bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5961: Inhibition of AURKA destabilizes glioblastoma primary cilia and sensitizes cells to tumor treating fields (TTFields) in vitro and ex vivo
Authors: Tian Jia , Mallinger Julianne , Shi Ping , Ling Dahao , Deleyrolle Loic , Lin Min , Khoshbouei Habibeh , Sarkisian Matthew 
Published: April 2023

Abstract
Tumor Treating Fields (TTFields) can prolong survival of patients with glioblastoma (GBM), but GBM cells still escape this therapy requiring an urgent need for new or adjunct treatments. Recently we reported that TTFields disrupt primary cilia on GBM cells which lowers resistance to standard of care temozolomide chemotherapy. Here we asked if concomitant or sequential treatment of TTFields with other agents that interfere with GBM ciliogenesis enhance TTFields efficacy. Aurora Kinase A (AURKA) plays both a role in promoting cilia disassembly and regulating GBM growth in xenograft models. We found that within 24hr of treatment with the AURKA inhibitor, Alisertib, there was surprising and significant decrease in GBM cilia frequency across multiple GBM patient derived lines. However, similar Alisertib treatment did not affect neuronal or glial cilia frequencies in mixed primary cell cultures from mouse forebrain. Notably, overnight treatment of patient GBM biopsy ex vivo with Alisertib also resulted in reduced cilia frequency within the tumor tissue. Next, we co- or sequentially treated several different GBM patient cell lines with 24hr of 200kHz TTFields together with or followed by 250nM or 1µM Alisertib and examined the subsequent expansion of cells in vitro. We observed a significant synergistic reduction in cell proliferation compared to either treatment alone. However, this synergistic effect does not appear to be cilia-dependent as dual treatment reduction in proliferation was still observed in cell lines lacking different key cilia genes or in GBM cells that naturally lack cilia. Considering Alisertib crosses the blood brain barrier and inhibits intracranial growth of tumors, our data warrant investigation of whether concomitant or sequential treatment of TTFields and Alisertib in vivo can further prolong survival.
Citation Format: Jia Tian, Julianne Mallinger, Ping Shi, Dahao Ling, Loic Deleyrolle, Min Lin, Habibeh Khoshbouei, Matthew Sarkisian. Inhibition of AURKA destabilizes glioblastoma primary cilia and sensitizes cells to tumor treating fields (TTFields) in vitro and ex vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5961.
Abstract 6095: A DNA damaging agent affects BRCA2 exon 3-skipping
Authors: Werner Jordan R. , Siddiqui Hiba , Castillo Ethan , Ansari Mohammed , Fackenthal James D. 
Published: April 2023

Abstract
Introduction: BRCA2 is a tumor suppressor that participates in DNA repair. Inherited pathogenic mutations in BRCA2 are associated with a 38%-84% risk for breast cancer and a 16.5%-27% risk for ovarian cancer. BRCA2 exon 3, which encodes a binding site for the interacting proteins EMSY and PALB2, is frequently skipped during alternative splicing events, often due to spliceogenic mutations, but also for as yet uncharacterized environmental conditions. A genomic deletion that results in complete allelic loss of exon 3 has been shown to be pathogenic, yet sequence variants that result in increased frequency of exon 3-skipping have been shown to be non-pathogenic. Thus, the allele-specific ability of BRCA2 to contribute to DNA repair is affected by the level of exon 3-skipping. In this study we explore the effects of the topoisomerase inhibitor doxorubicin on the frequency of BRCA2 exon 3-skipping. Isoform-specific end-point RT-PCR shows that doxorubicin increases the frequency of BRCA2 exon 3-skipping in the breast cancer cell line MCF7 at concentrations below those required for promoting apoptosis.
Methods: MCF7 cells were treated with doxorubicin according to standard protocols. IC50 levels of doxorubicin were determined using a crystal violet assay. Flow cytometry was performed using standard protocols. RNA was prepared using Qiagen reagents and amplified with Invitrogen One-Step RT-PCR reagents. Isoform-specific RT-PCR primers were designed to amplify the full-length region surrounding BRCA2 exon 3 independently from the region left by an exon-skipping event (∆3). RT-PCR products were visualized using agarose gel electrophoresis.
Results and Conclusion: Results suggest that doxorubicin increases the level of BRCA2 exon 3-skipping in MCF7 at concentrations below those required for promoting apoptosis. This suggests doxorubicin may compromise the DNA repair functions of BRCA2 and other alternatively spliced mRNAs, which may contribute to the therapeutic benefits and/or toxicity of the drug.
Citation Format: Jordan R. Werner, Hiba Siddiqui, Ethan Castillo, Mohammed Ansari, James D. Fackenthal. A DNA damaging agent affects BRCA2 exon 3-skipping [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6095.
Abstract 671: Exploiting macrophage differentiation and plasticity as an immunomodulating strategy
Authors: Castro Carla , Metzger Philipp , Obodozie Cynthia , Weber Holger 
Published: April 2023

Abstract
The infiltration of macrophages into the tumor microenvironment (TME) is a well-known characteristic of tumor progression and therapy resistance. Depending on the signals derived from TME, macrophage plasticity allows them to undergo a variety of polarization states. Two extremes of macrophage polarization are M1 and M2 phenotypes. M1 macrophages provide enhanced anti-tumor inflammatory reaction while M2 macrophages are usually associated with tumor progression due to their immune suppression, angiogenesis and neovascularization promoting capabilities. Modulating the macrophage differentiation or polarization has emerged as a therapeutic strategy to synergize or complement current treatments.
At Reaction Biology we have established human macrophage assay pipelines to evaluate the novel macrophage polarization compounds in vitro. Both, monocyte cell lines like THP-1, and and freshly isolated monocytes from donor derived PBMCs, are of great value. Donor derived differences could become an issue during the screening phase compared to the use of cell lines. Yet, after the screening phase, the evaluation across several healthy donors is crucial to assess the impact of a compound on the general population. Monocytes from primary human peripheral blood mononuclear cell (PBMC) are isolated, differentiated into naïve macrophages, and polarized. Effects of compounds modulating the differentiation and polarization of macrophages are analyzed by flow cytometry. Moreover, the functional reprogramming of monocytes and macrophages can be analyzed by measuring the phagocytic or tumor cell killing capacity via flow cytometry or using high content screening equipment. In the addition, a wide range of secreted cytokine can be quantified by MSD ELISA.
Our data shows that human primary monocytes grown under M1 polarizing conditions express high levels of CD86 and HLA-DR in contrast to those cultivated under M2 polarizing conditions, with high expression of CD163 and CD206. A pH-sensitive phagocytosis could be shown for M2 macrophages. Modulation of surface markers from polarized macrophages under different treatment conditions could be observed, as well as changes in their functionality, supporting the translational value to evaluate the efficacy of immunomodulating compounds in vitro.
Citation Format: Carla Castro, Philipp Metzger, Cynthia Obodozie, Holger Weber. Exploiting macrophage differentiation and plasticity as an immunomodulating strategy [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 671.
Abstract 6768: The potential predictive role of spatial phenotyping in non-small cell lung cancer
Authors: Ma Ning , Pratapa Aditya , Monkman James , O’Byrne Ken , Braubach Oliver , Kulasinghe Arutha 
Published: April 2023

Abstract
Background Lung cancers are the leading cause of cancer-related deaths with a 5-year-survival of only ~20%. Whilst immunotherapies have led to durable and prolonged survival, only a subset of patients remain responsive. Additional biomarkers are thus needed to better predict if patients will respond or develop resistance against immune checkpoint inhibitor (ICI) therapies. Spatial phenotyping of the tumor microenvironment (TME) is now recognized as a proxy for ICI therapy outcomes. Our study employs an end-to-end spatial biology strategy to survey non-small-cell lung cancer (NSCLC) tissues for new biomarkers that could guide immunotherapy treatments.
Methods We phenotyped pre-treatment biopsies from non-small-cell lung cancer (NSCLC) patients treated with single-agent Nivolumab. We first performed 57-plex whole-slide Single Cell Spatial Phenotyping on the PhenoCycler®-Fusion platform. Next, we developed the spatial analysis of a larger NSCLC cohort using customizable PhenoCode Signature Panels (PSP) for high-throughput immune profile (CD3/CD8/CD20/CD68/PanCK + CD4 add-in) and immuno-contexture (CD8/CD68/PD-L1/FoxP3/PanCK + PD-1 add-in) imaging. The PSP panel content combines the barcode-based antibody chemistry from the PhenoCycler platform with the signal amplification of Opal chemistry from the PhenoImager platform. PSP panels were profiled across n=27 NSCLC biopsies and provided a wider breadth of tissue profiling.
Results Our whole-slide single-cell spatial phenotyping analyses revealed high phenotypic diversity in the TME of patients responsive and resistant to ICI therapy. However, higher-throughput analyses of the same tissues using targeted PSP panels revealed no significant differences in quantities of immune cell lineages, including T-cells and macrophages. On the contrary, we discovered multiple quantifiable and statistically significant spatial signatures that appear to be predictive of treatment benefit, which confirms the potential biomarker value of spatial associations in patient tissues.
Conclusions This study amounts to a uniquely comprehensive Single Cell Spatial Phenotyping analysis of pre-treatment NSCLC biopsies from a single-agent Nivolumab study. Our data catalogue the diversity in the immune microenvironment of NSCLC but highlighted that immune cell quantification is insufficient to stratify patient cohorts. Single-cell spatial phenotyping, on the other hand, promises to reveal new biomarkers that may aid in better stratification of patients in pre-treatment evaluations.
Citation Format: Ning Ma, Aditya Pratapa, James Monkman, Ken O’Byrne, Oliver Braubach, Arutha Kulasinghe. The potential predictive role of spatial phenotyping in non-small cell 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 6768.
Abstract 677: Increasing plexity of imaging mass cytometry for tumor tissue analysis
Authors: Pfister Thomas D. , Ouladan Shaida , Yao Huihui , Majonis Daniel , Loh Christina , Zabinyakov Nick 
Published: April 2023

Abstract
Imaging Mass Cytometry™ (IMC™) is the leading platform for high-plex tissue imaging. IMC allows for detailed assessment of cell phenotype and function using 40-plus markers simultaneously at subcellular resolution on a single slide. A comprehensive IMC panel containing structural, functional, and immune markers enables us to reveal the complex heterogeneity of tumor tissues as well as the tumor microenvironment (TME). Driven by an increase in the number of antibody markers and the addition of mRNA markers, there is an increasing demand for larger panels. In addition, increasing the number of investigated target markers on a single tissue enriches spatial characterization that may facilitate a more accurate prediction of disease progression and preclinical outcome measures in clinical research projects using tumor biopsies or tissue microarrays (TMAs). Therefore, to increase the plexity of IMC panels, it is essential to expand the number of available metal channels. Here, we demonstrate the incorporation of conjugated antibodies with yttrium (89Y) and indium (115In), two low-mass metals, for IMC application. These metal tags have been previously tested as putative channels for IMC application.
We performed IMC analysis of various tissue types stained with panels of conjugated antibodies including the novel 89Y- and 115In-conjugated antibodies. At least 3 different regions of interest (ROIs) were assessed for each of the investigated tissue types. We compared images for the 89Y- and 115In-conjugated antibodies with the images generated using Maxpar® catalog antibodies of the same clones, with a focus on marker specificity and background signal. Compared with the lanthanide-conjugated catalog antibodies, the 89Y- and 115In-conjugated antibodies showed equivalent specificity and staining quality.
Our results open a new avenue to assign markers to 89Y and 115In, which enables a larger list of potential targets to be investigated in any IMC study. Expanding the number of markers to 40-plus in Imaging Mass Cytometry will improve the imaging results necessary to identify novel cell signatures (phenotype and interactions) in the TME.
Citation Format: Thomas D. Pfister, Shaida Ouladan, Huihui Yao, Daniel Majonis, Christina Loh, Nick Zabinyakov. Increasing plexity of imaging mass cytometry for tumor tissue analysis [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 677.
Abstract 682: Heterologous prime-boost viral vector vaccines for the treatment of a P1A-expressing BGL-1 glioblastoma model
Authors: Steffke Emily E. , McAuliffe James , Almeida Vinnycius Pereira , Wicki Amanda , Noblecourt Laurine , Kohanbash Gary , Okada Hideho , Leung Carol Sze Ki , Terabe Masaki , Eynde Benoit J. Van Den 
Published: April 2023

Abstract
Glioblastoma is an extremely aggressive and difficult cancer to treat, which may partly be due to its limited ability to induce T-cell responses. However, combining viral vector vaccines with other therapies to generate tumor-specific T cells may provide a meaningful benefit to patients. Here, we investigated whether heterologous prime-boost vaccination with chimpanzee-derived adenoviral vector ChAdOx1 and modified vaccinia Ankara (MVA) vaccines could generate therapeutically effective CD8+ T-cell responses against a model antigen P1A, a mouse homolog of human tumor-associated Melanoma Antigen GenE (MAGE)-type antigens, expressed by a BGL-1 mouse glioblastoma cell line. We demonstrated that heterologous prime-boost vaccination with ChAdOx1/MVA vaccines targeting P1A generated a high magnitude of CD8+ T cells specific for the P1A35-43 epitope presented by the MHC class I molecule H-2Ld. Prophylactic vaccination with ChAdOx1/MVA-P1A significantly prolonged the survival of syngeneic mice subcutaneously challenged with P1A-expressing BGL-1 tumors. Furthermore, different vaccination schedules significantly impact the magnitude of antigen-specific CD8+ T-cell responses and may impact protective efficacy. However, the substantial induction of myeloid-derived suppressor cells (MDSCs) by this tumor model presents a significant challenge in the therapeutic setting. Future work will investigate the efficacy of this vaccination strategy on intracranial P1A-expressing BGL-1 models.
Citation Format: Emily E. Steffke, James McAuliffe, Vinnycius Pereira Almeida, Amanda Wicki, Laurine Noblecourt, Gary Kohanbash, Hideho Okada, Carol Sze Ki Leung, Masaki Terabe, Benoit J. Van Den Eynde. Heterologous prime-boost viral vector vaccines for the treatment of a P1A-expressing BGL-1 glioblastoma model [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 682.
Abstract 692: OAd-mediated SA-4-1BBL fused to HPV-16 E7 elicits specific antitumor efficacy in syngeneic mouse lung cancer model
Authors: Gomez-Gutierrez Jorge G. , Garza-Morales Rodolfo , Martinez-Perez Alejandra G. , Perez-Trujillo Jose J. , Saucedo-Cardenas Odila , Loera-Arias Maria J. , Montes-de-Oca-Luna Roberto 
Published: April 2023

Abstract
Introduction: Human papillomaviruses (HPVs) are responsible for about 25% of cancer cases worldwide. HPV-16 E7 antigen is a tumor-associated antigen (TAA) commonly expressed in HPV-induced tumors; however, it has low immunogenicity. The interaction of 4-1BBL with its receptor induces pleiotropic effects on innate, adaptive, and regulatory immunity and, if fused to TAAs in DNA vaccines, can improve the antitumor response; however, there is low transfection and antitumor efficiency. Oncolytic virotherapy is promising for antitumor gene therapy as it can be selectively replicated in tumor cells, inducing cell lysis, and furthermore, tumor cell debris can be taken in by immune cells to potentiate antitumor responses.
Methods: The AdenoQuick 2.0 system was used to construct three OAds, expressing 1) SP-SA-E7-4-1BBL, 2) SA-E7-4-1BBL or 3) SP-SA-41BBL. The expression of transgenes was confirmed by western blot and subcellular localization of SP-SA-E7-4-1BBL was evaluated by immunofluorescence. The recombinant OAds-mediated cancer killing effect was evaluated by MTT assay and crystal violet staining, The recombinant OAds therapeutic efficacy was evaluated in a C57BL/6 mice bearing subcutaneous TC-1 lung tumor. Tumor growth and survival were monitored for a period of twenty-eight days.
Results: In this study, we expressed the immunomodulatory molecule SA-4-1BBL fused to E7 on an oncolytic adenovirus (OAd) system. In vitro infection of TC-1 tumor cells and NIH-3T3 non-tumor cells with SA/E7/4-1BBL OAd demonstrated that only tumor cells are selectively destroyed. Moreover, protein expression is targeted to the endoplasmic reticulum in both cell lines when a signal peptide (SP) is added. Finally, in an HPV-induced cancer murine model, the therapeutic oncolytic activity of OAd can be detected, and this can be improved when fused to E7 and SP.
Conclusions: In this study, we report the design of an oncolytic adenovirus expressing the SP/SA/E7/4-1BBL fusion gene, which is capable of infecting murine cancer and normal cells, and the expressed protein was able to be targeted to the endoplasmic reticulum. Most importantly, OAd induced a cell killing effect that is specific to cancer cells. Moreover, OAd treatment induced a potent antitumor effect in a mouse TC-1 cancer model.
Citation Format: Jorge G. Gomez-Gutierrez, Rodolfo Garza-Morales, Alejandra G. Martinez-Perez, Jose J. Perez-Trujillo, Odila Saucedo-Cardenas, Maria J. Loera-Arias, Roberto Montes-de-Oca-Luna. OAd-mediated SA-4-1BBL fused to HPV-16 E7 elicits specific antitumor efficacy in syngeneic mouse lung cancer model [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 692.
Abstract 70: Mechanisms of tumor dormancy induction mediated by abrogation of myeloid tgfβ signaling
Authors: Ahad Abdul , Leng Feng , Ichise Hiroshi , Gray Justin , Aprelikova Olga , O’Neill Maura , Holewinski Ronald , Kopardé4 Vishal N. , Moriwaki Yasuhiro , Hollander Christine , Germain Ronald , Andresson Thorkell , Li Yang 
Published: April 2023

Abstract
Persistence in a dormant state of residual tumor cells can occur following extended periods of clinical remission which may last decades. The disseminated tumor cells in distant organ sites eventually gain context dependent metastasis advantage during tumor progression. The mechanisms for dormancy induction or reactivation remain unclear. We found that the abrogation of myeloid TGF-β signaling induced tumor dormancy in breast cancer metastasis models. When given extended time, the TGFβRIImyeKO mice eventually developed similar number of metastatic nodules to that of control mice. We further validated our finding using an inducible system where reintroduction of TβRII diminished the tumor dormancy induction. RNAseq of dormant vs proliferative tumor cells identify 504 differential expressed genes, including those involving cell cycle arrest, IFNg response, MTOR signaling as well as MYC targeted genes. Immune cell profiling showed differential immune composition in micro and macro-tumor microenvironment between WT and TβRIImyeKO lung tissues. Of great interest, abrogation of myeloid specific TGF-β signaling increased the number of CD103+DCs that displayed elevated TNFa production, which in turn increased IFNg+ production in T cells leading to improved innate
and adaptive immunity. Importantly, depletion of CD103 DCs in tumor-bearing TβRIImyeKO mice diminished the dormancy phenotype, which was not observed for the pDC depletion in the WT control mice. These data demonstrate that abrogation of myeloid specific TGF-β signaling improved the immune microenvironment, which induced tumor dormancy. Our data provide mechanistic insight and resources to understand the tumor dormancy.
Citation Format: Abdul Ahad, Feng Leng, Hiroshi Ichise, Justin Gray, Olga Aprelikova, Maura O’Neill, Ronald Holewinski, Vishal N. Kopardé4, Yasuhiro Moriwaki, Christine Hollander, Ronald Germain, Thorkell Andresson, Yang Li. Mechanisms of tumor dormancy induction mediated by abrogation of myeloid tgfβ signaling [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 70.