bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2620: TRIM4 is the E3 ubiquitin-ligase for TPL2/MAP3K8 and is modulated by oncogenic KRAS
Authors: Bansod Sapana Prakashrao , Dodhiawala Paarth B. , Bulle Ashenafi , Liu Peng , Li Lin , Grierson Patrick M. , Held Jason , Lim Kian-Huat 
Published: April 2023

Abstract
TPL2 (tumor progression locus 2, a. k. a. Cot/MAP3K8) kinase becomes oncogenic via disruption of the C-terminus, which abrogates its proteasomal degradation. However, the mechanism underlying degradation of TPL2 remains unclear. Using proximity-dependent biotin identification (BioID), for the first time we have discovered that Tripartite Motif Containing 4 (TRIM4) is the E3-ligase that binds and degrades TPL2 via polyubiquitination of lysine 415 and 439. Notably, naturally-occurring oncogenic R442H and E188K mutations in TPL2 abrogated the binding and polyubiquitination by TRIM4. Furthermore, KRAS oncoprotein stabilizes TPL2 by promoting TRIM4 degradation. Using immunoprecipitation-mass spectrometry, we identified TRIM21 as a ligase that binds and stabilizes TRIM4, in part by reducing TRIM4 polyubiquitination. Therefore, oncogenic KRAS binds to and promotes TRIM21 degradation, destabilizing TRIM4 and prolonging the half-life of TPL2. Overall, we discovered novel mechanisms of TPL2 regulation which are influenced by oncogenic KRAS, providing opportunities for the development of novel therapies.
[S. Bansod and P.B. Dodhiawala contributed equally to this work.]
Citation Format: Sapana Prakashrao Bansod, Paarth B. Dodhiawala, Ashenafi Bulle, Peng Liu, Lin Li, Patrick M. Grierson, Jason Held, Kian-Huat Lim. TRIM4 is the E3 ubiquitin-ligase for TPL2/MAP3K8 and is modulated by oncogenic KRAS [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2620.
Abstract 2621: Characterizing MEK1/2 degradation in K-ras mutant cells
Authors: Herrera-Montavez Carlos , Kurimchak Alison M. , Jin Jian , Duncan James S. 
Published: April 2023

Abstract
Activating KRAS mutations frequently occur in several cancers including the majority of pancreatic ductal adenocarcinoma (96%), ~50% of colorectal cancers, and ~30% of lung adenocarcinoma. It has been shown that mutant K-ras preferentially signals through RAF-MEK-ERK signaling and that blockade of this pathway is essential for tumor regression. However, targeted MEK inhibitors (MEKi) have shown limited clinical benefit in the treatment of K-ras mutant cancers due to resistance caused by the feedback activation of the upstream kinase CRAF. Recently, PROTACs (Proteolysis-Targeting Chimeras) targeting MEK1 and MEK2 were developed, providing a new therapeutic strategy to degrade MEK1/2 in K-ras mutant cancer cells. Due to the fact that feedback-induced CRAF causes MEKi resistance in K-ras mutant cells, we postulated that degrading MEK1/2 could bypass CRAF-mediated resistance. Indeed, treatment of MEKi-sensitive or resistant K-ras mutant cell lines with MEK1/2 degraders rapidly reduced MEK1/2 protein levels and downstream ERK signaling causing growth inhibition and apoptosis. Proteomics characterization of K-ras mutant cell lines following MEK1/2 degrader or inhibitor treatment revealed MEK1/2 degradation exhibited distinct signaling pathway responses compared to inhibition. Together, our studies showed MEK1/2 degraders represent a promising avenue for the treatment K-ras mutant cancers that can by-pass resistance mechanisms observed with traditional MEK1/2 inhibitors.
Citation Format: Carlos Herrera-Montavez, Alison M. Kurimchak, Jian Jin, James S. Duncan. Characterizing MEK1/2 degradation in K-ras mutant 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 2621.
Abstract 2623: Identification of KRAS suppressor genes: an ORFeome library screening
Authors: Marcireau Christophe , Furge Philippine , Sennechael Fanny , Willart Alice , Blanchet Anne-Marie , Yaligara Veeranagouda , Didier Michel , Lacroix Frederic , Meunier Melina , Debussches Laurent 
Published: April 2023

Abstract
The RAS family genes encode small GTP-binding cytoplasmic proteins that are important signaling molecules. They regulate cell growth, survival and differentiation by coupling receptor activation to downstream effector pathways. Activating mutations of oncogenic RAS pathway genes are frequently detected in human cancers. KRAS is the most frequently mutated oncogene in human cancers. Most of these tumors are addicted to KRAS signaling. However, tumors can become resistant to RAS signaling pathway inhibitors through the activation of suppressor mechanisms. A possible explanation is that these tumors carry a second genomic perturbation that can compensate for the detrimental effects of the RAS signaling inhibitors, a phenomenon referred to as suppression. In model organisms, suppression interactions are generally divided into two classes: genomic suppressors which are secondary mutations in the genome that bypass a mutant phenotype, and dosage suppression interactions in which overexpression of a suppressor gene rescues a mutant phenotype. We decided to use the latter strategy in human cancer cells to identify KRAS suppressor genes. KRAS suppressor genes can be involved in resistance mechanisms of RAS pathway inhibitors and this approach could identify new targets for drug research programs and/or novel drug combinations. In this study we performed a screen on a human Open Reading Frame library (hORFeome collection) to identify potential KRAS suppressor genes. First, we engineered a SW620 cell line with a conditional expression of KRAS G12V by replacing the KRAS endogenous promoter by the doxycycline regulated TREG3 promoter. Second, we transduced this conditional cell line with a human ORFeome collection, and we looked for suppressors among the transduced cells that lead to cell proliferation in absence of KRAS G12V protein. Then candidate suppressors have been identified by NGS and validated after individual cloning and expression in SW620 KRAS conditional cells. Finally, two transcription factors and two orphan G protein-coupled receptors have been validated as KRAS suppressor genes when they are highly expressed.
Citation Format: Christophe Marcireau, Philippine Furge, Fanny Sennechael, Alice Willart, Anne-Marie Blanchet, Veeranagouda Yaligara, Michel Didier, Frederic Lacroix, Melina Meunier, Laurent Debussches. Identification of KRAS suppressor genes: an ORFeome library screening [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 2623.
Abstract 2633: Zanidatamab zovodotin (ZW49) induces hallmarks of immunogenic cell death and is active in patient-derived xenograft models of gastric cancer
Authors: Barnscher Stuart D. , Rojas Andrea Hernández , Hamblett Kevin J. , Escalante Nichole 
Published: April 2023

Abstract
Zanidatamab zovodotin (ZW49) is an antibody drug conjugate (ADC) comprised of a bispecific anti-HER2 IgG1 antibody (ZW25, zanidatamab) conjugated to a microtubule inhibitor auristatin payload (ZD02044) via a protease cleavable linker. Clinical studies of ZW49 in HER2-expressing advanced solid tumors are currently underway (NCT03821233). We have previously presented data illustrating the rapid internalization of ZW49 and the anti-tumor activity of ZW49 in multiple patient-derived xenograft models originating from breast carcinomas. Here we present ZW49’s ability to induce in vitro hallmarks of immunogenic cell death (ICD) including increased calreticulin exposure, increased high mobility group box-1 (HMGB1) exposure, and increased extracellular ATP secretion in a HER2 dependent manner. Additionally, we present the results from a study evaluating the anti-tumor activity of ZW49 in a panel of patient-derived xenografts of gastric cancer. HER2-positive SK-BR-3 and HER2-negative MDA-MB-468 tumor cell lines were treated with ZW49 and cell surface calreticulin was assessed by flow cytometry. ZW49 treatment of SKBR-3 cells induced a significantly higher percentage of cells staining positive for cell surface calreticulin compared to untreated cells. In contrast, ZW49 treatment of MDA-MB-468 cells resulted in no significant difference in the percentage of cells staining positive for calreticulin compared to untreated cells. ZW49 treated HER2-positive cancer cell lines SK-BR-3 and SK-OV-3 induced higher levels of extracellular ATP compared to untreated cells. In the HER2-negative cell line MDA-MB-468, treatment with ZW49 did not alter levels of extracellular ATP compared to untreated cells. ZW49 induced higher levels of HMGB1 in the HER2-positive cancer cell lines SKBR3 and N87 compared to untreated cells, whereas in the HER2-negative cell line MDA-MB-468, ZW49 induced similar levels of HMGB1 to untreated cells. Anti-tumor activity was observed in 5/7 (71%) PDX models of gastric cancer after a single i.v. dose of 6 mg/kg, including in models with moderate and weak HER2 expression. The strong anti-tumor activity of ZW-49 in vivo, together with its ability to induce ICD and potential adaptive immune responses, support ZW49 as a promising ADC for the treatment of HER2-expressing cancers warranting further investigation, including potential combination with checkpoint inhibitors.
Citation Format: Stuart D. Barnscher, Andrea Hernández Rojas, Kevin J. Hamblett, Nichole Escalante. Zanidatamab zovodotin (ZW49) induces hallmarks of immunogenic cell death and is active in patient-derived xenograft models of gastric 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 2633.
Abstract 2666: Preclinical investigations of concomitant tumor treating fields (TTFields) with cisplatin or paclitaxel for treatment of cervical cancer
Authors: Frechtel-Gerzi Roni , Gerasimova Daria , Zeevi Einav , Schlachet-Drukerman Inbar , Mumblat Helena , Martinez-Conde Antonia , Dor-On Eyal , Tzchori Itai , Haber Adi , Giladi Moshe , Weinberg Uri , Palti Yoram , Palmer Greg , Secord Angeles A. 
Published: April 2023

Abstract
Introduction: Cervical cancer is a serious health problem despite the fact it is highly preventable. Early-stage cervical cancer treatment often involves surgery; however advanced, recurrent, or metastatic cases require systemic therapy. Chemotherapy, mainly cisplatin, is the most commonly used systemic therapy for this cancer type. Combination regimens may be needed in the recurrent or metastatic settings, with first-line treatments including cisplatin with paclitaxel. TTFields are electric fields that disrupt cellular processes critical for cancer cell viability and tumor progression. Concomitant treatment with TTFields and cisplatin or paclitaxel has shown benefit in other tumor types. In the current in vitro study, we tested the effectiveness of TTFields for the treatment of cervical cancer, and the possible benefit of applying TTFields together with first-line treatments for cervical cancer.
Methods: Human cervical cancer cell lines - squamous cell carcinoma Ca Ski and SiHa cells, and adenocarcinoma HeLa cells - were treated with TTFields (72 h, 1 V/cm RMS) at frequencies of 100 to 400 kHz, and tested for cell count. For examining the efficacy of TTFields concomitant with cisplatin or paclitaxel, various doses of the drugs were applied together with TTFields (200 kHz), followed by measurements of cell count, colony formation, and apoptosis. Overall effect was defined as the product of percent reductions in cell count and colony formation.
Results: TTFields treatment reduced cell count in all tested cervical cancer cell lines. 200 kHz were found to be effective and were used throughout the experiments. Dose response effects were seen with cisplatin or paclitaxel, and were augmented when TTFields were co-applied to the cells.
Conclusions: These preclinical data suggest that TTFields may be an effective treatment against cervical cancer, and that applying them concomitantly with first-line treatment for this malignancy may provide enhanced effectiveness.
Citation Format: Roni Frechtel-Gerzi, Daria Gerasimova, Einav Zeevi, Inbar Schlachet-Drukerman, Helena Mumblat, Antonia Martinez-Conde, Eyal Dor-On, Itai Tzchori, Adi Haber, Moshe Giladi, Uri Weinberg, Yoram Palti, Greg Palmer, Angeles A. Secord. Preclinical investigations of concomitant tumor treating fields (TTFields) with cisplatin or paclitaxel for treatment of cervical 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 2666.
Abstract 2673: Several LSD1 inhibitors have a potential to disturb fatty acid metabolism necessary &It glycolysis-inhibited pancreatic cancers to survive
Authors: Zhang Zhiheng , Aoki Haruna , Umezawa Keitaro , Oshima Taichi , Hirao Takuya , Miura Yuri , Ito Kousei , Aoki Shigeki 
Published: April 2023

Abstract
Despite increased aerobic glycolysis is generally seen in human cancers, pancreatic ductal adenocarcinoma (PDAC) cells keep alive in glycolysis-suppressed conditions (1). Here, we sought there are potential therapeutic targets in glycolysis-suppressed PDAC cells. By screening anti-cancer metabolism compounds, we identified SP2509, a selective lysine specific demethylase 1 (LSD1) inhibitor (2). We also tested the effect of another three LSD1 inhibitors (Iadademstat, OG-L002 and T-3775440) on PDAC cells viability. The results showed that OG-L002, but not Iadademstat or T-3775440, lowered glycolysis-suppressed PDAC cells viability, like SP2509. Importantly, only knockdown of LSD1 or LSD2 failed to lower the viability of glycolysis suppressed PDAC cells. In our previous study, we proved that glucose-starvation make PDAC cells yield survival to mitochondrial oxidative phosphorylation (1). In this study, we cleared fatty acids metabolism played a crucial role for glycolysis suppressed PDAC cells survival. Then, we confirmed that SP2509 and OG-L002 can lead lipid droplets accumulation in glycolysis-suppressed conditions. In conclusion, our results showed that SP2509 and OG-L002 may involve regulatory effects on lipid droplets homeostasis, which control storage and release of fatty acids, not only by targeting LSD1 or LSD2. Moreover, SP2509 and OG-L002 significantly lowered viability of glycolysis-suppressed PDAC cells, supposing a novel therapeutic potential of approaches to override resistance of glucose-starvation in PDAC cells.
1.Shiratori R, Furuichi K, Yamaguchi M, Miyazaki N, Aoki H, Chibana H, et al. Glycolytic suppression dramatically changes the intracellular metabolic profile of multiple cancer cell lines in a mitochondrial metabolism-dependent manner. Sci Rep. 2019;9(1):18699. 2.Fiskus W, Sharma S, Shah B, Portier BP, Devaraj SGT, Liu K, et al. Highly effective combination of LSD1 (KDM1A) antagonist and pan-histone deacetylase inhibitor against human AML cells. Leukemia. 2017;31(7):1658.
Citation Format: Zhiheng Zhang, Haruna Aoki, Keitaro Umezawa, Taichi Oshima, Takuya Hirao, Yuri Miura, Kousei Ito, Shigeki Aoki. Several LSD1 inhibitors have a potential to disturb fatty acid metabolism necessary &It glycolysis-inhibited pancreatic cancers to survive [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 2673.
Abstract 2674: Synergistic combinations of lurbinectedin with irinotecan and ONC212 in pancreatic cancer
Authors: Tummala Tej , Cruz Arielle De La , Uruchurtu Ash , Liguori Nicholas R. , Abbas Abbas E. , Zhang Leiqing , Azzoli Christopher G. , Zhou Lanlan , El-Deiry Wafik S. 
Published: April 2023

Abstract
Pancreatic cancer is a devastating disease with a five-year survival rate below 10% according to the American Cancer Society. Novel chemotherapeutics and targeted therapies in pancreatic cancer have shown limited success, illustrating the urgent need for new treatments. Lurbinectedin is a chemotherapeutic synthetic tetrahydroisoquinoline alkaloid that inhibits active transcription by binding to guanine rich sequences in the minor groove of DNA. Lurbinectedin has been shown to reduce oncogenic transcription by stalling and degrading RNA Polymerase II while also inducing single- and double-stranded breaks to DNA, causing subsequent apoptosis. Lurbinectedin received accelerated FDA approval in 2020 for metastatic small cell lung cancer on or after platinum-based chemotherapy and is currently undergoing clinical trials in a variety of tumor types. We recently described a synergistic interaction between lurbinectedin and ONC201/TIC10, a compound that induces the TRAIL pathway, in killing SCLC cell lines associated with activation of p-Chk1 and the integrated stress response. We now demonstrate lurbinectedin’s efficient killing of pancreatic tumor cells as a single agent in PANC-1, BxPC-3, and HPAF-II cell lines, with IC-50s corresponding to sub-nanomolar concentrations. We also demonstrate that a combination of lurbinectedin and irinotecan, a topoisomerase I inhibitor with FDA approval for advanced pancreatic cancer, results in synergistic killing of pancreatic tumor cells in vitro. We further demonstrate a combination of lurbinectedin and ONC212, an imipridone, the same class of compounds as ONC201, also results in synergistic killing of pancreatic tumor cells. Cell viability was measured using the CellTiterGlo assay at varying drug concentrations. We hypothesize a combination therapy of lurbinectedin and irinotecan or ONC212 can enhance immune cell killing of pancreatic tumor cells. This is being explored by co-culturing CD8+ T lymphoblast cells with pancreatic tumor cells treated with lurbinectedin, irinotecan, ONC212, and combination, along with molecular mechanisms of cell death and effects on cytokines in the tumor microenvironment. Our results are developing insights regarding molecular mechanisms underlying therapeutic efficacy of a novel combination drug treatment for pancreatic cancer.
Citation Format: Tej Tummala, Arielle De La Cruz, Ash Uruchurtu, Nicholas R. Liguori, Abbas E. Abbas, Leiqing Zhang, Christopher G. Azzoli, Lanlan Zhou, Wafik S. El-Deiry. Synergistic combinations of lurbinectedin with irinotecan and ONC212 in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2674.
Abstract 2698: Improved calcium phosphate delivery system cNaD to aim clinical application
Authors: Yamamoto Hirofumi 
Published: April 2023

Abstract
Super carbonate apatite (sCA) nanoparticle is a superb calcium phosphate delivery system for miRNA and siRNA to tumors (PlosOne 2015). We reported that sCA delivered efficiently siRNA for survivin and PTBP as well as miR-340 (Mol Cancer Ther 2015, 2015) to colon tumors implanted into nude mice. MiR-4689 and miR-29b-1-5p suppressed KRAS mutated colon cancer cells (Mol Ther NA2015, 2018) and miR-302 and miR-369 realized in vivo cancer reprograming (PlosOne 2015). In every case the RNA function was fully realized. Moreover, effect of influenza vaccination was augmented by sCA plus CpG nucleic acid (NA) (Frontiers in Immunol 2018). sCA coupled with miR-29a or miR-29b prevented the DSS (dextran sodium sulfate)-induced inflammatory bowel disease (Mol Ther NA2018). We also applied the sCA-ICG complex to minimally invasive treatment of photodynamic therapy (Mol Cancer Ther 2018). Following these outstanding findings, we tried and succeeded to establish a clinically available novel calcium phosphate delivery system, named cNaD, which controls the nano-particle size and avoids accumulation in normal organs including liver and spleen. It is characterized by its ability to efficiently deliver siRNA and microRNA to cancers and provided prominent therapeutic effect in various mouse models. Safety test has been accomplished in rats and monkeys based on GLP standards. Also, the scale-up manufacturing system &gt;1000 folds has been attained. Our cNaD system will accelerate the development of NA medicine in the world.
Citation Format: Hirofumi Yamamoto. Improved calcium phosphate delivery system cNaD to aim clinical application [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 2698.
Abstract 2700: ORM-6151: A first-in-class CD33-antibody enabled GSPT1 degrader for AML
Authors: Palacino James , Lee Pedro , Jeong Hangyeol , Kim Yeonjoon , Song Yoojin , Permpoon Uttapol , Wong Wesley , Bai Chen , Fishkin Nathan , Takouri Khuloud , Yu Eunjun , Yi Yong , Skaletskaya Anna , Kim Min-Soo , Kim Da-Yeong , Choi Dong-Ki , Park Peter U. 
Published: April 2023

Abstract
Targeted protein degraders (TPDs) have expanded the breadth of therapeutic options through both their catalytic mechanism of action and ability to degrade previously “undruggable” target proteins. Prior reports of small-molecule GSPT1 degraders such as CC-90009 in AML demonstrate potent anti-tumor cytotoxicity, but with a potentially narrow therapeutic index. To increase the efficacy vs. tolerability window of TPDs and improve drug delivery, we introduce TPD-Squared (TPD2 TM), a dual-targeted protein degradation approach of combining the catalytic mechanism of targeted protein degradation with the precision of tumor-targeting therapeutic antibodies. We have previously shown in vitro and in vivo efficacy with a HER2-targeted TPD2 conjugate: ORM-5029. Following on that success, we generated conjugates using a CD33-targeting antibody (OR000283) produced by engineering the FAb (H&L) sequences from gemtuzumab onto an IgG1 Fc with N297A variant to inhibit Fc-γR binding. Medicinal chemistry optimization of linker-payloads led to the identification of ORM-6151, which is composed of SMol006, a highly potent GSPT1 degrader conjugated to OR000283 via a novel β-glucuronide releasable linker. ORM-6151 treatment in CD33-expressing cell lines showed picomolar activity with 10-1000-fold greater potency compared to several GSPT1 degrader molecules, including CC-90009 or Mylotarg, and had robust activity in Mylotarg-resistant lines (AML193 and Kasumi6). ORM-6151 also exhibited picomolar potency in in vitro cytotoxicity to primary relapsed/refractory AML patient blasts, with better potency than CC-90009 and Mylotarg. We evaluated ORM-6151 in several in vivo disseminated xenograft models and observed robust efficacy following a single treatment at doses as low as 0.1 mg/kg. Tumor growth inhibition correlated with the degree and duration of GSPT1 depletion and changes in the expression of previously described integrated stress response biomarker genes. In summary, ORM-6151 is a promising, potential therapy for AML and currently in preclinical development as a first-in-class targeted protein degrader therapy with CD33-targeted delivery.
Citation Format: James Palacino, Pedro Lee, Hangyeol Jeong, Yeonjoon Kim, Yoojin Song, Uttapol Permpoon, Wesley Wong, Chen Bai, Nathan Fishkin, Khuloud Takouri, Eunjun Yu, Yong Yi, Anna Skaletskaya, Min-Soo Kim, Da-Yeong Kim, Dong-Ki Choi, Peter U. Park. ORM-6151: A first-in-class CD33-antibody enabled GSPT1 degrader for AML [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 2700.