bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4837: Inhibition of mitochondrial fission activates glycogen storage to support cell survival in colon cancer
Authors: Hasani Sumati , Young Lyndsay E. , Banerjee Moumita , Rivas Dylan , Kim Jinhwan , Sun Ramon , Gentry Matthew , Xiong Xiaopeng , Gao Tianyan
Published: April 2023

Metabolic reprogramming has been increasingly recognized as one of the major mechanisms that fuels tumorigenesis and disease progression. Our previous studies have shown that in response to fatty acid uptake, colon cancer cells activate mitochondrial fission to support fatty acid oxidation and downstream Wnt signaling. Given the potential benefit of inhibiting mitochondrial fission, Dynamin-Related Protein 1 (Drp1), a pro-fission factor, has become an attractive target for developing anticancer agents. Here we investigated the role of Drp1 in promoting metabolic adaptation in colon cancer. We found that knockdown of Drp1 decreased mitochondrial respiration which resulted in increased glucose uptake and lactate production. In addition, downregulation of Drp1 increased AMP-activated protein kinase (AMPK) activity which coincided with glycogen accumulation. Consistently, results from GC/MS analysis of cellular metabolites revealed that the levels of glucose-6-phosphate, a precursor for glycogenesis, were significantly elevated in Drp1 knockdown cells whereas pyruvate and other TCA cycle metabolites remained unchanged. Mechanistically, AMPK transcriptionally activates the expression of glycogen synthase 1 (GYS1) and hexokinase 2 (HK2) genes and silencing GYS1 abolished the glycogen accumulation phenotype in Drp1 knockdown cells. Using APC-derived 3D organoids, we demonstrated that the glycogen levels were elevated in Apcf/f Drp1f/f tumor organoids upon deletion of Drp1. Similarly, increased GYS1 expression and glycogen levels were detected in xenograft tumors derived from Drp1 knockdown colon cancer cells compared to control. Functionally, increased glycogen storage allowed Drp1 knockdown cells to survive glucose starvation conditions suggesting an enhanced survival capacity compared to control cells. Taken together, our findings indicate that Drp1 inhibition by itself is unlikely to be sufficient to eradicate cancer cells as adaptive metabolic mechanisms are activated to promote cell survival. However, combining Drp1 inhibition may enhance the efficacy of chemotherapeutic agents for colon cancer treatment.

Citation Format: Sumati Hasani, Lyndsay E. Young, Moumita Banerjee, Dylan Rivas, Jinhwan Kim, Ramon Sun, Matthew Gentry, Xiaopeng Xiong, Tianyan Gao. Inhibition of mitochondrial fission activates glycogen storage to support cell survival in colon 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 4837.
Abstract 4964: SW-682: A novel TEAD inhibitor for the treatment of cancers bearing mutations in the Hippo signaling pathway
Authors: Chen Lei , de Marval Paula Milani , Powell Kendall , Johnson Mark , Falls Greg , Lawhorn Brian , Candi Aurélie , Kilonda Amuri , Vanderhoydonck Bart , Marchand Arnaud , Versele Matthias , Halder Georg , Gwaltney Stephen L. , Kamal Adeela
Published: April 2023

Many cancers harbor mutations in the Hippo pathway that lead to constitutive activation of the transcriptional co-activators YAP/TAZ that then bind the transcription factor TEAD and drive aberrant transcription of target genes involved in cell proliferation and tumor progression. Hyperactivation of YAP/TAZ has also been associated with resistance to targeted therapies, including MAPK pathway inhibitors. To target cancers that bear mutations in the Hippo pathway or are resistant to therapies due to YAP/TAZ activation, we developed SW-682, a pan-TEAD small molecule inhibitor that blocks TEAD-dependent transcription by binding to the palmitoylation pocket of all TEAD isoforms. In vitro, SW-682 inhibited the proliferation of human Hippo-mutant mesothelioma cells with nanomolar potency, with little to no effect on Hippo wild-type tumor cells. SW-682 down-regulated TEAD-dependent reporter gene expression in a dose-dependent manner, while having no effect on reporters monitoring other pathways. In vivo, daily oral administration of SW-682 to adult mice resulted in tumor regression in Hippo-mutant mesothelioma models and caused down-regulation of expression of the TEAD-dependent genes CCN1 and CCN2 and a YAP gene signature, as measured by qPCR or RNA-seq analysis. SW-682 has a favorable PK profile with good oral bioavailability in the mouse and was well tolerated with no signs of body weight loss. To test the hypothesis that TEAD inhibition can overcome YAP-driven resistance mechanisms, we explored SW-682 in combination with MEK inhibitors in several in vitro and ex vivo patient-derived tumor models including BRAF and NRAS mutated melanoma. Moreover, to identify new indications that may benefit from TEAD inhibition, we screened patient-derived 3D organoid tumor cells and matching patient-derived xenograft models that have been molecularly profiled. In summary, SW-682 is a potent and selective investigational TEAD inhibitor which demonstrated anti-tumor effects in models harboring aberrant expression of the Hippo pathway, suggesting therapeutic potential in multiple Hippo-mutant solid tumors.

Citation Format: Lei Chen, Paula Milani de Marval, Kendall Powell, Mark Johnson, Greg Falls, Brian Lawhorn, Aurélie Candi, Amuri Kilonda, Bart Vanderhoydonck, Arnaud Marchand, Matthias Versele, Georg Halder, Stephen L. Gwaltney, Adeela Kamal. SW-682: A novel TEAD inhibitor for the treatment of cancers bearing mutations in the Hippo signaling pathway. [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 4964.
Abstract 509: DM5167, a novel selective PARP1 inhibitor, efficiently reduces growth of triple-negative breast cancers
Authors: Nam Sujin , Chae Jiyoung , Cho Kyung-Ok , Koo Tae-Sung , Kim Jungho , Jung Myungeun , Kim Jeongmin , Kim Eunhee
Published: April 2023

Poly (ADP-ribose) polymerase 1 (PARP1) has been portrayed as a synthetic lethal target in cancers with homologous recombination deficiency. As all four PARP inhibitors on the market target both PARP1 and PARP2 isotypes, hematological toxicity issue emerged by PARP2 inhibition. To address the unmet need, we screened and optimized compounds exempt from the toxicity issue. Here, we present DM5167 as a new molecular entity with 0.41 nM of IC50 on the catalytic activity of PARP1. In addition, DM5167 exhibits an inhibitory profile against various PARP isotypes that differs from the four commercial PARP inhibitors. Specifically, DM5167 shows selectivity for PARP1 6.88 and 347.83 fold greater than PARP2 in the in vitro enzyme assay and DNA trapping assay, respectively. This contrasts with the corresponding PARP1/PARP2 selectivity values of 0.14 and 0.89 fold for olaparib, respectively. Moreover, DM5167 showed higher selectivity than AZD5305 in the enzymatic assay. The GLP toxicity studies assigned DM5167 as a safe compound based on its high NoAEL value, a general toxicity parameter. In addition, the safety margin of DM5167 was acceptable, indicating that DM5167 is a plausible drug candidate. To find suitable indications, we screened various human cancer cell lines using their inhibitory potentials against proliferation and colony formation. Several hits were seen in triple-negative breast cancer cell lines including MDA-MB-231, MDA-MB-436, MDA-MB-468 and HCC1395. In an orthotopic xenograft experiment in which MDA-MB-231 cells were implanted, oral administration of 50mpk of DM5167 inhibited tumor growth by 45.8%. In contrast, olaparib failed to reduce xenograft tumors derived from MDA-MB-231 cells. Collectively, these studies present DM5167 as an attractive anti-TNBC drug candidate.

Citation Format: Sujin Nam, Jiyoung Chae, Kyung-Ok Cho, Tae-Sung Koo, Jungho Kim, Myungeun Jung, Jeongmin Kim, Eunhee Kim. DM5167, a novel selective PARP1 inhibitor, efficiently reduces growth of triple-negative breast cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 509.
Abstract 5093: Chemically inducible splice-neoantigens for cancer immunotherapy
Authors: Matsushima Shingo , Ajiro Masahiko , Iida Kei , Chamoto Kenji , Honjo Tasuku , Hagiwara Masatoshi
Published: April 2023

Although immune checkpoint blockade therapies have dramatically improved the survival rate of patients with cancer compared with other treatment approaches, a substantial number of patients show poor response or resistance to these therapies. Meta-analyses for programmed cell death 1 (PD-1) blockade responses highlight the importance of neoantigens, derived from non-synonymous mutations in cancer cells, for antitumor immune responses. Intriguingly, recent transcriptome and genomics studies predict the presence of putative neoantigens as a result of an irregular pre-mRNA splicing regulation in cancer cells, indicating that alterations in RNA splicing in cancer cells might induce antitumor immune responses. Here, we report that induction of serine/arginine-rich splicing factor (SRSF)-dependent splicing boosts the production of splicing-associated neoantigens (splice-neoantigens) and potentiates the response to PD-1 blockade. Administration of a synthetic SRSF activator RECTAS suppressed tumor growth in a host CD8+ T cell- and tumor major histocompatibility complex class I-dependent manner and promoted the antitumor effect of anti-PD-L1 antibody without detectable autoimmunity. Subsequent transcriptome analysis and validation for immunogenicity identified six splice-neoantigen candidates whose expression was induced by RECTAS treatment. Importantly, vaccination of the identified neoepitopes elicited T cell responses capable of killing cancer cells in vitro, in addition to suppression of tumor growth in vivo upon sensitization with RECTAS. Collectively, these results provide support for the further development of splice variant-inducing treatments for cancer immunotherapy.

Citation Format: Shingo Matsushima, Masahiko Ajiro, Kei Iida, Kenji Chamoto, Tasuku Honjo, Masatoshi Hagiwara. Chemically inducible splice-neoantigens for cancer immunotherapy. [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 5093.
Abstract 512: Identification of SS008871, a novel Polθ inhibitor that effectively inhibits tumors with homologous recombination deficiency in vitro and in vivo
Authors: Zhou Feng , Liu Lu , Jiang Lei , Tang Feng , Li Zhen , Yang Wenqing , Xue Liting , Chen L , Tang Renhong
Published: April 2023

The repair of DNA double strand break (DSB) is crucial for genome stability and cell survival. There are three main DSB repair pathways: homologous recombination (HR), non-homologous end joining (NHEJ) and microhomology-mediated end joining (MMEJ). HR is a high-fidelity, error-free DSB repair pathway, and the dysfunction of HR confers cell genome instability and leads to tumorigenesis. HR deficiency is especially prevalent in gynecologic tumors, sharing about 10% population of all the gynecologic tumor patients. In situation of HR deficiency, MMEJ, in which DNA polymerase theta (Polθ) plays an essential role, is up-regulated to serve as a backup pathway for DSB repair. Several studies have proved that the inhibition of Polθ causes synthetic lethality with HR deficiency. Hence, Polθ emerges as a potential DNA damage repair target for the treatment of HR deficient tumors. Here we report a novel small molecular Polθ inhibitor, SS008871, which inhibits Polθ activity with an IC50 of 22 nM, and strongly inhibits cellular MMEJ pathway with an IC50 of single-digital nanomolar level. SS008871 strongly inhibits proliferation of HR deficient BRCA2-/- DLD-1 cells, and shows a >125× selectivity folds over DLD-1 parent cells as well as non-malignant cells. Besides, SS008871 elicits synergetic anti-proliferation activities in combination of a PARP inhibitor, olaparib on BRCA2-/- DLD-1 and MDA-MB-436 cells. In the BRCA2-/- DLD-1 xenograft model, SS008871 shows tumor growth inhibition as a single agent, and the combination of SS008871 and olaparib further results in tumor regression. Accordingly, the level of γH2AX, a common DSB marker, correlates well to the anti-tumor efficacy. In a human hematopoietic stem cells based in vitro hematotoxicity assay, SS008871 shows no significant inhibition on lineage-specific (myeloid, erythroid and megakaryocytic) cell differentiation and survival, suggesting the low hematotoxicity risk. In comparison, olaparib significantly attenuated hematopoietic stem cells on both differentiation and survival in the parallel assay, which is consistent with the hematological toxicity observed in human. Furthermore, there is no clinical abnormalities observed after a high dose treatment of SS008871 in mice, demonstrating that SS008871 is well-tolerated. Taken together, SS008871 is proved to be an encouraging Polθ inhibitor with good safety.

Citation Format: Feng Zhou, Lu Liu, Lei Jiang, Feng Tang, Zhen Li, Wenqing Yang, Liting Xue, L Chen, Renhong Tang. Identification of SS008871, a novel Polθ inhibitor that effectively inhibits tumors with homologous recombination deficiency in vitro and in vivo [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 512.
Abstract 5149: Age-associated remodeling of anti-tumor T cell immunity and metabolism
Authors: Han SeongJun , Georgiev Peter , Ringel Alison , Sharpe Arlene , Haigis Marcia
Published: April 2023

Aging results in remodeling of T cell immunity and is associated with poor clinical outcome in age-related diseases such as cancer. Amongst the hallmarks of aging, changes in host and cellular metabolism are critically involved in the development, maintenance, and functional responses of T cells. Although metabolic perturbations impact anti-tumor T cell responses, the link between age-associated metabolic dysfunction and anti-tumor immunity remains unclear. First, we utilized B16, MC38 and E0771 tumor-bearing young (2-4 months) and aged (18-24 months) C57BL/6 mice to examine the effects of aging on tumor growth. Aged mice exhibited accelerated tumor growth in B16, MC38 and E0771 tumor models. Flow cytometric analyses of tumor infiltrating lymphocytes (TILs) revealed distinct changes in CD8+ T cell population, decreased activation status and tumor-specificity in aged mice. Consistent with these findings, CD8+ T cells functionally played a limited role in controlling tumor growth in aged mice. Here, we will explore the mechanisms of age-associated metabolic dysfunction in T cells and its implications in anti-tumor immunity. As the average life expectancy continues to increase, understanding the interplay between age-related metabolic reprogramming and maladaptive T cell immunity will be instrumental in the development of novel therapeutic strategies for older patients.

Citation Format: SeongJun Han, Peter Georgiev, Alison Ringel, Arlene Sharpe, Marcia Haigis. Age-associated remodeling of anti-tumor T cell immunity and metabolism. [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 5149.
Abstract 515: Discovery of potent and orally available small molecule inhibitors of the SOS1-KRAS interaction
Authors: Yu Ha Na , Ki Dong Hyuk , Nam Joonwoo , Kim Eun-Jung , Kim Sungeun , Choi Hunmi , Kim Jieun , Yu Jihyun , Kim Donggeon , Park Dohyun , Yoon Kyeong Jin , Jo Seongin , Hwang So-Hyeon , Lim Sang Kyun , Shin Young Sook , Han Wooseok
Published: April 2023

Abnormal activating mutation of KRAS is frequently found in many human cancers, including pancreatic ductal adenocarcinoma (PDAC), colorectal cancer (CRC), and non-small cell lung cancer (NSCLC). KRAS activating mutations lead to hyperactivation of the MAPK/ERK signaling pathway, resulting in promotion of cell proliferation and growth. SOS1 is one of the major guanine nucleotide exchange factors (GEFs) that regulates RAS proteins including KRAS. Since SOS1 plays a critical role in converting the GDP-bound inactive KRAS “off” state to the GTP-bound active KRAS “on” state, disruption of SOS1 and KRAS protein-protein interaction would be effective to block KRAS-driven oncogenic signaling regardless of its mutation status. Importantly, SOS1 activity is crucial during the reactivation of the KRAS/MAPK signaling upon the treatment of RAS/MEK/ERK inhibitors, thus a SOS1 inhibitor would be an effective therapeutic option to treat KRAS-driven tumors in combination with RAS pathway inhibitors. We developed potent, selective, and orally available small molecules that effectively disrupt the interaction between SOS1 and KRAS. Current lead compounds originated from a virtual screening displayed excellent ADME and PK profiles. In cellular assays, a robust reduction of phospho-ERK level and cancer cell growth were shown. We also observed excellent in vivo antitumor activity in the mouse xenograft models. Moreover, combination with Sotorasib synergistically inhibited tumor cell growth both in vitro and in vivo. Overall, our SOS1 inhibitors demonstrate great therapeutic potential for cancer patients with KRAS mutations.

Citation Format: Ha Na Yu, Dong Hyuk Ki, Joonwoo Nam, Eun-Jung Kim, Sungeun Kim, Hunmi Choi, Jieun Kim, Jihyun Yu, Donggeon Kim, Dohyun Park, Kyeong Jin Yoon, Seongin Jo, So-Hyeon Hwang, Sang Kyun Lim, Young Sook Shin, Wooseok Han. Discovery of potent and orally available small molecule inhibitors of the SOS1-KRAS interaction [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 515.
Abstract 5408: AI powered-platform to predict gene modifications from prostate and breast cancer whole slide images
Authors: Huang Wei , Jain Parag , Xie Chensu , Muhammad Hassan , Basu Hirak , Wilding George , Roy Rajat
Published: April 2023

There is little published evidence of predicting cancer genotypes directly from tissue histology, especially for breast and prostate cancers. Artificial intelligence (AI) enables discriminating and extracting morphological features reflective of the underlying genomic alterations at visual and subvisual levels. We have built a morphology-based and AI-powered platform for cancer genotyping, risk stratification and outcome prediction that addresses the needs for treatment decision-making in a cost effective and timely fashion. A cohort of 390 prostate and 742 invasive breast cancer patients with known molecular status of key genes, such as TP53, PIK3CA, MYC, ERBB2, TMPRSS2-ERG and PTEN from The Cancer Genome Atlas were included in this study. Hematoxylin and eosin (H&E) stained whole slide images (WSI) of the cancer tissue sections were available at 20x or 40x. The WSI from the two cancer cohorts were split 2:1 into a training and test dataset, respectively. Our platform involved two different deep Convolutional Neural Network (DCNN) architectures. The platform first divided each WSI into multiple tiles. Each tile was then analyzed using a DCNN that graded the tile and generated a high dimensional vector to provide a mathematical representation of the morphology. The combination of high dimensional vectors across the WSI was then fed into a second DCNN that generated a morphological score, which predicted whether the gene under consideration was wild type or modified. Our platform has achieved 70 - 80% accuracy as defined by the Area under the Curve for the receiver operating characteristics curves for the genetic markers on the test cohorts (Table 1). Our platform can predict genotypes/molecular alterations directly from H&E stained WSI with high accuracy. This technology presents a novel, practical and cost-effective approach for cancer molecular classification and risk stratification, enabling timely and optimal treatment decision-making for positive clinical outcomes.

Genotype Prediction for Breast Cancer (BrCa) and Prostate Cancer (PCa) from WSI Cohort Molecular Biomarker Training Dataset (n) Test Dataset (n) Test ROC AUC Score (%) Gene Modification/Loss Intact/Wild type Gene Modification/Loss Intact/Wild type BrCa TP53 196 302 96 149 80 PIK3CA 153 344 75 170 74 MYC 68 414 33 204 77 ERBB2 66 416 32 205 78 PCa TMPRSS2-ERG 104 157 51 78 70 PTEN 57 201 28 99 73

Citation Format: Wei Huang, Parag Jain, Chensu Xie, Hassan Muhammad, Hirak Basu, George Wilding, Rajat Roy. AI powered-platform to predict gene modifications from prostate and breast cancer whole slide images. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5408.
Abstract 5410: Predictive biomarker discovery method to bridge the gap between preclinical disease model dose-response and clinical trials
Authors: Futorian David , Fischman Oren , Arad Gali , Simchi Nitzan , Erez Omri , Seger Eran , Groen Rozanne , Pevzner Kirill
Published: April 2023

The translational gap of drug and biomarker discovery remains one of the biggest challenges in the pharmaceutical industry. On one hand high throughput screening and omics methods facilitate the generation of in-vitro and in-vivo data of dose-response for a particular drug or combinations. On the other hand, clinical omics techniques allow for the creation of large scale treatment-naive clinical datasets, with the TCGA and CPTAC as prominent examples. However, the question of which disease model best represents the response/resistance mechanisms of an oncology patient remains challenging.

In this study we present a machine learning (ML) based computational technique for integrating omics from preclinical dose-response studies with clinical treatment-naive samples to create putative predictive biomarkers, and exemplify its application 2 case studies - PARP or AKT inhibitors.

We utilize the Cancer Cell Line Encyclopedia (CCLE) and Genomics of Drug Sensitivity in Cancer (GDSC) as a resource for in-vitro dose response data for PARPi and AKTi, coupled with multi-omic molecular data. We utilize the treatment naive molecular characterization from the Clinical Proteomic Tumor Analysis Consortium (CPTAC) data of Breast cancer patients. Multi-omic analysis of these datasets derived 2 putative predictive biomarkers for PARPi and AKTi. The multi-omic analysis resembles the data availability at the critical drug development stage of transition from preclinical response models to a clinical trial. These biomarkers are tested on a validation dataset obtained from the iSPY adaptive clinical trial and achieve superior results compared to the original enrolment biomarkers and even the retrospectively derived biomarkers.

This study presents a novel computational approach to bridge the gap between preclinical dose-response and clinical datasets and suggests an efficient way to discover predictive biomarkers based on data accessible in a preclinical setting.

Citation Format: David Futorian, Oren Fischman, Gali Arad, Nitzan Simchi, Omri Erez, Eran Seger, Rozanne Groen, Kirill Pevzner. Predictive biomarker discovery method to bridge the gap between preclinical disease model dose-response and clinical trials. [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 5410.
Abstract 5791: Autophagy & PIM1 kinase inhibition in breast cancer stem cells & bulk cancer cells attenuates breast cancer & metastasis
Authors: Manupati Kanakaraju , Hao Mingang , Haas Michael , Yeo Syn Kok , Guan Jun-Lin
Published: April 2023

Autophagy is a self-degradation process, it is essential for growth of heterogenous of breast cancer cells. Breast tumor heterogeneity majorly comprises bulk cancer cells & cancer stem cells (CSCs). Targeting either CSCs or cancer cells could lead to interconversion between these cells due to EMT/MET transitions. Therefore, it is important to identify a common molecular targets & its mechanism of regulation in these populations for improvement of breast cancer therapy. Here, we utilized a PCR based screen & identified NUMA1 downregulation in FIP200 KO (autophagy gene) cells of three major breast cancer subtypes, FF99 (luminal), N418 (Her2+) & BF3M (basal). Further, NUMA1 KO in these cells reduced tumor formation & CSCs indicating the role of NUMA1 on regulation of CSCs. Therefore, we sorted breast CSCs, ALDH+ and CD29hi CD61+ from WT & NUMA1 KO cells, tail-vein injected nude mice depicted reduced metastatic potential with NUMA1 KO than WT CSCs. Mechanistically, NUMA1 regulation by PIM1 kinase was observed in CSCs but not in bulk cancer cells. Cell viability assays revealed that combination treatment of autophagy & PIM1 inhibitor showed higher efficacy against cancer cells of three breast cancer subtypes than individual treatments alone. Finally, the combination treatment of autophagy inhibitor, Lys05 & PIM1 inhibitor, SMI-4a reduced tumor formation & metastasis of breast cancer. Human tissue microarray results depicted high expression of NUMA1 & PIM1 in different breast cancer subtypes than non- tumor tissues. Our study demonstrated that inhibition of autophagy & PIM1 in bulk cancer cells & CSCs showed attenuation of tumor formation & metastasis, suggesting this combination treatment serves as an important therapeutic approach for treating breast cancer.

Citation Format: Kanakaraju Manupati, Mingang Hao, Michael Haas, Syn Kok Yeo, Jun-Lin Guan. Autophagy & PIM1 kinase inhibition in breast cancer stem cells & bulk cancer cells attenuates breast cancer & metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5791.
Abstract 5794: Targeting CD133+/CD90+ liver cancer stem cells by piperlongumine: A novel strategy for liver cancer treatment
Authors: Abdelaziz Basma A. , Sadek Omayma M. , Sultan Ahmed S.
Published: April 2023

Liver cancer is an aggressive malignant disease. It’s the most common cancer worldwide and the leading cause of cancer death. Because of the atypical early symptoms of liver cancer, the majority of liver cancer patients are diagnosed with advanced-cancer stage, decreasing the chance of resection and resulting in a poor survival rate. Liver cancer stem cells (LCSCs) are a small subpopulation inside the tumor bulk, characterized by the maintenance of self-renewal and pluripotency. Thus, LCSCs are responsible for cancer initiation, metastasis, relapse, and chemoresistance. Elucidating LCSC characteristics and disclosing their regulatory mechanism might facilitate the development of prognostic, diagnostic, and therapeutic approaches to improve the clinical management of liver cancer. Piperlongumine (PL) an active biological pyridine alkaloid has anti-angiogenic, anti-diabetic, and anti-bacterial activities. The present study aimed to isolate highly tumorigenic and aggressive subpopulation of LCSCs (CD133+/CD90+) from primary tumors and different cell lines and investigate the effect of (PL), a novel targeting therapy, on LCSCs proliferation. The isolated LCSCs were treated with different concentrations of PL and its effect was examined by cell viability, cell morphological changes, western blot, caspase-3 activity, apoptotic induction, and clonogenic formation. By using Immunocytochemistry (ICC) analysis, the localization of nuclear and cytoplasmic Aldehyde dehydrogenase (ALDH6A1) was detected. PL inhibited the viability of CD133+/CD90+LCSC-sub-popupation and caused morphological changes after 48 hours while enhancing the pre-apoptotic signals and caspase-3 activity. In addition, PL decreased the colonies’ formation ability compared to control group. Furthermore, PL suppressed the protein expression levels of ALDH6A1, Notch-1, β-catenin, ERK1/2, and Cyclin-D1 in the dose and time-dependent manners. Besides, PL decreased the localization and intensity of nuclear and cytoplasmic ALDH6A1 compared to the control group, indicating that PL decreased the proliferation and stemness markers of LCSCs. Overall, Piperlongumine as a novel targeting therapy has an important role in targeting CD133+/CD90+ LCSCs sub-population proliferation.

Citation Format: Basma A. Abdelaziz, Omayma M. Sadek, Ahmed S. Sultan. Targeting CD133+/CD90+ liver cancer stem cells by piperlongumine: A novel strategy for liver cancer treatment [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 5794.
Abstract 5816: Tensin1 tension sensor reveals novel features associated with actomyosin, focal adhesion dynamics, and mechanosensitivity
Authors: Jang Yoon-Kwan , Suh Jung-Soo , Choi Gyuho , Ahn Sanghyun , Han Ki Seok , Kim EunHye , Lee Yerim , Hu Xiaoqi , Hwang Eunbae , Kim Tae-Jin
Published: April 2023

Cellular responses to mechanical stimuli play an imperative role in the regulation of physiological and pathological functions. Unlike other integrin-mediated adhesion proteins, the force transmission mechanism of tensin1 is not well understood. In this study, we describe the development and visualization of a tensin1 tension sensor. A signal from this sensor indicates that tensin1 is under greater tension at peripheral adhesions than at central adhesions, and that it is controlled by actomyosin in fibroblasts. In a bidirectional manner, tension in tensin1 is regulated by focal adhesion kinase (FAK) activity. Our findings demonstrate that tensin1 is capable of sensing the extracellular matrix through alterations in the tension it receives from the extracellular matrix. Furthermore, tensin1 depletion increases the tension of talin1, but the absence of talin1 decreases the tension applied to tensin1. These results indicate that tensin1 is complementary and dependent on the tension received by talin1. Overall, our data, together with the prognosis following tensin1 expression in colorectal and urothelial cancer, elucidate the tension-receiving function of tensin1 and highlight the necessity of tensin1-mediated mechanotransduction studies.

Citation Format: Yoon-Kwan Jang, Jung-Soo Suh, Gyuho Choi, Sanghyun Ahn, Ki Seok Han, EunHye Kim, Yerim Lee, Xiaoqi Hu, Eunbae Hwang, Tae-Jin Kim. Tensin1 tension sensor reveals novel features associated with actomyosin, focal adhesion dynamics, and mechanosensitivity [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 5816.
Abstract 593: Bispecific antibody against TIGIT and PVRIG enhances antitumor immunity
Authors: Liang Jiabei , Gao Xiao , Zhang Cong , Zhao Hui , Peng Hao , Hao Feng , Ning Jinying , Wu Guojin
Published: April 2023

T cell immunoreceptor with Ig and ITIM domains (TIGIT) and poliovirus receptor-related immunoglobulin domain-containing protein (PVRIG) are two immune checkpoint receptors. They compete with CD226 for binding to CD155 or CD112 and suppress the activity of NK cells and T cells. Here we developed a bispecific antibody KA-1874 targeting TIGIT with an anti-PVRIG scFv fused to the C-terminus. KA-1874 is a fully human IgG4 with high binding affinity to TIGIT (KD=0.08nM) and PVRIG (KD=0.17nM). KA-1874 blocks both bindings of PVRIG-mFc to PVRL2 and TIGIT-mFc to CD155. In cell-based luciferase reporter assays, KA-1874 blocks PVRIG and TIGIT-mediated inhibitory signaling and enhances CD3 signaling with EC50 of 1.1nM and 1.4nM respectively, while COM701 against PVRIG and EOS488 against TIGIT both with EC50 above 2nM. In a CMV recall assay, PBMC increases INF-γ secretion when cultured with KA-1874, suggesting KA-1874 enhances primary T cell function. Immunodeficient mice humanized with PBMC are transplanted with A374 tumor cells. Co-injection of KA-1874 and pembrolizumab, which against PD-1, suppress established tumor growth. In comparison, KA-1874 or pembrolizumab alone fails to inhibit tumor growth. The above results indicate KA-1874 is an excellent candidate therapeutical antibody for cancer immunotherapy.

Citation Format: Jiabei Liang, Xiao Gao, Cong Zhang, Hui Zhao, Hao Peng, Feng Hao, Jinying Ning, Guojin Wu. Bispecific antibody against TIGIT and PVRIG enhances antitumor immunity [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 593.
Abstract 5940: Enhanced mast cell abundance is associated with the enrichment of CCR2+ cytotoxic T cells and favorable prognosis in lung adenocarcinoma
Authors: Fan Fan , Zhang Xiaoming , Chen Haiquan
Published: April 2023

Mast cells constitute indispensably immunoregulatory sentinel cells in tumor microenvironment, however, their functions are poorly characterized in lung adenocarcinoma (LUAD). Here, we performed flow cytometry and single-cell RNA sequencing to comprehensively characterize LUAD infiltrating MCs. MCs exhibited remarkably functional heterogeneity and were enriched in ground-glass opacity featured LUAD (gLUAD). The MCs in gLUAD exhibited proinflammatory and chemotactic properties while the MCs in radiologically solid LUAD (sLUAD) were associated with tumor angiogenesis. MCs were an important source of CCL2 that correlated with the recruitment of CCR2+ cytotoxic T lymphocytes (CTLs), which represent a specific subcluster of pre-exhausted T cells with tissue-resident memory phenotype and enhanced cytotoxicity. Increased infiltration of MCs-CCR2+ CTLs and their colocalization showed a strong association with favorable prognosis after surgery but were not associated with improved survival after chemotherapy. Collectively, these findings reveal a key role of MCs in LUAD and their potential crosstalk with CTLs, suggesting that targeting MCs may be a novel immunotherapeutic strategy for LUAD.

Citation Format: Fan Fan, Xiaoming Zhang, Haiquan Chen. Enhanced mast cell abundance is associated with the enrichment of CCR2+ cytotoxic T cells and favorable prognosis in lung adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5940.
Abstract 5957: Chemotherapy-induced extracellular matrix remodeling in HGSOC
Authors: Kotantaki Panoraia , Laforêts Florian , Maniati Eleni , Berlato Chiara , Malliouri Anna , Devlin Michael John , Malacrida Beatrice , Elorbany Samar , Manchanda Ranjit , Balkwill Frances R.
Published: April 2023

In this study we explored the effects of chemotherapy on the extracellular matrix of high-grade serous ovarian cancer metastasis (HGSOC) at structural-textural protein level and transcriptionally, in order to identify the microenvironmental features that accompany a successful response. We studied two murine orthotopic, transplantable syngeneic models of HGSOC, one that displays excellent response to carboplatin/paclitaxel (60577) and one that is resistant to treatment (HGS2)1. Using immunohistochemistry, we stained omental tumors from both models for the ECM components fibronectin (FN1), collagen 1A1 (COL1A1) and versican (VCAN), also using Masson’s Trichrome to stain for collagen. We analyzed their abundance, textural features (Haralick features) and structure with the image analysis softwares, QuPath and TWOMBLI respectively. A compilation of 44 ECM metrics, which we named the “structure index”, correlated with response to chemotherapy, as measured by reduction in omental tumor weight. In parallel, we performed RNAseq on the omental tumors, identifying clusters of genes that change over time with chemotherapy treatment of the responsive model 60577, while the chemoresistant model HGS2 displayed limited alteration of gene expression. We integrated RNAseq and ECM structure and texture data, and showed the structure index was correlated with 144 core matrisome genes. To identify potential therapeutic targets, we then focused our study on genes that remained highly expressed in resistant tumors post chemotherapy, but were originally low or decreased with treatment in the sensitive tumors. Members of the LOX and P4HA family were found amongst these genes. We confirmed by immunohistochemical analysis that expression of LOX, LOXL2, P4HA1 and P4HA2 was downregulated by chemotherapy in the 60577 chemo-sensitive tumors while their levels remained relatively unaffected in the resistant HGS2 tumors. This study refines our understanding of the mechanisms of microenvironmental chemoresistance in HGSOC, highlighting putative therapeutic targets to increase the efficacy of platinum-based chemotherapy in ovarian cancer.

1 Maniati, E. et al. Mouse Ovarian Cancer Models Recapitulate the Human Tumor Microenvironment and Patient Response to Treatment. Cell Rep 30, 525-540.e527, doi:10.1016/j.celrep.2019.12.034 (2020).

Citation Format: Panoraia Kotantaki, Florian Laforêts, Eleni Maniati, Chiara Berlato, Anna Malliouri, Michael John Devlin, Beatrice Malacrida, Samar Elorbany, Ranjit Manchanda, Frances R. Balkwill. Chemotherapy-induced extracellular matrix remodeling in HGSOC [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 5957.
Abstract 5958: Analysis of MDSCs as a diagnostic and therapeutic target for gastric cancer
Authors: Yamauchi Suguru , Iwasawa Takumi , Ito Tomoaki , Brock Malcom V. , Fukunaga Tetsu , Orita Hajime , Kato Kazunori
Published: April 2023

Background: Gastric cancer (GC) is the second most common cancer in Japan in terms of incidence and the third most common in terms of deaths. The onset and progression of cancer are generally suppressed by the action of immune cells, which is an inflammatory response. MDSCs are thought to promote cancer progression and invasion by suppressing the function of immune cells, making them potential therapeutic and diagnostic targets. We measured MDSCs in GC patients and observed more monocyte-derived MDSCs in peripheral blood in advanced GC than in early GC. However, granulocyte-derived MDSCs showed no difference in GC progression when markers for classical MDSCs were used. We considered that granulocyte-derived MDSCs were much more abundant than monocyte-derived MDSCs in peripheral blood and were involved in cancer progression. We hypothesized that we had a new granulocyte-derived marker for advanced GC that did not show differences in classical markers, but may have different functions, and aimed to identify a novel subset that may be a therapeutic and diagnostic target.

Method: For GC patients, we used heparin blood samples provided by patients who consented to Juntendo University. We analyzed the expression of various cell surface proteins against populations of CD33high, CD66bhigh, and HLA-DRlow as classical markers of MDSCs derived from granulocytes of GC patients using FACS (Melody). MDSCs were isolated by a cell sorter and used for experiments such as RNA-Seq. Novel markers and involved cytokines were quantified in plasma components by cytometric bead array.

Result: The chemokine receptors CCR2, CCR5, CXCR4, and CXCR5 were highly expressed in MDSCs of advanced GC patients. The ability to suppress T-cell activation was also found to be stronger in MDSCs with advanced GC. In addition, IL-10 produced by MDSCs was also more abundant in the plasma of patients with advanced GC, as well as chemokines according to chemokine receptors.

Conclusion: The high expression of chemokine receptors on MDSCs in advanced GC may indicate that chemokines produced at the cancer site facilitate the migration of MDSCs. These novel subsets of MDSCs found in advanced GC suggest that they may be new therapeutic targets.

Citation Format: Suguru Yamauchi, Takumi Iwasawa, Tomoaki Ito, Malcom V. Brock, Tetsu Fukunaga, Hajime Orita, Kazunori Kato. Analysis of MDSCs as a diagnostic and therapeutic target for 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 5958.
Abstract 5961: Inhibition of AURKA destabilizes glioblastoma primary cilia and sensitizes cells to tumor treating fields (TTFields) in vitro and ex vivo
Authors: Tian Jia , Mallinger Julianne , Shi Ping , Ling Dahao , Deleyrolle Loic , Lin Min , Khoshbouei Habibeh , Sarkisian Matthew
Published: April 2023

Tumor Treating Fields (TTFields) can prolong survival of patients with glioblastoma (GBM), but GBM cells still escape this therapy requiring an urgent need for new or adjunct treatments. Recently we reported that TTFields disrupt primary cilia on GBM cells which lowers resistance to standard of care temozolomide chemotherapy. Here we asked if concomitant or sequential treatment of TTFields with other agents that interfere with GBM ciliogenesis enhance TTFields efficacy. Aurora Kinase A (AURKA) plays both a role in promoting cilia disassembly and regulating GBM growth in xenograft models. We found that within 24hr of treatment with the AURKA inhibitor, Alisertib, there was surprising and significant decrease in GBM cilia frequency across multiple GBM patient derived lines. However, similar Alisertib treatment did not affect neuronal or glial cilia frequencies in mixed primary cell cultures from mouse forebrain. Notably, overnight treatment of patient GBM biopsy ex vivo with Alisertib also resulted in reduced cilia frequency within the tumor tissue. Next, we co- or sequentially treated several different GBM patient cell lines with 24hr of 200kHz TTFields together with or followed by 250nM or 1µM Alisertib and examined the subsequent expansion of cells in vitro. We observed a significant synergistic reduction in cell proliferation compared to either treatment alone. However, this synergistic effect does not appear to be cilia-dependent as dual treatment reduction in proliferation was still observed in cell lines lacking different key cilia genes or in GBM cells that naturally lack cilia. Considering Alisertib crosses the blood brain barrier and inhibits intracranial growth of tumors, our data warrant investigation of whether concomitant or sequential treatment of TTFields and Alisertib in vivo can further prolong survival.

Citation Format: Jia Tian, Julianne Mallinger, Ping Shi, Dahao Ling, Loic Deleyrolle, Min Lin, Habibeh Khoshbouei, Matthew Sarkisian. Inhibition of AURKA destabilizes glioblastoma primary cilia and sensitizes cells to tumor treating fields (TTFields) in vitro and ex vivo [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 5961.
Abstract 6095: A DNA damaging agent affects BRCA2 exon 3-skipping
Authors: Werner Jordan R. , Siddiqui Hiba , Castillo Ethan , Ansari Mohammed , Fackenthal James D.
Published: April 2023

Introduction: BRCA2 is a tumor suppressor that participates in DNA repair. Inherited pathogenic mutations in BRCA2 are associated with a 38%-84% risk for breast cancer and a 16.5%-27% risk for ovarian cancer. BRCA2 exon 3, which encodes a binding site for the interacting proteins EMSY and PALB2, is frequently skipped during alternative splicing events, often due to spliceogenic mutations, but also for as yet uncharacterized environmental conditions. A genomic deletion that results in complete allelic loss of exon 3 has been shown to be pathogenic, yet sequence variants that result in increased frequency of exon 3-skipping have been shown to be non-pathogenic. Thus, the allele-specific ability of BRCA2 to contribute to DNA repair is affected by the level of exon 3-skipping. In this study we explore the effects of the topoisomerase inhibitor doxorubicin on the frequency of BRCA2 exon 3-skipping. Isoform-specific end-point RT-PCR shows that doxorubicin increases the frequency of BRCA2 exon 3-skipping in the breast cancer cell line MCF7 at concentrations below those required for promoting apoptosis.

Methods: MCF7 cells were treated with doxorubicin according to standard protocols. IC50 levels of doxorubicin were determined using a crystal violet assay. Flow cytometry was performed using standard protocols. RNA was prepared using Qiagen reagents and amplified with Invitrogen One-Step RT-PCR reagents. Isoform-specific RT-PCR primers were designed to amplify the full-length region surrounding BRCA2 exon 3 independently from the region left by an exon-skipping event (∆3). RT-PCR products were visualized using agarose gel electrophoresis.

Results and Conclusion: Results suggest that doxorubicin increases the level of BRCA2 exon 3-skipping in MCF7 at concentrations below those required for promoting apoptosis. This suggests doxorubicin may compromise the DNA repair functions of BRCA2 and other alternatively spliced mRNAs, which may contribute to the therapeutic benefits and/or toxicity of the drug.

Citation Format: Jordan R. Werner, Hiba Siddiqui, Ethan Castillo, Mohammed Ansari, James D. Fackenthal. A DNA damaging agent affects BRCA2 exon 3-skipping [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 6095.