bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3250: Targeting CD38 on exhausted T cells overcomes resistance to cancer immunotherapy
Authors: Revach Or-Yam , Cicerchia Angelina M. , Sade-Feldman Moshe , Anderson Seth , Manguso Robert T. , Hacohen Nir , Jenkins Russell W.
Published: April 2023

Despite the unprecedented success of immune checkpoint blockade (ICB) in melanoma and other cancers, therapeutic resistance remains a major challenge. CD38, an ecto-enzyme involved in NAD+ catabolism, is upregulated in dysfunctional/exhausted CD8+ T cells in human melanoma. As enrichment of dysfunctional CD8+ T cells is associated with lack of response to ICB, and NAD+ depletion via enhanced CD38 activity has been associated with diminished immune response, we hypothesized that CD38 might represent an attractive target to overcome resistance to ICB. While CD38 blockade has been shown to augment response to ICB in murine tumor models, the specific role of CD38 signaling in CD8 T cell exhaustion, and the therapeutic relevance of CD38 in human cancer have not been defined. Here, we confirm and extend previous observations that CD38 is enriched in exhausted CD8+ T cells during tumor progression and in response to ICB. Further, we demonstrate that disrupting CD38 signaling via neutralizing antibodies or pharmacological inhibition enhances response to ICB using established murine tumor models and patient-derived tumor explant models. Notably, we observed 54% response rate to combination PD-1/CD38 blockade using a cohort (n=35) of patient-derived organotypic tumor spheroids (PDOTS) from patients with ICB-refractory melanoma (11.4% response rate to single-agent PD-1 blockade). Supplementation of nicotinamide riboside (NR) to boost NAD+ levels mimicked the effects CD38 blockade/inhibition on PD-1 response, whereas FK866, an inhibitor of NAMPT a key step in NAD+ biosynthesis, blunted the response to combined PD-1/CD38 blockade, suggesting a major role for NAD+ in the efficacy of CD38 blockade/inhibition. Lastly, we demonstrate that in vitro CD38 inhibition/blockade in CD8+ T cells increases the levels of TCF7 (an effector/memory T cell transcription factor), decreases surface expression of co-inhibitory receptors (e.g., TIM-3, CD39, PD-1), and improves effector CD8 T cell function. Taken together, these data confirm a role for CD38 in CD8+ T cell exhaustion in melanoma and support further pre-clinical and clinical development of this novel therapeutic strategy to enhance anti-tumor immune responses.

Citation Format: Or-Yam Revach, Angelina M. Cicerchia, Moshe Sade-Feldman, Seth Anderson, Robert T. Manguso, Nir Hacohen, Russell W. Jenkins. Targeting CD38 on exhausted T cells overcomes resistance to 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 3250.
Abstract 3367: Tumor-derived Extracellular Vesicles as a potential PD biomarker for TEAD central pocket binder activity in liquid biopsy
Authors: Nunes Manoel , Spanakis Emmanuel , Dos-Santos-Bele Wilson , Rabia Emilia , Soubigou Stephane , Muzard Gaelle , Dos-Santos Odette , Pollard Jack , Hadjipanayis Angela , Delahaye Fabien , Venier Olivier , Debussche Laurent , Jackson Don , Dib Colette , Valtingojer Iris , Cesaroni Matteo
Published: April 2023

We assessed the utility of mRNAs in extracellular vesicles as biomarkers for inhibition of YAP1/TEAD signaling by the TEAD central pocket inhibitor. The field of liquid biopsies is of enormous interest for noninvasive monitoring of cancer progression and response to treatment. Extracellular Vesicles (EVs) such as exosomes, microvesicles, and apoptotic bodies, are nanoparticles found in all biological fluids. These cell-derived, small secreted vesicles convey biological information, either by surface-to-surface interaction or by shuttling bioactive molecules to a recipient cell’s cytoplasm. Because EVs harbor the cargos of their original cells, their contents may be useful as biomarkers to follow drug activity. (F. Urabi et al., 2020). We compared the effects of YAP1/TEAD inhibitor treatment on intracellular and EV-encapsulated mRNAs in three compound-sensitive tumor cell lines (and one insensitive cell line) based on TEAD inhibitor IC50 assay profiles. Cell lines were treated at four doses of YAP1/TEAD inhibitor (0.0 µM, 0.3 µM, 1.0 µM & 3.0 µM) for 24h. EVs and intracellular RNAs were isolated and profiled by RNAseq. To estimate YAP1/TEAD pathway activity, we used a transcriptomic signature identified previously by our group (L. Calvet et al., 2022) and we showed that the effects of YAP1/TEAD inhibitor treatment on the YAP1/TEAD score in intracellular mRNAs corresponded to a decrease of the score in EV-secreted RNAs. In addition, we ran an unbiased analysis to identify single transcripts in the EVs that were modulated by YAP1/TEAD inhibitor. One of the best markers identified by regression analysis was CYR61, that is part of the signature and is a well-established downstream target of YAP/TEAD pathway. In conclusion, these results suggest that the YAP1/TEAD signature in EV mRNAs represents a relevant PD biomarker to monitor YAP1/TEAD inhibitor activity and could potentially serve as a noninvasive liquid biopsy-based biomarker detection in the clinic.

Citation Format: Manoel Nunes, Emmanuel Spanakis, Wilson Dos-Santos-Bele, Emilia Rabia, Stephane Soubigou, Gaelle Muzard, Odette Dos-Santos, Jack Pollard, Angela Hadjipanayis, Fabien Delahaye, Olivier Venier, Laurent Debussche, Don Jackson, Colette Dib, Iris Valtingojer, Matteo Cesaroni. Tumor-derived Extracellular Vesicles as a potential PD biomarker for TEAD central pocket binder activity in liquid biopsy [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 3367.
Abstract 3416: Suppression of AXL inhibits tumor growth through promotion of dendritic cell migration in lung cancer
Authors: Kim Da-Som , Lee Cha Won , Im Kyungtaek , Choi Yun Jung , Kim Dong Ha , Choi Chang-Min , Ji Wonjun , Lee Jae Cheol , Rho Jin Kyung
Published: April 2023

AXL is a receptor tyrosine kinase (RTK) that has a various role in tumor progression such as proliferation, metastasis, angiogenesis, drug resistance and immune tolerance. Although targeting AXL exhibits an anti-cancer effects partially depending on the immune system, their molecular mechanisms have not been fully elucidated. To search the roles of AXL in immune tolerance of tumor, we generated AXL knockout (KO) mice. These AXL KO mice showed the reduced tumor growth in syngeneic LLC-1 model compared to wild type mice. To validate the mechanisms underlying the inhibition of AXL KO-derived tumor growth, we examined tumor-infiltrating lymphocytes (TIL). Compared with tumor-bearing wild type mouse, CD45-positive cells were significantly enhanced in tumor-bearing AXL KO mouse. Especially, these results were dominant in CD8-positive cells and dendritic cells (DCs). Tumor-infiltrating DCs showed the reduction of PD-L1 and Tim3 expression. In addition, the inhibition of AXL led to induce the DCs migration. In conclusion, our data demonstrated that targeting AXL may inhibit tumor growth through the activation of DCs including the enhancement of migration and reduced expression of immune check point proteins.

Citation Format: Da-Som Kim, Cha Won Lee, Kyungtaek Im, Yun Jung Choi, Dong Ha Kim, Chang-Min Choi, Wonjun Ji, Jae Cheol Lee, Jin Kyung Rho. Suppression of AXL inhibits tumor growth through promotion of dendritic cell migration 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 3416.
Abstract 3429: Beyond cereblon IMIDs - biophysics-based discovery of novel molecular glue chemotypes
Authors: Zhang Yujia , Bates Jessica , Gourdet Benoit , Birch Louise , Addis Philip , Hjerpe Roland , Jordan Allan M.
Published: April 2023

Molecular glue degraders are compact, low molecular weight molecules that can efficiently induce specific and potent degradation of a target protein. This class of degraders function by inducing interactions between a target of interest and a ubiquitin-ligase, either by stabilization of weak pre-existing interactions, or by generation of entirely novel interactions. These molecules offer significant opportunity beyond heterobifunctional degraders such as PROTACs, not least in terms of improved molecular properties.

However, beyond the IMID molecular glues, typified by thalidomide, pomalidomide and lenalidomide, novel molecular glue chemotypes remain scarce.

To address this need, we have developed biophysics-based molecular glue screening platform, exploiting our internal, high quality fragment library and proximity-based screening platforms to rapidly identify promising new molecular glues for further optimization. A potential advantage of utilizing cell-free biophysical systems is the opportunity to select both the target and the desired ligase, opening up for development of degraders that capitalize upon differential expression of ligases in different tissues.

As proof of concept, we have applied this platform to find new molecular glues to degrade CK1α. This Ser/Thr kinase has been found to be over-expressed in metastatic colorectal cancer, and this over-expression correlates with poor overall survival. The kinase has also been implicated as an oncogenic driver in tumors such as B-Cell lymphomas and non-Hodgkin lymphomas, suggesting a potential therapeutic application for novel CK1α molecular glues.

Utilizing known IMID-derived molecular glues between CK1α and CRBN as benchmark controls, we identified several non-IMID derived chemotypes as tentative stabilizers of the CRBN/CK1α interaction. Further studies on these novel candidate degrader templates are now underway.

Citation Format: Yujia Zhang, Jessica Bates, Benoit Gourdet, Louise Birch, Philip Addis, Roland Hjerpe, Allan M. Jordan. Beyond cereblon IMIDs - biophysics-based discovery of novel molecular glue chemotypes [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 3429.
Abstract 3527: WNT5B drives osteosarcoma stemness, metastasis and chemoresistance
Authors: Perkins Rachel S. , Suthon Sarocha , Miranda-Carboni Gustavo A. , Krum Susan A.
Published: April 2023

Osteosarcoma is a pediatric bone cancer that has no targeted therapy and has had no treatment advances for the last three decades. Osteosarcoma frequently metastasizes to the lungs, reducing patient survival with metastatic disease to 20% after 5 years. The goal of this study is to describe the function of WNT5B and its signaling pathway in osteosarcoma stem cells, metastasis and chemoresistance and its potential as a therapeutic target. Using RNA sequencing from publicly available datasets and immunohistochemistry on tumor microarrays, we reveal that WNT5B is the most expressed WNT in osteosarcoma patients and correlates with both metastasis and survival. Osteosarcoma is thought to result from a block in differentiation, and based on WNT5B’s role in inhibiting osteoblast differentiation of mesenchymal stem cells from normal bone, we hypothesized a role for WNT5B in osteosarcoma stem cells. In spheroids, we show that both protein and mRNA levels of WNT5B are enhanced in the stem cell population compared to adherent cells. We found that WNT5B upregulates the expression of the stemness gene SOX2 and directs stemness phenotypes, such as sphere forming efficiency, proliferation and migration. We show a reduction in sphere forming efficiency with 143B-WNT5B-knockdown cells compared to parental control cells which can be rescued with re-introduction of WNT5B. Additionally, we show that WNT5B drives proliferation and migration of osteosarcoma stem cells through increasing sphere size and capability to migrate on collagen matrix. Further, WNT5B enhances osteosarcoma chemoresistance to methotrexate. Through revealing a novel role for WNT5B in osteosarcoma cancer stem cells and therapy resistance, we present the WNT5B pathway as a candidate for therapeutically targeting osteosarcoma stem cells in patients.

Citation Format: Rachel S. Perkins, Sarocha Suthon, Gustavo A. Miranda-Carboni, Susan A. Krum. WNT5B drives osteosarcoma stemness, metastasis and chemoresistance. [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 3527.
Abstract 3534: Interrogating the immune microenvironment of a novel mouse model of fusion positive rhabdomyosarcoma
Authors: Resch Erin E. , Oristian Kristianne M. , Mendes Elizabeth , Linardic Corinne M. , Ladle Brian H.
Published: April 2023

Immunotherapies have been largely ineffective for pediatric soft tissue sarcomas, in particular for fusion-positive rhabdomyosarcoma (FP-RMS), characterized by the PAX-FOXO1 gene fusion. We are hampered by a paucity of predictive, immune-competent mouse models of this cancer with which to develop novel and effective immune-based therapies. To overcome this barrier, we analyzed the immune microenvironment of one of the few genetically-engineered mouse models (GEMM) of FP-RMS. Briefly, this model is driven by conditional knock-in expression of Pax3:Foxo1 at the endogenous Pax3 locus, with or without conditional loss of MST1/2 (gene name Stk4/3), a Hippo pathway mediator that is modulated by PAX3-FOXO1. These two genotypes demonstrate different disease penetrance and tumor location, with mice bearing loss of MST1/2 exhibiting higher tumor penetrance and favoring head and neck disease sites. We hypothesize that these models exhibit differences in tumor-immune interactions in the microenvironment that correlate with these phenotypic differences. We aim to characterize the immune microenvironment differences between these tumors and to establish this GEMM as a predictive, syngeneic model of FP-RMS. Tumors were generated spontaneously from a GEMM of FP-RMS. Tumors expressed PAX3-FOXO1 and either carried a conditional loss of MST1/2 (MSTnull) or native MST1/2 (MSTwt). We analyzed 8 formalin-fixed, paraffin-embedded (FFPE) tumors and 2 tumor-derived cell lines from these novel models. FFPE tumor samples were analyzed via immunohistochemistry (IHC) for CD3 (T cell marker) and CD11b (myeloid cell marker) and were scored for immune cell infiltration. Overall, tumors demonstrate poor CD3+ T cell infiltration with significantly higher infiltration of CD11b+ myeloid cells (p=0.0002); this trend is also seen within the individual MSTwt (p=0.0286) and MSTnull cohorts (p=0.0286). There was no significant difference in CD3+ T cell or CD11b+ myeloid cell density between the two genotypes. The drivers of this immune phenotype and the role of Hippo pathway signaling in mediating RMS tumor immune cell infiltration warrants further investigation. Additional ongoing studies are utilizing a spatial gene expression platform to visualize the spatially-preserved transcriptome of intact tumors to reveal potential pathways mediating the immune microenvironment.

Citation Format: Erin E. Resch, Kristianne M. Oristian, Elizabeth Mendes, Corinne M. Linardic, Brian H. Ladle. Interrogating the immune microenvironment of a novel mouse model of fusion positive rhabdomyosarcoma. [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 3534.
Abstract 3610: Prenylated PALM2 promotes the migration of esophageal squamous cancer cells through activating ezrin
Authors: Deng Danxia , Li Chengyu , Zheng Zhenyuan , Wen Bing , Liao Liandi , Li Enmin , Xu Liyan
Published: April 2023

Proteins containing a CAAX motif at the C-terminus can occur prenylation modification and regulate the localization and activity of a series of key regulatory proteins, including RAS superfamily members, heterotrimeric G proteins, nuclear lamina protein, and several protein kinases and phosphatases. However, studies of prenylated proteins in esophageal cancer are limited. Here, we found that paralemmin-2 (PALM2), a potential prenylated protein, was up-regulated and associated with a poor prognosis of patients, through research on large-scale proteomic data of esophageal cancer in our laboratory. The low throughput verification also showed that the expression of PALM2 in esophageal cancer tissues was higher than that in their paired normal esophageal epithelial tissues, and it was generally expressed in the membrane and cytoplasm of esophageal squamous cancer cells (ESCC). PALM2 interacted with the two subunits of farnesyl transferase (FTase), FNTA and FNTB. The FTase inhibitor weakened the membranous location of PALM2 and mutation in the CAAX motif of PALM2 (PALM2C408S) impaired its membranous localization, indicating PALM was prenylated by FTase. Overexpressed PALM2 promoted the migration of cancer cells, whereas PALM2C408S lost this ability. Mechanistically, PALM2 interacted with the N-terminal FERM domain of ezrin of Ezrin/Radixin/Moesin (ERM) family dependent on its prenylation. The mutagenesis indicated that lysine residues K253/K254/K262/K263 in ezrin’s FERM domain and cysteine residue C408 in PALM2’s CAAX motif were important for their interaction and ezrin activation. The knockout of ezrin prevented enhanced cancer cell migration by PALM2 overexpression. PALM2, depending on its prenylation, made ezrin get more membranous distribution and increased the phosphorylation of ezrin Y146 site. In summary, prenylated PALM2 promoted the migration of cancer cells through activating ezrin. (This work was supported by the National Natural Science Foundation of China (82173034, 81172264), and Innovative Team Grant of Guangdong Department of Education (2021KCXTD005)).

Citation Format: Danxia Deng, Chengyu Li, Zhenyuan Zheng, Bing Wen, Liandi Liao, Enmin Li, Liyan Xu. Prenylated PALM2 promotes the migration of esophageal squamous cancer cells through activating ezrin. [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 3610.
Abstract 3631: Infiltrating Bruton’s tyrosine kinase expressed B cells effects on prostate cancer metastasis
Authors: Byrd Crystal , Liou Geou- Yarh
Published: April 2023

Prostate cancer is the most common type of cancer found in senior males and is the 2nd leading cause of cancer death in American males. Although prostate cancer grows slowly within the prostate glands, because of a lack of efficient treatments at the appropriate timing, cancer cells eventually become aggressive and metastatic. The progression of cancer is supported by many factors, such as signals received from surrounding cells, like B cells and other effector cells. B cells play a role as a part of the adaptive immune system to produce antibodies. To become mature B cells, they rely on activation of a non-receptor kinase, Bruton’s Tyrosine Kinase (BTK). Preliminary data from our lab showed that in the prostate cancer tissues, more infiltrated B cells expressing BTK were present nearby the prostate cancer cells. Upon this finding, we hypothesize that BTK expressed B cells will support the survival of prostate cancer cells through BTK signaling. In this study, we will overexpress BTK in B cells to study the proliferation and migration of prostate cancer cells through a co-culture system. Our findings will show an increase in migration of prostate cancer cells when cultured with BTK expressed B cells. In the future, we will validate the importance of BTK in regulating prostate cancer migration and invasion; and identify their mechanisms. Our overall findings from this project would reveal key elements of the signaling pathways of BTK expressed B cells to promote prostate cancer migration.

Citation Format: Crystal Byrd, Geou- Yarh Liou. Infiltrating Bruton’s tyrosine kinase expressed B cells effects on prostate 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 3631.
Abstract 3666: A human skeletal muscle stem/myotube model reveals multiple signaling targets of cancer secretome in skeletal muscle
Authors: Wang Ruizhong , Kumar Brijesh , Bhat-Nakshatri Poornima , Khatpe Aditi Sanjay , Murphy Michael P. , Wanczyk Kristen E. , Doud Emma H. , Mosley Amber L. , Liu Yunlong , Chen Duojiao , Simpson Ed , Gao Hongyu , Nakshatri Harikrishna
Published: April 2023

Skeletal muscle dysfunction due to the effects of cancer secretome is observed in multiple cancer types and extreme dysfunction is manifested as cachexia. Major preclinical studies on cancer-associated muscle defects utilized mouse models or mouse C2C12 mouse myoblast cell line for in vitro studies. Because of species specificity of certain cytokines/chemokines in the secretome, a human model system is required to fully comprehend the effects of cancer secretome on skeletal muscle. Here, we report a simple method to establish skeletal muscle stem cell line (hMuSC), which can be differentiated into myotubes. Using single nuclei ATAC-seq (snATAC-seq) and RNA-seq (snRNA-seq), we document chromatin accessibility and transcriptomic changes associated with hMuSCs to myotube transition. Cancer cell line derived factors accelerated stem to myotube differentiation with accompanying changes including an increase in PAX7+/MyoD+ myogenic progenitor cells. Among the pathways activated by cancer-derived factors include inflammatory pathway involving CXCL8 (also called IL-8), glucocorticoid receptor (GR) pathway, and wound healing pathway. Furthermore, cancer-derived factors significantly altered splicing machinery in hMuSCs. Additionally, AKT and p53 pathways that function in metabolic/survival pathways of the skeletal muscle were adversely affected when hMuSCs were exposed to cancer cell-derived factors. Cancer-derived factors increased the expression levels of previously known cachexia-associated genes such as MT-2, ZIP14, and PDK4. Thus, the model system not only recapitulates results of previous mouse studies but also provides much needed human model system that can easily be adapted for large scale studies to explore the epigenomic changes during hMuSC differentiation and to screen for drugs that restore skeletal muscle function in various diseases.

Citation Format: Ruizhong Wang, Brijesh Kumar, Poornima Bhat-Nakshatri, Aditi Sanjay Khatpe, Michael P. Murphy, Kristen E. Wanczyk, Emma H. Doud, Amber L. Mosley, Yunlong Liu, Duojiao Chen, Ed Simpson, Hongyu Gao, Harikrishna Nakshatri. A human skeletal muscle stem/myotube model reveals multiple signaling targets of cancer secretome in skeletal muscle. [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 3666.
Abstract 3700: Isocitrate dehydrogenase 1 sustains a hybrid cytoplasmic-mitochondrial tricarboxylic acid cycle in prostate cancer
Authors: Gonthier Kevin , Weidmann Cindy , Berthiaume Line , Jobin Cynthia , Lacouture Aurélie , Lafront Camille , Harvey Mario , Neveu Bertrand , Loehr Jérémy , Bergeron Alain , Fradet Yves , Lacombe Louis , Riopel Julie , Latulippe Éva , Atallah Chantal , Shum Michael , Lambert Jean-Philippe , Pouliot Frédéric , Pelletier Martin , Audet-Walsh Étienne
Published: April 2023

Background: The androgen receptor (AR) is an established orchestrator of cell metabolism in prostate cancer (PCa), notably by inducing an oxidative mitochondrial program. Intriguingly, AR regulates cytoplasmic isocitrate dehydrogenase 1 (IDH1) but not its mitochondrial counterparts IDH2 and IDH3. Here, we aimed to understand the functional role of IDH1 in PCa.

Methods: Mouse models, in vitro human PCa cell lines, and human prostate organoids were used to study the expression and activity of IDH enzymes in the normal prostate and PCa. Genetic and pharmacological inhibition of IDH1 was then combined with extracellular flux analysis and gas chromatography-mass spectrometry for metabolomic analyses and cancer cell proliferation in vitro and in vivo.

Results: In PCa cells, more than 90% of the total IDH activity is mediated through IDH1 rather than its mitochondrial counterparts. This profile seems to originate from the specialized prostate metabolic program, as observed using mouse prostate and human patient-derived organoids. Pharmacological and genetic inhibition of IDH1 impaired mitochondrial respiration, suggesting that this cytoplasmic enzyme contributes to the mitochondrial tricarboxylic acid cycle (TCA) in PCa. Mass spectrometry-based metabolomics confirmed this hypothesis, showing that inhibition of IDH1 impairs carbon flux into the TCA cycle. Consequently, inhibition of IDH1 decreased PCa cell proliferation in vitro and in vivo.

Conclusions: These results demonstrate that PCa cells have a hybrid cytoplasmic-mitochondrial TCA cycle that depends on IDH1. This metabolic enzyme represents a metabolic vulnerability of PCa cells and a potential new therapeutic target.

Citation Format: Kevin Gonthier, Cindy Weidmann, Line Berthiaume, Cynthia Jobin, Aurélie Lacouture, Camille Lafront, Mario Harvey, Bertrand Neveu, Jérémy Loehr, Alain Bergeron, Yves Fradet, Louis Lacombe, Julie Riopel, Éva Latulippe, Chantal Atallah, Michael Shum, Jean-Philippe Lambert, Frédéric Pouliot, Martin Pelletier, Étienne Audet-Walsh. Isocitrate dehydrogenase 1 sustains a hybrid cytoplasmic-mitochondrial tricarboxylic acid cycle in 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 3700.
Abstract 3704: The essential role of an alternate mRNA translation initiation in the regulation of breast cancer cell metabolism
Authors: Rosenstock Hanna , Mitaishvili Erna , de la Parra Columba
Published: April 2023

Translational control and metabolic reprogramming are hallmarks of advanced cancers. Important genes involved in cancer progression express mRNAs that are selectively translated, including regulators of cancer cell metabolism.

Cancer cells acquire an altered metabolism, switching from oxidative phosphorylation (OXPHOS) to a glycolytic phenotype (Warburg effect), to increase reliance on alternate metabolic pathways to support growth, proliferation, and metastasis. Triple-negative breast cancer (TNBC), one of the most aggressive and highly metastatic subtypes with the poorest outcome, is characterized by elevated glycolysis and low OXPHOS. Still, the exact mechanism for this metabolic switch is largely unknown. Using a TNBC cell model it has been shown an alternate mechanism of cap-dependent but mTORC1/eIF4Eindependent mRNA translation by the eIF4G homolog, DAP5, which directly binds the cap-binding protein eIF3d. Our research indicates that this alternate translation initiation is essential in regulating several mRNAs involved in breast cancer metabolism. DAVID analysis from genome-wide transcriptomic and translatomic studies, metabolomic assays including Glycolysis Assay [Extracellular Acidification], Oxidative Phosphorylation, and Mitotracker Assay, as well as RT-PCR and western blot analysis was performed from well-characterized triple-negative aggressive breast cancer cell lines,4T1 and MDA-231, Non-silencing (Nsi) control and silencing DAP5 (shDAP5) samples.

Several mRNAs related to glucose metabolism decreased after silencing DAP5 and on the opposite, critical mRNAs associated with OXPHOS increased, independent of their steady-state mRNA abundance. Silencing DAP5 increased Oxidative Phosphorylation activity and decrease the glycolytic rate in TNBC breast cancer cells. Proteins correlated with aberrant glycolytic metabolism including the E-cadherin transcriptional repressors Snail (SNAI1) were reduced after silencing of DAP5. Our data indicates that the alternate mRNA translation initiation mechanism by DAP5 of specific mRNAs correlated to glycolysis and OXPHOS could play an essential role in the mechanism by which TNBC breast cancer cells orchestrate a metabolic switch required for malignant progression and metastasis.

Citation Format: Hanna Rosenstock, Erna Mitaishvili, Columba de la Parra. The essential role of an alternate mRNA translation initiation in the regulation of breast cancer cell 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 3704.
Abstract 375: Smoking induces Wee1 expression through miRNA deregulation, promoting docetaxel resistance in esophageal adenocarcinoma
Authors: Thangaretnam Krishnapriya , Obaidul Islam MD , Lu Heng , Peng Dunfa , Bhat Nadeem Sidiq , Soutto Mohammed , Chen Zheng
Published: April 2023

Esophageal Adenocarcinoma (EAC) is the most common subtype of Esophageal Cancer in Western countries, and its incidence rate has increased over the past few decades. The overall five-year survival rate of EAC is less than 20%. Smoking is a major risk factor for EAC development. WEE1 kinase plays a crucial role in the cell cycle by regulating the G2/M checkpoint, providing a time frame for the cells to repair DNA damage. Thus, targeting WEE1 using an inhibitor could potentiate the effect of chemotherapeutic drugs. This study investigated how smoking induces WEE1 protein expression, promoting docetaxel resistance in EAC.

IHC staining for WEE1 in the normal esophagus and EAC tissue sections revealed significant over-expression of WEE1 protein in EAC. Nicotine, Nicotine derived Nitrosamine Ketone (NNK), and 2% cigarette smoke extract (CSE) treatment induced WEE1 expression with a concomitant increase in CDC2 phosphorylation (p-CDC2, Y15), a well-established read-out of WEE1 activity in EAC cells. In addition, we found that smoking upregulates WEE1 expression by decreasing miR-195-5p expression levels in EAC.

Based on the differential gene expression signatures in EAC cell lines FLO1 and OE33 following WEE1 knockdown, drug and small molecule induced gene expression signatures analysis, L1000 fireworks display (L1000FWD), predicted that docetaxel is one of the best drug candidates which can synergize with the WEE1 inhibitor MK1775. Smoking-induced docetaxel resistance in EAC cells measured by cell viability assay. Down-regulating WEE1 expression with siRNA or pharmacological inhibition using MK1775 significantly sensitized EAC cells to docetaxel treatment, as evidenced by a remarkable decrease in the IC50 value of docetaxel. Inhibition of WEE1 combined with docetaxel can be considered an ideal therapeutic strategy due to its superior anti-tumor efficacy compared to the standard single-agent treatment. In conclusion, our data provide a firm rationale for the clinical combination of docetaxel with MK1775 in EAC patients.

Citation Format: Krishnapriya Thangaretnam, Islam MD Obaidul, Heng Lu, Dunfa Peng, Nadeem Sidiq Bhat, Mohammed Soutto, Zheng Chen. Smoking induces Wee1 expression through miRNA deregulation, promoting docetaxel resistance in esophageal 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 375.
Abstract 3753: miR-567 modulates the progression of melanoma cells and functions of macrophages
Authors: Nguyen Mai-Huong Thi , Lin Chen-Huan , Huang Yu-Chi , Tsai Mu-Shiun , Chen Ming-Hong , Miyashita Azusa , Fukushima Satoshi , Ma Nianhan
Published: April 2023

Purpose: Target therapy and immunotherapy are advancements in the treatment of melanoma; unfortunately, a subset of patients does not benefit or produces drug resistance. Therefore, an alternative strategy should be investigated to further improve clinical benefits. Macrophages are the abundant immune cells related to the progression of melanoma and resistance to drugs. In this study, we explored the novel roles of miR-567 in the progression of melanoma and effects of macrophages.

Methods: miR-567 expression in nevus and melanoma tissues was analyzed by in situ hybridization. miR-567 was transfected into melanoma cells to investigate its biological effects through functional assays such as proliferation, colony formation, soft agar, and migration assays. In addition, miR-567-related signal transduction pathways in the melanoma cells were studied by western blot and next-generation sequencing analyses. Different phenotypes of macrophages polarized from THP-1 monocyte cells were used as in vitro model to investigate the effects of miR-567 on macrophages.

Results: Our results showed that miR-567 expression levels were decreased in melanoma cells compared to melanocyte cells. Consistently, expression of miR-567 was 0.403-fold lower in melanoma tissues (n=118) as compared to nevus tissues (n=40). In addition, the receiver operating characteristic presented that the area under the curve value of miR-567 is 0.9495. Importantly, higher expression of miR-567 enhanced the overall survival of melanoma patients. Overexpression of miR-567 significantly reduced proliferation, survival, anchorage-independent growth, migratory, and invasive abilities of melanoma cells and BRAF-inhibitor resistant cells. Furthermore, our results demonstrated that IGF1R, E2F1, and Cyclin B2 are direct targets of miR-567 and knockdown of these genes attenuated proliferation and survival of melanoma cells. Interestingly, introduction of miR-567 downregulated MAPK/ERK and PI3K/AKT pathways in melanoma cells and regulated multiple pathways related to immune response. Of note, overexpression of miR-567 reduced the promoting melanoma growth induced by M2 or melanoma-associated macrophages, indicating the involvement of miR-567 in regulation of macrophages.

Conclusion: miR-567 could be served not only as biomarkers but also as potential molecular target for prevention of melanoma progression.

Citation Format: Mai-Huong Thi Nguyen, Chen-Huan Lin, Yu-Chi Huang, Mu-Shiun Tsai, Ming-Hong Chen, Azusa Miyashita, Satoshi Fukushima, Nianhan Ma. miR-567 modulates the progression of melanoma cells and functions of 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 3753.
Abstract 3757: miR-200 inhibits the activation of cancer-associated fibroblasts in lung tumor microenvironment
Authors: Cheon Inyoung , Lee Sieun
Published: April 2023

Cancer-associated fibroblasts (CAFs) are key components of the tumor microenvironment and promote cancer progression and metastasis. CAFs are regulated by diverse processes including regulation by miRNAs. To identify miRNAs involved in the regulation of CAF activation, we profiled miRNA expression within mouse lung normal fibroblasts (LFs) and CAFs using the NanoString nCounter method. Among numerous miRNAs differentially expressed between CAFs and LFs, miR-200 was selected for further investigation, which is more highly downregulated in CAFs. Overexpression of miR-200 in CAFs suppressed the migration and invasion of lung cancer cells in co-culture systems and inhibited metastasis in a mouse model. In addition, we verified that lung cancer patients with high miR-200 levels showed a good prognosis than those with low miR-200 levels. Next, we tried to identify novel miR-200 targets based on RNA sequencing data, lung adenocarcinoma TCGA, and TargetScan. NRP2 was confirmed to be a target of miR-200 by 3΄-UTR assay. NRP2 knockdown by siRNAs in CAFs suppressed cancer cell invasion, and NRP2 addback in CAF-200 restored cancer cell invasion inhibited by miR-200. NRP2 functions as a co-receptor with VEGFR, thus, CAF-200 was less responsive to VEGF than control CAFs. Collectively, this study suggests that downregulation of miR-200 in CAFs promotes lung cancer progression in tumor microenvironment; therefore, miR-200 will be a good therapeutic target or a biomarker in lung adenocarcinoma.

Citation Format: Inyoung Cheon, Sieun Lee. miR-200 inhibits the activation of cancer-associated fibroblasts in lung tumor microenvironment. [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 3757.
Abstract 3784: miR-489 induces apoptosis and immunogenic cell death through targeting FOXM1 in triple negative breast cancer cells
Authors: Gupta Gourab , Titus Ryan , Patel Yogin , Sarvanan Shakthika , Chen Hexin
Published: April 2023

It has been well established that microRNAs (miRNAs) have an important role in cancer sustenance and progression. Our previous studies have established the role of miR-489 as a tumor suppressor miRNA in breast cancer. However, the multiple targets of this miRNA have diversified its mechanism from preventing tumor cell proliferation to promoting cell death pathways. In this study we have aimed to establish the role of miR-489 in cell cycle inhibition, leading to Endoplasmic Reticulum stress (ER stress) and ultimately immunogenic cell death (ICD). Firstly, we found that overexpression of miR-489 in triple negative breast cancer (TNBC) cell lines including MDA-MB-231, BT 549, drastically reduced cell proliferation using colony formation assay and real-time cell analysis. Furthermore, studies like GO analysis, sequence analysis and cell cycle analysis demonstrated that miR-489 induces cell cycle arrest and apoptosis in TNBC by directly targeting FOXM1 and regulating other kinases like CDK1. It was also shown for the first time that miR-489 overexpression induces ER stress and the release of damage associated molecular patterns (DAMPs), consistent with hallmarks of ICD like Calreticulin exposure on cellular surface and ATP release, triggering phagocytosis. It was also established that miR-489-induced apoptosis lead to the elevation of cleaved caspase 3, which was responsible for the activation of PANX1 and the consequent release of ATP. In conclusion, we tried to mechanistically understand the role of miR-489 in cell cycle arrest leading to cellular stress. Stress induced apoptosis and consequent ICD was also a possible outcome and it turned out miR-489 overexpression without any other ICD inducer was good enough to trigger the release of DAMPs and elucidate an immunogenic response.

Citation Format: Gourab Gupta, Ryan Titus, Yogin Patel, Shakthika Sarvanan, Hexin Chen. miR-489 induces apoptosis and immunogenic cell death through targeting FOXM1 in triple negative 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 3784.
Abstract 3785: Telomerase RNA component lncRNA regulates hsa-miR-320 family and promotes prostate cancer metastasis
Authors: Mbeje Mandisa , Kandhavelu Jeyalakshmi , Penny Clement , Dlamini Zodwa , Marima Rahaba
Published: April 2023

Telomerase RNA component (TERC) is a long non-coding (lncRNA) associated with prostate cancer progression and has novel prognostic biomarker potential. Differential methylation patterns of lncRNAs affect their expression levels, leading to cancer. This study aimed to profile and elucidate differential lncRNA expression patterns in prostate cancer (PCa). A 384-well plate of PCa associated lncRNA gene array panel was used in castration resistant PC-3 PCa cells vs androgen-sensitive PCa LNCaP cells. Annotation and enrichment analysis of lncRNA differential gene expression was performed using a human lncRNA sets database, LncSEA. Furthermore, digital droplet PCR (ddPCR) was used to verify the PCR array results. Up or down-regulation of ±2 and p <0.05 were considered significant. Decreased hsa-miR-320 gene family expression has been reported in various tumors including PCa. However, the precise mechanisms of hsa-miR-320 gene family in PCa remain to be elucidated. Thirty six of 84 lncRNAs were shown to be upregulated, while 14% of the lncRNA genes were downregulated. Notably, TERC lncRNA was upregulated in high metastatic PC-3 cells. Hypermethylation patterns of this lncRNA in high metastatic PC-3 cells was also shown by enrichment analysis. Furthermore, TERC lncRNA was demonstrated to regulate the hsa-miR-320 family, suggesting a correlation between TERC hypermethylation and downregulation of hsa-miR-320 gene family expression in PCa, as revealed by bioinformatics analysis. TERC lncRNA promotes PCa cell proliferation, migration, invasion and metastasis by sponging hsa-miR-320 family. This TERC/hsa-miR-320 interaction holds potential to understanding PCa prognostic mechanisms by lncRNAs and may also be targeted for novel therapeutics.

Citation Format: Mandisa Mbeje, Jeyalakshmi Kandhavelu, Clement Penny, Zodwa Dlamini, Rahaba Marima. Telomerase RNA component lncRNA regulates hsa-miR-320 family and promotes prostate 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 3785.
Abstract 3826: Potency of cardiac glycosides by structure in organotypic models of pancreatic and ampullary carcinomas
Authors: Hossan Md Shahadat , Lin Ethan , Stram Austin , Warner Jamie , Koeppel Luke , Riedl Ellie , Kratz Jeremy
Published: April 2023

Background: Organotypic models have emerged as an important preclinical model for developmental therapeutics in pancreatic and biliary cancer. Naturally derived cardiac glycoside in 2D monoculture exquisite potency, here we describe their activity in cancer and normal liver with patient-derived cancer organoids (PCOs).

Methods: PCOs of pancreatic and ampullary cancers and normal hepatic tissues were generated. Response was assayed with four structurally different cardiac glycoside (cerberin, neriifolin, digitoxin, digoxin; 72h). Response was characterized by growth inhibition (GI50) using 3D CellTiterGlo (CTG) and subclonal population response by high content imaging using normalized change in diameter (n=1038).

Results: Dose response revealed improved potency for cerberin and neriifolin, versus digoxin and digitoxin across models (see table). In a normal liver organoid model, there was no convergence in dose response observed when extended to 1uM of each cardiac glycoside. Ampullary PCO response was assayed using subclonal population analysis. Cerberin at 10nM achieved growth arrest across the population as assayed by median Δ diameter (growth +4.0±32.0, GΔ=0.94, p<0.005). Neriifolin at 10nM had subclonal with residual growth representing 72% of overall population as assayed by Δ diameter (growth +18.3%, GΔ=0.65). Traditional cardiac glycosides of digoxin and digitoxin at 10nM had significant portions with residual growth (94.9% and 84.7%, respectively) as well as did not achieve meaningful effect (GΔ=-0.10 and GΔ=0.05, respectively).

Conclusions: CGs have varied potency for anticancer activity between different structural motifs. Cerberin induce population growth arrest in cancer models which differed from normal liver. This work requires dedicated mechanistic modeling to explain differences in anti-cancer activity that may confer sensitivity and in considering models of cardiac toxicity.

Potency of Cardiac Glycosides in Organoid Models. GI50 (nM) Ampullary GI50 (nM) Pancreatic GI50 (nM) Normal Liver Cerberin 7.4 7.20 71.0 (NC) Neriifolin 10.2 7.4 62.0 (NC) Digoxin 29.6 47.9 55.5 (NC) Digitoxin NC 40.0 85.5 (NC)

Citation Format: Md Shahadat Hossan, Ethan Lin, Austin Stram, Jamie Warner, Luke Koeppel, Ellie Riedl, Jeremy Kratz. Potency of cardiac glycosides by structure in organotypic models of pancreatic and ampullary carcinomas. [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 3826.
Abstract 3949: CYR61, a member of the CCN protein family, regulates IGF-1 in metastatic prostate cancer
Authors: Ortiz-Hernandez Greisha L. , Walker Mya , Woods-Burnham Leanne , Kittles Rick A. , Casiano Carlos A. , Neuhausen Susan L.
Published: April 2023

The cysteine-rich angiogenic inducer 61 (CYR61) is a member of the cellular communication network (CCN) protein family that can be induced by growth factors. CYR61-specific functions are dependent on cellular context. In prostate cancer (PCa), CYR61 contributes to cell growth and survival through its interactions with extracellular ligands, such as integrins avb3 and a6b4, in the extracellular matrix space. Interestingly, CYR61 contains an insulin-like growth factor-binding protein domain suggesting that it may interact with insulin-like growth factor-I (IGF-1). High serum levels of IGF-1 are directly linked to PCa development and recently have been studied as a predictor of metastatic disease. Given the important roles that IGF-1 and CYR61 play in PCa and their potential interaction, it is critical to investigate their molecular interplay. This study is designated to specifically determine and characterize the molecular interaction between CYR61 and IGF-1 in metastatic PCa and to determine if this interaction contributes to aggressive tumor properties (e.g., proliferation, tumorsphere formation). Using immunoblotting, we demonstrated that CYR61 is upregulated in the metastatic cell line PC3 and downregulated in the docetaxel-resistant cell line PC3-DR. Furthermore, optimized knockdown of CYR61 using siRNA significantly reduced PC3 cell proliferation, viability, and prostasphere formation. To examine the underlying mechanisms associated with IGF-1 signaling, we assessed the activation of either the PI3/Akt and MAPK pathways in PC3 cells with CYR61 silencing. CYR61 siRNA-mediated knockdown decreased activation of the PI3/Akt pathway but did not affect the MAPK pathway. We also initiated studies to determine the co-localization of CYR61 and IGF-1 using immunofluorescence microscopy (IF). Based on the initial IF observations, we will proceed to examine if their interaction is primarily intra- or extra-cellular. For intracellular interactions, we will co-immunoprecipitate CYR61 with IGF-1 and for extracellular interactions, we will use the AVidity-based EXtracellular Interaction Screen (AVEXIS) system. Our goal is to establish the contribution of CYR61-IGF1 protein-protein interaction to PCa cell proliferation. Specifically, the correlation between PCa aggressiveness and circulating levels of CYR61 and IGF-1 is poorly studied. Their combined role may explain a proportion of aggressive PCa.

Citation Format: Greisha L. Ortiz-Hernandez, Mya Walker, Leanne Woods-Burnham, Rick A. Kittles, Carlos A. Casiano, Susan L. Neuhausen. CYR61, a member of the CCN protein family, regulates IGF-1 in metastatic 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 3949.