bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4602: Differences in tumor vascular density between smoker and non-smoker cancer patients
Authors: Singh Shubhangi , Khan Mohammad Aslam , Herrera Elba T. , Singh Seema , Singh Ajay P. , Dasgupta Santanu
Published: April 2023

Smoking is among the most significant preventable risk factors associated with cancer and other benign diseases. Cancer development is a multi-step process in which transforming cells gradually gain malignant properties through a series of oncogenic genetic alterations. Successful cancer progression also depends on the remodeling of the surrounding stroma, which feeds the cancer cells in a host-parasite interaction. Tumor angiogenesis, the process of blood vessel formation, is an essential feature of solid tumors that ensures that they have the needed supply of food and oxygen to sustain their proliferative growth. We previously demonstrated that nicotine, an addictive component of tobacco smoke, promoted tumor angiogenesis by impacting the growth of endothelial cells. In the present work, we examined the differences in tumor vasculature between smoker and non-smoker patients to find clinical support for our laboratory findings. After searching the available tumor cases in the repository for the patient’s smoking history, I selected 27 smokers (12 lung and 15 prostate adenocarcinomas) and 19 non-smokers (4 lung and 15 prostate adenocarcinomas) cases. The tissue blocks were sectioned and subjected to hematoxylin and eosin staining and immunohistochemistry using an antibody against CD31, a biomarker for endothelial cells. Following digital scanning, all slides were reviewed to identify tumor areas and CD31+ tubular structures (blood vessels). We observed that lung and prostate tumor tissues from smoker patients had a significantly higher number of blood vessels than those from non-smoker patients. Thus, increased tumor vascularization appears to be a common feature of smoking cancer patients providing clinical support for a novel mechanism by which smoking may contribute to tumor pathobiology.

Citation Format: Shubhangi Singh, Mohammad Aslam Khan, Elba T. Herrera, Seema Singh, Ajay P. Singh, Santanu Dasgupta. Differences in tumor vascular density between smoker and non-smoker cancer patients. [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 4602.
Abstract 4607: TRPS1 inhibits angiogenesis in triple negative breast cancer down regulating JAG1 and TYMP genes involved in angiogenesis pathways
Authors: Guzman Liliana
Published: April 2023

Background. Triple-negative breast cancer (TNBC), the most difficult subtype to treat, is defined as estrogen receptor, HER2, and progesterone receptor negative. TNBC constitutes 10-20% of all breast cancer and has a higher rate of distal recurrence and a poorer prognosis than other breast cancer subtypes. Less than 30% of women with metastatic TNBC survive 5 years and almost all die from their disease despite adjuvant chemotherapy. Angiogenesis is a critical factor in the development of tumors and metastases in numerous cancers. Clinical trials with anti-angiogenic drugs showed to be inadequate, with moderate response rates and insignificant survival gains for patients. To understand the genetic forces involved in TNBC, we performed a transposon mutagenesis screen in Pten mutant mice that identified several candidate trunk drivers and progression genes. A major finding of our screen was the discovery and functional validation of, Trichorhinophalangeal syndrome type 1 (TRPS1), a GATA-like transcription factor, which functions as a transcriptional repressor or activator, depending on cell type, stage of development, or pathological conditions. Based on this data, we explored the role of TRPS1 in angiogenesis.

Methods and Results. Tube formation and sprouting assays were performed using overexpression and inactivation of TRPS1 in MDA-MD-231 and HCC70 cells, respectively. Interestingly, inactivation of TRPS1 expression accelerates tube formation structures compared to the vector control as well as cell branching in the sprouting assay. Overexpression of TRPS1 prevents tubing and branching formation in vitro assays. Moreover, immunohistochemistry staining of CD31 detected a reduced number of blood vessels in MDA-MB-231 tumor xenografts overexpressing TRPS1, and an increase of angiogenic vasculature in HCC70 TRPS1-shRNA tumor xenografts. In vitro and in vivo assays demonstrate the role of TRPS1 in tumor angiogenesis. Furthermore, human ChIP qPCR angiogenesis array identified 10 top candidate genes potentially regulated by TRPS1 transcription factor. Of these candidates, JAG1 and TYMP showed to have a higher fold enrichment compared to other angiogenic related genes. Finally, we validated its direct functional binding by using luciferase reporter assay, such demonstrated that TRPS1 is a direct transcriptional regulator of JAG1 and TYMP.

Conclusion. TRPS1 is a tumor suppressor involved in the mechanisms regulating tumor angiogenesis by repressing the expression of JAG1 and TYMP genes in TNBC.

Citation Format: Liliana Guzman. TRPS1 inhibits angiogenesis in triple negative breast cancer down regulating JAG1 and TYMP genes involved in angiogenesis pathways. [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 4607.
Abstract 461: Targeting glucose metabolism in lung cancer
Authors: Telang Sucheta , Chang Simone , Sanders Nicole
Published: April 2023

Positron emission tomographic studies with 2-18F-fluoro-deoxy-glucose (FDG-PET) consistently demonstrate that non-small cell lung cancers (NSCLC) take up significantly more glucose than normal lung. NSCLCs preferentially utilize glucose to support their rapid growth- a phenotype driven by several oncogenic alterations among which mutations of the oncoprotein Ras are frequently observed. Oncogenic Ras is shown to activate downstream effectors (e.g. HIF-1α) that stimulate glycolysis - in part by activation of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase family of enzymes (PFKFB1-4). The PFKFBs produce fructose-2,6-bisphosphate (F26BP) which activates a key rate-limiting glycolytic enzyme, 6-phosphofructo-1-kinase (PFK1). In preliminary studies, we found that the PFKFB4 family member was highly expressed relative to other PFKFB isoforms (by multiplex RT-PCR) in a series of patient-derived NSCLCs relative to paired normal lung tissue. We further explored PFKFB4 protein expression by immunohistochemistry in NSCLCs derived from a cohort of patients and found markedly elevated PFKFB4 in the majority of tumors relative to adjacent normal tissue. We then determined that silencing PFKFB4 markedly reduced F26BP, glycolysis and cell survival in NSCLC cell lines and inhibited xenograft lung tumor growth in vivo. In order to target PFKFB4, we conducted a virtual screen of compounds that fit into the active site of PFKFB4 and discovered a first-in-class small molecule selective PFKFB4 inhibitor, termed 5MPN, that selectively inhibits recombinant PFKFB4 activity and decreases F26BP, glycolysis and proliferation in multiple NSCLC cell lines, with significantly higher anti-proliferative effects in Ras-driven cell lines. 5MPN also caused marked suppression of growth in established NSCLC xenograft tumors, importantly without systemic toxicity. We have developed an inducible, pan-tissue homozygous floxed Pfkfb4 knockout mouse and crossed it with a Cre recombinase mouse driven by a tamoxifen-inducible β-actin promoter to produce Cre/Pfkfb4fl/fl mice. We crossed Cre/Pfkfb4fl/fl mice with a Ras-driven lung cancer model, induced Pfkfb4 deletion by tamoxifen administration and found significantly lower total lung tumor burden in these mice accompanied by decreased PFKFB4 expression in tumors and lung tissue. Taken together, these data provide strong rationale for the further evaluation of small molecule inhibitors of PFKFB4 as a potential therapeutic strategy against NSCLC.

Citation Format: Sucheta Telang, Simone Chang, Nicole Sanders. Targeting glucose metabolism in lung 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 461.
Abstract 4613: Inhibition of myosin II in triple-negative breast cancer cells limits macrophage recruitment in a 3D environment
Authors: Cho Youngbin , Li Ruxuan , Zervantonakis Ioannis
Published: April 2023

Among all breast cancer subtypes, triple-negative breast cancer (TNBC) exhibits the least favorable outcomes with a complex tumor microenvironment that facilitates metastatic progression. Remodeling of the extracellular matrix at the tumor invasive front is associated with increased cancer cell motility and poor outcomes. Myosin II, a key cytoskeletal regulator, plays a central role in cancer cell metastatic potential. In the clinic, high expression of myosin II in breast tumors has been associated with distant metastasis and lower survival rates. Interestingly, breast tumors overexpressing myosin II exhibited a higher density of tumor-associated macrophages and an immunosuppressive microenvironment. However, the relationship between myosin II activation in TNBC cancer cells and macrophage recruitment remains poorly understood. Here we employed a novel 3D microfluidic model of tumor-macrophage co-culture to study macrophage infiltration toward cancer cells with high spatiotemporal resolution. Using a panel of TNBC cell lines we found that high myosin II expression in cancer cells correlated with higher macrophage recruitment in a 3D matrix. Pharmacologic or genetic inhibition of myosin II reduced macrophage recruitment. Furthermore, using our microfluidic devices, we controlled the spatial distribution of cancer cells in a 3D matrix and separated the role of cancer cell-secreted paracrine factors from cancer cell-mediated matrix remodeling on macrophage recruitment. Analysis of matrix remodeling over time revealed that targeting myosin II in cancer cells limited the establishment of migratory tracks that facilitated macrophage recruitment. In sum, our studies revealed the critical role of cancer cell-driven matrix remodeling in promoting macrophage infiltration that can be therapeutically targeted to reverse the establishment of an immunosuppressive microenvironment.

Citation Format: Youngbin Cho, Ruxuan Li, Ioannis Zervantonakis. Inhibition of myosin II in triple-negative breast cancer cells limits macrophage recruitment in a 3D environment. [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 4613.
Abstract 4630: Deep Immuno-profiling of syngeneic tumor mouse models for preclinical studies
Authors: Joachim Anais , Maturin Emilie , Mello Marielle , Hadjem Lillia , Grange Magali , Deas Olivier , Zarubica Ana , Malissen Bernard , Luche Hervé
Published: April 2023

Identification of major cellular players involved in progression of cancer is key to success of new immunotherapies. It is however a daunting challenge due to the complex interplay between tumor cells and the immune system. Component of innate and adaptive immunity may be directed to a pro- or anti-tumor function. In order to cope with the complexity of the tumor microenvironment (TME), it is necessary to use an experimental approach characterizing the heterogeneity of cell types, the evolution of their relative proportion and their fine-tuned functional specificity. This approach should lead to the identification of new cellular biomarkers characteristics of different stages of tumor cancer progression.

To decipher the impact of immunotherapy treatments, cellular phenotyping of leukocytes infiltrating TME but also those present in peripheral organs is necessary. In order to increase our understanding in the precise mode of action of anti-PD1 treatment at the cellular level in sensitive and unsensitive syngeneic models, we investigated immunophenotypes and responses to immune checkpoint inhibitor (ICI) of several hallmark tumor models (MC38, CT26, B16F10, B16-OVA, RENCA, EMT6) in immunocompetent mouse models by flow and mass cytometry. We compared growth kinetics and profiled the immune cell composition of tumor microenvironment (TME), draining lymph node (dLN) and blood in order to establish immune-phenotypic cell signatures that correlates with treatment efficacy. Supervised, unsupervised and integrative data analysis arewere used to identify significant changes across different experimental settings. Our results indicate that each model possesses a unique tumor-immune infiltrate profile that can be modulated with immunotherapies. Overall, these studies provide an important resource of highly well-characterized syngeneic tumor model and highlight the importance of tumor immune landscape changes across models that will drive selection of the most appropriate model to test novel immunotherapeutic agents and enhance our translation of knowledge from syngeneic models to human tumors.

Citation Format: Anais Joachim, Emilie Maturin, Marielle Mello, Lillia Hadjem, Magali Grange, Olivier Deas, Ana Zarubica, Bernard Malissen, Hervé Luche. Deep Immuno-profiling of syngeneic tumor mouse models for preclinical studies. [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 4630.
Abstract 4667: Establishment of patient-derived human cell lines using PDX mouse model for intrahepatic cholangiocarcinoma
Authors: Matsumoto Takashi
Published: April 2023

Background: In recent years, patient tumor transplant models (PDX models) have attracted method that can predict the effects of anticancer drugs on patients. The PDX model is a technique in which the patient's tumor tissue is directly transplanted into immunodeficient mice. Tumors can grow three-dimensionally in a living mouse, so compared to culturing in petri dishes, the situation is like the growth of tumors in the patient's body. In addition, there are reports that the efficacy of anticancer drugs in patients and the results of administration of anticancer drugs in PDX show a high concordance rate, and it is attracting attention as a model that can predict efficacy in patients. In this time, we report on the establishment of cell lines using PDX mice for resected cases of intrahepatic cholangiocarcinoma in our hospital.

Methods: Twelve patients who underwent surgical resection between September 2019 and December 2021 were included. After resected the tumor, the tumor was transplanted into up to 10 PDX mice, and cell lines were established from the formed tumor tissue.

Results: Of the 12 cases, tumors formed in PDX mice after transplantation in 6 cases, and the median time to engraftment after the first transplantation was 133 days (75-188). Furthermore, among the 6 cases, 3 cases achieved the establishment of cell lines.Case1:moderately differentiated adenocarcinoma, tumor diameter 50mm, single, vascular invasion positive, no lymph node metastasis, Stage IIIA (UICC 8th)Case2:well differentiated adenocarcinoma, tumor diameter 60mm, single, vascular invasion negative, no lymph node metastasis, Stage II (UICC 8th)Case 3:moderately differentiated adenocarcinoma, tumor diameter 55mm, single, vascular invasion positive, no lymph node metastasis, Stage IIIA (UICC 8th)

Conclusion: The cell line we established this time is a case of advanced cancer, and we would like to continue to increase the number of cases and continue to establish cell lines, which will lead to future research.

Citation Format: Takashi Matsumoto. Establishment of patient-derived human cell lines using PDX mouse model for intrahepatic cholangiocarcinoma. [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 4667.
Abstract 474: Preclinical development of an oral selective estrogen receptor degrader (SERD) TFX06 for the treatment of ER+HER2− breast cancer
Authors: Wu Jinping , An Ke , Fang Douglas D. , Lu Jianyu , Hu Lihong , Wang Jing , Zhang Kaili , Xia Yuanfeng , Ding Charles , Chen Shuhui , Wei Sha
Published: April 2023

Breast cancer (BC) has recently surpassed lung cancer to become the most commonly diagnosed cancer in women worldwide. Approximately 70% of BC was ER+, in which estrogen receptor α (ERα, encoded by ESR1 gene) drives dysregulated cell proliferation. In clinic, ER+ BC has been effectively treated by endocrine therapy targeting estrogen or ER. As the first approved ERα degrader, fulvestrant has proven to be effective for locally advanced or metastatic BC. However, its clinical utilization is hampered by inconvenience of intramuscular injection, as well as its poor PK/PD profile and limited efficacy, especially in the patient population that developed ESR1 mutation-conferring drug resistance after the earlier line of treatment with an aromatase inhibitor. Thus, developing an oral and more effective therapy for ER+ BC remains an unmet medical need. We report herein discovery and preclinical investigation of a novel orally bioavailable SERD TFX06. TFX06 demonstrated potent binding affinity to ERα with a Ki of 0.10 nM and preferably antagonized ERα to ERβ with an IC50 value of 0.086 nM and 2.83 nM, respectively. TFX06 potently induced ERα degradation in ER+ BC cells and substantially inhibited cell proliferation in BC cell lines with ER wild type (WT), and MCF-7 cells expressing Y537S or D538G mutants. Oral administration of TFX06 demonstrated impressive antitumor efficacy in ER WT (TGI = 101%) and ER D538G-expressing (TGI = 99%) MCF-7 cell line-derived xenograft (CDX) models in mice. Similarly, in an ER+ BC patient-derived xenograft (PDX) model (HCI-013) with ER Y537S mutation, TFX06 also exerted excellent antitumor activity (TGI = 99%). Furthermore, TFX06 in combination with palbociclib achieved synergistic activity in MCF-7 CDX model. Moreover, TFX06 exhibited an excellent correlation between systemic/local drug exposure, pharmacodynamic modulation (i.e., ERα downregulation) and antitumor activity. Collectively, these findings demonstrate that TFX06 is a novel, orally bioavailable SERD. TFX06 demonstrates substantial antitumor activity in both in vitro and in vivo preclinical tumor models, including those expressing ESR1 mutations, through downregulation of ERα. The preclinical data warranted the clinical evaluation of TFX06 in human with an IND application submitted to the China NPMA.

Citation Format: Jinping Wu, Ke An, Douglas D. Fang, Jianyu Lu, Lihong Hu, Jing Wang, Kaili Zhang, Yuanfeng Xia, Charles Ding, Shuhui Chen, Sha Wei. Preclinical development of an oral selective estrogen receptor degrader (SERD) TFX06 for the treatment of ER+HER2− breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 474.
Abstract 4751: The function of bromodomain containing protein 4 (BRD44) in hematopoiesis
Authors: Nihozeko Francine
Published: April 2023

Bromodomain containing protein 4 (BRD4) is an epigenetic transcription regulator that reads acetylated chromatin and is involved in the recruitment of transcriptional factors to initiate gene transcription. BRD4 is functionally versatile and has extensively been studied for its role in cell cycle progression, gene transcription regulation, chromatin organization and remodeling, as well as DNA replication and repair. In addition, BRD4 has been implicated in many solid and liquid tumors, and its inhibition shows promising preclinical activity, especially in hematopoietic malignancies. BRD4 is overexpressed in Leukemia and has been reported to play a role in hematopoietic stem cell expansion and progenitor development. However, the mechanisms of BRD4 in hematopoietic stem cell functions and disease development remain poorly understood. Using BRD44 knockout (BRD44 Δ/Δ, driven by Mx1 Cre to delete BRD44 in hematopoietic system) mouse model, we found that BRD4 is required for HSCs self-renewal and the loss of BRD44 impaired erythroid differentiation and skewed myeloid differentiation. BRD44 Δ/Δ HSPCs showed G0/G1 arrest, decreased S-phase, and increased senescence compared to wild type (WT) HSPCs. Our study demonstrates that BRD4 is essential for normal hematopoiesis. To further investigate the role of BRD4 in hematopoiesis, we generated BRD44 transgenic mice (driven by Vav1 promoter, to assure all BRD44 transgene expression is instructed in the hematopoietic system). Ongoing studies focus on determining the impact of BRD4 overexpression on hematopoietic stem cell functions.

Citation Format: Francine Nihozeko. The function of bromodomain containing protein 4 (BRD44) in hematopoiesis. [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 4751.
Abstract 4848: Effects of cinnamon extracted under acidic conditions on proliferation of MCF7 cells
Authors: Anderson Madison , Stockert Amy
Published: April 2023

Recent evidence suggests that MCF7 cells utilize free fatty acids as a primary energy source by increasing lipase activity in neighboring adipocytes. Research in our lab suggests that lipase activity is increased by cinnamon extract in differentiated 3T3-L1 adipocytes. We hypothesized that since co-cultures between adipocytes and MCF7 cells demonstrated an increase in lipase activity and increased proliferation, cinnamon extract may increase proliferation of MCF7 cells, making cinnamon an important ingredient to avoid in patients with breast cancer. Cells were plated and treated with varying amounts of cinnamon and monitored for proliferation using the MTT assay. Cells consistently, albeit not significantly, showed an increase in proliferation when treated with cinnamon. Additional studies need to be completed in order to confirm these effects and look for significance, although the importance of this finding warrants rapid reporting. Our lab will examine the effects of cinnamon extract at varying concentrations on MCF7 cell proliferation under variable media conditions as well as monitoring the lipase activity in the MCF7 cells.

Citation Format: Madison Anderson, Amy Stockert. Effects of cinnamon extracted under acidic conditions on proliferation of MCF7 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 4848.
Abstract 4850: High salt diet induced tumor initiating stem cells mediate breast cancer progression
Authors: Tucker Lisa , Reid Sonya , Rathmell Jeffrey C. , Tiriveedhi Venkataswarup
Published: April 2023

High-salt (sodium chloride) diets have been associated with several chronic inflammatory diseases. While the role of chronic inflammation in cancer is well established, the specific role of high salt diet in carcinogenesis is unknown. Previous studies with syngeneic murine breast cancer models, both in our laboratory and others, have shown that high salt (HS) diet induced tumor regression through inflammatory activation of anti-tumor adaptive immune responses. We tested this counter-intuitive and suspiciously beneficial role of HS diet by performing sequential passage studies in preclinical murine models. Six week old mice were placed on high salt diet for 2 weeks prior to injection with syngeneic 4T1 and Py230 triple negative breast cancer cells (into BALB/c and C57Bl/6J mice, respectively, referred to as passage-1). Tumors were harvested after four weeks of injection. The cancer cells depleted of immune cells were collected from these harvested tumors and reinjected into new non-tumor bearing mice. This cycle was repeated three times. In passage-1 cohort, by day 28, HS diet induced reduced tumor progression in both 4T1-BALB/c (267 ± 59 mm3) and Py230-C57Bl/6J (238 ± 54 mm3) as compared to regular salt (RS) diet cohort (611 ± 94 mm3 and 473 ± 69 mm3, respectively; p<0.05 for both). However, in passage-4 cohort, by day 28, HS diet induced significantly higher tumor progression in both 4T1-BALB/c (806 ± 91 mm3) and Py230-C57Bl/6J (743 ± 81 mm3) models, as compared to regular salt (RS) diet cohort (577 ± 83 mm3 and 462 ± 77 mm3, respectively; p<0.05 for both). Cellular analysis by flow cytometry revealed that there was a 12-19 fold increase in tumor initiating stems cells (TISCs) in passage-4 HS diet cohort compared to RS diet cohort. Mechanistic studies have demonstrated that there was increased TGFβ expression on TISCs obtained from passage-4 HS diet cohort which correlated with enhanced exhaustion phenotype (CTLA4+) of tumor infiltrating adaptive immune cells (CD4 and CD8 T cells) in this cohort. Co-treatment with anti-TGFβ and anti-CTLA4 monoclonal antibodies (mAb) in passage-4 HS diet cohort, significantly reduced tumor progression (p<0.05), as compared to treatment with either mAb alone. Taken together, these data demonstrated that chronic HS diet leads to expansion of TGFβ expressing TISCs leading to host immune exhaustion and tumorigenesis. Importantly, anti-TGFβ mAb exerted a favorable synergistic effect to enhance the anti-tumor efficacy of immune check-point inhibitors.

Citation Format: Lisa Tucker, Sonya Reid, Jeffrey C. Rathmell, Venkataswarup Tiriveedhi. High salt diet induced tumor initiating stem cells mediate breast cancer progression. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4850.
Abstract 4874: Hyperforin induces apoptosis and inhibits invasion via inhibiting AKT/NF-ΚB signaling pathway in colorectal cancer cells
Authors: Chen Ying-Tzu , Chiang I-Tsang , Chang Yuan , Hsu Fei-Ting
Published: April 2023

Recently, the incidence of colorectal cancer (CRC) has been increasing year by year and has become the first incidence cancer in Taiwan. Epidermal growth factor receptor inhibitor (EGFR inhibitor) has been recognized as an important target of CRC. Therefore, targeting EGFR and its related signaling could be a potential strategy to overcome this disease. Hyperforin is a natural prenylated phloroglucinol and has been identified as the major molecule responsible for the antidepressant effects. Previous studies indicated that hyperforin has other potentially pharmacologically meaningful biological properties, including antibacterial ability and inhibition of inflammatory mediators. Numerous studies have also pointed out the anti-tumor potential of hyperforin is associated with the induction of apoptosis signaling. However, whether hyperforin may regulate CRC progression and its underlying mechanism is unclear. In our study, we used the MTT assay to prove the cytotoxicity of CRC cells (HT29 and HCT116 cells) was induced by hyperforin. We then used the flow cytometry to confirmed that hyperforin may induce both extrinsic/intrinsic apoptotic pathways in CRC cells. We further demonstrated hyperforin-induced apoptosis pathways is associated with mitochondrial-dependent apoptosis and death receptor dependent apoptosis. Then, transwell invasion, migration, and wound healing assay results proved the inhibition effect by hyperforin in HT29 and HCT116 cells. Additionally, we suggested that hyperforin can effectively inhibit EGFR downstream genes MEK-ERK and AKT signaling, as well as inhibit the phosphorylation of NF-ΚB and the nuclear translocation in CRC cells. In sum, our results show that hyperforin can inhibit the apoptosis and metastasis ability of CRC cells by inhibiting both AKT/NF- ΚB and ERK/NF-ΚB signaling pathway.

Citation Format: Ying-Tzu Chen, I-Tsang Chiang, Yuan Chang, Fei-Ting Hsu. Hyperforin induces apoptosis and inhibits invasion via inhibiting AKT/NF-ΚB signaling pathway in colorectal cancer cells. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4874.
Abstract 4913: Enhancer RNA profiles predict sensitivity to a novel GPX4 inhibitor
Authors: Read Timothy , Simpson David , Rimel Jenna , Rhine Christy , Pham Casey , Liechty Cole , Martin Eric , Azofeifa Joey
Published: April 2023

Inhibition of the GPX4 enzyme leads to the rapid accumulation of toxic lipid peroxides and cell death through a process known as ferroptosis. This mechanism has recently garnered significant attention in the field of oncology because many difficult-to-treat tumor types are particularly susceptible to this non-canonical form of cell death. However, robust biomarkers that predict sensitivity to ferroptosis are needed to help select well-defined tumor subtypes and patient segments that are likely to respond to GPX4-targeting. Our nascent RNA platform measures the active transcription of not only protein-coding transcripts, but also transcripts initiating from enhancer regions (enhancer RNAs) critical for defining cell type. We reasoned that these data could afford the unique opportunity to identify non-coding transcripts that could serve as novel biomarkers of sensitivity to ferroptosis. We recently discovered a potent and selective GPX4 inhibitor and profiled the compound for its ability to induce ferroptosis across a diverse group of cancer cell types. We chose several of the most sensitive and resistant cell lines for further examination using our nascent RNA platform and identified novel relationships between sensitivity to ferroptosis and cell-specific enhancer RNA profiles. Importantly, enhancer RNA profiles were more effective at stratifying sensitive and resistant cell lines than protein-coding gene expression. Using enhancer RNA expression profiles as biomarkers of drug sensitivity represents a novel strategy that could ultimately improve our ability to select patient cohorts more likely to benefit from emerging drug candidates and established therapeutics.

Citation Format: Timothy Read, David Simpson, Jenna Rimel, Christy Rhine, Casey Pham, Cole Liechty, Eric Martin, Joey Azofeifa. Enhancer RNA profiles predict sensitivity to a novel GPX4 inhibitor. [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 4913.
Abstract 4931: Novel cancer suppressing role of REIC/DKK3 protein through a downregulation of cell surface PD-L1 in breast cancer cells
Authors: Gohara Yuma , Tomonobu Nahoko , Kinoshata Rie , Audebert Lena , Chen Youyi , Komalasari Ni Luh Gede Yoni , Jiang Fan , Yoshizawa Chikako , Murata Hitoshi , Yamamoto Ken-ichi , Sakaguchi Masakiyo
Published: April 2023

Adenovirus vector carrying REIC gene (Ad-REIC) is currently in clinical evaluation. The anti-cancer effect of Ad-REIC is mainly caused by two ways, cancer specific apoptosis via endoplasmic reticulum stress with an abundant REIC protein and indirect immune effects by the secreted REIC protein. The former had been well studied that gave us general comprehension of its mechanisms, however latter has still not been fully elucidated. To clear this, we have aimed to identify putative receptor(s) to the extracellularly secreted REIC protein, and have fortunately succeeded to identify five candidates. Surprisingly, the identified receptors all worked to bind with PD-L1 and hold it to stabilize on the plasma membrane, and the REIC binding to the receptors induced liberation of PD-L1 from the receptors’ lock, resulting in an accelerated degradation of PD-L1. This novel insight may lead to the elucidation in important part of the REIC-mediated indirect anti-cancer role.

Citation Format: Yuma Gohara, Nahoko Tomonobu, Rie Kinoshata, Lena Audebert, Youyi Chen, Ni Luh Gede Yoni Komalasari, Fan Jiang, Chikako Yoshizawa, Hitoshi Murata, Ken-ichi Yamamoto, Masakiyo Sakaguchi. Novel cancer suppressing role of REIC/DKK3 protein through a downregulation of cell surface PD-L1 in breast cancer cells. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4931.
Abstract 5015: Wee1 inhibition and its synthetic lethal combination with Chk1 inhibition in mouse model of neuroendocrine prostate cancer
Authors: Chao Yapeng , Chen Yuzhou , Liu Yu , Connelly Jaclyn A. , Wang Hong , Wang Qiming Jane
Published: April 2023

The mitotic regulator Wee1 is a critical regulator of G2/M cell cycle checkpoint and DNA damage response pathways. Wee1-targeted agents are under active clinical investigation in multiple cancer types. However, Wee1 and its inhibitors have not been well studied in prostate cancer, particularly neuroendocrine prostate cancer (NEPC). In this study, we investigated the therapeutic potential of Wee1 inhibitor AZD1775 and its synthetic lethal combination with Chk1 inhibitor in the context of NEPC. Our data demonstrated increased Wee1 expression in NEPC cells and tumors. Treatment of NEPC cells with AZD1775 inhibited NEPC tumor cell survival with greater potency as compared to other prostate cancer cells, demonstrating increased vulnerability of NEPC cells to Wee1 inhibition. We further identified synthetic lethality between the inhibition of Wee1 and several protein kinases, particularly Chk1, another key G2/M checkpoint kinase. Using an autochthonous adenocarcinoma mouse prostate (TRAMP) model that gives rise to NEPC tumors, we assessed the efficacy of AZD1775 and a highly selective Chk1 inhibitor SRA737 as single agents and in combination on the progression, survival, and metastatic spread of NEPC tumors in vivo. AZD1775 and SRA737 when used as single agents significantly suppressed tumor growth, improved overall survival, and incidence of distant metastases of mice with NEPC tumors. The combination of AZD1775 and SRA737 on a sequential 2-wk treatment schedule was more efficacious in suppressing tumor growth and metastasis than each drug alone in castrated TRAMP NEPC tumors. Mechanistically, the combination of Chk1 and Wee1 inhibitors synergized to reduce the inhibitory phosphorylation of CDK1 at Y15 and Cdc25c at S216, resulting in enhanced activation of CDK1/cyclin B complex, abrogation of G2/M checkpoint, and consequent premature mitotic entry, which ultimately led to mitotic catastrophe and apoptosis. Our findings provide promising preclinical evidence supporting the therapeutic values of Wee1 and Chk1 inhibitors as single agents and in combination for the treatment of NEPC.

Citation Format: Yapeng Chao, Yuzhou Chen, Yu Liu, Jaclyn A. Connelly, Hong Wang, Qiming Jane Wang. Wee1 inhibition and its synthetic lethal combination with Chk1 inhibition in mouse model of neuroendocrine prostate 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 5015.
Abstract 5191: Modeling age-related changes in the tumor microenvironment in an orthotopic immunocompetent murine pancreatic cancer model - a novel approach
Authors: Green Morgan M. , Fujimoto Tara , Taniguchi Cullen , Fowlkes Natalie
Published: April 2023

Pancreatic cancer is the third leading cause of cancer-related deaths in the U.S. and is most frequently diagnosed between the ages of 65-74 years. Only 11.5% of patients are expected to survive 5 years after diagnosis. There is extensive evidence that tumor microenvironment (TME) plays a critical role in cancer progression and metastasis. Despite the fact that most patients are diagnosed over the age of 60, most preclinical murine tumor models utilize young, healthy, adolescent mice, failing to account for structural and/or functional changes in stroma and immune cells that occur in tissues over time and contribute to tumor progression. Marked differences in lifespan between mice and humans preclude natural development of many age-related physiologic changes in mice that occur in humans over the course of decades. We performed a pilot study to test our hypothesis that KPC pancreatic tumors grown orthotopically in the pancreas of genetically engineered, C57BL/6-Tg (LMNA*G608G) HClns/J mice, a model of Hutchinson-Gilford progeria syndrome with features of accelerated aging, would have altered tumor growth in comparison to tumors in age-matched wild type controls. We found that tumor growth in LMNA mice outpaced that of WT mice. Quantitative digital image analysis showed significantly reduced intratumoral infiltration of CD45+ leukocytes in LMNA mice in comparison to WT (p=0.0098). There was a significant reduction in tumor-infiltrating CD3+ T cells (p=0.038) and F4/80+ macrophages (p=0.0102). Our data supports the hypothesis that functional, age-related, immune and/or stromal changes in the TME impact pancreatic tumor growth and progression. Innovative strategies that better model age-related changes in the TME may better recapitulate the aggressive course of pancreatic cancer in aged patients.

Citation Format: Morgan M. Green, Tara Fujimoto, Cullen Taniguchi, Natalie Fowlkes. Modeling age-related changes in the tumor microenvironment in an orthotopic immunocompetent murine pancreatic cancer model - a novel approach. [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 5191.
Abstract 5197: NCG-hIL2 mice: a novel model to support the development of T regulatory and NK cells
Authors: Zhang Xinxin , Wu Meirong , Wang Huiyi , Yu Weiwei , Zhu Fang , Xu Jianming , Sun Hongyan , Wang Hongyu , Ju Cunxiang , Chen Santi Suryani , Li Zhiying , Moore Mark Wade , Zhao Jing , Gao Xiang
Published: April 2023

Interleukin 2 is a key molecule that promotes the expansion and activation of lymphocytes, including T regulatory cells (Tregs) and natural killer (NK) cells. Tregs are specialized subpopulation of T cells that suppress the immune response by inhibiting T cell proliferation and cytokine production. NK cells are the predominant innate immune subset that mediates anti-tumor and anti-viral responses. Both Tregs and NK cells are two critical immune cells that play an important role in cancer immunotherapy, however, there is a lack of models that supports the in vivo status of these two immune subsets. To address this, we developed a severe immunodeficient mouse model, NOD/ShiLtJGpt-Prkdcem26Cd52Il2rgem26Cd22/Gpt (NCG-hIL2), by knocking in the human IL-2 gene on the NCG background. The ability of this model to support Treg and NK cells was then compared to NCG and tested with the reconstitution of human peripheral blood mononuclear cells (PBMC) and human hematopoietic stem cells (HSC).

In the PBMC engrafted mice, the presence of IL-2 supported the development of T cells, especially the Tregs, with 53.74% in Tregs in NCG-IL2 compared to 4.81% in NCG mice (test at 2 weeks of reconstruction). Similarly, the NK cells are also supported with those cohort engrafted with human HSC (91.63% in NK cells in NCG-IL2 compared to 0.53% in NCG mice, test at 10 weeks of reconstruction). Notably, reconstituted NK cells expressed various NK receptors such as NKp30, NKp44, NKp46, NKG2D, and CD94 in NCG-IL2 mice. They produced comparable levels of granzyme when compared with human peripheral blood-derived NK cells, and a considerable amount of perforin protein was detected in the plasma of huHSC-NCG-hIL2 mice.

In conclusion, compared with NCG mice, humanized NCG-hIL2 mice can reconstruct more abundant immune cells and have a wider application in anti-tumor research. It is an ideal model for the preclinical study of anti-tumor therapeutics.

Citation Format: Xinxin Zhang, Meirong Wu, Huiyi Wang, Weiwei Yu, Fang Zhu, Jianming Xu, Hongyan Sun, Hongyu Wang, Cunxiang Ju, Santi Suryani Chen, Zhiying Li, Mark Wade Moore, Jing Zhao, Xiang Gao. NCG-hIL2 mice: a novel model to support the development of T regulatory and NK 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 5197.
Abstract 520: Investigation the function and application of spermine in castration-resistant prostate cancer
Authors: Sun Jingkai , Chiu Peter Ka-Fung , Xie Tingting , Teoh Jeremy Yuen-Chun , Ng Chi-Fai
Published: April 2023

Introduction & Objectives: Spermine (Spm) is a typical metabolite that is highly concentrated in healthy prostate epithelium maintaining cell growth and functions. Our previous studies found that Spm was significantly decreased in PCa tissue than in benign prostate tissue, as was the urinary Spm in PCa. This study aimed to reveal the specific role of Spm in PCa.

Materials & Methods: In this study, Spm was resupplied to castration-resistant prostate cancer (CRPC) cells (PC3 and DU145). Following Spm treatment, MTT (methyl thiazolyl tetrazolium) and EdU (5-ethynyl-2’-deoxyuridine) assay were used to determine the cell proliferation of CRPC cell lines. Cellular apoptosis, cell cycle distribution and mitochondrial membrane potential were analyzed by flow cytometry. Cellular reactive Oxygen species (ROS) level was measured by fluorescence microscope. mRNA and ATAC sequencing in spermine-treated CRPC cells were performed to reveal the related mechanism for Spm effect.

Results: In the in vitro studies, Spm was found to inhibit the cell viability of CRPC cells. Transwell assays demonstrated that Spm impaired migration and invasion ability. And Spm blocked tumor progression by inducing G2/M arrest and cellular apoptosis in CRPC cells dose-dependently. mRNA seq revealed that a series of genes were altered after Spm treatment and indicated the potential functional pathway of Spm might be mitophagy. We integrated mRNA seq data with ATAC seq profiles and accessed those genes regulated by polyamine metabolism and Spm-induced mitophagy. Results showed the sequencing tracks for the HMOX1 locus contain higher ATAC-seq peaks at the promoter under Spm treatment. Spm could affect oxidative phosphorylation which will lead to mitophagy and mitochondrial metabolism by modulating the transcriptional accessibility of genes. After Spm treatment, cellular ROS was upregulated while the mitochondrial membrane potential collapsed. Subsequently, western blot results showed that PINK1-mediated mitophagy related-proteins were significantly regulated.

Conclusion: Our results indicated that Spm treatment led to mitochondrial dysfunction and subsequently degraded through mitophagy in CRPC cells. Our findings could provide a theoretical basis and have preliminary clinical implications in developing Spm as a promising agent against CRPC by inducing cellular mitophagy in CRPC cells.

Citation Format: Jingkai Sun, Peter Ka-Fung Chiu, Tingting Xie, Jeremy Yuen-Chun Teoh, Chi-Fai Ng. Investigation the function and application of spermine in castration-resistant prostate 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 520.
Abstract 5209: Generation of humanized TREM2 mice for preclinical evaluation of therapeutics targeting tumor-associated macrophages
Authors: Lv Ruili , Yu Jia , Shen Zhiyuan , Jin James
Published: April 2023

In recent years, immune checkpoint inhibitors (ICIs) have improved treatment for several types of cancer. However, the overall response rates remain low, and drug resistance can occur in patients after initial responses. This resistance is thought to be due in part to tumor-associated macrophages (TAMs), which can deploy a number of mechanisms to suppress anti-tumor immunity, resulting in tumor immune escape. Triggering receptor expressed on myeloid cells-2 (TREM2), a transmembrane receptor that is part of the immunoglobulin superfamily, is highly expressed on the surface of TAMs. Moreover, the expression level of TREM2 is reported to be higher in cancerous tissues vs normal tissues, with higher expression associated with poorer patient outcomes across multiple solid tumor types. Conversely, mice that lack Trem2 experience slower tumor cell expansion and are more responsive to anti-PD-1 immunotherapy, suggesting that TREM2 may be a promising therapeutic target for improving responses to ICI immunotherapies. To better assist the efficacy evaluation of anti TREM2 antibodies, Biocytogen has generated a humanized TREM2 mouse model. In these mice, exons 1-5 and the 3’UTR of mouse Trem2 gene, which encodes the full-length protein, were replaced by its human counterpart. In homozygous mice, human TREM2 protein expression is detected. The distribution of basal leukocyte subpopulations of blood, spleen, and lymph nodes in humanized B-hTREM2 mice were similar and comparable to those in wild-type C57BL/6 mice. Using the MC38 tumor model, we showed modest efficacy of an anti-human TREM2 antibody in inhibiting tumor growth in vivo. In summary, TREM2 humanized mice provide a powerful preclinical model for in vivo evaluation of anti-human TREM2 antibodies for cancer treatment.

Citation Format: Ruili Lv, Jia Yu, Zhiyuan Shen, James Jin. Generation of humanized TREM2 mice for preclinical evaluation of therapeutics targeting tumor-associated macrophages. [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 5209.