bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5384: Cell growth of sunitinib-resistant renal cell carcinoma was inhibited by Shikonin through necroptosis induction and Akt/mTOR signaling inhibition
Authors: Juengel Eva , Markowitsch Sascha D. , Vakhrusheva Olesya , Akele Yasminn , Kitanovic Jovanna , Schupp Patricia , Slade Kimberly S. , Thomas Anita , Tsaur Igor , Mager Rene , Efferth Thomas , Haferkamp Axel
Published: June 2022

Despite the establishment of new therapy options during the last decades, advanced renal cell carcinoma (RCC) remains an uncurable disease, due to evolving drug-resistance. New, more efficient treatment strategies are still in urgent demand. Shikonin (SHI) from Traditional Chinese Medicine has exhibited anti-tumor properties in several tumor entities. Thus, in the current investigation we evaluated the impact of SHI on progressive growth and metastatic behavior, not only in therapy-sensitive (parental) but also in therapy-resistant RCC cells. Parental and sunitinib-resistant RCC cells, Caki-1, 786-O, KTCTL-26, and A498, were treated with SHI. Tumor cell growth, proliferation, clonogenic capacity, cell cycle phase distribution, induction of cell death (apoptosis and necroptosis) and the expression and activity of regulating and signaling proteins were evaluated. Moreover, the adhesion and chemotactic activity of the RCC cells after exposure to SHI were investigated. SHI did not affect cell motility but led to significant inhibition of progressive tumor cell growth in both therapy-sensitive and therapy-resistant RCC cells. Thereby, SHI significantly inhibited the growth, proliferation and clone formation in parental and sunitinib-resistant RCC cells by G2/M phase arrest through down-regulation of cell cycle activating proteins, blocking cell division. Furthermore, SHI induced apoptosis and necroptosis by activating necrosome complex proteins. Concomitantly, SHI impaired the Akt/mTOR pathway, pivotal for cell survival and growth. Thus, SHI may hold promise as an additive option in treating patients with advanced and therapy-resistant RCC.

Citation Format: Eva Juengel, Sascha D. Markowitsch, Olesya Vakhrusheva, Yasminn Akele, Jovanna Kitanovic, Patricia Schupp, Kimberly S. Slade, Anita Thomas, Igor Tsaur, Rene Mager, Thomas Efferth, Axel Haferkamp. Cell growth of sunitinib-resistant renal cell carcinoma was inhibited by Shikonin through necroptosis induction and Akt/mTOR signaling inhibition [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 5384.
Abstract 5385: Multiple modes of action of the CD38 x ICAM-1 bispecific antibody
Authors: Chen Xiaocheng , Wong Oi Kwan , Post Leonard
Published: June 2022

The cluster of differentiation 38 (CD38) is a well validated target for treating multiple myeloma. Although the anti-CD38 monoclonal antibody treatment has resulted in outstanding responses in patients with multiple myeloma, a significant portion of patients still failed to respond and nearly all patients eventually relapsed. The mechanism of resistance has been partially associated with downregulation of CD38 expression. In addition, CD38 is broadly expressed on normal lymphoid, myeloid, as well as red blood cells (RBCs). To effectively target CD38 on tumor cells, we hypothesize that a bispecific antibody against a tumor associated guide antigen (TAG) and CD38 could achieve tumor specificity and improve potency. ICAM-1, an Ig-like cell adhesion molecule, is highly expressed in multiple myeloma, lymphoma and a subset of solid tumors. Antibody against ICAM-1, bersanlimab (BI505, BioInvent), was well-tolerated, but only showed limited clinical efficacy in MM patients. We generated bispecific CD38 x ICAM-1 antibodies with different formats and geometries and compared their in-vitro activities on ICAM1high tumor cells with low to medium CD38 expression. Our results showed that the asymmetric 1+1 format has potent ADCC and ADCP activities on ICAM-1+CD38medium or ICAM-1+CD38low cells. Fc modifications further improved ADCC activities of the bispecific antibody. To further understand the impact of antigen densities, we tested ADCC activities on a selected set of myeloma and lymphoma cell lines with a range of CD38 and ICAM-1 expressions. We observed superior ADCC activity on cells with extremely low CD38 expression. Moreover, we showed that the bispecific antibody reduced NK fratricide and RBC binding comparing to the bivalent CD38 antibody. Most importantly, the CD38 x ICAM1 bispecific antibody demonstrated potent tumor inhibition activities in in-vivo myeloma and lymphoma models. These studies suggested that CD38 x ICAM-1 bispecific antibody has multiple modes of action for treating lymphoma and multiple myeloma.

Citation Format: Xiaocheng Chen, Oi Kwan Wong, Leonard Post. Multiple modes of action of the CD38 x ICAM-1 bispecific antibody [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 5385.
Abstract 5389: Exploring potential predictive biomarkers for the responsiveness to CTX-712 by in vivo drug sensitivity analysis on PDX models
Authors: Morishita Daisuke , Yagishita Shigehiro , Mizutani Akio , Yamakawa Hiroko , Ebara Shunsuke , Hamada Akinobu
Published: June 2022

Cancer’s remarkable diversity had been understood through a logical framework of six key hallmarks. Additionally, several equally prevalent hallmarks have been identified in recent analyses of cellular phenotypes as the stress phenotypes of cancers. A novel hallmark of cancer RNA deregulation has been newly proposed as another hallmark of cancer within this group of stress phenotypes. However, the development of cancer therapies targeting RNA deregulation has only just begun. Indeed, small molecules that attack the RNA maturation processes and produce aberrant RNA are currently under development. We developed orally available, selective, and a first-in-class pan- CDC-like kinase (CLK) inhibitor, CTX-712, currently in phase 1 clinical trials for advanced, relapsed or refractory malignant cancers. CTX-712 dephosphorylates serine and arginine-rich proteins and induces primarily skipped exon type of splicing changes, resulting in the generation of RNA deregulation stress. In multiple preclinical models, excessive RNA deregulation stress caused cancer cell death and tumor growth inhibition. To explore the patient selection biomarkers for CTX-712, in vivo efficacy survey analysis has been conducted on breast, ovarian, colorectal, pancreatic, gastric and lung cancer models from patient derived xenograft (PDX) library with a detailed clinical information. Besides the efficacy investigation, the preclinical pharmacokinetics (PK) and pharmacodynamics (PD) analysis on tested PDX models were also performed to confirm plasma/tumor PK and PD (target CLK inhibition in xenografted tumors). Importantly, tested PDX models could be divided to sensitive (T/C on the most sensitive model = -71.1) and insensitive (T/C on the least sensitive model = 67.7), and the correlated tendency was observed between the efficacy and the inhibition magnitude of intratumor PD marker, production of exon skipped aberrant mRNA. To clarify the differences of sensitivity among tested PDXs will lead us to identity patient selection biomarkers of CTX-712. Therefore, whole transcriptome RNA sequencing, exome sequencing, and other approaches have been applied to reveal the differences of sensitivity among tested PDXs. Through this study on multiple PDXs, integration of drug responsiveness, and genomic/transcriptomic profiling may provide prediction markers for treatment responsiveness of CTX-712.

Citation Format: Daisuke Morishita, Shigehiro Yagishita, Akio Mizutani, Hiroko Yamakawa, Shunsuke Ebara, Akinobu Hamada. Exploring potential predictive biomarkers for the responsiveness to CTX-712 by in vivo drug sensitivity analysis on PDX 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 5389.
Abstract 5395: DNA damage repair gene mutations predict the efficacy of platinum-based chemotherapy in colorectal cancer
Authors: Lin Yan , Zhang Jinyan , Liao Xiaoli , Zhang Yumei , Luo Min , Li Qian , Xie Mingzhi , Liang Chaoyong , Liao Sina , Zheng Yating , Hu Xue , Huang Mengli , Liang Rong , Li Yongqiang
Published: June 2022

Background: DNA damage repair (DDR) mutations are known to predict response to platinum-based chemotherapy in multiple solid tumors. However, their predictive value remained unknown in patients with colorectal cancer.

Methods: The genomic and survival datas from the TCGA-COAD and TCGA-READ cohorts of patients receiving platinum-based chemotherapy were used to analyze the predictive value of DDR mutations on platinum-based chemotherapy.

Result: The DDR genes were commonly mutated (85.82%) in the TCGA-COAD and TCGA-READ cohorts. The objective response rates (ORRs) were 80% for the patients with DDR mutations (DDRmut) subgroup and 56% for the DDR wild-type (DDRwt) subgroup (P<0.05), and the disease control rates (DCRs) were 86% for the DDRmut subgroup and 56% for the DDRwt subgroup (P<0.05). In patients with stage I, II and III colorectal cancer, there was no significant difference in the overall survival (OS) between DDRmut subgroup and DDRwt subgroup (Hazard Ratio=0.48, 95%CI 0.1−2.31, log-rank P=0.35). In patients with stage IV colorectal cancer, the OS was significantly better among the DDRmut patients than in the DDRwt subgroup (Hazard Ratio=0.21, 95%CI 0.06−0.8, P= 0.011).

Conclusions: DDR mutations may serve as a positive predictor of platinum-based chemotherapy therapy in patients with CRC and their clinical value warrants further investigation.

Citation Format: Yan Lin, Jinyan Zhang, Xiaoli Liao, Yumei Zhang, Min Luo, Qian Li, Mingzhi Xie, Chaoyong Liang, Sina Liao, Yating Zheng, Xue Hu, Mengli Huang, Rong Liang, Yongqiang Li. DNA damage repair gene mutations predict the efficacy of platinum-based chemotherapy in colorectal 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 5395.
Abstract 5400: Detection of satellite tumor cells in peritumoral edema in myxofibrosarcoma and undifferentiated pleomorphic sarcoma using targeted gene sequencing
Authors: Principe Miguel Alfonso V. , Gokgoz Nalan , Prochazka Patrick , Ferguson Peter C. , Dewji Simin , Wunder Jay S. , Andrulis Irene A. , Dickson Brendan C. , Tsoi Kim M.
Published: June 2022

Myxofibrosarcoma (MFS) and undifferentiated pleomorphic sarcomas (UPS) are soft tissue sarcomas with high local recurrence rates following resection. Common with these cancers is peritumoral edema, termed tumor tails, which our group has found could contain satellite tumor cells. A precise and accurate method of determining the presence of satellite tumor cells in this region is required to improve surgical planning, as the standard morphology-based technique is prone to misclassification. An emerging technology is targeted gene sequencing (t-NGS), a combination of whole genome or exome sequencing (WGS/WXS) comparing primary tumor and patient-matched blood, and digital droplet PCR (ddPCR). The objective of this study was to determine the viability of t-NGS in the detection of satellite tumor cells in the areas of edema surrounding MFS and UPS. A cohort of patients diagnosed with extremity MFS and UPS at Mount Sinai Hospital (Toronto, Canada) who underwent resection were retrospectively studied. Cases with available WGS/WXS data were used (n=8) to identify case-specific tumor-specific variants (TSV) to detect with the QX200 ddPCR platform. (n=4-5/case). Cases with a major TSV-positive droplet population in the tumor tails tissue indicated the presence of satellite tumor cells. Our quality analysis showed DNA above the ddPCR threshold of detection was extracted from the tumor tails of 6 of 8 cases. The ddPCR assay showed that a TSV-positive droplet population was detected within the tumor tails in 5 of 6 cases, while 1 of 6 cases showed no such population in the tumor tails. Overall, our results indicates that t-NGS has potential for TSV detection within tumor tails. A prospective study is underway to further test its validity and compare it with standard histology. The project aims to improve surgical planning by ensuring removal of satellite tumor cells while lowering the risk of wound complications.

Targeted gene-sequencing results of all cases within this study Case # Histology Pre-op Radiation DNA Extraction Successful TSV Probes (n) TSV+ Tumor TSV+ Tails TSV+ Normal 1 MFS N Y 5 4 4 0 2 MFS N Y 3 2 2 0 3 MFS Y Y* (normal insufficient) 3 3 0 N/A 4 UPS Y Y 3 2 0 1 5 MFS N Y 4 4 1 0 6 UPS N Y 3 3 3 1 7 MFS Y N N/A N/A N/A N/A 8 MFS Y N N/A N/A N/A N/A

Citation Format: Miguel Alfonso V. Principe, Nalan Gokgoz, Patrick Prochazka, Peter C. Ferguson, Simin Dewji, Jay S. Wunder, Irene A. Andrulis, Brendan C. Dickson, Kim M. Tsoi. Detection of satellite tumor cells in peritumoral edema in myxofibrosarcoma and undifferentiated pleomorphic sarcoma using targeted gene sequencing [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 5400.
Abstract 5401: Fluoxetine inhibit osteosarcoma progression via RANK/RANKL signaling pathway
Authors: Tan Peggy , Hsu Fei-Ting , Chiang I-Tsang , Chang Yuan
Published: June 2022

Osteosarcoma (OS) is an aggressive malignant bone cancer commonly occurs on adolescent. OS was mostly found at sites of bone growth. Sixty percent OS cases happen at knee, 15% around hip, 10% shoulder and 8% jaw. Non-stop osteoclast cells differentiation by which osteoblasts secrete RANKL bind to RANK receptor on osteoclasts’ cell surface, stimulate rapid bone metabolism and considerable as an important signaling pathway correlate with OS progression.

Depressant is usually occurring in cancer patients. Several reports suggested that several anti-depressant drugs presented anti-tumour potential. Previous study has reported, fluoxetine, a prescription antidepressant can inactivate NF-κB and thus suppress hepatocellular carcinoma and non-small cell lung cancer progression. Furthermore, fluoxetine may inhibit DNA repair in non-small cell lung cancer.

In our study, the MTT assay demonstrated fluoxetine significantly reduced viability of U-2 OS cells. Next, target protein RANK/RANKL expression were also downregulated by fluoxetine in dose dependent manner. Besides, STAT3 mediated NF-κB activity, were also inactivated by fluoxetine. Additionally, the expression of PARP1 (DNA damage related, and several oncogenes such as XIAP and Cyclin D1 were also downregulated by fluoxetine in dose dependent manner. Moreover, the inhibition of tumour angiogenesis by fluoxetine has also proven by using aorta ring assay. Further in vivo experiment may need to be performed to identify anti-tumor efficacy of fluoxetine.

In sum, fluoxetine can be used anti-tumour adjuvant of OS therapy and the detailed mechanism or further combination strategy may be worth exploring.

Citation Format: Peggy Tan, Fei-Ting Hsu, I-Tsang Chiang, Yuan Chang. Fluoxetine inhibit osteosarcoma progression via RANK/RANKL signaling pathway [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 5401.
Abstract 5406: Novel quatramer nano-formulation of dual-targeted PI3-Kδ/HDAC6 inhibitor, HSB-510, suppresses growth of triple negative breast cancer
Authors: Tiwari Sachchidanand , Liu Suiyang , Anees Mohammad , Mehrotra Neha , Thakur Ashish , Tawa Gregory , Grewal Gurmit , Stone Richard M. , Kharbanda Surender M. , Singh Harpal
Published: June 2022

Triple negative breast cancer (TNBC) is an aggressive subtype with limited treatment options and a worse clinical outcome as compared with other breast cancer subtypes. Multiple studies have shown that progression of TNBC depends on more than one signalling pathways, suggesting that the therapies with multi-targeted anti-cancer agents will offer improved therapeutic benefit through synergistic effects in inhibiting cancer growth. Dual-targeted inhibitors of phosphoinositide 3-kinase (PI3-K) and histone deacetylase (HDAC) have emerged as promising cancer therapy candidates. However, poor aqueous solubility and bioavailability limited their efficacy in cancer and accordingly, improved delivery systems are needed. The present studies have investigated the encapsulation of PI3-Kδ/HDAC6 dual inhibitor into hybrid block co-polymers (polylactic acid-methoxy polyethylene glycol; polylactic acid-polyethylene glycol-polypropylene glycol-polyethylene glycol-polylactic acid) (HSB-510) as a delivery system to target PI3-Kδ and HDAC6 pathways in TNBC cells. The prepared HSB-510 Quatramer nano-formulation showed an average diameter of 96 ±3 nm, the zeta potential of -17±2 mV, and PDI of ˂0.1 with a slow and sustained release profile of PI3-Kδ/HDAC6 inhibitors in a non-physiological buffer. In vitro studies with HSB-510 have demonstrated substantial growth inhibition of TNBC cell lines, MDA-MB-468, SUM-149 and Ehrlich ascites carcinoma (EAC) as well as downregulation of phospho-AKT, phospho-ERK and c-MYC levels. Importantly, bi-weekly treatment of BALB/c wild-type mice harbouring EAC cells (syngeneic mouse model) with HSB-510 at a dose of 25 mg/kg resulted in significant (76%) tumor growth inhibition (TGI). The treatment with HSB-510 was without any significant effect on the body weights of the mice. These results demonstrate that a novel Quatramer nano-formulation of a novel PI3-Kδ/HDAC6 dual inhibitor (HSB-510) represents an approach for successful targeting of TNBC and potentially other cancer types.

Citation Format: Sachchidanand Tiwari, Suiyang Liu, Mohammad Anees, Neha Mehrotra, Ashish Thakur, Gregory Tawa, Gurmit Grewal, Richard M. Stone, Surender M. Kharbanda, Harpal Singh. Novel quatramer nano-formulation of dual-targeted PI3-Kδ/HDAC6 inhibitor, HSB-510, suppresses growth of triple negative breast 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 5406.
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

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

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 >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

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 >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

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

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

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<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<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<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<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

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

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

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.
Abstract 55: A genome-wide atlas of recurrent repeat expansions in human cancer genomes
Authors: Erwin Graham S. , Gursoy Gamze , Al-Abri Rashid , Hoerner Christian , Dolzhenko Egor , Eberle Michael , Fan Alice , Leppert John , Gerstein Mark , Snyder Michael P.
Published: June 2022

Expansion of a single repetitive DNA sequence, termed a tandem repeat (TR), is known to cause more than 40 neurodegenerative diseases. However, repeat expansions have not been systematically examined in contexts beyond neurological disorders. In a specific form of cancer, termed microsatellite instability (MSI), mutations accumulate in short (<50 bp) tracts of TRs, suggesting that cancer genomes may also harbor larger repeat expansions. Here, we developed a new method to systematically identify TR alterations and applied it to the analysis of 2,622 cancer genomes spanning 29 cancer types. We identified 160 recurrent repeat expansions (rREs) in 7 cancers; most (155/160) rREs were cancer-subtype specific. Surprisingly, rREs were not correlated with MSI status, demonstrating that they arise by a distinct process. We also found that rREs are situated closer to candidate cis-regulatory elements than expected by chance, suggesting a role in gene regulation. Interestingly, an rRE in the intron of UGT2B7, detected in 34% of renal cell carcinoma samples, is associated with a decrease in gene expression. Our results demonstrate that rREs are an important and unexplored source of genetic variation in human cancer and provide a large catalog for further study.

Citation Format: Graham S. Erwin, Gamze Gursoy, Rashid Al-Abri, Christian Hoerner, Egor Dolzhenko, Michael Eberle, Alice Fan, John Leppert, Mark Gerstein, Michael P. Snyder. A genome-wide atlas of recurrent repeat expansions in human cancer genomes [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 55.
Abstract 5504: Adoptive cell therapy with tumor-specific Th9 cells induces viral mimicry to restrain acquired resistance from antigen-loss-variant tumor cells
Authors: Lu Yong
Published: June 2022

Adoptive cell therapy (ACT) with tumor-specific T cells expressing T-cell-receptors (TCR) or chimeric antigen receptors (CAR) has not mediated sustained responses in solid tumors. Several factors limit the efficacy of ACT in solid tumors and lead to primary resistance and acquired resistance (refers to recurrence after the initial response). These include but not limited to the insufficient proliferation of T cells, the low capacity of T cells to penetrate stroma-rich solid tumors, and heterogeneity of antigen expression in solid tumors, which allows the escape of T cell attack by tumor cells when becoming antigen-loss-variant cancer cell (ALV). Hence, it is urgent to develop novel ACT strategies to eradicate solid tumors and prevent acquired resistance.

We have previously reported (Yong Lu, Cancer Cell, 2018) that tumor-specific IL-9-producing CD4+ Th9 cells represent a novel T cell paradigm for ACT - they are less exhausted, fully cytolytic, and hyperproliferative. This hyperproliferation of Th9 cells is driven by a unique Th9 master transcriptional factor Pu.1 mediated Pu.1-Traf6-NFκB signaling to ensure Th9 cells persist as long as “stem cell-like” T cells, which is not presented in other known CD4+ T cells subsets. In our most recent study, we demonstrate that murine and human Th9 cells, but not Th1/Tc1 or Th17 cells, expressing tumor-specific T-cell-receptors (TCR) or chimeric antigen receptors (CAR), eradicate advanced tumors that contain ALVs. This unprecedented antitumor capacity of Th9 cells is attributed to both enhanced direct tumor cell killing and bystander antitumor effects promoted by intratumor release of IFNα/β. Mechanistically, tumor-specific Th9 cells increase the intratumor accumulation of extracellular ATP (eATP, released from dying tumor cells), because of a unique feature of Th9 cells that lack the expression of ATP degrading ectoenzyme CD39. Intratumor enrichment of eATP promotes the monocyte infiltration and stimulates their production of IFNα/β by inducing eATP-endogenous retrovirus-TLR3/MAVS pathway activation. These results identify tumor-specific Th9 cells as a unique T cell subset endowed with the unprecedented capacity to eliminate ALVs for curative responses.

Citation Format: Yong Lu. Adoptive cell therapy with tumor-specific Th9 cells induces viral mimicry to restrain acquired resistance from antigen-loss-variant tumor cells [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 5504.