN assays and RNA-seq in HPV-negative HNSCC cell lines. Syngeneic and xenograft mouse models are also planned to evaluate the effect of SUV420H1 depletion on flank tumor growth in vivo. These studies may elucidate the oncogenic effects and mechanisms of SUV420H1, and may provide evidence to support SUV420H1 as a potential novel target in the treatment of HPV-negative HNSCC tumors with SUV420H1 amplification or overexpression.

Citation Format: Arfa Moshiri, Hui Cheng, Sohyoung Kim, Vassiliki Saloura. SUV420H1 as a novel target in HPV-negative 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 6284.
Abstract 6307: A novel pegylated bispecific antibody-drug conjugate (P-BsADCpb-adc) targeting cancers co-expressing PD-L1 and CD47
Authors: Liu Shumin , Lyu Weidong , Yin Shuqiang , Lei Yang , Zhuo Qiudong , Zheng Liling , Sun Bin , Tan Shuangyu , Jiang Lidong , Zhang Teng , Gao Bo , Xu Rui , Huang Dechang , Li Yong , Wu Zibin , Wu David , Wen Yvonne (Yu)
Published: June 2023

ADCs have demonstrated improved efficacy and target selectivity comparing to the non-specific small molecule cytotoxicity drugs in cancer treatment. Yet, in solid tumor therapies, effort for further improvement of efficacy and safety has been hindered by the poor tumor penetration of conventional IgG based ADC, low internalization efficiency, undesired efflux of ADC from tumor cells, narrow therapeutic window due to the on-target/off tumor and Fc induced toxicity, etc. To address those issues, we previously reported that the compound JY201, a Polyethylene Glycol (PEG)-based bispecific ADC (P-BsADC) targeting two epitopes of Her2, demonstrated advantages in tumor penetration, internalization efficiency, lysosome trafficking, no Fc related toxicity, and better efficacy in tumor inhibition than transtuzumab deruxtecan (Ds-8201). In this study, we reveal another novel pegylated P-BsADC (JY207) formed by site-specific conjugation of a bispecific single chain fusion protein targeting PD-L1 and CD47 with PEG-MMAE (a pegylated cytotoxic payload MMAE). As expected, the compound JY207 retains all the advantages that JY201 possesses. Furthermore, JY207 does not bind to human red blood cells, but preferentially binds to CD47/PD-L1 double positive tumor cells, thus reduces the possibility of on-target toxicities. In vitro cytotoxicity studies showed that JY207 has strong potencies in CD47/PD-L1 double positive tumor cells, while showing almost no killing effect to CD47 or PD-L1 single positive tumor cells. In CDX models and a PDX model of transplanted tumor tissues from lung cancer patients, the compound demonstrated excellent tumor inhibition at low doses. In an in vitro plasma stability test, JY207 displays high stability in cynomolgus monkey and human serums. Preliminary repeated-dosing toxicological study has found the maximum tolerated dose of JY207 in CD47/PD-L1 double transgenic mice is 50mg/kg. In vivo pharmacokinetics and toxicological studies of JY207 are being conducted in cynomolgus monkeys and are expected to show desirable results. All those findings in this study warrant JY207 as a promising candidate for the clinical development for patients with CD47/PD-L1 double positive cancers.

Citation Format: Shumin Liu, Weidong Lyu, Shuqiang Yin, Yang Lei, Qiudong Zhuo, Liling Zheng, Bin Sun, Shuangyu Tan, Lidong Jiang, Teng Zhang, Bo Gao, Rui Xu, Dechang Huang, Yong Li, Zibin Wu, David Wu, Yvonne (Yu) Wen. A novel pegylated bispecific antibody-drug conjugate (P-BsADCpb-adc) targeting cancers co-expressing PD-L1 and CD47. [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 6307.
Abstract 6334: VBI-002, a CD47xICAM-1 bispecific antibody for the treatment of hepatocellular carcinoma, melanoma and non-small cell lung cancers
Authors: Wong Oi Kwan , Wang Xinhua , Post Leonard , Chen Xiaocheng
Published: June 2023

Intercellular Cell Adhesion Molecule 1 (ICAM-1, CD54) is a type I transmembrane protein and a member of the immunoglobulin superfamily. ICAM-1 is involved in many key processes such as cell-cell interactions, signal transduction and leukocytes trans-endothelial migration. ICAM-1 is constitutively present at low levels on healthy tissues but highly expressed in some hematological cancers and solid tumors, making it an attractive tumor target.

VBI-002 is a novel human IgG1 based CD47xICAM-1 bispecific antibody with excellent safety profile and potent single agent activities in xenograft models of lymphoma, multiple myeloma and some solid tumors (Cancer Res 2022;82(12_Suppl):Abstract nr 3430). The bispecific design allows VBI-002 to selectively block CD47/SIRPα binding in tumor cells with high ICAM-1 expression. In contrast to the benchmark CD47 monoclonal antibody, VBI-002 exhibits minimal red blood cell binding and does not cause hemagglutination. In non-human primate (NHP) safety study, four weekly doses of 60 mg/kg VBI-002 were well-tolerated with only mild and reversible reduction in hemoglobin and platelets.

Here we further investigate the anti-tumor activities of VBI-002 in solid tumors. Gene expression analysis suggested that ICAM-1 is highly expressed in non-small cell lung cancers (NSCLC), melanoma and a subset of hepatocellular carcinoma. Flow cytometry studies confirmed the frequent co-expression of ICAM-1 and CD47 in these tumors. VBI-002 has potent ICAM-1 dependent SIRPα blocking, antibody-dependent cellular phagocytosis and antibody-dependent cellular cytotoxicity activities against cell lines of NSCLC, hepatocellular carcinoma, and melanoma. Furthermore, in vivo efficacy studies demonstrated that VBI-002 has potent single agent activities in cell line derived xenograft (CDX) models of NSCLC, hepatocellular carcinoma, and melanoma. Weekly dose of 20 mg/kg VBI-002 is sufficient to stop tumor growth or drive tumor regression in most models. Notably, VBI-002 is highly effective against NSCLC CDX models carrying Kras mutation and it synergizes with paclitaxel to provide additional anti-tumor activity. In conclusion, VBI-002 is a versatile novel CD47xICAM-1 bispecific antibody with potent single agent activity in various solid tumors and combined well with standard of care chemotherapies for added activity. Thus, VBI-002 warrants clinical evaluations as a single agent and in combination with standard of care chemotherapies.

Citation Format: Oi Kwan Wong, Xinhua Wang, Leonard Post, Xiaocheng Chen. VBI-002, a CD47xICAM-1 bispecific antibody for the treatment of hepatocellular carcinoma, melanoma and non-small cell lung 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 6334.