bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2760: Evaluation of functional heterogeneity in the potent responses of single CAR-T effector cells
Authors: Yellen Benjamin 
Published: April 2023

Abstract
Over the last decade, chimeric antigen receptor (CAR)-T cell immunotherapies have produced some miraculous cures in advanced stage, aggressive leukemias and lymphomas in both children and adults. However, many barriers continue to limit the availability and effectiveness of these therapies, including the lack of reliable potency assays for these cell-based “living” medicines. Unlike molecular medicines, which have well established manufacturing processes, cell based medicines are inherently heterogeneous, and each engineered lot of cells will inevitably have a distribution of growth properties, cytolytic activity, gene expression patterns, and secretion profiles, and it is not clear which cell subpopulations are most correlated with positive clinical outcomes.Here we report results of an evaluation of phenotypic heterogeneity in a population of anti-CD19 CAR-T cells co-cultured with target cells in an array of &gt;300,000 microwells. Our approach uses high throughput time lapse imaging to continually monitor the cell-cell interactions between effector and target cells. We maintained cell viability in the microwells for more than 5 days, which gave us sufficient time to observe the full range of antigen-mediated cell killing phenotypes, and also evaluate growth rates of the potent effector cells. We tracked thousands of individual co-cultures through time lapse imaging, and cell behavior was evaluated and sorted by deep learning algorithms. We were able to quickly identify and retrieve clones that exhibited desirable phenotypes for full molecular characterization.Our analysis shows that three fundamental subpopulations of effector cells can be clearly distinguished and quantified. These are 1) effector cells that kill the targets and then die, 2) effector cells that kill the target and then proliferate, and 3) effector cells that do not kill the target. We further dissect these populations by characterizing their secretion and transcriptomic profiles, to identify further subtype classifications.
Citation Format: Benjamin Yellen. Evaluation of functional heterogeneity in the potent responses of single CAR-T effector cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2760.
Abstract 278: Role of ATPIF1 in glioblastoma cell survival and metabolism
Authors: Guda Maheedhara Reddy , Mnatsakanyan Nelli , Morris Daniel , Asuthkar Swapna , Tsung Andrew J. , Velpula Kiran 
Published: April 2023

Abstract
The ATPase inhibitory factor 1 (ATPIF1) is a 12 kDa protein that inhibits the mitochondrial F1Fo-ATP synthase in the F1domain. It reverses the mitochondrial ATP synthase complex's function, contributing to the onset of oncogenesis. The overexpression of ATPIF1 in various cancers has recently attracted greater attention; however, its biological significance in human pathology, specifically cancer, remains unknown. Based on immunohistochemistry and Western blot analysis of glioblastoma (GBM) patient specimens, we found ATPIF1 is highly expressed in GBMs. Here we showed that mitochondrial fractions of silencing ATPIF1 in U-87 and U251 cells showed decreased expression of ATPIF1, ATP5A, and ATPG1-G2-G3. Next, we showed that manipulation of ATPIF1 showed increased Reactive Oxygen Species (ROS) and Caspase3/7 activity levels compared to untreated cells. ATPIF1 knockdown further caused cell cycle arrest at the G₀/G1stage, and FITC Annexin V analysis showed increased apoptosis. Increased glucose uptake is associated with aggressive oncological behavior, and here we found elevated levels of TCA metabolites in ATPIF1 knockdown cells. ATPIF1 silenced cells had a shift in extracellular acidification rate, accompanied by a simultaneous increase in proton leak, according to the metabolic analysis performed by Seahorse bio-analyzer. Overall, these observations suggest ATPIF1 plays a key role in the progression of GBM. However, further research is needed to verify the precise mechanisms by which ATPIF1 functions in GBM.
Citation Format: Maheedhara Reddy Guda, Nelli Mnatsakanyan, Daniel Morris, Swapna Asuthkar, Andrew J. Tsung, Kiran Velpula. Role of ATPIF1 in glioblastoma cell survival and metabolism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 278.
Abstract 2782: Small molecule H182 suppresses Stat3 activation in tumor cells and combines with radiation therapy to block breast tumor growth in mouse syngeneic models
Authors: Chen Yue , Yue Peibin , Fu Wenzhen , Chen Weiliang , Kershaw Kathleen M. , Shiao Stephen L. , Tius Marcus A. , Lopez-Tapia Francisco , Turkson James 
Published: April 2023

Abstract
Signal transducer and activator of transcription 3 (Stat3) is a latent transcription factor that contributes to tumor cell growth and survival in constitutively-active form in several types of human cancers, and hence, serves as a therapeutic target. The azetidine-based compound, H182 irreversibly binds to Stat3. In cell-free DNA-binding assay, H182 selectively inhibited Stat3 DNA-binding activity (IC50 0.38-0.66 μM) over Stat1 or Stat5 (IC50&gt;15.8 μM) in vitro. In treated pancreatic cancer cells, H182 specifically blocked the association of Stat3 with gp130 and JAK2, and inhibited Stat3 tyrosine phosphorylation and DNA-binding activity. Coimmunoprecipitation and colocalization studies of hemagglutinin (HA)-tagged Stat3 and EGFP-tagged Stat3 expressed in prostate cancer cells showed that treatment with H182 blocked the HA-Stat3:EGFP-Stat3 interactions in intact cells. Immunofluorescence staining with laser-scanning confocal microscopy analysis for the intracellular localization of Stat3 showed treatment of H182 disrupted Stat3 nuclear accumulation and promoted the aggregation of Stat3 at the perinuclear region. H182 consequently suppressed Stat3-dependent transcriptional activity and the expression of Stat3 downstream genes, including Cyclin A, Bcl-2, Cyclin B1, and Mcl-1. Moreover, H182 significantly inhibited the colony survival, migration, and invasion in vitro of breast, pancreatic and prostate cancer cells harboring aberrant Stat3 activation. Significantly, in vivo administration of H182 in combination with radiation induced a strong antitumor response against mouse triple-negative breast cancer in syngeneic models and prolonged survival. Thus, our study provides a novel Stat3 inhibitor with significant antitumor activity against human tumors cancer harboring persistently active STAT3.
Citation Format: Yue Chen, Peibin Yue, Wenzhen Fu, Weiliang Chen, Kathleen M. Kershaw, Stephen L. Shiao, Marcus A. Tius, Francisco Lopez-Tapia, James Turkson. Small molecule H182 suppresses Stat3 activation in tumor cells and combines with radiation therapy to block breast tumor growth in mouse syngeneic models [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 2782.
Abstract 2783: TET2-STAT3-CXCL5 nexus promotes triglyceride transfer from neutrophils to cancer cells to fuel lung adeno-to-squamous transdifferentiation
Authors: Xue Yun , Chen Yuting , Sun Sijia , Ji Hongbin 
Published: April 2023

Abstract
Phenotypic plasticity is a rising cancer hallmark and lung adeno-to-squamous transdifferentiation (AST) triggered by LKB1 inactivation is significantly associated with drug resistance. Mechanistic insights into AST are urgently needed to identify therapeutic vulnerability in LKB1-deficient lung cancer. Here we find that Ten-Eleven Translocation (TET)-mediated active DNA demethylation is elevated during AST in KrasLSL-G12D/+;Lkb1L/L(KL) mice, and knockout of individual Tet gene reveals that Tet2 is required for squamous transdifferentiation. STAT3 recruits TET2 to CXCL5 promoter region for demethylation and promotes neutrophil infiltration. Targeting the STAT3-CXCL5 nexus effectively inhibits squamous transdifferentiation through reducing neutrophil infiltration. Interestingly, tumor-infiltrating neutrophil is laden with triglycerides and transfer lipid to tumor cells to promote cell proliferation and squamous transdifferentiation. Pharmacological inhibition of macropinocytosis dramatically inhibits neutrophil-to-cancer cell lipid transfer and blocks squamous transdifferentiation. These data uncover an epigenetic mechanism orchestrating phenotypic plasticity through regulating immune microenvironment and metabolic communication, and identify therapeutic strategies to inhibit AST.
Citation Format: Yun Xue, Yuting Chen, Sijia Sun, Hongbin Ji. TET2-STAT3-CXCL5 nexus promotes triglyceride transfer from neutrophils to cancer cells to fuel lung adeno-to-squamous transdifferentiation [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 2783.
Abstract 2826: Electromagnetic fields and hyperthermia induce lysosomal permeabilization and death in different cancer cell models: in vitro and in vivo studies
Authors: Obrador Elena , Jihad-Jebbar Ali , Salvador-Palmer Rosario , López-Blanch Rafael , Oriol-Caballo María , Moreno-Murciano María Paz , Navarro Enrique , Cibrián Rosa , Estrela Jose M. 
Published: April 2023

Abstract
Introduction: Non-ionizing radiations (NIR), e.g. tumor-treating fields, high intensity focused ultrasounds (HIFU) or high frequency microwaves, have potential applications in Oncotherapy. Nevertheless, as today, their efficacy is limited. We investigated whether externally applied magnetic fields and non-ablative hyperthermia affect cancer cell viability and growth.
Methods: Under in vitro conditions, human A2058 melanoma, AsPC1 pancreas carcinoma, and MDA-MB-231 breast carcinoma cells were treated with electromagnetic fields (EMFs, using solenoids) and/or hyperthermia (HT, using a thermostated water bath). The effect of both energies was also studied in combination with standard chemotherapy and targeted therapy. An experimental system combining EMFs and HIFU-induced HT was developed and tested in vivo.
Results: EMFs (25 μT, 4h) or HT (52ºC, 40 min) quite moderately affect cancer cell viability. However, their combination decreased viability to approx. 16%, 50%, and 21% of controls values in A2058, AsPC1, and MDA-MB-231 cells, respectively). Increased lysosomal permeability, release of cathepsins into the cytosol and activation of the mitochondria-dependent cell death are the underlying mechanisms. Cancer cells were completely eliminated combining EMFs, HT and standard chemotherapy or EMFs, HT and anti-Hsp70-targeted therapy. In vivo experiments, using pancreatic cancer xenografts growing in nude mice, show that combination of EMFs, HIFU-induced HT and gemcitabine induces a drastic cancer regression (to approx. 3-4% of control values).
Conclusion: Non-invasive NIR (EMFs and HIFU-induced HT) can complement current Oncotherapies. This strategy, associated to current chemotherapy or anti-Hsp70-targeted therapy, may facilitate the elimination of tumors that can be previously located using standard imaging techniques (CT scan and NMR). These results may help to overcome the limitations in the use of different NIR in Oncotherapy.
Citation Format: Elena Obrador, Ali Jihad-Jebbar, Rosario Salvador-Palmer, Rafael López-Blanch, María Oriol-Caballo, María Paz Moreno-Murciano, Enrique Navarro, Rosa Cibrián, Jose M. Estrela. Electromagnetic fields and hyperthermia induce lysosomal permeabilization and death in different cancer cell models: in vitro and in vivo studies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2826.
Abstract 2827: Application of AZD2281 combine with radiation therapy in glioblastoma
Authors: Liao Tsal-Lan , Chiang I-Tsang , Chang Yuan , Hsu Fei-Ting 
Published: April 2023

Abstract
Glioblastoma (GBM), can be divided into primary tumors from brain tissue (&gt;90% of all cases) and secondary tumors that progress from low-grade gliomas. The current GBM treatment strategy is chemotherapy drug temozolomide (TMZ) combined with radiotherapy (RT), which has limited treatment effect, poor prognosis, high recurrence rate, and short prognosis survival time. Recently, studies indicated that breast cancer susceptibility gene 1/2 (BRCA1/2) mutations are frequently occur in GBM, while few studies have explored this pathway in depth. At present, AZD2281 (PARP Inhibitor) has been used to treat breast cancer and ovarian cancer patients with BRCA1/2 mutation...etc. Whether AZD2281 can play a role in affecting the relevant pathways after BRCA1/2 mutate GBM patients, and the possible mechanism and benefit of combined RT are the focus of this study. AZD2281 is the first Poly (ADP-ribose) polymerase (PARP) inhibitor approved by the U.S. Food and Drug Administration (FDA) for the treatment of germline BRCA mutation-associated ovarian cancer. AZD2281 is a targeted therapy that used to block DNA Damage Response (DDR) in cells/tumors deficient in homologous recombination repair (HRR). After RT, there may be a phenomenon of DNA repair; therefore, AZD2281 can be used to disrupt the repair process. At the beginning, we used RSI analysis to confirm that patient with higher expression level of BRCA1 may relatively resistance to RT. To overcome this issue, we identified the effect of AZD2281 combined with RT on GBM cells (BRCA1/2 higher and lower expression cells) by (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay), and colony formation. The internal/external apoptosis pathways (Caspase-3/Caspase-8/Caspase-9) and TUNEL assay were used to prove the superior apoptosis effect of AZD2281 combined with RT as compared to single treatment. Additionally, cells lower BRCA1 expression level (U87 and U251 siBRCA1) may sensitive to RT treatment. In our DNA damage, DNA fragmentation, and transwell invasion/migration results, we showed that the combination of AZD2281 and RT may effective induce DNA damage. In our animal data, we proved that combination therapy can effective suppression tumor growth and apoptosis-related protein signaling increased in DNA damage by IHC (p-ATM/p-CHK2/Caspase-3/Caspase-8/Caspase-9). In conclude, we suggested that AZD2281 can be used as a radiosensitizer to sensitize GBM patient to RT, which is dependent to BRCA1 expression.
Citation Format: Tsal-Lan Liao, I-Tsang Chiang, Yuan Chang, Fei-Ting Hsu. Application of AZD2281 combine with radiation therapy in glioblastoma [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 2827.
Abstract 2836: H2S donors as radiosensitizers for glioblastoma multiforme (GBM)
Authors: Modi Morni , Weber Camila , Xu Qinqin , Chen Kuanling , Cheong Ana , Nagel Zachery , Harrison Lynn 
Published: April 2023

Abstract
Glioblastoma multiforme (GBM) is an extremely aggressive CNS cancer in adults. Radiation is a cornerstone therapy but poses high risk of healthy tissue necrosis. Despite maximal treatment, GBM results in death within 15-21 months of diagnosis. These poor statistics suggest the need for a novel therapy. The use of radiosensitizing agents is a novel approach to minimizing radiation mediated healthy tissue necrosis by allowing use of lower doses to achieve comparable cytotoxic effects. Our lab has shown that sodium sulfide (Na2S), a hydrogen sulfide (H2S) donor, selectively kills GBM (T98G) cells while sparing normal human brain endothelial (hCMEC/D3) cells. Na2S also selectively radiosensitizes GBM cells and enhances cell killing with photon or proton radiation. We are currently exploring Diallyl Trisulfide (DATS), another H2S donor, as a radiosensitizing agent and have demonstrated that DATS can kill GBM cells. At a given dose of 2 Gy, radiation alone decreases T98G survival by 10%. With DATS, survival decreases by 70%, suggesting an additive effect. To determine if DATS increases DNA damage mediated cell death, T98G cells were treated with DATS, radiation, or combined DATS + radiation treatment and then analyzed for γH2AX foci, an indicator of double stand breaks (DSB). Our results show that compared to no treatment, DATS increased the number of γH2AX foci per cell by 1.5-fold, by 3-fold with radiation, and by 4-fold with combined DATS + radiation, an additive response. To elucidate the mechanism of increased DNA damage, a novel fluorescence multiplex host cell reactivation (FM-HCR) reporter system was used to assess repair activity of DSBs in live cells. T98G cells pre-treated with DATS were transfected with reporter plasmids containing site-specific lesions, mimicking radiation-like DNA damage. Repair resulted in expression of a fluorescent reporter protein, which was quantitated using flow cytometry. Our results show that DATS did not alter non-homologous end joining (NHEJ) or homologous repair (HR) activity of DSB. However, DATS did alter base excision repair (BER) activity by decreasing repair of abasic lesions and increasing repair 8oxoG lesions. We hypothesize that this attempted BER via simultaneous strand cleavage by AP endonuclease (Ape1) or glycosylase (Ogg1) may lead to build up of repair intermediates, threatening genomic stability and inducing DSB. Pursuing specific DNA repair biochemical assays is required to further understand this effect of DATS on DNA damage and repair inhibition.
Citation Format: Morni Modi, Camila Weber, Qinqin Xu, Kuanling Chen, Ana Cheong, Zachery Nagel, Lynn Harrison. H2S donors as radiosensitizers for glioblastoma multiforme (GBM) [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 2836.
Abstract 2838: Tumor-targeted interleukin-2 enhances the anti-cancer activity of multivalent OncoFAP radioligand therapeutics
Authors: Galbiati Andrea , Bocci Matilde , Gilardoni Ettore , Zana Aureliano , Neri Dario , Cazzamalli Samuele 
Published: April 2023

Abstract
Radioligand Therapeutics (RLTs) are products composed of a tumor-targeting small organic ligand and a biocidal radioactive payload. The therapeutic potential of RLTs strongly depends on their uptake and residence time in tumors and in the healthy structures after administration. Fibroblast Activation Protein (FAP) is a cell surface antigen abundantly expressed in the microenvironment of the majority of solid human cancers of epithelial origin. We have recently described OncoFAP, an ultra-high affinity ligand of FAP whose targeting performances have been validated by Nuclear Medicine in patients with a wide variety of solid malignancies. Multivalent OncoFAP radioligand therapeutics (OncoFAP-RLTs) show high and prolonged tumor uptake with favorable tumor-to-organ ratios. Here we describe the development of novel multivalent OncoFAP-RLTs and their combination with a clinical-stage tumor-targeted interleukin-2 (L19-IL2). We designed and synthesized a set of multivalent OncoFAP derivatives, aiming at maximizing their affinity for the target protein and their residence time in the catalytic pocket of FAP. The multivalent ligands were coupled to the DOTAGA chelator, suitable for radiolabeling with 177Lu for therapeutic application. The tumor targeting properties of the multivalent OncoFAP ligands were investigated in a comparative biodistribution study in SK-RC-52.hFAP tumor-bearing mice. 177Lu-OncoFAP-23 emerged for its superior uptake and residence time in tumors and for its favorable tumor-to-kidney ratio. Moreover, when administered at very low dose 177Lu-OncoFAP-23 exhibited an enhanced anticancer activity compared to a monovalent (177Lu-OncoFAP-DOTAGA) and bivalent (177Lu-BiOncoFAP-DOTAGA) version of OncoFAP. When OncoFAP-based RLTs were combined with L19-IL2, a strong synergistic anticancer activity was observed. This synergy is driven by NK-cell mediated immune response directed to the tumor, with the concomitant release of granzymes and other pro-apoptotic proteins in the tumor microenvironment. Our findings heighten OncoFAP-based RLTs as promising anti-cancer drugs for treatment of FAP-positive tumors, alone or in combination with targeted interleukin-2.
Citation Format: Andrea Galbiati, Matilde Bocci, Ettore Gilardoni, Aureliano Zana, Dario Neri, Samuele Cazzamalli. Tumor-targeted interleukin-2 enhances the anti-cancer activity of multivalent OncoFAP radioligand therapeutics [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 2838.