bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3008: Induction of necroptosis through caspase-independent mechanisms in MCF-7 breast cancer cells following HDAC inhibition
Authors: Natarajan Umamaheswari , Waldron Grace , Venkatesan Thiagarajan , Rathinavelu Appu 
Published: June 2022

Abstract
Cell proliferation and cell death are essential for maintaining homeostatic balance in eukaryotic organisms. Many diseases develop when cellular homeostasis is disturbed. Currently, several anti-cancer drugs are known to induce apoptosis through activation of intrinsic or extrinsic pathways. However, many types of cancer cells have acquired resistance due to quick adaptation to resist pro-apoptotic mechanisms. Therefore, simultaneous induction of other forms of cell deaths such as necroptosis, autophagy, etc., in addition to apoptosis, is becoming an attractive strategy for controlling tumor growth and metastasis. For this purpose, we have explored the role of XIAP in caspase-independent mechanisms of cell death during HDAC (Histone Deacetylase) inhibition. Our experiments with HDAC inhibitor SAHA (Suberoylanilide hydroxamic acid) have revealed that the anti-apoptotic biomarkers, including survivin and XIAP, exhibit important regulatory functions for inducing necroptosis in cancer cells. Interestingly, many of the signal adaptors of apoptosis and necroptosis are known to show dual functions during drug-induced cancer cell death. For example, the formation of the survivin-XIAP complex prevents XIAP from undergoing proteasomal degradation and polyubiquitination, thereby stabilizing XIAP in cancer cells. When the survivin level is increased, it is also known to stop the inhibition of XIAP by Smac. In addition, since XIAP can inhibit caspase -9 and the executioner caspases -3 and -7, the apoptosis mechanisms will be blocked, and cell proliferation will continue to promote cell survival and tumorigenesis. Our western blot data revealed that the caspase -9, -3, -7, and -8 levels were significantly downregulated in MCF-7 breast cancer cells following SAHA treatment. Furthermore, SAHA treatment also increased the levels of necroptosis inducers such as phospho-RIP3 and MLKL in MCF-7 cells. However, treatment of the same MCF-7 cells with other drugs such as RG-7388, an MDM2 inhibitor, was able to induce apoptosis through activation of caspases. Our current results show that MCF-7 cells may switch to the necroptosis pathway due to the inhibition of caspases and increased levels of survivin and XIAP. (This research was supported by the Royal Dames of Cancer Research Inc., Ft. Lauderdale, Florida).
Citation Format: Umamaheswari Natarajan, Grace Waldron, Thiagarajan Venkatesan, Appu Rathinavelu. Induction of necroptosis through caspase-independent mechanisms in MCF-7 breast cancer cells following HDAC inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3008.
Abstract 3011: Development of mitochondrial fission inhibitors in inducing non-apoptotic cell death in triple-negative breast cancer
Authors: Malla Saloni , Kumari Shikha , Terrero David , Raman Dayanidhi , Tiwari Amit K. 
Published: June 2022

Abstract
Triple-negative breast cancer (TNBC) is the most aggressive subtype of metastatic breast cancer with poor clinical prognosis. Presently used neoadjuvant chemotherapy against TNBC, such as taxanes, and/or anthracyclines show an initial treatment response but follows with drug resistance, tumor relapse, side effects and high proclivity to develop metastases. Recent studies point out that targeting dysregulated mitochondrial dynamics, that is now linked to the tumor initiation, progression, and metastasis in TNBC, as an effective therapeutic strategy. In particular, mitochondrial fission factors such as dynamin-related protein 1 (Drp1) is upregulated in TNBC and is known to affect mitochondrial energetics and alter metabolism which favors tumor progression. We have designed and developed small molecule inhibitors to specifically target Drp1 as a viable therapeutic strategy for metastatic TNBC (mTNBC). We recently identified a new class of structurally constrained thienopyridine (TPH) probes, and particularly a new analog TPH104c that showed higher binding affinity in silico to Drp1 as compared to Mdivi-1, a weak inhibitor of Drp1. Surface plasmon resonance studies suggest that TPH104c binds with Drp1 with high affinity. Similarly, mechanistic studies using Western blotting and immunofluorescence analysis confirmed that TPH104c significantly downregulated the levels of total and phosphorylated form of Drp1 at serine 616 in MDAMB-231 and BT-20 TNBC cells. Interestingly, Drp1 inhibition by TPH104c led to a hitherto unreported form of caspase- and reactive oxygen species (ROS)-independent non-apoptotic form of cell death. Rather than shrinking in size, or formation of apoptotic blebs which are classical features of apoptosis, TPH104c treated TNBC cells were enlarged, rounded, and swollen in size. These cells eventually lost their proliferative ability and were significantly less confluent than control cells. Additionally, in TPH104c-treated cells, neither the mitochondrial membrane potential was lost, nor the caspases were activated as also indicated by a lack of rescue of cell death when incubated with pan-caspase inhibitor. Further studies are in progress to understand the role of Drp1 inhibition in the induction of non-apoptotic cell death which will further help to optimize an effective lead compound for the future pre-clinical development of promising anticancer agents targeting mTNBC.
Citation Format: Saloni Malla, Shikha Kumari, David Terrero, Dayanidhi Raman, Amit K. Tiwari. Development of mitochondrial fission inhibitors in inducing non-apoptotic cell death in triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3011.
Abstract 3053: Using individual or a consortium of bacteria for immuno-oncology research
Authors: Maubant Sylvie , Leblanc Marie , Bertrand Elisabeth , Bertaux Audrey , Morgand Loic , Ramelet Maxime , Lux Marie , Duchamp Olivier , Vivani Fabrice 
Published: June 2022

Abstract
Preclinical and clinical studies have shed light on the beneficial role of bacteria for cancer therapy. Indeed, these studies have demonstrated that these microorganisms have beneficial properties that allow them to selectively colonize tumor and that they could also be considered as predictive drug efficacy biomarkers. Based on these results, bacteria are now used for delivering therapeutic components or for shaping the gut microbiota. Ultimately these approaches lead to the activation of an immune response against the tumor. Owing to our scientific and technological expertise in manipulating microbes, we propose tailor-made strategies for investigating the efficacy of bacteria-based treatments and/or the effect of therapies on the microbiota both in vitro and in vivo. Different methods/analyses can be used for culturing, detecting, quantifying, identifying and localizing live bacteria (e.g. counting of CFUs, PMA-qPCR, 16 rRNA gene sequencing, mass spectrometry, bioluminescence) in simple or complex samples (e.g. from culture of single bacterial species to rodent/human stools or other tissues). In addition, a continuum of assays allows us to evaluate the impact of bacteria or derived products directly on tumor and/or immune cells (e.g. immune infiltrate and phenotyping, cytokine/chemokine profiling, tumor burden). We will highlight some results obtained in a cancer context such as the immunostimulatory properties of bacteria or their derivatives, the selective colonization of tumor tissue by bacteria, the benefits in delivery of therapeutic proteins or antigens by bacteria, the impact of tumor engraftment/growth on gut microbiota, the effects of chemotherapeutic agents on intestinal microflora, the consequences of supplementation with bacteria or antibiotics treatment on the response to immune checkpoint inhibitors. Altogether these data demonstrate that bacteria are now allies in the treatment of cancer and that our comprehensive platform is suitable for evaluating both in vitro and in vivo therapeutics developed from bacteria (individuals, consortium or derived products; native or modified). All these technologies can be also applied to develop novel therapeutic strategies for inflammatory and infectious diseases known to increase the risk of cancer development.
Citation Format: Sylvie Maubant, Marie Leblanc, Elisabeth Bertrand, Audrey Bertaux, Loic Morgand, Maxime Ramelet, Marie Lux, Olivier Duchamp, Fabrice Vivani. Using individual or a consortium of bacteria for immuno-oncology research [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3053.
Abstract 3156: Tumor Treating Fields reduce cellular survival of human mesenchymal stromal cells via apoptosis and senescence induction
Authors: Rühle Alexander , Strack Maren , Grosu Anca-Ligia , Nicolay Nils H. 
Published: June 2022

Abstract
Background Tumor Treating Fields (TTFields), alternating electric fields with intermediate field intensity, have shown clinical efficacy in several tumor entities, leading to clinical approval for glioblastoma and mesothelioma treatment. As mesenchymal stromal cells (MSCs) belong to the tumor microenvironment and exhibit - depending on the tumor type and microenvironment - either pro-tumorigenic or anti-tumorigenic abilities, we aimed to study the effects of TTFields on human MSCs.
Methods Human MSCs were isolated from the bone marrow of three different healthy donors and were exposed to TTFields for 72 hours. MSCs’ clonogenic survival, proliferation and viability was analyzed after different TTFields frequencies. Boyden chamber assays were carried out to investigate the influence of TTFields on the migratory potential of MSCs. Flow cytometry analyses of caspase-3 and annexin V were conducted to determine MSCs’ apoptosis rates after TTFields treatment. TTFields-induced senescence was quantified using beta-galactosidase stainings. MSCs’ characteristic surface marker expression (CD73, CD90 and CD105) was studied after TTFields treatment.
Result TTFields led to significantly reduced clonogenic survival (70-90% reduction compared to controls) and cellular proliferation (approximately 50% reduction) in all analyzed MSC specimens. However, MSCs’ metabolic viability was largely unaffected after TTFields exposure. Within a range between 100 and 500 kHz, there was no frequency dependence of TTFields’ anti-proliferative effect on human MSCs. As determined by Bodyen Chamber assays, TTFields resulted in a considerable reduction of MSCs’ migratory potential. TTFields increased both cellular apoptosis and senescence in all MSC samples, whereas there was no alteration of MSCs’ characteristic surface marker pattern.
Conclusion As TTFields considerably reduce MSCs’ survival and migratory potential, TTFields could attenuate the pro-tumorigenic effects of tumor-associated MSCs that were recently described in glioblastoma patients. Co-culture experiments of MSCs and tumor cells after TTFields treatment as well as in vivo experiments are planned in the future.
Citation Format: Alexander Rühle, Maren Strack, Anca-Ligia Grosu, Nils H. Nicolay. Tumor Treating Fields reduce cellular survival of human mesenchymal stromal cells via apoptosis and senescence induction [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3156.
Abstract 3259: TAS3351 is a 4th-generation EGFR-TKI overcoming T790M and C797S-mediated resistance in NSCLC with EGFR common mutations
Authors: Kasuga Hidefumi , Kataoka Yuki , Yamamoto Fuyuki , Mizutani Takashi , Tsuji Shingo , Tanaka Sakiho , Mizuarai Shinji 
Published: June 2022

Abstract
Background: Common mutations (exon 19 deletion; ex19del or L858R point mutation; L858R) of the epidermal growth factor receptor (EGFR) are found in patients with non-small cell lung cancer (NSCLC) and are sensitive to EGFR-tyrosine kinase inhibitors (TKIs), including osimertinib, a third-generation, irreversible EGFR-TKI. Although osimertinib is widely utilized as the standard of care in first-line therapy of NSCLC, resistance inevitably occurs during the treatment. Among EGFR-dependent/on-target resistant mechanisms of osimertinib, the C797S mutation is reported to emerge as the most frequent alteration. The cysteine residue at position 797 of EGFR is covalently bound with osimertinib, and its alteration to serine causes an inability to form a covalent bond, making osimertinib resistant to NSCLC with a C797S mutation. While first-generation, reversible EGFR-TKIs (gefitinib and erlotinib) might be effective for patients with C797S mutation, the T790M mutation is reported to emerge in almost half of patients with EGFR common mutations during treatment with first-generation EGFR-TKIs. Here, we report TAS3351 as a novel, next-generation EGFR-TKI designed to overcome C797S and T790M-mediated resistance in NSCLC patients.
Methods: The inhibitory potency of TAS3351 on cellular phospho-EGFR was analyzed by In-cell western assay. In addition, inhibition of cell viability in Ba/F3 cells transformed by human wild type or mutated EGFR, as well as in human NSCLC cell lines whose EGFR harbors common mutations, was tested. Anti-tumor activity of TAS3351 was evaluated in NIH/3T3-EGFR allograft models and human NSCLC xenograft models.
Results: TAS3351 exhibited almost comparable inhibitory potency against cellular phosphorylation of EGFR harboring ex19del or L858R with or without C797S and/or T790M while sparing wild type EGFR activity. TAS3351 also showed similar trends in the cell viability assay for Ba/F3-EGFR cells and human NSCLC cell lines. Finally, TAS3351 demonstrated favorable anti-tumor activity in NIH/3T3 allografts transformed by human EGFR harboring ex19del/C797S/T790M.
Conclusion: TAS3351 is a fourth-generation EGFR-TKI overcoming T790M and C797S-mediated resistance in NSCLC with common mutations. The present results support the clinical development of TAS3351 as monotherapy in NSCLC patients with EGFR mutations harboring C797S and/or T790M EGFR-TKI resistance mutations.
Citation Format: Hidefumi Kasuga, Yuki Kataoka, Fuyuki Yamamoto, Takashi Mizutani, Shingo Tsuji, Sakiho Tanaka, Shinji Mizuarai. TAS3351 is a 4th-generation EGFR-TKI overcoming T790M and C797S-mediated resistance in NSCLC with EGFR common mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3259.
Abstract 3287: Baseline CD166 expression determines CRISPR-mediated ALCAM gene knockout effects on migratory phenotype in metastatic melanoma cells
Authors: Kaminski Rafal , Luongo Francesca P. , Dabrowski Konrad , Bellizzi Anna , Mancuso Pietro , Zaidi M. Raza , McMillan Johanna , Patel Meha , Coleman Tyrone , Arosarena Oneida A. 
Published: June 2022

Abstract
Advanced stage melanoma is a mostly incurable, highly aggressive tumor whose incidence is rising. Metastatic dissemination of primary tumors involves the repeated crossing of tissues barriers, which requires dynamic modulation of tumor cell adhesion, presumably through mechanisms like those utilized by trafficking leukocytes. Activated leukocyte cell adhesion molecule (ALCAM/CD166) has been shown to play an important role in the metastatic process, including detachment from the primary tumor, adhesion to endothelium, and extravasation. ALCAM expression correlates with melanoma progression, and it has been identified as a marker of pluripotent stem cells. Furthermore, its expression in primary melanoma strongly correlates with tumor thickness and level of invasiveness. shRNA silencing of ALCAM in cells with high levels of ALCAM resulted in reduced motility and transmigration. In the current study, we tested the effect of CRISPR-mediated ALCAM knockout (KO) on migration and invasion of two human melanoma cell lines representing opposite phenotypic states of melanoma. MEL501 (proliferative type, SOX10/MITF-high and AP1/TEAD1-low) cells express low levels of ALCAM at baseline, while the Rosi cell line (invasive type, SOX10/MITF-low and AP1/TEAD1-high) expresses high baseline ALCAM levels. ALCAM KO promoted migration of MEL501 cells (p&lt;0.000001) but inhibited migration of Rosi cells (p=0.005050) as compared to the parent cell lines. In addition, ALCAM KO promoted the invasion of MEL501 cells (p=0.019768) but had no impact on Rosi cell invasion. While ALCAM has been shown to regulate matrix metalloproteinase-2 (MMP-2) activation, zymograms revealed no difference in MMP-2 activity between MEL501 and Rosi ALCAM-KO cells and the parent cell lines. These data indicate that ALCAM may play a tumor-suppressive role in melanoma cells with low ALCAM expression and are consistent with a previous study which demonstrated that introduction of an amino-terminally truncated ALCAM in early stage melanoma cells promoted metastasis. Our data also provide in vitro evidence that gene therapy may reverse the migratory phenotype of melanoma cells with high ALCAM expression.
Citation Format: Rafal Kaminski, Francesca P. Luongo, Konrad Dabrowski, Anna Bellizzi, Pietro Mancuso, M. Raza Zaidi, Johanna McMillan, Meha Patel, Tyrone Coleman, Oneida A. Arosarena. Baseline CD166 expression determines CRISPR-mediated ALCAM gene knockout effects on migratory phenotype in metastatic melanoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3287.
Abstract 3291: Anti-HER2 scFv antibody-directed lentiviral delivery of myrosinase for eradicating HER2+ cancer cells after dosing prodrug sinigrin
Authors: Tarar Ammar Ahmad , Peng Ching-An 
Published: June 2022

Abstract
Chemotherapy is widely used for the treatment of cancers, but it lacks selectivity and therefore results in collateral damage to healthy cells and drug resistance of cancer cells. Gene-directed enzyme prodrug therapy (GDEPT) has been adopted to mitigate these issues. In this study, we demonstrated the specific targeting of myrosinase-sinigrin paired GDEPT to treat HER2+ (human epidermal growth factor 2) cancer cells. Sinigrin excessively present in cruciferous vegetables can be catalyzed by myrosinase to produce cytotoxic ally isothiocyanate (AITC), which has shown anticancer effectiveness. The myrosinase gene was packed in biotinylated lentiviral vectors produced by co-transfection of HEK293T cells. The single chain variable fragment (scFv) gene sequence of anti-HER2 antibody and core streptavidin (coreSA) gene sequence were cloned in pET-30a(+) plasmid, which was then transformed in Lemo21 (DE3) competent cells for protein expression. The anti-HER2-coreSA chimeric protein was purified using immobilized metal affinity chromatography and characterized by SDS-PAGE and Western blot. The purified chimeric protein was tagged on biotinylated lentivirus via streptavidin-biotin binding. Our results showed that myrosinase-encoding lentivirus tagged with anti-HER2 selectively delivered myrosinase to HER2+ cancer cells. When treated with prodrug sinigrin, the targeted cancer cells underwent apoptosis and ensuing cell death. In summary, the targeted delivery of myrosinase to HER2+ cancer cells using lentivirus functionalized with anti-HER2 ScFv antibody can eradicate malignant cells when treated with enzymatically catalyzed prodrug sinigrin.
Citation Format: Ammar Ahmad Tarar, Ching-An Peng. Anti-HER2 scFv antibody-directed lentiviral delivery of myrosinase for eradicating HER2+ cancer cells after dosing prodrug sinigrin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3291.
Abstract 3298: Propagation competence of a self-antigen-targeting arenavirus vector based cancer therapy determines antitumor efficacy in mouse melanoma
Authors: Purde Mette-Triin , Hartmann Fabienne , Cupovic Jovana , Schmidt Sarah , Bomze David , Stemeseder Felix , Lercher Alexander , Besse Lenka , Berner Fiamma , Tüting Thomas , Bergthaler Andreas , Schietinger Andrea , Kochanek Stefan , Ludewig Burkhard , Orlinger Klaus K. , Bald Tobias , Ring Sandra S. , Flatz Lukas 
Published: June 2022

Abstract
Breaking self-tolerance and inducing a robust immune response targeting tumor cell derived self-antigens by therapeutic vaccines has remained challenging. We have generated a replicating viral vector based on the arenavirus lymphocytic choriomeningitis virus expressing the melanoma tissue-specific self-antigen Trp2 (artLCMV-Trp2) and show that a single immunization of mice bearing established melanoma with this vector can break tolerance against this self antigen and induces high levels of tumor-infiltrating TRP2+ CD8+ T cells. These T cells express the effector cytokines IFNγ and TNFα, lack markers of T cell exhaustion, and their appearance coincides with marked inhibition of tumor growth and significant survival benefit. artLCMV-TRP2 induced a strong type I interferon response that resulted in transient type I-dependent Treg inhibition that was necessary for the generation of optimal antitumoral T cell responses. Moreover, artLCMV-TRP2 vaccination proved able to induce complete tumor remission when combined with infusion of exogenous TRP2-specific T cells into mice. In conclusion, replication competence is a key property of artLCMV-TRP2 that enabled effective anti-cancer immunity and therapeutic effect.
Citation Format: Mette-Triin Purde, Fabienne Hartmann, Jovana Cupovic, Sarah Schmidt, David Bomze, Felix Stemeseder, Alexander Lercher, Lenka Besse, Fiamma Berner, Thomas Tüting, Andreas Bergthaler, Andrea Schietinger, Stefan Kochanek, Burkhard Ludewig, Klaus K. Orlinger, Tobias Bald, Sandra S. Ring, Lukas Flatz. Propagation competence of a self-antigen-targeting arenavirus vector based cancer therapy determines antitumor efficacy in mouse melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3298.