bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5412: Role of her3 mutations on breast cancer oncogenesis
Authors: Mishra Rosalin , Kilroy Mary Kate , Patel Hima , Alanazi Samar , Garrett Joan T. 
Published: June 2022

Abstract
We sought to determine if naturally occurring mutations in HER3 could drive oncogenic growth of HER3KO HER2+ HCC1569 cells in which endogenous HER3 has been eliminated via CRISPR-Cas9. A series of HER3 mutations identified in breast cancer patients (F94L, V104L, G284R, D297Y, T355I, and E928G) were introduced using lentiviral transduction and stable cell lines were generated in HER3KOHCC1569 cells via puromycin selection. We identified HER3V104L mutation to have higher cell proliferation and higher p-HER3 expression compared to wild-type (wt) and empty-vector (EV) HER3. We observed that HCC1569HER3KO cells stably expressing WT and V104L were sensitive to increasing doses of neratinib (0.1-0.5 µM) concentration. Next we analyzed if this mutation rendered resistance to the recently FDA- approved irreversible HER2 tyrosine kinase inhibitor, tucatinib. Our data indicated that both that V104L cells were sensitive to higher concentration of tucatinib compared to neratinib. In parallel experiments, we utilized COS7 cells to examine the signaling properties of HER3V104L. Our data indicated that transient transfection of COS7 cells with HER3V104L mutant significantly induces p-HER3/p-AKT and p-HER2 expression compared to WT HER3 in a ligand dependent manner. In addition, we observed that the V104L mutation stabilizes HER3 protein expression independent of HER2 and ligand stimulation. Experiments are ongoing to determine whether V104L induced HER3 stabilization and downstream signaling activation is dependent on HER3 binding partners including EGFR, HER2 or HER4. We also aim to understand how the V104L mutation stabilizes HER3 expression. Structural modeling of V104L mutation will provide insight about the mechanism of stabilization of the HER3 protein. We will use MCF10A and HEK293 cells to determine the effect of the V104L mutation on HER3 expression and downstream signaling. We also aim to decipher the signaling mechanism that drives the oncogenic potential of V104L mutation. In addition, we are using various PDXs with different HER3 mutations to determine other driver HER3 mutations in breast cancer and how this can be targeted in the clinic using HER targeted therapy.
Citation Format: Rosalin Mishra, Mary Kate Kilroy, Hima Patel, Samar Alanazi, Joan T. Garrett. Role of her3 mutations on breast cancer oncogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5412.
Abstract 5420: DPYD enhancer variant confers increased risk of severe toxicity in 5-FU chemotherapy
Authors: Zhang Ting , Ambrodji Alisa , Huang Huixing , Bouchonville Kelly , Etheridge Amy , Schmidt Remington , Nunez Jose Cardiel , Temesgen Zoey , Innocenti Federico , Diasio Robert , Largiadèr Carlo , Offer Steven M. 
Published: June 2022

Abstract
Genetic variants in the DPYD gene that are deleterious to dihydropyrimidine dehydrogenase (DPD) enzyme function have demonstrated clinical validity as predictive biomarkers for severe (i.e., CTCAE grade ≥3) and potentially lethal toxicity to fluorouracil (5-FU) and capecitabine chemotherapies. Variants within the coding region only explain 5-10% of 5-FU-related toxicities; and GWAS approaches have been unable to identify validated predictive biomarkers to date. Our previous studies demonstrated that DPD expression was controlled by multiple epigenetic processes; considerable variation was also noted in liver DPYD/DPD levels. Using CRISPR-interference (CRISPRi) and CRISPR-activation (CRISPRa), we identified functional enhancer regions within liver tissues and cells. Epigenetic status at these regions was shown to correlate with DPYD expression in cell models and human liver tissues. Chromosome conformation capture (3C) confirmed direct promoter-enhancer interactions. A multi-omics analysis of &gt;1000 human liver specimens identified a proximal eQTL for DPYD expression within an enhancer, with a global minor allele frequency of 27%. This variant was subsequently shown to directly alter DPYD expression in knock-in cell line models. The variant disrupts transcription factor binding at the DPYD enhancer, leading to reduced enhancer-promoter interaction, reduced DPD expression, and increased cellular sensitivity to 5-FU. Collectively, our data suggest that the variant could represent a novel predictive biomarker for severe 5-FU toxicity. The evaluation of this marker in the clinical setting is currently underway.
Citation Format: Ting Zhang, Alisa Ambrodji, Huixing Huang, Kelly Bouchonville, Amy Etheridge, Remington Schmidt, Jose Cardiel Nunez, Zoey Temesgen, Federico Innocenti, Robert Diasio, Carlo Largiadèr, Steven M. Offer. DPYD enhancer variant confers increased risk of severe toxicity in 5-FU chemotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5420.
Abstract 5452: Discovery of a novel seleno-leonurine analog as a potential therapeutic for ovarian cancer
Authors: Raza Asif , Krzeminski Jacek , Amin Shantu , Sharma Arun 
Published: June 2022

Abstract
Leonurine is an active alkaloid that is extracted from Traditional Chinese Medicine Herba leonuri. It has been reported to be beneficial for cardiovascular diseases, including atherosclerosis, acute and chronic myocardial infarction (MI), and ischemic stroke. Mao et al. illustrated that leonurine could up-regulate the expression of phosphorylated p38 and down-regulate phosphorylated Akt expression through inhibiting mitochondrial pathway, thus suppressing the proliferation and promoting lung cancer cell apoptosis. Accumulating evidence suggest that the extract of Leonurus sibiricus transgenic roots could induce apoptosis in different grades of human glioma cells and significantly promotes cisplatin sensitivity in cervical cancer cells via repressing cell proliferation, improving intrinsic cell apoptosis and inhibiting the expression of P-Gp and MRP1 protein, which has potential therapeutic value in cervical cancer treatment. In the present study, we designed and synthesized several selenium analogs of leonurine and their cytotoxicity were assessed using four different ovarian cancer cell lines viz. OVCAR-3, OVCAR-10, ES2, and drug resistant HeyA8 cells. Compared to the control and leonurine group, compound 834 inhibited the ovarian cancer cell viability in dose- and time-dependent manner. The IC50 values of compound 834 was 4.6 µM, 3.8 µM, 6.2 µM, and 5.7 µM for OVCAR-3, HeyA8, OVCAR-10, and ES2 cells, respectively, while for leonurine the IC50 value was &gt;500 µM for 48 h treatment. To further acquaint the effect of 48 h treatment on cell proliferation, we performed trypan blue cell viability assay, trypan blue dye exclusion assay, Annexin V/7-AAD, Caspase 3/7 apoptosis assays and Western blotting of apoptotic proteins involved in apoptosis. Compound 834 significantly induced apoptosis in OVCAR-3 and HeyA8 cells in dose dependent manner. Upregulation of several apoptotic proteins, as evident by Western blot, suggested that 834 causes induction of apoptotic pathways efficiently. Further, the cell cycle analysis showed a significant arrest of OVCAR-3 and HeyA8 cells in the S-phase, which was further confirmed by the analysis of cyclin A2, cyclin-E1, p21 and p27 proteins expression. Conclusively, our research revealed that leonurine analog 834 significantly induces apoptosis in ovarian cancer cells via repressing cell proliferation, improving intrinsic cell apoptosis and inhibiting the expression of several signaling proteins, which has potential therapeutic value in ovarian cancer treatment.
Citation Format: Asif Raza, Jacek Krzeminski, Shantu Amin, Arun Sharma. Discovery of a novel seleno-leonurine analog as a potential therapeutic for ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5452.
Abstract 5459: Synergistic effects of WX-0593 in combination with panitumumab on osimertinib-resistant EGFR-mutantnon-small cell lung cancer in vitro and in vivo
Authors: Zheng Shansong , Zheng Qingmei , Chen Dongmei , Wang Xinmei , Dong Xin , Zhang Xin , Ma Cuicui , Duan Huicheng , Zhang Chenwei 
Published: June 2022

Abstract
Objective: To preliminarily demonstrate the synergistic effect of WX-0593 (anaplastic lymphoma kinase/ROS proto-oncogene 1 receptor tyrosine kinase [ALK/ROS1] tyrosine kinase inhibitor) in combination with panitumumab on osimertinib-resistant cis EGFR triple-mutant non-small cell lung cancer (NSCLC) both in vitro and in vivo.
Methods: The tumor cell inhibition rates of WX-0593 and panitumumab alone or combined were assessed in osimertinib-resistant cis EGFR triple-mutant cell lines, including PC9 (EGFR Del19/T790M/C797S), NCI-H1975 (EGFR L858R/T790M/C797S), Ba/F3 (EGFR L858R/T790M/C797S and EGFR Del19/T790M/C797S). The in vivo tumor suppressive efficacy of single-agent WX-0593, panitumumab, and their combination was evaluated in H1975 (EGFR del19/T790M/C797S) and Ba/F3 (EGFR L858R/T790M/C797S) cell line-derived xenograft (CDX) models of BALB/c nude mice. The tolerability was also assessed.
Results: In vitro studies showed that the combination of WX-0593 and panitumumab had a potent synergistic effect on proliferation inhibition in all the four cell lines with cis EGFR triple mutations resistant to osimertinib. WX-0593 in combination with panitumumab demonstrated a delaying effect on tumor growth without increasing drug toxicity in the H1975 CDX model. WX-0593 and panitumumab alone had tumor suppressive effects in the Ba/F3 CDX model. And WX-0593 plus panitumumab significantly enhanced the antitumor activity in the Ba/F3 model.
Conclusion: The data from the study suggested that WX-0593 and panitumumab had a promising synergistic effect on osimertinib-resistant EGFR-mutant NSCLC both in vitro and in vivo. The combination regimen may have important clinical implications for the future treatment of patients with EGFR triple-mutant NSCLC resistant to osimertinib.
Citation Format: Shansong Zheng, Qingmei Zheng, Dongmei Chen, Xinmei Wang, Xin Dong, Xin Zhang, Cuicui Ma, Huicheng Duan, Chenwei Zhang. Synergistic effects of WX-0593 in combination with panitumumab on osimertinib-resistant EGFR-mutantnon-small cell lung cancer in vitro and in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5459.
Abstract 5466: Efficacy of ASC60, an oral fatty acid synthase inhibitor, in two tumor mouse models
Authors: Wu Jinzi J. , He Handan 
Published: June 2022

Abstract
Background: Fatty acid synthase (FASN) is a key enzyme controlling endogenous lipid biosynthesis and overexpressed in many cancers. Studies have shown that FASN inhibition can disrupt lipid synthesis in tumor cells and disrupt tumor-associated signal transduction, suggesting FASN inhibition can serve as a potential treatment for cancer. Here we report the in vivo efficacy data of ASC60, a small molecular FASN inhibitor, either as a single agent or in combination with mouse programmed cell death-1 (mPD-1) antibody in two tumor mouse models.
Methods: In the MC38 mouse model, mice were inoculated subcutaneously with MC38 cells. When the average tumor volume reached approximately 62 mm3, mice were treated with ASC60 (60, and 100 mg/kg) or vehicle once a day (QD) for 3 weeks in combination mPD-1 antibody twice a week (1 mg/kg, BIW) for 2 weeks. In patient derived orthotopic xenograft (PDOX) brain metastasis mouse model, luciferase labelled breast cancer cells (LD1-2009-362541-Luc) were transplanted into the brain of nude mice. Tumor volumes were estimated by the luciferase reporter signal. When the average signal reached 4.12X107 photon/sec, mice were treated with ASC60 (20, 60, and 100 mg/kg) or vehicle QD for 4 weeks. In both mouse models, body weights and tumor volumes were measured regularly, and tumor growth inhibitions (TGI) of different groups were compared.
Results: All animals showed a gradually increase in body weight during the study and no significant difference of body weight or body weight change was found among different groups. In the MC38 colon cancer mouse model, mice treated with ASC60 (60 or 100 mg/kg) in combination with mPD-1 antibody showed higher TGI values compared to those treated with mPD-1 alone (91.86%, 99.88% vs 89.84%) on Day 13, indicating combination with ASC60 would enhance the antitumor activity of mPD-1 antibody. In PDOX mouse model, mice treated with ASC60 (20, 60, and 100 mg/kg) showed dose dependent TGI values of 10.83%, 36.95% and 49.21% on Day 13 and of 6.46%, 21.27% and 30.47% on Day 27, respectively. ASC60 also showed a significant improvement in life span (51.5 days in ASC60 60 mg/kg vs 39 days in vehicle control).
Conclusion: Previous in vitro testing showed that inhibition of palmitate synthesis by ASC60 was 137-fold lower in mouse than in human (IC50: 2.05 µM in mouse vs 0.015 µM in human). However, results of the present studies showed that ASC60 could suppress tumor growth and/or expand lifespan in the two tumor mouse models either alone or in combination with mPD-1 antibody. ASC60 will possibly demonstrate better efficacies in human.
Citation Format: Jinzi J. Wu, Handan He. Efficacy of ASC60, an oral fatty acid synthase inhibitor, in two tumor mouse models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5466.
Abstract 5478: EC-18 suppressed tumor growth and alleviated side effects caused by cisplatin in the HNSCC implantation model
Authors: KIM Guentae , Kim Eun Young , Shin Su-Hyun , Lee Hyowon , Yoon Sun Young , Park Kaapjoo , Sohn Ki-young , Kim Jae Wha 
Published: June 2022

Abstract
Introduction: Head and neck squamous cell carcinomas (HNSCC) are very unfavorable carcinoma that lowers the quality of life. In addition, side effects associated with treatment such as oral mucositis also deteriorate enough to be classified as a disease. This study verified the synergistic antitumor effect of EC-18 (PLAG, 1-Palmitoyl-2-linoleoyl-3-acetyl-roc-glycerol) with or without cisplatin as a chemotherapy and side effects alleviation effects in the metastatic mouse oral squamous carcinoma (MOSCC) orthotopic model.
Method: After inserting mouse-derived squamous oral carcinoma into the right side of the tongue (n=12), it was treated with EC-18 alone or with cisplatin for three weeks. The changes in feed rate and body weight were quantitatively verified on a 2-day interval. Changes in tumor size and oral mucositis symptoms (toluidine-blue positive) were analyzed on the sacrifice day. In addition, changes in the amount of damage-associated molecular pattern (DAMP) increased by tumor and cisplatin, and changes in related active factors were quantitatively analyzed.
Result: Compared to the positive control group, the tumor size was reduced by 42% in the group treated with EC-18 alone (P&lt;0.05). In addition, the tumor size was further reduced by 25% in the group co-treated with EC-18 and cisplatin compared to the cisplatin alone (P&lt;0.05), and it was observed that the tumor was disappeared in 3 mice in the co-treated group. In the positive control group, feed rate and body weight gradually decreased after about 12 days. Also, in the cisplatin-treated group, feed rate and body weight decreased rapidly after six days. On the other hand, in the group treated with EC-18 alone, there was no decrease in body weight and diet, and in the treatment with cisplatin, the diet and body weight gradually recovered after 14 days. Unlike the occurrence of oral mucositis symptoms independent of tumor growth inhibition by cisplatin, simultaneous treatment with EC-18 recovered oral mucositis symptoms along with the reduction of tumor growth (P&lt;0.05). This effect could be explained by the fact that the expression of inflammation-related factors (TNFalpha, IL-1beta, IL-6) and DAMP (HMGB1, Adenosine) were effectively controlled by EC-18 (P&lt;0.05).
Conclusion: EC-18 can effectively inhibit the growth of HNSCC and alleviate the side effects caused by existing anticancer drugs at the same time. Therefore, it can be a desirable therapy for HNSCC patients.
Citation Format: Guentae KIM, Eun Young Kim, Su-Hyun Shin, Hyowon Lee, Sun Young Yoon, Kaapjoo Park, Ki-young Sohn, Jae Wha Kim. EC-18 suppressed tumor growth and alleviated side effects caused by cisplatin in the HNSCC implantation model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5478.
Abstract 5484: Preclinical evaluation of CAP-0121 for multidrug resistant cancers
Authors: Kelner Michael J. , Kotamraju Venkata R. 
Published: June 2022

Abstract
CAP-0121 is a second generation illudofulvene agent that produces an unusual type of DNA damage and is being developed to treat ovarian cancer by Califia Pharma. CAP-0121 DNA damage, like other illudofulvenes, can only be recognized and repaired by the Transcription-Coupled Repair pathway, and is ignored by the Global Genome Repair Pathway. The first generation illudofulvene, irofulven, displayed significant activity in clinical trials against ovarian, prostate and hepatocellular tumors. While irofulven has proven to be a potent antitumor agent against drug-resistant tumors, its efficacy is limited by hematologic toxicity that reduced the maximum dosage that can be administered to patients. CAP-0121 is as cytotoxic, or even more cytotoxic, than irofulven when tested in the NCI 60 cell line panel. Despite this increased cytotoxicity noted in vitro, CAP-0121 is less toxic to animals and the maximum tolerated dose is approximately 3 times higher than irofulven (mg/kg basis). CAP-0121 also retains activity against multidrug resistant cell lines including MRP1 and MDR1 expressing variants. As a result CAP-0121 is capable of inducing tumor remission in MDR xenograft models that are unresponsive to 35 conventional and experimental agents, and in which irofulven is only capable of slowing the rate of tumor growth. This response of CAP-0121 in drug resistant xenografts is observed at nontoxic doses. Synergistic action is observed when CAP-0121 is administered in combination with traditional drugs used for treating ovarian cancer, including taxanes, platinum agents, and PARP inhibitors.
Citation Format: Michael J. Kelner, Venkata R. Kotamraju. Preclinical evaluation of CAP-0121 for multidrug resistant cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5484.
Abstract 549: The IgG4 hinge with CD28 transmembrane domain mediates CAR dimerization and improves the activity of glypican 1-targeted CAR T cells in pancreatic cancer
Authors: Li Nan , Li Dan , Pan Jiajia , Cachau Raul , Ho Mitchell 
Published: June 2022

Abstract
Low antigen density has been suggested as a major cause of insufficient antitumor activity of chimeric antigen receptor (CAR) T cells. The impact of the hinge and the transmembrane (TM) domain on modulating CAR T cell reactivity is important to investigate. Here, we isolated a camel nanobody D4 recognizing glypican 1 (GPC1), a cell surface proteoglycan that is expressed in pancreatic cancer. We constructed a series of D4 4-1BBζ CARs differing only by their hinge (CD8 or IgG4) and TM (CD8 or CD28). D4 CARs containing a mutated IgG4 hinge and the CD28 TM, but not CD8 hinge with either TM, formed dimers in human T cells and resulted in improved stability of the CAR extracellular domain in a 3D structure model. The CAR dimerization increased phosphorylation of T cell signaling molecules, activated non-canonical NF-κB signaling after antigen stimulation. Consequently, the D4-IgG4 hinge-CD28TM CAR T cells triggered the most cytokine secretion and showed the best reactivity against low GPC1-expressing pancreatic tumor cells among all the D4 CARs in vitro and in vivo. Conversely, a CAR with mutations of the two cysteine residues in the IgG4 hinge disrupted CAR dimerization, damaged the formation of TM, and resulted in the loss of improved reactivity. Furthermore, adding extra spacers of CH3 or CH2CH3 between IgG4 hinge and CD28TM CAR attenuated the CAR activity. By optimizing the hinge and TM, our data indicated that 4-1BBζ CAR T cell activity can be optimized against low antigen density tumors.
Citation Format: Nan Li, Dan Li, Jiajia Pan, Raul Cachau, Mitchell Ho. The IgG4 hinge with CD28 transmembrane domain mediates CAR dimerization and improves the activity of glypican 1-targeted CAR T cells in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 549.
Abstract 5491: Targeting importin-YAP axis in pancreatic ductal adenocarcinoma
Authors: Zeng Renya , Dong Jixin 
Published: June 2022

Abstract
Background: To date, pancreatic ductal adenocarcinoma (PDAC) remains to have a dismal prognosis, with a 5-year survival rate of only 10%, which brings out an imperative to develop new therapeutic strategies to improve patient outcome. Recently, aberrant nucleocytoplasmic transport machinery in cancer cells has emerged as a rational therapeutic target. We aim to validate the nuclear importin complex involved in the import of macromolecules across the nuclear membrane as a potential therapeutic target in PDAC.
Methods: Tet-inducible shRNA and FDA-approved agent (Ivermectin) were used for the inhibition of importins. Cell death (apoptosis and pyroptosis) was detected by western blot. In vivo tumor growth of human and mouse PDAC cells was evaluated in immunocompetent and immunodeficient animal models. MTT assay was employed to evaluate the patient-derived organoid viability following drug treatment.
Results: We found that PDAC cells/patients harbored unusually higher levels of importins. Inducible knockdown of importin-α2/α5 (KPNA2/KPNA1) or importin-β1 (KPNB1) strongly suppressed PDAC tumor growth and induced apoptosis and pyroptosis. Interestingly, the importin inhibitor Ivermectin (an FDA-approved antiparasitic agent) worked effectively in PDAC cells, patient-derived organoids and animal models. Mechanistically, we show that importins associate with YAP (the transcriptional coactivator of the Hippo pathway) and control its expression at mRNA levels. In line with these observations, the expression levels of importin-αs, importin-β1, and YAP are positively correlated in PDAC patients.
Conclusion: Our study identifies the importin complex as a prognostic marker and therapeutic target for pancreatic cancer and is expected to provide preclinical evidence for the evaluation of Ivermectin in PDAC patients.
Citation Format: Renya Zeng, Jixin Dong. Targeting importin-YAP axis in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5491.