bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4993: Label-free metabolic imaging of T cell response to the tumor microenvironment
Authors: VanDommelen Ava , Gillette Amani , Patankar Manish S. , Skala Melissa 
Published: September 2020

Abstract
Numerous factors including low pH, low glucose, and high lactic acid levels in tumors create an immunosuppressive tumor microenvironment. This immunosuppression is partially due to the metabolic requirements of T cell activation, which are difficult to meet in the tumor microenvironment. The goal of this study is to use label-free optical metabolic imaging (OMI) of the metabolic co-factors NAD(P)H and FAD in CD3-conjugated (activated) and control T cells to monitor metabolic changes of T cells within tumor micro environmental conditions. Single-cell measurements include the optical redox ratio (fluorescence intensity of NAD(P)H divided by FAD), which measures the oxidation-reduction state of the cell, and the mean fluorescence lifetimes (τm) of NAD(P)H and FAD, which measure enzyme-binding activity in the cell. Jurkat T cells were conjugated with CD3 antibody over a two-day incubation period to achieve activation. Control and CD3-conjugated cells were exposed to environments with increasing concentrations of lactic acid (0-20 mM), hydrochloric acid (HCl; pH 6.4- 7.4), and glucose deprivation to mimic the tumor microenvironment. OMI monitored metabolic changes on a single-cell level without confounding factors associated with cell surface marker labeling. Treatment with lactic acid resulted in a significant dose-dependent decrease in the FAD τm and optical redox ratio (p&lt;0.005). Along with changes in metabolism, OMI also resolved impaired mobility of the T cells with increasing lactic acid concentrations. Lactic acid affects both pH and metabolism, so cells were separately exposed to different concentrations of HCl (pH 6.4 - pH 7.4) to monitor pH effects alone. In control cells, the optical redox ratio decreased with decreasing pH (p&lt;0.005). Conversely, in the presence of CD3, the optical redox ratio increased with decreasing pH (p&lt;0.005). CD3 conjugation in glucose deprived environments also resulted in increases in both optical redox ratio and NAD(P)H τm. Cellular viability assays indicate apoptosis only in the lowest pH environment (pH 6.4, p&lt;0.005). These studies indicate that OMI is a promising label-free approach to investigate the effects of the tumor microenvironment on T cell metabolism, activation, and drug response.
Citation Format: Ava VanDommelen, Amani Gillette, Manish S. Patankar, Melissa Skala. Label-free metabolic imaging of T cell response to the tumor microenvironment [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4993.
Abstract 500: Discovering epigenetic regulators of cells states in triple negative breast cancers
Authors: Burnett Joseph Patrick , Johnson Garrett , Truchan Nathan , Brooks Michael , Wicha Max S. , Sun Duxin 
Published: September 2020

Abstract
The implementation of targeted therapies have been instrumental in achieving clinical responses for ER, PR, and HER2 receptor expressing breast cancers. However, triple negative breast cancers in themselves do not respond to such therapies and are highly heterogeneous diseases with significantly worse progression and survival. Our preliminary data and literature have also shown that in TNBC different cancer cell subpopulations are present in any given tumor, each with its own unique chemo-response and resistance. To discover novel targets regulating cancer cell states in TNBCs, we have created a reporter cell line using CRISPR/Cas9 mediated knock-in of fluorescent reporter proteins after the endogenous loci of major cell type markers in SUM149. Using this reporter line allows for the direct visualization of ALDH1A3+/- and CD24+/- cells, facilities tracking of distinct cell state transitions in a given cell, and allows for lineage tracing following cell divisions. These reporter cells were subjected to HTS-FACS analysis after exposure to a siRNA library of known epigenetic regulators. Following secondary screening, three siRNA targets are able to uniquely inhibit distinct subpopulations of cells within the cell line DCAF1 (ALDH-/CD24- cells), MLL2 (ALDH-/CD24+ cells), and TRIM24 (ALDH+ cells). In combination, these siRNA's proportionally eliminated all cell types. Due to the inherent heterogeneity of TNBCs we then sought to determine if efficacy was consistent or unique across different cell lines with varying percentages of CD44, CD24, and Aldefluor+ cell populations. Most notably, in MDA-MB-468 which contains similar cell population percentages to SUM149 each siRNA was capable of reducing cell proliferation. Similarly, TRIM24 was also capable of reducing cellular proliferation in BT20, MLL2 in MDA-MB-231, and DCAF1 in HCC1937. Knockdown of TRIM24 consistently reduced Aldefluor+ cells in MDA-MB-468 and BT20 cells. Finally, coupled with sc-Omics technologies we utilized this efficacy data identify a subset of TNBC patients likely to respond to therapies which will target these genes in the future.
Citation Format: Joseph Patrick Burnett, Garrett Johnson, Nathan Truchan, Michael Brooks, Max S. Wicha, Duxin Sun. Discovering epigenetic regulators of cells states in triple negative breast cancers [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 500.
Abstract 5031: Salmonella-mediated suppression of Akt/mTOR signaling inhibits metastasis via downregulation of Snail expression in melanoma
Authors: Pangilinan Christian R. , Lee Che-Hsin 
Published: September 2020

Abstract
The expression of EMT inducer Snail in many cancer types has been implicated in high-grade malignancies and correlated with poor prognosis. Tumor-intrinsic Snail not only has direct effects on metastasis but also impacts tumor microenvironment by altering secretomes. Our previous reports demonstrated that systemic administration of Salmonella significantly reduced tumor growth and metastatic nodules of tumor-bearing mice, in part, by coaxing changes in the tumor microenvironment. These led us to hypothesize that Salmonella targets metastasis inducers. Here, we use B16F10 and K1735 mouse melanoma cells to investigate the anti-metastatic potentials of Salmonella against melanoma. Immunoblotting revealed that the expression of Snail in both B16F10 and K1735 melanoma cell lines was significantly downregulated following Salmonella treatment in a dose-dependent manner. Likewise, the phosphorylation of Akt and mTOR was significantly reduced after Salmonella treatment. Our findings further revealed that in vitro metastatic activities of Salmonella-treated melanoma cells were significantly inhibited as demonstrated in the wound-healing and Transwell assays. When transfected with constitutively active AKT plasmid, B16F10 and K1735 markedly increased the phosphorylation of Akt and mTOR accompanied by an elevated Snail expression. Interestingly, Salmonella treatment reversed the Akt/mTOR and Snail upregulation of cells overexpressing Akt. Our data therefore suggest that Salmonella significantly reduces melanoma metastasis by downregulating Snail.
Citation Format: Christian R. Pangilinan, Che-Hsin Lee. Salmonella-mediated suppression of Akt/mTOR signaling inhibits metastasis via downregulation of Snail expression in melanoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5031.
Abstract 5034: Effect of estrogen on heteronemin-induced anti-proliferation in breast cancer cells
Authors: Li Zi-Lin , Pan Yi-shin , Huang Tung-Yung , Shih Ya-Jung , Chen Yi-Ru , Tang Heng-Yuan , Whang-Peng Jacqueline , Lin Hung-Yun , Davis Paul J. , Wang Kuan 
Published: September 2020

Abstract
Estrogen, 17β-estradiol (E2) has multiple functions in breast cancers including stimulating cancer growth and interfering with chemotherapeutic efficacy. Heteronemin, a marine sesterterpenoid-type natural product, processes anti-proliferative effect in cancer cells. In the current study, we investigate the antitumor mechanism of heteronemin in breast cancer cells and further explore its molecular targets. Heteronemin exhibited potent cytotoxic effects against breast cancer cells. On the other hand, E2 stimulated cancer cell growth. Heteronemin and E2 showed different effects on gene expression of proliferation, angiogenesis or growth factor receptors in breast cancer cells. NanoString® analysis was further used to detect mRNAs, heteronemin repressed gene expression of transforming growth factor (TGF)-β, mothers against decapentaplegic homolog (SMAD) and PCNA, whose protein products play important roles in regulating cell growth, angiogenesis, and metastasis. The results indicate that heteronemin was able to modulate cell adhesion, expression of extracellular matrix (ECM) receptors, TGF-β pathway, cell motility, membrane integration, metastasis response, matrix metalloproteinase (MMP) remodeling, regulation of metabolism, sprouting angiogenesis, transcription factors, and vasculogenesis in breast cancer cells. However, those effects were partially reversed by E2. Furthermore, Heteronemin and E2 altered several signaling transduction pathways. In summary, this study provides insight into the complex pathways by which anti-proliferation is induced by heteronemin in E2-depleted and -repleted environments.
Citation Format: Zi-Lin Li, Yi-shin Pan, Tung-Yung Huang, Ya-Jung Shih, Yi-Ru Chen, Heng-Yuan Tang, Jacqueline Whang-Peng, Hung-Yun Lin, Paul J. Davis, Kuan Wang. Effect of estrogen on heteronemin-induced anti-proliferation in breast cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5034.
Abstract 5060: Establishment and characterization of humanized mouse NPC-PDX model for testing immunotherapy
Authors: LIU Wai Nam , FONG Shin Yie , TAN Wilson Wei Sheng , TAN Sue Yee , LIU Min , IYER Narayanan Gopalakrishna , LIM Darren Wan-Teck , CHEN Qingfeng 
Published: September 2020

Abstract
Epstein-Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC) is common in Southern China and South-East Asia. Immune checkpoint blockade (ICB) is being studied in NPC and initial monotherapy ICB studies show early promise. Currently, lack of representative NPC models limit the preclinical studies to evaluate the efficacy and safety of novel ICB and combination regimens. Hence, there is a need to develop and characterize a mouse model with human immune system for testing immunotherapy. Here, we engrafted NPC-PDX in immunodeficient NSG mice, with a successful growth rate of 16.7% (6 out of 36), and humanized mouse NPC-PDX model was established subsequently. Of note, EBV latent membrane protein-1 was retained in the NPC in both NSG and humanized mice as revealed by IHC staining. There was an increase in tumor-infiltrating CD4+ T cells and a decrease in CD8+ cytotoxic T cells in humanized mice, when compared to the immune cells profile in circulation. Intriguingly, the expression levels of various exhaustion markers, namely PD-1, CTLA4 and TIM-3 were upregulated, particularly in the tumor-infiltrating T cells. Therefore, the use of ICB, either alone or in combination, may restore the function of anti-tumor immune effector cells. Taken together, we have established a humanized mouse NPC-PDX model, which plausibly provides a robust platform to test for the efficacy and safety of ICB and combination therapy regimens.
Citation Format: Wai Nam LIU, Shin Yie FONG, Wilson Wei Sheng TAN, Sue Yee TAN, Min LIU, Narayanan Gopalakrishna IYER, Darren Wan-Teck LIM, Qingfeng CHEN. Establishment and characterization of humanized mouse NPC-PDX model for testing immunotherapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5060.
Abstract 5062: Modeling NK cells activities in full human immune system mouse models
Authors: Yin Kiave Yune Ho Wang 
Published: September 2020

Abstract
The cytotoxic potential of NK-cells for the elimination of cancer cells represents the basis for many important ongoing clinical efforts. These efforts can now be supported by the humanized immune system mouse models (Hu-Mice), which offer an improved in vivo system for translating therapies to the clinic.
Using hydrodynamic injection of human IL-15, TransCure bioServices has developed a NK boosted hu-Mice (CD34+ engrafted NCG) providing sufficient and functional human NK cells. The transient and physiological expression of IL-15 allow the expansion of human NK cells in the context of a full and stable human immune system bringing numerous advantages compared to the NK infused mouse model in IL15 transgenic mouse models. One week after boost, three times more NK cells are present in peripheral blood of IL15-boosted hu-Mice and this level is stable up to five weeks, providing sufficient time to engraft tumor cells and test drug efficacy. A better infiltration of the tumor by human NK cells is also visible in IL-15 boosted mice. Furthermore, the efficacy of ADCC dependent drugs (administered prophylactically or therapeutically) have been highlighted in IL-15 boosted hu-Mice engrafted with melanoma and multiple myeloma cells.
In conclusion, the IL-15 boosted hu-Mice provide a reliable and validated in vivo model to better assess the efficacy and MOA of drug depending on NK cell activity.
Citation Format: Kiave Yune Ho Wang Yin. Modeling NK cells activities in full human immune system mouse models [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5062.
Abstract 5079: Immuno-competent 3d in vitro tumor models as novel platform to assess combinatorial biologics therapy
Authors: Agarkova Irina , Buschmann Nicole , Strebel Silvan , Wolf Armin , Chiovaro Francesca 
Published: September 2020

Abstract
Background: Immunotherapy has revolutionized cancer treatment, but clinical success is still limited to a subset of patients, underlining the need for novel therapeutic targets. Available ex vivo animal platforms for drug screening show limitations due to only partial recapitulation of tumor-intrinsic features and of the interactions of tumors with stromal/immune cells. Here, we review our 3D InSightTM tumor models, a complex in-vitro platform allowing to study the efficacy of immune-targeting agents on tumor infiltration and killing by peripheral blood mononuclear cells (PBMCs) and to acquire insights on the modulatory capacity of immune checkpoint inhibitors.
Aim: In the present study we have developed a 3D tumor - immune cell coculture model and evaluated its application to study the effect of immune checkpoint inhibitors on tumor viability, cytokine secretion, and immune cell infiltration.
Material and Methods: Lung carcinoma GFP-A549 cells were cultured with dermal fibroblasts and PBMCs in Akura 96/384-well plates. Immune cells were pre-treated using different activation protocols to induce priming and pT-cell exhaustion. We assessed the effect of PD-1 inhibitors in combination with cytokines for potential synergistic anti-cancer effects. Killing activity and effector function of T cells were evaluated by measuring tumor microtissue size by automatic stage fluorescence microscopy and measuring release of IL-6, TNF, IFNγ and GM-CSF with a MAGPIXTM Luminex system over time. Immune tumor infiltration was quantified using fluorescent staining of CD3+ cells and the effect of the immune cell attack on the tumor cells was assessed by IHC staining.
Results: Combination treatment with cytokines and anti-PD-1 led to higher release of IL-6/TNF/IFNγ/GM-CSF and increased tumor microtissue infiltration by CD3+ lymphocytes, stronger expression of Granzyme B, resulting in enhanced tumor killing and apoptosis.
Conclusion: Our 3D InSightTM tumor models can be successfully employed for optimal high throughput screening of biologics to unveil the efficacy and anti-tumor efficacy of immune-targeting therapeutic agents.
Citation Format: Irina Agarkova, Nicole Buschmann, Silvan Strebel, Armin Wolf, Francesca Chiovaro. Immuno-competent 3d in vitro tumor models as novel platform to assess combinatorial biologics therapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5079.
Abstract 510: Novel glutamine antagonist JHU-395 suppresses MYC-driven medulloblastoma growth and induces apoptosis
Authors: Pham Khoa , Maxwell Micah , Sweeney Heather , Alt Jesse , Rais Rana , Slusher Barbara S. , Eberhart Charles G. , Raabe Eric H. 
Published: September 2020

Abstract
Medulloblastoma is the most common malignant pediatric brain tumor. Genomic amplification of MYC is a hallmark of a subset of poor-prognosis medulloblastoma. MYC upregulates glutamine metabolism, and medulloblastoma tumors with evidence of high glutamate have a poor prognosis. We modified the naturally occurring glutamine analog 6-diazo-5-oxo-l-norleucine (DON) to increase its lipophilicity and its stability in plasma, creating a prodrug termed JHU-395. We hypothesized that this prodrug would have enhanced cell penetration compared to DON and would kill MYC-expressing medulloblastoma. JHU-395 treatment caused decreased growth and increased apoptosis in human MYC-expressing medulloblastoma cell lines at lower concentrations than DON. In D283MED for example IC50 for DON is 20 uM while JHU 395 IC50 is 1 uM (p value &lt; 0.05). Similar results were obtained with other high-MYC medulloblastoma cell lines, including D425MED and D341MED. JHU-395 induced apoptosis at low micromolar concentrations as measured by cleaved PARP Western blot and cleaved caspase 3 (CC3) immunofluorescence (60% CC3+ cells compared to 30% CC3+ in control p&lt;0.05). Parenteral administration of JHU-395 in Nu/Nu mice led to the accumulation of micromolar concentrations of DON in brain as measured by mass spectrometry. These data suggest that JHU-395 may have activity against high MYC medulloblastoma in vivo, and testing this prodrug against MYC-driven medulloblastoma orthotopic xenografts is currently underway.
Citation Format: Khoa Pham, Micah Maxwell, Heather Sweeney, Jesse Alt, Rana Rais, Barbara S. Slusher, Charles G. Eberhart, Eric H. Raabe. Novel glutamine antagonist JHU-395 suppresses MYC-driven medulloblastoma growth and induces apoptosis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 510.
Abstract 5139: An atlas of perturbed functional proteomics profiles of cancer cell lines
Authors: Zhao Wei , Li Jun , Chen Mei-Ju , Akbani Rehan , Lu Yiling , Mills Gordon , Liang Han 
Published: September 2020

Abstract
In recent years, tremendous efforts have been made to systematically characterize the molecular profiles of tumor tissues from individuals with cancer, laying a critical foundation for elucidating the molecular basis of tumorigenesis and developing biomarker-based diagnostic, prognostic and therapeutic approaches. In particular, cancer genomic data at the DNA or RNA level are being accumulated at an unprecedented speed. However, it remains to be a big challenge in cancer research to systematically understand causality and mechanisms underlying the behaviors of cancer cells. To address it, perturbation experiments are a very powerful approach in which the cells are first modulated by perturbagens and the downstream consequences are then monitored. Recently, large-scale compendia of the phenotypic and cellular effects of perturbed cancer cell lines have been established. However, similar resources for the proteomic responses of perturbed cancer cell lines have yet to be established. Reverse-phase protein arrays (RPPAs) is a powerful targeted functional proteomics approach to studying cancer mechanisms, biomarkers and therapies. This quantitative antibody-based assay is able to assess a large number of protein markers in many samples in a cost-effective, sensitive manner. More recently, we have applied this technology to quantify the protein expression levels of large patient cohorts and cancer cell lines (&gt;8,000 patient samples of 32 cancer types from The Cancer Genome Atlas, &gt;650 cell lines across 19 lineages). Here, using RPPAs, we have generated and compiled the perturbed functional proteomic profiles of &gt;12,000 cancer cell line samples in response to &gt;150 drug compounds and other perturbagens using reverse-phase protein arrays. We show that integrating protein response signals substantially increases the predictive power for drug sensitivity and gains insights into the mechanisms of drug resistance. We build a comprehensive map of “protein-drug” connectivity and develop an open-access, user-friendly data portal for community use. Our study provides a valuable proteomic resource for a broad range of quantitative modeling and biomedical applications.
Citation Format: Wei Zhao, Jun Li, Mei-Ju Chen, Rehan Akbani, Yiling Lu, Gordon Mills, Han Liang. An atlas of perturbed functional proteomics profiles of cancer cell lines [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5139.
Abstract 5161: Validation of drug candidates targeting dormant cancers in live cells as a key for successful translation into the clinic
Authors: Belenky Alexei , Kuznetsova Alexandra , Frid Michael , Chapovsky Felix , Duey Marc , Vilenchik Maria 
Published: September 2020

Abstract
Thermal shift assay was utilized to demonstrate target engagement of drug candidate compounds to their specific targets. We explored novel cancer dormancy targets: DYRK1B and AXL kinases and assessed interaction with corresponding small molecule inhibitors. We demonstrated the correlation of direct target binding with ASMS (affinity selected mass spectrometry) and mechanism of action with cell-based phenotypic assays (using flow cytometry) for previously identified and current clinical candidates.
We demonstrated that the above described approach can characterize and potentially predict clinical activity of novel molecules. Herein we investigated DYRK1B inhibitors from an azaindole-quinoline series. It was shown earlier that these molecules have picomolar activity in in vitro assays against DYRK1 kinases (DYRK1B and DYRK1A) and now we confirmed their DYRK1B specificity in cells as well as their activity against dormant cancers.
We suggest that described inhibitors are valid clinical candidates for targeting cancer dormancy based on direct target binding and engagement in live cancer cells. These compounds in combination with chemotherapeutic agents, have the potential to improve patient outcomes and prolong survival.
Citation Format: Alexei Belenky, Alexandra Kuznetsova, Michael Frid, Felix Chapovsky, Marc Duey, Maria Vilenchik. Validation of drug candidates targeting dormant cancers in live cells as a key for successful translation into the clinic [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5161.