bluuebunny/crossref_metadata_2025 huggingface dataset:
Figure S13 from ERBB4-Mediated Signaling Is a Mediator of Resistance to PI3K and BTK Inhibitors in B-cell Lymphoid Neoplasms
Authors: Arribas Alberto J. , Napoli Sara , Cascione Luciano , Barnabei Laura , Sartori Giulio , Cannas Eleonora , Gaudio Eugenio , Tarantelli Chiara , Mensah Afua A. , Spriano Filippo , Zucchetto Antonella , Rossi Francesca M. , Rinaldi Andrea , Castro de Moura Manuel , Jovic Sandra , Bordone Pittau Roberta , Stathis Anastasios , Stussi Georg , Gattei Valter , Brown Jennifer R. , Esteller Manel , Zucca Emanuele , Rossi Davide , Bertoni Francesco
Published: March 2024

Immunoblotting for total and p-ERBB4, p-AKT and p-ERK in parental (left) and resistant (right) Karpas1718 cells (A-B), and TMD8 cells (C). All blots are representative of two independent experiments. Karpas1718 or TMD8 cells were stimulated with PBS or HBEGF (30ng/mL) for 4 hours, washed-up and treated with DMSO (0.1%), idelalisib (1µM), lapatinib (1µM) or the combination. Levels of phospho proteins were normalized to vinculin and total protein. Data on the barplots represent the mean of two experiments, errors bars represent standard deviation of the mean. * for p<0.05, ** for p<0.01 and ns for not significant from a t-test.
Figure S1 from Genetic Silencing of AKT Induces Melanoma Cell Death via mTOR Suppression
Authors: Parkman Gennie L. , Turapov Tursun , Kircher David A. , Burnett William J. , Stehn Christopher M. , O'Toole Kayla , Culver Katie M. , Chadwick Ashley T. , Elmer Riley C. , Flaherty Ryan , Stanley Karly A. , Foth Mona , Lum David H. , Judson-Torres Robert L. , Friend John E. , VanBrocklin Matthew W. , McMahon Martin , Holmen Sheri L.
Published: March 2024

Supplementary Figure S1 shows that pharmacological inhibition of AKT has little effect on melanoma cells. Cell confluency assay under pharmacological inhibition. A375, HT144, SK-MEL28, WM793, MTG001, and MTG004 cells were largely resistant to AKT inhibitors MK2206 (2.5 µM) and GDC0068 (1µM) while A375, HT144, WM793, SK-MEL 28 and MTG001 cells were largely sensitive to PI3Kα inhibition by GDC0941 (1µM) (P < 0.001). Error bars indicate standard error of the mean of triplicate wells.
Supplementary Table S3 from Palazestrant (OP-1250), A Complete Estrogen Receptor Antagonist, Inhibits Wild-type and Mutant ER-positive Breast Cancer Models as Monotherapy and in Combination
Authors: Parisian Alison D. , Barratt Susanna A. , Hodges-Gallagher Leslie , Ortega Fabian E. , Peña Guadalupe , Sapugay Judevin , Robello Brandon , Sun Richard , Kulp David , Palanisamy Gopinath S. , Myles David C. , Kushner Peter J. , Harmon Cyrus L.
Published: March 2024

RNASeq dataset of gene pathway changes following treatment with OP-1250, palbociclib and ribociclib in an MCF7 xenograft model.
Supplementary Legends from Palazestrant (OP-1250), A Complete Estrogen Receptor Antagonist, Inhibits Wild-type and Mutant ER-positive Breast Cancer Models as Monotherapy and in Combination
Authors: Parisian Alison D. , Barratt Susanna A. , Hodges-Gallagher Leslie , Ortega Fabian E. , Peña Guadalupe , Sapugay Judevin , Robello Brandon , Sun Richard , Kulp David , Palanisamy Gopinath S. , Myles David C. , Kushner Peter J. , Harmon Cyrus L.
Published: March 2024

Supplementary Legends
Supplementary Figure S7 from Palazestrant (OP-1250), A Complete Estrogen Receptor Antagonist, Inhibits Wild-type and Mutant ER-positive Breast Cancer Models as Monotherapy and in Combination
Authors: Parisian Alison D. , Barratt Susanna A. , Hodges-Gallagher Leslie , Ortega Fabian E. , Peña Guadalupe , Sapugay Judevin , Robello Brandon , Sun Richard , Kulp David , Palanisamy Gopinath S. , Myles David C. , Kushner Peter J. , Harmon Cyrus L.
Published: March 2024

OP-1250 demonstrates potent activity in an ER+ breast cancer intracranial xenograft and outperforms ribociclib. A, Schematic of intracranial xenograft experiment with data shown in B–D. B, Mean and SEM intracranial tumor volume over time with listed treatments as assessed by MRI in an ESR1Y537S ST941 PDX intracranial xenograft model. C, Percent change in intracranial tumor volume of individual animals in listed treatment groups at day 101 or at the last recorded timepoint in the ST941 intracranial xenograft study, normalized to day 1 tumor volume. D, Kaplan–Meier graphs of animal survival over time in listed treatment groups of mice implanted intracranially with the ST941 PDX xenograft. Dosing ended on day 100 of the study in the OP-1250 10 mg/kg group, as indicated by the shaded area. E, Representative intracranial MRIs from ST941-implanted mice with data shown in Figure 6 (see main article) at indicated timepoints and treatment groups. PO=orally.
Supplementary Figure S3 from Palazestrant (OP-1250), A Complete Estrogen Receptor Antagonist, Inhibits Wild-type and Mutant ER-positive Breast Cancer Models as Monotherapy and in Combination
Authors: Parisian Alison D. , Barratt Susanna A. , Hodges-Gallagher Leslie , Ortega Fabian E. , Peña Guadalupe , Sapugay Judevin , Robello Brandon , Sun Richard , Kulp David , Palanisamy Gopinath S. , Myles David C. , Kushner Peter J. , Harmon Cyrus L.
Published: March 2024

OP-1250 acts as a selective ER degrader (SERD) in wild-type and ESR1 mutant cell lines. A, B, Representative blot images from the simple western assay shown in Figure 3 (see main article) after 24-h treatment with 300 nM compound or 1 nM E2 with indicated compound treatments and cell lines. C, D, Representative blot images from simple western assay of MCF7 cells treated for 2 h, 4 h, or 8 h with the listed concentrations of compounds in the presence or absence of MG-132 and simple western analysis of ERα protein levels after treatment with listed compounds. Samples were normalized to untreated control, with mean and SD across at least three independent experiments shown. E, F, ERα degradation in CAMA-1 cells with the indicated homozygous mutant as assessed by simple western assay after 24-h treatment with 300 nM compound or 1 nM E2. Samples were normalized to untreated control, represented by a dotted line, with mean and SD across at least three independent experiments shown. G, H, Representative blot images from simple western data shown in panels E and F with indicated compound treatments and cell lines.
Table S2 from Bifunctional Inhibitor Reveals NEK2 as a Therapeutic Target and Regulator of Oncogenic Pathways in Lymphoma
Authors: McCrury Mason , Swafford Kennith , Shuttleworth Sydnye L. , Mehdi Syed Hassan , Acharya Baku , Saha Debasmita , Naceanceno Kevin , Byrum Stephanie D. , Storey Aaron J. , Xu Ying-Zhi , Doshier Claire , Patel Vijay , Post Ginell R. , De Loose Annick , Rodriguez Analiz , Shultz Leonard D. , Zhan Fenghuang , Yoon Donghoon , Frett Brendan , Kendrick Samantha
Published: March 2024

Cox proportional hazard model of patients with DLBCL adjusted for prognostic factors.
Supplementary Figure S3 from Dual CAR-T Cells Targeting CD19 and CD37 Are Effective in Target Antigen Loss B-cell Tumor Models
Authors: Imai Kanae , Takeuchi Yuki , Terakura Seitaro , Okuno Shingo , Adachi Yoshitaka , Osaki Masahide , Umemura Koji , Hanajiri Ryo , Shimada Kazuyuki , Murata Makoto , Kiyoi Hitoshi
Published: March 2024

Supplemental Figure S3. Analysis of differentiation markers of CAR-T cells. Each CAR-T cells and tEGFR-T cells were stimulated with CD19/CD37 positive Raji cells at a 1:5 ratio every 7 days and stained for CD62L, CD45RA, CCR7, CD27 and CD28. A, B, Summary data of the percentage of CD45RA-/CD62L+ central memory T (TCM) cell fraction (A) and CD45RA-/CD62L- effecter memory T (TEM) cell fraction (B).
C–E, Summary data of the percentage of CCR7-/CD27+/CD28+ early TEM cell fraction (C), CCR7-/CD27+/CD28- late TEM cell fraction (D) and CCR7-/CD27-/CD28- effecter T cell fraction (E). Data represent mean ± SEM from five different donors (two-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001).
Figure S3 from Raludotatug Deruxtecan, a CDH6-Targeting Antibody–Drug Conjugate with a DNA Topoisomerase I Inhibitor DXd, Is Efficacious in Human Ovarian and Kidney Cancer Models
Authors: Suzuki Hirokazu , Nagase Shotaro , Saito Chiemi , Takatsuka Atsuko , Nagata Motoko , Honda Kokichi , Kaneda Yuki , Nishiya Yumi , Honda Tomoyo , Ishizaka Tomomichi , Nakamura Kensuke , Nakada Takashi , Abe Yuki , Agatsuma Toshinori
Published: March 2024

CDH6 expression in cell lines and cytotoxic activity in vitro.
Fig. S6 from Radiation Synergizes with IL2/IL15 Stimulation to Enhance Innate Immune Activation and Antitumor Immunity
Authors: Li Xuefeng , Huntoon Kristin , Wang Yifan , Lee DaeYong , Dong Shiyan , Antony Abin , Walkey Carl , Kim Betty Y.S. , Jiang Wen
Published: March 2024

Fig. S6
Fig. S4 from Radiation Synergizes with IL2/IL15 Stimulation to Enhance Innate Immune Activation and Antitumor Immunity
Authors: Li Xuefeng , Huntoon Kristin , Wang Yifan , Lee DaeYong , Dong Shiyan , Antony Abin , Walkey Carl , Kim Betty Y.S. , Jiang Wen
Published: March 2024

Fig. S4
Figure S7 from Improving Intracellular Delivery of an Antibody–Drug Conjugate Targeting Carcinoembryonic Antigen Increases Efficacy at Clinically Relevant Doses In Vivo
Authors: Nessler Ian , Rubahamya Baron , Kopp Anna , Hofsess Scott , Cardillo Thomas M. , Sathyanarayan Nalini , Donnell Jennifer , Govindan Serengulam V. , Thurber Greg M.
Published: March 2024

Spider plots of tumor growth. Nude mice bearing LS174T xenografts were injected with 4 doses of 10 mg/kg (marked with arrows on day 0, 4, 8, and 12) of either LG, XAb-SN-38, or vehicle. Individual growth curves for all mice are shown.
Figure S10 from Improving Intracellular Delivery of an Antibody–Drug Conjugate Targeting Carcinoembryonic Antigen Increases Efficacy at Clinically Relevant Doses In Vivo
Authors: Nessler Ian , Rubahamya Baron , Kopp Anna , Hofsess Scott , Cardillo Thomas M. , Sathyanarayan Nalini , Donnell Jennifer , Govindan Serengulam V. , Thurber Greg M.
Published: March 2024

Binding of T84.66 on LS174T cells pre-incubated in a saturating antibody solution. While the binding of fluorescent dye labelled T84.66 is blocked on LS174T cells pre-incubated with unlabeled T84.66, pre-incubation with hMN-14 does not block the binding of labelled T84.66.
Fig. S17 from Radiation Synergizes with IL2/IL15 Stimulation to Enhance Innate Immune Activation and Antitumor Immunity
Authors: Li Xuefeng , Huntoon Kristin , Wang Yifan , Lee DaeYong , Dong Shiyan , Antony Abin , Walkey Carl , Kim Betty Y.S. , Jiang Wen
Published: March 2024

Fig. S17
Fig. S12 from Radiation Synergizes with IL2/IL15 Stimulation to Enhance Innate Immune Activation and Antitumor Immunity
Authors: Li Xuefeng , Huntoon Kristin , Wang Yifan , Lee DaeYong , Dong Shiyan , Antony Abin , Walkey Carl , Kim Betty Y.S. , Jiang Wen
Published: March 2024

Fig. S12
Data from PUMA/RIP3 Mediates Chemotherapy Response via Necroptosis and Local Immune Activation in Colorectal Cancer
Authors: Chen Dongshi , Ermine Kaylee , Wang Yi-Jun , Chen Xiaojun , Lu Xinyan , Wang Peng , Beer-Stolz Donna , Yu Jian , Zhang Lin
Published: March 2024

Abstract
Induction of programmed cell death (PCD) is a key cytotoxic effect of anticancer therapies. PCD is not confined to caspase-dependent apoptosis, but includes necroptosis, a regulated form of necrotic cell death controlled by receptor-interacting protein (RIP) kinases 1 and 3, and mixed lineage kinase domain-like (MLKL) pseudokinase. Necroptosis functions as a defense mechanism against oncogenic mutations and pathogens and can be induced by a variety of anticancer agents. However, the functional role and regulatory mechanisms of necroptosis in anticancer therapy are poorly understood. In this study, we found that RIP3-dependent but RIP1-independent necroptosis is engaged by 5-fluorouracil (5-FU) and other widely used antimetabolite drugs, and functions as a major mode of cell death in a subset of colorectal cancer cells that express RIP3. We identified a novel 5-FU–induced necroptosis pathway involving p53-mediated induction of the BH3-only Bcl-2 family protein, p53 upregulated modulator of apoptosis (PUMA), which promotes cytosolic release of mitochondrial DNA and stimulates its sensor z-DNA-binding protein 1 (ZBP1) to activate RIP3. PUMA/RIP3-dependent necroptosis mediates the in vitro and in vivo antitumor effects of 5-FU and promotes a robust antitumor immune response. Our findings provide a rationale for stimulating necroptosis to enhance tumor cell killing and antitumor immune response leading to improved colorectal cancer treatments.
Data from Dual CAR-T Cells Targeting CD19 and CD37 Are Effective in Target Antigen Loss B-cell Tumor Models
Authors: Imai Kanae , Takeuchi Yuki , Terakura Seitaro , Okuno Shingo , Adachi Yoshitaka , Osaki Masahide , Umemura Koji , Hanajiri Ryo , Shimada Kazuyuki , Murata Makoto , Kiyoi Hitoshi
Published: March 2024

Abstract
Chimeric antigen receptor T (CAR-T) cells targeting multiple antigens (Ag), may reduce the risk of immune escape following the loss of the target Ag and further increase the efficacy of treatment. We developed dual-targeting CAR-T cells that target CD19 and CD37 Ags and evaluated their antitumor effects. CD19/CD37 dual CAR-T cells were generated using cotransduction and simultaneous gene transfer of two types of lentiviral vectors transferring CD19CAR or CD37CAR genes, including the intracellular domains of CD28 and CD3ζ signaling domains. These dual CAR-T cells contained three fractions: CD19/CD37 bispecific CAR-T cells, single CD19CAR-T cells, and single CD37CAR-T cells. In the functional evaluation of CAR-T cells in vitro, CD19/CD37 dual CAR-T cells showed adequate proliferation and cytokine production in response to CD19 and CD37 antigen stimulation alone or in combination. Evaluation of intracellular signaling revealed that dual CAR-T cell–mediated signals were comparable with single CAR-T cells in response to CD19- and CD37-positive B-cell tumors. Although the cytotoxicity of CD19/CD37 dual CAR-T cells in both CD19- and CD37-positive B-cell tumors was similar to that of single CD19 and CD37CAR-T cells, against CD19 and CD37 Ag-heterogeneous tumor, dual CAR-T cells demonstrated significantly superior tumor lysis compared with single CAR-T cells. Furthermore, CD19/CD37 dual CAR-T cells effectively suppressed Ag-heterogeneous Raji cells in a xenograft mouse model. Collectively, these results suggest that CD19/CD37 dual CAR-T cells may be effective target-Ag-loss B-cell tumor models in vitro and in vivo, which represents a promising treatment for patients with relapsed/refractory B-cell malignancies.
Abstract B039: Identification of sex-specific molecular features in glioma transcriptome
Authors: Huang Yingbo , Shan Yuting , Zhang Weijie , Printzis Christina , Pesce Lorenzo , Stranger Barbara Elaine , Huang R. Stephanie
Published: March 2024

Glioma is a lethal disease with limited treatment options available. Sex differences in glioma are well-documented in clinical settings, with males having twice the risk of developing the disease and increased mortality compared to female counterparts. To investigate the molecular basis of sex differences in glioma, we analyzed transcriptomic sex differences in a collection of three independent glioma datasets and validated them in additional independent studies. The impact of differentially expressed genes were assessed through Gene Set Enrichment Analysis (GSEA) on approximately 5000 gene sets to interpret their biological relevance. Given the well-recognized heterogeneity of glioma, we also integrated three single-cell RNA (scRNA) sequencing datasets to decipher the sex differences in cellular composition and cell-type specific differential gene expression between male and female tumors. The effects of sex on gene expression are modest and primarily driven by the X chromosome (14 out of 16 differentially expressed genes were on the X chromosome). These genes are associated with escape from X-chromosome inactivation (e.g., KDM5C, KDM6A) and are also sex-differentially expressed (DE) in the healthy human brain according to Genotype-Tissue Expression (GTEx) data. GSEA results suggest increased activity in cell proliferation and neuronal signaling in male GBM samples, whereas females show enrichment in glycoprotein/lipid metabolism. By deconvoluting the cell-type composition, we identified a higher percentage of pericytes in the male glioma tumor microenvironment compared to those in females, which is consistent with recent spatial-omic discoveries in glioma. By integrating scRNA sequencing data, we found that both differentially expressed genes and pathways show cell-type-specific enrichment. For example, the female-biased gene GBGT1 is exclusively expressed in the myeloid lineage, while male-biased neuronal pathways are enriched in cancer stem cells. Integrating with the inference of cell trajectories, we observed that male tumors harbor more unconnected, invasive malignant cells than females, which might explain the poor prognosis in male glioma patients. In summary, our results provide a comprehensive multi-level characterization of sex differences in glioma tumors, and these findings will provide novel insights into glioma disease progression and development of new therapy.

Citation Format: Yingbo Huang, Yuting Shan, Weijie Zhang, Christina Printzis, Lorenzo Pesce, Barbara Elaine Stranger, R. Stephanie Huang. Identification of sex-specific molecular features in glioma transcriptome [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr B039.
Abstract PR-008: Modeling epigenetic lesions that cause gliomas
Authors: Rahme Gilbert J. , Javed Nauman M. , Puorro Kaitlyn L. , Xin Shouhui , Hovestadt Volker , Johnstone Sarah E. , Bernstein Bradley E.
Published: March 2024

Epigenetic lesions that disrupt gene regulatory elements and expression are increasingly recognized as potential drivers of human cancers. However, we currently lack the in vitro and in vivo models required to functionally validate such lesions and their tumorigenic impact. Here we model aberrations that arise in Isocitrate Dehydrogenase (IDH) mutant lower-grade gliomas, which exhibit profound DNA hypermethylation as a direct consequence of mutant IDH. DNA hypermethylation may promote gliomagenesis by silencing tumor suppressor genes or, alternatively, by activating proto-oncogenes through disruption of CCCTC-binding factor (CTCF) insulators. CTCF insulator sites define the three-dimensional shape of the genome by dictating the boundaries of topologically associated domains (TADs). Enhancers and promoters can interact when located in the same TAD but are restricted from interacting across different TADs. In IDH mutant gliomas, CpG sites around CTCF binding sites are frequently methylated, effectively compromising CTCF binding and thus TAD organization, allowing for cross-TAD interactions and aberrant activation of genes. We discovered a CTCF insulator downstream of the PDGFRA proto-oncogene that is recurrently disrupted in IDH mutant gliomas. We demonstrate that disruption of the syntenic insulator in mouse oligodendrocyte progenitor cells (OPCs) allows an OPC-specific enhancer to contact and induce Pdgfra, thereby increasing proliferation. In contrast, insulator disruption did not affect Pdgfra expression in neural progenitor cells (NPCs), which lack the enhancer. We also model a second recurrent epigenetic lesion in IDH mutant gliomas, the methylation-dependent silencing of the CDKN2A tumor suppressor. We show that inactivation of Cdkn2a/p19ARF by de novo promoter methylation or mutation drives OPC proliferation and cooperates with Pdgfra insulator loss. Finally, we use lentiviruses to coordinately inactivate the Pdgfra insulator and Cdkn2a in mouse corpus callosum, resulting in low-grade gliomagenesis in vivo with histological features reminiscent of human IDH mutant gliomas. Our study recapitulates recurrent epigenetic lesions in mouse models and demonstrates that the combination of Pdgfra activation and Cdkn2a silencing can transform OPCs in vitro and drive gliomagenesis in vivo.

Citation Format: Gilbert J. Rahme, Nauman M. Javed, Kaitlyn L. Puorro, Shouhui Xin, Volker Hovestadt, Sarah E. Johnstone, Bradley E. Bernstein. Modeling epigenetic lesions that cause gliomas [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr PR-008.
Abstract A021: The beta-arrestin1/endothelin axis bolsters a CAF-dependent invadosome activity and ovarian cancer metastatic potential
Authors: Rio Danila Del , Masi Ilenia , Ottavi Flavia , Salvati Erica , Caprara Valentina , Bagnato Anna , Giannitelli Sara , Spadaro Francesca , Rosanò Laura
Published: March 2024

The recruitment of fibroblasts to tumors and their activation to cancer-associated fibroblasts (CAFs) is a strategy used by tumor cells to guide extracellular matrix (ECM) remodeling, invasion, and metastasis, highlighting the need to dissect molecular mechanisms involved in CAF function. Endothelin-1 (ET-1) regulates cancer/stroma crosstalk, supporting serous ovarian cancer (SOC) progression. By binding its receptors ETA and ETB, ET-1 allows the recruitment of β-arrestin1 (β-arr1) and the generation of signaling complexes regulating tumor progression. However, how ET-1 receptors might “educate” ovarian fibroblasts (HOFs) to supply altered ECM needs and bolster metastasis is to be investigated. This study identifies ET-1 as a critical factor in activating CAFs capable of proteolytic ECM remodeling by invadosome and creating heterotypic spheroids with cancer cells with favorable metastatic abilities. An autocrine/paracrine ET-1/ETA/BR/β-arr1 loop promotes HOF proliferation, an increased CAF marker expression, secretion of proinflammatory cytokines, collagen contraction ability, and motility. Moreover, ET-1 allows ECM remodeling by supporting the lytic activity of invadosome and activation of integrin beta1. Of note, ET-1 signaling favors the formation of heterotypic HOF/SOC spheroids with enhanced ability to survive, migrate through mesothelial monolayer mesothelial, and invade, representing metastatic units. Blockade of ETA/BR or β-arr1 silencing prevents CAF activation, invadosome function, mesothelial clearance, and invasive ability of heterotypic spheroids. In vivo, therapeutical inhibition of ETA/BR with bosentan markedly reduced the metastatic potential of HOF/SOC spheroids. Our findings support a model in which ET-1/β-arr1 amplifies tumor/stroma interaction via CAF activation and fuels HG-SOC metastatic properties, which might be hampered by ETA/BR antagonists.

Citation Format: Danila Del Rio, Ilenia Masi, Flavia Ottavi, Erica Salvati, Valentina Caprara, Anna Bagnato, Sara Giannitelli, Francesca Spadaro, Laura Rosanò. The beta-arrestin1/endothelin axis bolsters a CAF-dependent invadosome activity and ovarian cancer metastatic potential [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr A021.
Abstract A032: The role of ASIC2 and calcium influx in epithelial ovarian cancer pathogenesis
Authors: Joshi Tanvi , Wilhite Annelise , Chokshi Sagar , Singleton Mary Howard , Scalici Jennifer , Lee Kevin
Published: March 2024

Objective: Inflammation associated with incessant ovulation plays a key role in epithelial ovarian cancer (EOC) pathogenesis. Ion channels, such as acid-sensing ion channel-2 or ASIC2 is known to be upregulated in inflammatory conditions and may play a role in calcium ion dysregulation. Here, we describe a potential relationship between ASIC2 and intracellular calcium levels in EOC development and progression. Methods: EOC cell lines OVCAR-8, ES-2 and SKOV-3 were grown to sub-confluence prior to experimentation. Global extracellular pH was changed with the use of hydrochloric acid (HCl) over various time points at hours (h): 2 h, 4 h, 8 h, 12 h and 24 h. ASIC2 expression was subsequently measured via western blot. Intracellular pH was measured with the use of pH-sensitive pHrodo red dye. Cytosolic pH was determined across all cell lines at baseline and after treatment with a known ASIC2 inhibitor: Diminazene (10 μM) over 30 minutes. Intracellular calcium levels were measured with a Fluo Calcium Indicator at baseline and with similar ASIC2 inhibition. Live cell imaging was utilized on a Nikon A1r confocal microscope. Results: Changing global, extracellular pH to 6.0 and 6.5 across various time points from 2-24 h did not affect ASIC2 protein expression via western blot analysis. We then utilized a pH-sensitive assay to determine whether intracellular pH changes were associated with ASIC2 expression. Interestingly, intracellular pH was noted to increase across most cell lines with ASIC2 inhibition via Diminazene treatment. There was a statistically significant increase in pH in ES2 and OVCAR8 cell lines. Because ASIC2 is known to result in influx of various ions, assessment of intracellular calcium levels was performed first after 30 minutes of Diminazene treatment. There was a corresponding significant increase in intracellular calcium levels in ES2 cells after ASIC2 inhibition. And there was a small trend, albeit not significant, toward a similar increase across SKOV-3 and OVCAR-8 cells. Conclusion: There seems to be a role for ASIC2 in EOC, particularly in the context of inflammation as noted by changes in intracellular pH levels. It is likely that ASIC2 may mediate changes in intracellular calcium signaling and promote tumorigenesis in ovarian cancer.

Citation Format: Tanvi Joshi, Annelise Wilhite, Sagar Chokshi, Mary Howard Singleton, Jennifer Scalici, Kevin Lee. The role of ASIC2 and calcium influx in epithelial ovarian cancer pathogenesis [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr A032.
Abstract A063: What can we learn from a retrospective evaluation of HER2, P53 and P16INK4A protein expression in rare ovarian tumors?
Authors: Advikolanu-Rao Kavitha M. , Magliocco Anthony M. , Maksymiuk Andrew W.
Published: March 2024

In this retrospective study, we examined the incidence and prevalence of HER2, P53, and P16INK4A protein accumulation patterns in patients diagnosed with rare ovarian cancer. The population-based study included a total of thirteen patients diagnosed at the Saskatoon Cancer Centre during the years 1983-1995. Immunohistochemistry for evaluation of HER2, P53 and P16INK4A protein expression and ploidy status in paraffin-embedded tissue (PET) specimens were correlated with clinicopathologic variables. Immunoexpression studies were with the following rabbit monoclonal antibodies: Erb-B2, 485 and P16INK4A, G175-405, 13251A, except for P53, mouse monoclonal antibody, D07. DNA status was assessed by flow-cytometric analysis. The most prevalent histologic subtype was sex-cord stromal, followed by three germ-cell, while the remaining two tumors were malignant Brenner. The patients’ ages ranged between (15-77 years). The data on stage was unavailable in five patients, two patients had undifferentiated tumor, one had grade 2-3, and data on grade was unavailable in the remaining six cases. Two of the 13 patients died: one patient in stage IV, grade 3, granulosa-theca, P53 immunonegative and near-tetraploid DNA status; the other patient with malignant Brenner histologic subtype, P53 overexpression, and diploid DNA status. Tumor tissues were available in nine of 13 randomly selected patients. Except for one case with P53 overexpression, the remaining 89% tumor specimens had low to zero P53, P16INK4A, and Erb-B2 expression. The nonsignificance of the immunoexpression studies and preclusion from prognostic prediction is likely due to the restricted sample size. Thus, immunoexpression of P53, P16INK4A, and Erb-B2 have not been associated with rare ovarian tumors. Moreover, the observation that malignant Brenner had P53 overexpression and diploid DNA status in one of the patients is a significant finding of this retrospective study. Additionally, these findings argue for routine use of P53 protein expression and ploidy status in the assessment of malignant Brenner tumors. The vast majority of tumors with P53 mutations progress rapidly. Therefore, the subset of patients with malignant Brenner tumors that have P53 protein disruption will likely benefit from therapy using PARP inhibitors. A. Geissel and J. L. Griffin, “Preparation of nuclei for flow cytometry,” in AFIP Advances in Laboratory methods in Histology and Pathology, ed. Mikel UV (Washington DC: American Registry of Pathology, 1944), 111-121.

Citation Format: Kavitha M. Advikolanu-Rao, Anthony M. Magliocco, Andrew W. Maksymiuk. What can we learn from a retrospective evaluation of HER2, P53 and P16INK4A protein expression in rare ovarian tumors? [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr A063.
Abstract A112: Mechanisms underlying platinum-induced ovarian cancer stem cell plasticity
Authors: Metcalfe Tara X. , Zhang Shu , Hassel Christiane , O'Hagan Heather M. , Nephew Kenneth P.
Published: March 2024

Most women diagnosed with late-stage high grade serous ovarian cancer (HGSOC) develop recurrent, platinum-resistant tumors. Ovarian cancer stem cells (OCSCs) are hypothesized to contribute to the emergence of these resistant tumors. CSCs have been postulated to reside in a plastic state, which can allow for the conversion of non-CSC to CSC. This process of dedifferentiation continues during tumor development and chemotherapeutic agents like platinum can exaggerate CSC plasticity. We have previously demonstrated that acute platinum treatment enriched for OCSCs. However, whether platinum transforms non-OCSCs into OCSCs to contribute to this subpopulation of cells remains unclear, and the underlying mechanism remains incompletely understood. To examine OCSC plasticity, aldehyde dehydrogenase (ALDH; functional marker) and fluorescence activated cell sorting were used to isolate OCSCs (ALDH+) and non-OCSCs (ALDH-) from HGSOC cell lines, OVCAR5 and OVCAR3. To determine the stability of the non-OCSC phenotype, ALDH- cells were cultured for 3 and 5 days and ALDH activity was measured using flow cytometry. At both timepoints examined, ALDH- cells remained ALDH-, with approximately less than 1% being ALDH+ (p<0.05). To determine if platinum can induce conversion of non-OCSC to OCSC, ALDH- cells were treated with cisplatin (12µM or 15 µM for 16h), and the percent of ALDH+ cells was measured using flow cytometry. Treatment of ALDH- cells with cisplatin resulted in conversion of approximately 4% and 10% of ALDH- cells into ALDH+ cells (p<0.05) in OVCAR5 and OVCAR3 cells, respectively. Furthermore, both cell lines displayed increased expression (p<0.05) of stemness genes BMI1, NANOG, OCT4, and SOX2 and spheroid formation was observed in converted OCSCs compared to parental ALDH+ and whole cell populations treated with cisplatin, suggesting that platinum induced the observed differences in the stemness phenotype. With the goal of targeting key genes and pathways to inhibit platinum-induced OCSC conversion, RNA-sequencing was performed using converted OCSCs. Activated pathways in converted OCSCs were identified, including protein kinase signaling, inhibition of matrix metalloproteases, SPINK1 pathway, and VDR/RXR activation. We are further investigating if these pathways contribute to the mechanism involved in OCSC plasticity and whether targeting these pathways can reduce the OCSC population, which is critical to blocking the persistence of OCSCs and ultimately reduce mortality in patients.

Citation Format: Tara X. Metcalfe, Shu Zhang, Christiane Hassel, Heather M. O'Hagan, Kenneth P. Nephew. Mechanisms underlying platinum-induced ovarian cancer stem cell plasticity [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr A112.
Abstract B024: Maintenance therapy inhibition of ptk2 yields decreased disease in preclinical HRP/HRD models of recurrent HGSOC
Authors: Hill Breana , Bondre Ioana , Soylu H Meryem , Antoine Juliette , Chen Xiao Lei , Tahon Elise , Zhang Yichi , Olajill Marjaana , Kinkel Sarah , Cock Terrie-Anne , Binder Pratibha , Burns Chris , Schlaepfer David , McHale Michael McHale T. , Stupack Dwayne G
Published: March 2024

High grade serous ovarian cancer remains the most lethal of the gynecologic malignancies, in part, because of the high incidence of recalcitrant and rapidly recurring disease. Disease recurrence is now better controlled due to maintenance therapy with PARP inhibitors, particularly in homologous repair deficient tumors. Efficacy in homologous repair proficient tumors represents an unmet need. We previously demonstrated that inhibitors of protein tyrosine kinase 2 (ptk2), a gene that is frequently gained in HGSOC, compromises the expression of tumor repair genes, decreases stemness, and limits immune evasion. The low toxicity of ptk2 inhibitors appears to be well suited for a maintenance role. Here, we tested the effect of six weeks of ptk2 inhibitor maintenance treatment, post-chemotherapy, in two preclinical models. In a homologous repair proficient model (KMF), and in a PARP inhibitor-resistant model (HGS2), ptk2 inhibitor treatment yielded lower tumor burden, fewer solid tumors, and decreased bloody ascites accumulation relative to treatment with Niraparib. The low toxicity and overall efficacy provide strong support for future clinical trials with ptk2 inhibitors in the maintenance role, for patients with HGSOC.

Citation Format: Breana Hill, Ioana Bondre, H Meryem Soylu, Juliette Antoine, Xiao Lei Chen, Elise Tahon, Yichi Zhang, Marjaana Olajill, Sarah Kinkel, Terrie-Anne Cock, Pratibha Binder, Chris Burns, David Schlaepfer, Michael McHale T. McHale, Dwayne G Stupack. Maintenance therapy inhibition of ptk2 yields decreased disease in preclinical HRP/HRD models of recurrent HGSOC [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr B024.
Abstract B048: Evaluating the impact of pulmonary diseases on the use of microRNAs as biomarkers for early detection of ovarian cancer
Authors: Moore Monica , Wollborn Laura , Mishra Sudhanshu , Webber James , Elias Kevin M.
Published: March 2024

Background: Efforts are ongoing to develop new biomarkers to diagnose ovarian cancer at earlier, more curable stages. microRNAs (miRNAs) have emerged as a promising new class of circulating biomarkers for ovarian cancer diagnosis and prognosis. As other biomarkers, such as CA-125, are non-specifically elevated in pulmonary diseases, evaluating how miRNA expression is affected by various lung conditions will inform the interpretation of miRNAs as a potential clinical test. Asthma and bronchitis are of particular interest because miRNA expression in the lung is markedly affected by environmental respiratory irritants such as cigarette smoke and air pollution. Moreover, as the burden of asthma and bronchitis in the United States falls disproportionately on minority communities, so a test confounded by asthma might be less generalizable to a diverse patient population. Methods: Serum samples from 1,614 women enrolled in the Mass General Brigham Biobank were analyzed for miRNAs using a focused panel of 179 miRNA Fireplex® probes optimized for serum profiling. In addition to the miRNA data, electronic health records were reviewed to pair the biomarker data with demographics such as age, race, ethnicity, zip code, reproductive health histories, and comorbidities, including pulmonary diseases like asthma/bronchitis. miRNAs associated with pulmonary diseases were identified by univariate analysis, adjusting for multiple comparisons using Bonferroni correction. Results: The analysis compared the circulating miRNA profiles of 446 asthma/bronchitis subjects vs. 1168 non-asthma/non-bronchitis subjects. Among the miRNAs examined, there was only one miRNA, miR-133a-3p, that showed a significant difference between the two groups. We further focused on 8 specific miRNAs that have been proposed as diagnostic biomarkers for ovarian cancer and found no significant difference in expression between asthma/bronchitis subjects vs. non-asthma/non-bronchitis subjects.Conclusion: Asthma and bronchitis do not significantly confound measurements of miRNAs that are potential biomarkers for ovarian cancer. This further supports miRNAs as biomarkers for early detection of ovarian cancer and suggests that miRNAs are likely to generalize well if deployed as part of a screening test among the general population.

Citation Format: Monica Moore, Laura Wollborn, Sudhanshu Mishra, James Webber, Kevin M. Elias. Evaluating the impact of pulmonary diseases on the use of microRNAs as biomarkers for early detection of ovarian cancer [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr B048.
Abstract IA003: Advancing novel therapeutics through reverse translation
Authors: Adams Sarah F.
Published: March 2024

Background: The established paradigm for cancer drug development was designed to advance cytotoxic agents targeting tumor cells. An increased appreciation of the role of the tumor microenvironment in treatment efficacy and disease outcomes has identified tumor-extrinsic therapeutic targets. The different mechanisms of action employed by anti-angiogenic drugs or immunotherapeutics highlight the need for updated study endpoints to identify the maximally effective dose for clinical applications. These advances, and the translation of novel therapeutics, can be accelerated through reverse translation, a term used to describe research using patient samples to inform mechanistic studies – going from bedside-to-bench. Objective: To present an example of reverse translation in the development of an immune therapy combination for ovarian cancer. Preclinical studies demonstrated therapeutic synergy of CTLA4 immune checkpoint blockade and poly(ADP ribose) polymerase (PARP) inhibition in high grade serous ovarian cancer models. These results were translated to two clinical trials for the treatment of recurrent ovarian cancer. Tumor samples from study subjects have been collected to support ongoing mechanistic investigations and exploratory biomarker studies. Both targeted and unbiased approaches identified candidate biomarkers of response to treatment for further development. Ongoing mechanistic studies are examining the significance of top prognostic markers. Conclusions: Strategies are needed to support the integration of translational studies with clinical research to inform next generation trials and to optimize clinical applications of novel therapeutics.

Citation Format: Sarah F. Adams. Advancing novel therapeutics through reverse translation [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr IA003.
Abstract PR-011: Role of cancer associated fibroblasts in ovarian cancer relapse
Authors: Fang Yiming , Ghosh Argha , Wang Ji , Dasari Subramanyam , Zamarin Dimitriy , Mitra Anirban Kumar
Published: March 2024

Frequent relapse and development of chemoresistance leads to poor prognosis in ovarian cancer (OC). Cancer stem cells (CSCs) consist of a small subpopulation in the tumor that are typically resistant to cytotoxic chemotherapy and are a potential cause of relapse and chemoresistance. Using patient specimens as well as in vitro models, we determined that CSCs are enriched in a niche adjacent to cancer associated fibroblasts (CAFs) after chemotherapy. Cancer associated fibroblasts (CAFs) are a major constituent of the ovarian cancer tumor microenvironment and are highly enriched in the residual tumors following chemotherapy. Therefore, we studied the role of CAFs in promoting ovarian cancer chemoresistance and disease relapse by providing an optimal microenvironment for CSCs. CAFs isolated from ovarian cancer patient tumors were used in heterotypic 2D or 3D coculture systems with high-grade serous ovarian cancer cell lines or with patient-derived ovarian cancer cells to study their effect on CSCs and chemoresistance. Matched pre-and post-chemotherapy patient tumors were used to confirm our findings. CAFs significantly increased resistance to carboplatin and enriched CSCs by increasing their symmetric division as well as dedifferentiation of bulk ovarian cancer cells. Pre-coculture with CAFs increased in vivo tumor initiation capacity of the ovarian cancer cells by 10-fold. The CSC-CAF crosstalk responsible for CSC induction was found to be mediated by Wnt5a signaling. CRISPR knockdown of Wnt5a in CAFs or treatment with a specific Wnt5a inhibitor abrogated the induction of CSCs by CAFs. Only cancer cells with ROR2 Wnt coreceptor respond to Wnt5a signaling triggered by CAFs and developed into CSCs. Responders were found to signal through a non-canonical Wnt pathway involving the coreceptor ROR2, protein kinase C (PKC), and CAMP Responsive Element Binding Protein 1 (CREB1). Inhibition of each of them prevented CSC induction and functional rescue experiments confirmed the sequence of the Wnt5a-ROR2-PKC-CREB1 axis. Treatment of mouse xenografts was established by co-injection of CAFs and ovarian cancer cells, with the Wnt5a inhibitor sensitized them to carboplatin, and eliminated the CSCs in the residual tumors. Our results indicate that CAF-derived Wnt5a is instrumental in ovarian cancer CSC growth and maintenance. In the long term, our studies will broaden the understanding of the mechanism of CSC maintenance by the tumor microenvironment and contribute towards the development of novel therapeutic approaches to prevent ovarian cancer chemoresistance and relapse.

Citation Format: Yiming Fang, Argha Ghosh, Ji Wang, Subramanyam Dasari, Dimitriy Zamarin, Anirban Kumar Mitra. Role of cancer associated fibroblasts in ovarian cancer relapse [abstract]. In: Proceedings of the AACR Special Conference on Ovarian Cancer; 2023 Oct 5-7; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_2):Abstract nr PR-011.
Figure S6 from Neutralizing Antibodies Impair the Oncolytic Efficacy of Reovirus but Permit Effective Combination with T cell–Based Immunotherapies
Authors: Groeneveldt Christianne , Kinderman Priscilla , Griffioen Lisa , Rensing Olivia , Labrie Camilla , van den Wollenberg Diana J.M. , Hoeben Rob C. , Coffey Matt , Loghmani Houra , Verdegaal Els M.E. , Welters Marij J.P. , van der Burg Sjoerd H. , van Hall Thorbald , van Montfoort Nadine
Published: March 2024

Figure S6. B-cell depletion during preexposure does not completely abrogate the presence of Reo-specific neutralizing antibodies.
Figure S5 from Neutralizing Antibodies Impair the Oncolytic Efficacy of Reovirus but Permit Effective Combination with T cell–Based Immunotherapies
Authors: Groeneveldt Christianne , Kinderman Priscilla , Griffioen Lisa , Rensing Olivia , Labrie Camilla , van den Wollenberg Diana J.M. , Hoeben Rob C. , Coffey Matt , Loghmani Houra , Verdegaal Els M.E. , Welters Marij J.P. , van der Burg Sjoerd H. , van Hall Thorbald , van Montfoort Nadine
Published: March 2024

Figure S5. Depletion efficiency during Reo preexposure.
Supplementary Figure 9 from An NFAT1-C3a-C3aR Positive Feedback Loop in Tumor-Associated Macrophages Promotes a Glioma Stem Cell Malignant Phenotype
Authors: Zhang Yaochuan , Song Yifu , Wang Xiaoliang , Shi Mengwu , Lin Yibin , Tao Dongxia , Han Sheng
Published: March 2024

M2-like glioma-associated macrophages promoted MES transition of GSCs and angiogenesis.
Table S2 from Intragenic Rearrangement Burden Associates with Immune Cell Infiltration and Response to Immune Checkpoint Blockade in Cancer
Authors: Zhang Han , Lee Sanghoon , Muthakana Renee R. , Lu Binfeng , Boone David N. , Lee Daniel , Wang Xiao-Song
Published: March 2024

Table S2. Gene signatures used in the study
Figure S5 from Intragenic Rearrangement Burden Associates with Immune Cell Infiltration and Response to Immune Checkpoint Blockade in Cancer
Authors: Zhang Han , Lee Sanghoon , Muthakana Renee R. , Lu Binfeng , Boone David N. , Lee Daniel , Wang Xiao-Song
Published: March 2024

Figure S5. IGR burden correlates with tumor-infiltrating lymphocytes and pro-inflammatory pathways in WGS560 TNBC samples. (a) The distributions of IGR burden in TNBC subtypes. Wilcoxon rank sum tests were conducted to test if the median of IGR burdens in a certain subtype is significantly different than the other subtypes. `*` p<0.05. (b) Scatter plot revealing the association between HRD score (X-axis) and inflame signature (Y-axis), colored according to IGR burden. (c) Boxplots showing the distribution of inflaming signature, total mitoses and HRD score in IGRhigh and IGRlow tumors (samples outside the 10%-90% range are considered outliers). (d) Scatter plot showing the correlation between SCNA and HRD score (Pearson R=0.263, p=0.0008), and between SCNA and T-inflamed signature (Pearson R=-0.037, p=0.76) (e) The p-value for each marker in the multivariate model containing all markers including HRD and different types of genetic markers against T inflamed signature in TNBC tumors, when the confounding effects from other variables are removed. (f) Heatmap displays the normalized expression level of genes important in tumor immunology in TNBC samples. Samples are annotated by mitotic score, tumor grade, TIL and IGR level. (g) Enrichment plots for the pathways of interest as revealed by GSEA.
Figure S3 from Intragenic Rearrangement Burden Associates with Immune Cell Infiltration and Response to Immune Checkpoint Blockade in Cancer
Authors: Zhang Han , Lee Sanghoon , Muthakana Renee R. , Lu Binfeng , Boone David N. , Lee Daniel , Wang Xiao-Song
Published: March 2024

Figure S3. Correlations between IGR burden, TMB, SCNA and Fusions in the ICGC Pan-cancer dataset. Scatter plots of ICGC tumors: (a) SCNA burden vs TMB, (b) SCNA burden vs IGR burden, (c) Fusion burden vs IGR burden, (d) Fusion burden vs SCNA burden, colored by T-inflamed signature are shown. Pearson’s correlation coefficients in a-d are 0.023, 0.22, 0.298 and 0.852 respectively.
Figure S5 from Immune-Related Colitis Is Associated with Fecal Microbial Dysbiosis and Can Be Mitigated by Fecal Microbiota Transplantation
Authors: Elkrief Arielle , Waters Nicholas R. , Smith Natalie , Dai Angel , Slingerland John , Aleynick Nathan , Febles Binita , Gogia Pooja , Socci Nicholas D. , Lumish Melissa , Giardina Paul A. , Chaft Jamie E. , Eng Juliana , Motzer Robert J. , Mendelsohn Robin B. , Markey Kate A. , Zhuang Mingqiang , Li Yanyun , Yang Zhifan , Hollmann Travis J. , Rudin Charles M. , van den Brink Marcel R.M. , Shia Jinru , DeWolf Susan , Schoenfeld Adam J. , Hellmann Matthew D. , Babady N. Esther , Faleck David M. , Peled Jonathan U.
Published: March 2024

Figure S5
Figure S2 from Immune-Related Colitis Is Associated with Fecal Microbial Dysbiosis and Can Be Mitigated by Fecal Microbiota Transplantation
Authors: Elkrief Arielle , Waters Nicholas R. , Smith Natalie , Dai Angel , Slingerland John , Aleynick Nathan , Febles Binita , Gogia Pooja , Socci Nicholas D. , Lumish Melissa , Giardina Paul A. , Chaft Jamie E. , Eng Juliana , Motzer Robert J. , Mendelsohn Robin B. , Markey Kate A. , Zhuang Mingqiang , Li Yanyun , Yang Zhifan , Hollmann Travis J. , Rudin Charles M. , van den Brink Marcel R.M. , Shia Jinru , DeWolf Susan , Schoenfeld Adam J. , Hellmann Matthew D. , Babady N. Esther , Faleck David M. , Peled Jonathan U.
Published: March 2024

Figure S2