tbuckley/cabot-search huggingface dataset:
Macrophage‐Specific Hypoxia‐Inducible Factor‐1α Contributes to Impaired Autophagic Flux in Nonalcoholic Steatohepatitis
Authors: Xiaojing Wang, Marcelle de Carvalho Ribeiro, Arvin Iracheta‐Vellve, Patrick Lowe, Aditya Ambade, Abhishek Satishchandran, Terence Bukong, Donna Catalano, Karen Kodys, Gyongyi Szabo
Journal: Hepatology (2018)

Inflammatory cell activation drives diverse cellular programming during hepatic diseases. Hypoxia‐inducible factors (HIFs) have recently been identified as important regulators of immunity and inflammation. In nonalcoholic steatohepatitis (NASH), HIF‐1α is upregulated in hepatocytes, where it induces steatosis; however, the role of HIF‐1α in macrophages under metabolic stress has not been explored. In this study, we found increased HIF‐1α levels in hepatic macrophages in methionine‐choline‐deficient (MCD) diet‐fed mice and in macrophages of patients with NASH compared with controls. The HIF‐1α increase was concomitant with elevated levels of autophagy markers BNIP3, Beclin‐1, LC3‐II, and p62 in both mouse and human macrophages. LysM Cre HIF dPA fl/fl mice, which have HIF‐1α levels stabilized in macrophages, showed higher steatosis and liver inflammation compared with HIF dPA fl/fl mice on MCD diet. In vitro and ex vivo experiments reveal that saturated fatty acid, palmitic acid (PA), both induces HIF‐1α and impairs autophagic flux in macrophages. Using small interfering RNA–mediated knock‐down and overexpression of HIF‐1α in macrophages, we demonstrated that PA impairs autophagy via HIF‐1α. We found that HIF‐1α mediates NF‐κB activation and MCP‐1 production and that HIF‐1α—mediated impairment of macrophage autophagy increases IL‐1β production, contributing to MCD diet‐induced NASH. Conclusion : Palmitic acid impairs autophagy via HIF‐1α activation in macrophages. HIF‐1α and impaired autophagy are present in NASH in vivo in mouse macrophages and in human blood monocytes. We identified that HIF‐1α activation and decreased autophagic flux stimulate inflammation in macrophages through upregulation of NF‐κB activation. These results suggest that macrophage activation in NASH involves a complex interplay between HIF‐1α and autophagy as these pathways promote proinflammatory overactivation in MCD diet‐induced NASH.
Abstract 3586: A universal killer T-cell for adoptive cell therapy of cancer
Authors: Pierre Dillard, Nadia Mensali, June Helen Myklebust, Mickael Hebeisen, Gjertrud Skorstad, Marit Renee Myrhe, Anne Fåne, Gustav Gaudernack, Gunnar Kvalheim, Else Marit Inderberg, Sebastien Wälchli
Journal: Cancer Research (2018)

Abstract T cell-mediated immunotherapy of cancer has achieved remarkable results in hard-to-beat cancers. The main challenge of adoptive T-cell transfer (ACT) is its labor intensive and costly production and logistics as well as its dependency on the quality of the patient's T cells. To overcome these hurdles we have designed a universal cell line for TCR expression by modifying the FDA-approved NK cell line, NK-92. Advantages of using this cell line is that it is easy to expand and can readily be genetically engineered. However, tumor cell recognition and killing by NK-92 is not antigen specific. This can be controlled by introducing an antigen receptor, such as a chimeric antigen receptor (CAR) or, as in the current work, a TCR. We herein present evidence that NK-92 can be modified to become a T cell-like lymphocyte which we named UK-92 cell (Universal Killer). UK-92 expressing a therapeutic TCR showed conserved binding capacity to the cognate pMHC. Phosphoflow cytometry results indicated that the introduced TCR was able to mediate intracellular signaling upon either crosslinking or cognate pMHC binding. Our data showed that both early and late TCR signalling players were activated in a TCR-specific manner (anti-CD3/anti-CD28 stimulation) and further in a pMHC specific manner. In vitro functional assays using TCRs isolated from both CD8 and CD4 T cells demonstrated that UK-92-TCR could be stimulated in a pMHC-specific manner and, importantly, could kill tumor cells specifically even in a tumor-like microenvironment such as spheroids. We have now shown in vitro that UK-92 cells are as specific and potent as redirected T cells to kill target cells. Finally, encouraging in vivo data showed that mice receiving UK92 cells expressing a therapeutic TCR experienced reduction in tumor load and enhanced survival compared with control mice. If confirmed, the use of UK92 as a universal cell line might pave the way to truly off-the-shelf therapeutic effector cells for cancer immunotherapy and leading to drastic reduction of cell production time, logistic and cost. Citation Format: Pierre Dillard, Nadia Mensali, June Helen Myklebust, Mickael Hebeisen, Gjertrud Skorstad, Marit Renee Myrhe, Anne Fåne, Gustav Gaudernack, Gunnar Kvalheim, Else Marit Inderberg, Sebastien Wälchli. A universal killer T-cell for adoptive cell therapy of cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3586.
Abstract 3713: Targeting β-catenin pathway by nanoparticle-mediated delivery of norcantharidin impairs the stemness of triple negative breast cancer cells
Authors: Yunfei Li, Zhishan Wang, Yajuan Xiao, Chengfeng Yang
Journal: Cancer Research (2018)

Abstract Background Norcantharidin (NCTD) is known as an inhibitor of serine/threonine protein phosphatase types 1 and 2A (PP1/PP2A) and exhibits potent anticancer efficacies. Our preliminary study identified NCTD interfering with the interaction between phospho-β-catenin and PP1/PP2A could downregulate the levels of non-phospho β-catenin in the cytoplasm in Triple Negative Breast Cancer (TNBC) cells. As β-catenin pathway plays an important role in cancer stemness, we hypothesize that NCTD could prevent β-catenin nuclear translocation, impair the stemness and reduce the metastasis of TNBC. However, the inherent features of short half-life and nephrotoxicity hinder NCTD's further in vivo application. The goal of this study was to test the therapeutic effect of targeting β-catenin pathway by nanoparticle-mediated delivery of NCTD against TNBC. Experimental Design NCTD was loaded into an integrin α5 targeting nanoparticle (NP) (RGD-LPH) via complexing with polyethyleneimine. The targeting efficiency of RGD-LPH was tested by the in vivo imaging of RGD-LPH labeled by cyanine 7.5 (cy7.5). The effects of NP-mediated delivery NCTD on TNBC cell malignant behaviors and tumor metastasis were tested in various in vitro and in vivo models. Results We found that RGD-LPH NP-mediated delivery of NCTD significantly reduced the active and total β-catenin levels as evidenced by Western Blot. The self-renewal capacity was remarkably blocked by RGD-LPH, as examined by sphere formation. RGD-LPH also suppressed features of cancer cells related to proliferation, as examined by MTT, clonogenic assay and soft agar colony formation. In particular, compared to free NCTD, RGD-LPH demonstrated extended-release profile and long-last effect in the long-term experiments such as clonogenic assay and soft agar colony formation assay. In vivo, systemic delivery of RGD-LPH(cy7.5) indicated a preferential uptake in primary tumor and lung of the tumor-bearing mice. Efficacy studies indicated reduced metastasis of mice from RGD-LPH treatment. Conclusion: Collectively, our results strongly suggest that NCTD being encapsulated in RGD-LPH, which warrants further development for preclinical and clinical studies as a novel TNBC therapeutic approach, is a robust modulator of Wnt signaling with anti-metastasis activity. Citation Format: Yunfei Li, Zhishan Wang, Yajuan Xiao, Chengfeng Yang. Targeting β-catenin pathway by nanoparticle-mediated delivery of norcantharidin impairs the stemness of triple negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3713.
Vasoconstrictor Potential of Coronary Aspirate From Patients Undergoing Stenting of Saphenous Vein Aortocoronary Bypass Grafts and Its Pharmacological Attenuation
Authors: PetraKleinbongard, DirkBöse, TheodorBaars, StefanMöhlenkamp, ThomasKonorza, SandraSchöner, MiriamElter-Schulz, HolgerEggebrecht, HubertusDegen, MichaelHaude, BodoLevkau, RainerSchulz, RaimundErbel, GerdHeusch
Journal: Circulation Research (2011)

Rationale:Stent implantation into atherosclerotic plaques releases, apart from particulate debris, soluble substances that contribute to impaired microvascular perfusion. Objective:To quantify the release of vasoconstrictors and to determine the efficacy of coronary dilators to attenuate their action. Methods and Results:Using a distal protection/aspiration device, coronary arterial blood was retrieved before and during stenting in 22 patients with severe saphenous vein aorto-coronary bypass stenoses. The release of catecholamines, endothelin, serotonin, thromboxane B2, and tumor necrosis factor (TNF)α was measured. The response of rat mesenteric arteries with intact (+E) and denuded (−E) endothelium to aspirate plasma was normalized to that by KCl. Responses to selective receptor blockade, adenosine, nitroprusside, and verapamil against the aspirate-induced constriction were determined. The coronary arterial plasma withdrawn before stenting induced 21±5% and the aspirate plasma after stenting induced 95±...
Switchable CAR-T cells mediate remission in metastatic pancreatic ductal adenocarcinoma
Authors: Deepak Raj, Ming-Hsin Yang, David Rodgers, Eric N Hampton, Julfa Begum, Arif Mustafa, Daniela Lorizio, Irene Garces, David Propper, James G Kench, H M Kocher, Travis S Young, Alexandra Aicher, Christopher Heeschen
Journal: Gut (2018)

Objective Pancreatic ductal adenocarcinoma (PDAC) is a disease of unmet medical need. While immunotherapy with chimeric antigen receptor T (CAR-T) cells has shown much promise in haematological malignancies, their efficacy for solid tumours is challenged by the lack of tumour-specific antigens required to avoid on-target, off-tumour effects. Switchable CAR-T cells whereby activity of the CAR-T cell is controlled by dosage of a tumour antigen-specific recombinant Fab-based ‘switch’ to afford a fully tunable response may overcome this translational barrier. Design In this present study, we have used conventional and switchable CAR-T cells to target the antigen HER2, which is upregulated on tumour cells, but also present at low levels on normal human tissue. We used patient-derived xenograft models derived from patients with stage IV PDAC that mimic the most aggressive features of PDAC, including severe liver and lung metastases. Results Switchable CAR-T cells followed by administration of the switch directed against human epidermal growth factor receptor 2 (HER2)-induced complete remission in difficult-to-treat, patient-derived advanced pancreatic tumour models. Switchable HER2 CAR-T cells were as effective as conventional HER2 CAR-T cells in vivo testing a range of different CAR-T cell doses. Conclusion These results suggest that a switchable CAR-T system is efficacious against aggressive and disseminated tumours derived from patients with advanced PDAC while affording the potential safety of a control switch.
Regulated Expression and Role of c-Myb in the Cardiovascular-Directed Differentiation of Mouse Embryonic Stem Cells
Authors: MasayoshiIshida, OmarEl-Mounayri, StevenKattman, PeterZandstra, HiroshiSakamoto, MinetaroOgawa, GordonKeller, MansoorHusain
Journal: Circulation Research (2012)

Rationale:c-myb null (knockout) embryonic stem cells (ESC) can differentiate into cardiomyocytes but not contractile smooth muscle cells (SMC) in embryoid bodies (EB). Objective:To define the role of c-Myb in SMC differentiation from ESC. Methods and Results:In wild-type (WT) EB, high c-Myb levels on days 0–2 of differentiation undergo ubiquitin-mediated proteosomal degradation on days 2.5–3, resurging on days 4–6, without changing c-myb mRNA levels. Activin-A and bone morphogenetic protein 4–induced cardiovascular progenitors were isolated by FACS for expression of vascular endothelial growth factor receptor (VEGFR)2 and platelet-derived growth factor receptor (PDGFR)α. By day 3.75, hematopoesis-capable VEGFR2+ cells were fewer, whereas cardiomyocyte-directed VEGFR2+/PDGFRα+ cells did not differ in abundance in knockout versus WT EB. Importantly, highest and lowest levels of c-Myb were observed in VEGFR2+ and VEGFR2+/PDGFRα+ cells, respectively. Proteosome inhibitor MG132 and lentiviruses enabling induci...
Abstract 5664: Chemical proteomic platforms to probe metabolic heterogeneity in the tumor microenvironment
Authors: Raymond E. Moellering
Journal: Cancer Research (2018)

Abstract Proteomic platforms provide quantitative snapshots of the proteins present in a biological sample. Despite widespread use, these methods typically require homogenization of samples, signal-averaging over thousands-to-millions of cells and provide no information on protein function. This talk will focus on several novel technologies to circumvent these limitations and to interrogate protein activity within the complex architecture of biological samples, and on spatial scales ranging from single cells to whole organisms. At the single cell level we have developed a chemical proteomic technology, activity-dependent proximity ligation, which enables multiplexed quantification of enzyme activity with high spatial resolution in cellular co-culture and in patient tissue samples. Application of this platform has enabled detection of several metastasis-associated biomarker activities in primary patient samples, as well as simultaneous detection of activity and mRNA expression profiles. These data provide insight into the progression lineage of metastatic ovarian cancers. On the other end of the spectrum, we have recently developed a first-in-class activity-dependent PET imaging probe that specifically and irreversibly labels one of these enzyme biomarkers in vivo. This provides an opportunity to detect aggressive tumor populations, both at sites of primary occurrence and metastasis, with this novel PET radiotracer. Together, these approaches offer new paradigms for studying protein function in basic and translational biology and medicine, ideally providing new tools and biological hypotheses to understand and target tumor malignancy. Citation Format: Raymond E. Moellering. Chemical proteomic platforms to probe metabolic heterogeneity in the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5664.
Abstract 3665: Longitudinal monitoring of prostate cancer evolution by plasma genome sequencing
Authors: Naveen Ramesh, Emi Sei, Ruli Gao, Pei Ching Tsai, Christopher Logothetis, Amado J. Zurita, Nicholas E. Navin
Journal: Cancer Research (2018)

Abstract Castration-resistant prostate cancer (CRPC) is associated with poor survival and frequent metastasis to the bone. Disease progression and evolution of therapy resistance have been challenging to study, in part due to the difficulty in collecting invasive core biopsy samples longitudinally over time. While liquid biopsy methods hold great potential for longitudinal monitoring, they are currently limited to targeted measurements of single genes, or small cancer gene panels. To address this challenge, we developed an unbiased whole-genome sequencing method called PEGASUS (Plasma Exome and Genome Analysis by Size-Selection and Unbiased Sequencing) that enables the detection of copy number aberrations and exome mutations from circulating-tumor DNA (ctDNA). We applied PEGASUS to 15 CRPC patients, which identified common driver mutations and copy number aberrations in plasma DNA including AR, MYC, RB1 and PTEN. We compared matched ctDNA and metastatic tumor samples in 8 CRPC patients, which showed a high correlation (r=0.79) of copy number profiles and moderate concordance of exome mutations (~56%), but also revealed a number of ctDNA and metastasis-specific mutations. We further applied PEGASUS to analyze longitudinal samples from 10 prostate cancer patients that were treated with chemotherapy or androgen inhibition to delineate clonal evolution in response to therapy. Clonal subpopulations and dynamics were inferred from mutation frequencies, revealing both chemoresistant and chemosensitive clones. These data showed that while a subset of point mutations were under positive and negative selection; most copy number aberrations remained stable and did not change in response to therapy. Our data show the technical feasibility of performing whole-genome and exome profiling of plasma DNA and pave the way for future clinical applications in biomarker discovery and informing clinical treatment decisions in prostate cancer patients. Citation Format: Naveen Ramesh, Emi Sei, Ruli Gao, Pei Ching Tsai, Christopher Logothetis, Amado J. Zurita, Nicholas E. Navin. Longitudinal monitoring of prostate cancer evolution by plasma genome sequencing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3665.
Abstract 4367: The small GTPase ARF6 is necessary for melanomagenesis
Authors: Lehi Acosta, Aaron Rogers, Jingfu Peng, Alan Mueller, Zongzhong Tong, Donghan Shin, Jae Hyuk Yoo, Dean Y. Li, Shannon J. Odelberg, Sheri L. Holmen, Allie H. Grossmann
Journal: Cancer Research (2018)

Abstract ARF6 is a member of the adenosine diphosphate (ADP)-ribosylation factor (ARF) family of small GTPases and is part of the RAS superfamily. We have shown that pharmacologic inhibition of ARF6 can inhibit proliferation of uveal melanoma and invasion/metastasis of cutaneous melanoma. Most cutaneous melanomas are defined by mutually-exclusive driver mutations in BRAF, NRAS, or NF1. These subtypes share similar mechanisms of oncogenesis because they align along a continuum of RAS signaling. Mutations in the PI3K pathway occur in a fraction of all melanoma subtypes, indicating a selective advantage for PI3K/AKT activation beyond BRAF or RAS activation. We and others have reported ARF6-dependent ERK activation occurring upon either extracellular ligand stimulation or mutant oncoprotein signaling. Recently, ARF6 has also been shown to be upstream of PI3K signaling. Based on these findings, we hypothesized that ARF6 is critical for both the RAF and PI3K arms of the RAS pathway and that loss of ARF6 will impede tumor growth driven by these oncogenes. We tested this hypothesis using human melanoma cell lines and with an inducible, genetically engineered mouse model (GEMM) of melanoma. Our data reveal that pharmacologic inhibition of ARF6 can reduce proliferation of BRAF, NRAS and NF1-mutated melanoma cells. Furthermore, loss of Arf6 causes a reduction in mean tumor size and improves survival in BRAFV600E/CDKN2ANULL/PTENNULL/AKTE17K and BRAFV600E/CDKN2ANULL/PTENNULL mice. These results indicate that ARF6 is necessary for melanomagenesis and for oncogenic BRAF and/or PI3K signaling. Our data suggest that ARF6 may be an actionable node in the RAS pathway and that therapeutic targeting of ARF6 has the potential to improve current treatment methods for progressive disease. Citation Format: Lehi Acosta, Aaron Rogers, Jingfu Peng, Alan Mueller, Zongzhong Tong, Donghan Shin, Jae Hyuk Yoo, Dean Y. Li, Shannon J. Odelberg, Sheri L. Holmen, Allie H. Grossmann. The small GTPase ARF6 is necessary for melanomagenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4367.
Long-Term Beneficial Effect of Late Reperfusion for Acute Anterior Myocardial Infarction With Percutaneous Transluminal Coronary Angioplasty
Authors: HajimeHorie, MasayukiTakahashi, KazuoMinai, MasafumiIzumi, AtsushiTakaoka, MasatoNozawa, HiroshiYokohama, TamotsuFujita, TaizoSakamoto, OsamuKito, HiroshiOkamura, MasahikoKinoshita
Journal: Circulation (1998)

Background—Although the short-term and long-term beneficial effects of early coronary revascularization by primary PTCA or thrombolytic therapy have been established for acute myocardial infarction, thrombolytic therapy >24 hours after the onset of acute myocardial infarction has not been shown to improve clinical outcome. The purpose of this study was to assess the effect of late revascularization by primary PTCA over a 5-year period. Methods and Results—Eighty-three patients with initial Q-wave anterior myocardial infarction >24 hours after onset were randomized into a PTCA group (n=44) and a no-PTCA group (n=39). Long-term follow-up was conducted with regard to end points, which included cardiac death, nonfatal recurrence of myocardial infarction, and development of congestive heart failure. Left ventricular ejection fraction and regional wall motion at 6 months after myocardial infarction were similar in the 2 groups. Left ventricular end-diastolic and end-systolic volume indexes were significantly sm...
Quantitative approaches to variant classification increase the yield and precision of genetic testing in Mendelian diseases: the case of hypertrophic cardiomyopathy
Authors: Roddy Walsh, Francesco Mazzarotto, Nicola Whiffin, Rachel Buchan, William Midwinter, Alicja Wilk, Nicholas Li, Leanne Felkin, Nathan Ingold, Risha Govind, Mian Ahmad, Erica Mazaika, Mona Allouba, Xiaolei Zhang, Antonio de Marvao, Sharlene M. Day, Euan Ashley, Steven D. Colan, Michelle Michels, Alexandre C. Pereira, Daniel Jacoby, Carolyn Y. Ho, Kate L. Thomson, Hugh Watkins, Paul J. R. Barton, Iacopo Olivotto, Stuart A. Cook, James S. Ware
Journal: Genome Medicine (2019)

International guidelines for variant interpretation in Mendelian disease set stringent criteria to report a variant as (likely) pathogenic, prioritising control of false-positive rate over test sensitivity and diagnostic yield. Genetic testing is also more likely informative in individuals with well-characterised variants from extensively studied European-ancestry populations. Inherited cardiomyopathies are relatively common Mendelian diseases that allow empirical calibration and assessment of this framework. We compared rare variants in large hypertrophic cardiomyopathy (HCM) cohorts (up to 6179 cases) to reference populations to identify variant classes with high prior likelihoods of pathogenicity, as defined by etiological fraction (EF). We analysed the distribution of variants using a bespoke unsupervised clustering algorithm to identify gene regions in which variants are significantly clustered in cases. Analysis of variant distribution identified regions in which variants are significantly enriched in cases and variant location was a better discriminator of pathogenicity than generic computational functional prediction algorithms. Non-truncating variant classes with an EF ≥ 0.95 were identified in five established HCM genes. Applying this approach leads to an estimated 14–20% increase in cases with actionable HCM variants, i.e. variants classified as pathogenic/likely pathogenic that might be used for predictive testing in probands' relatives. When found in a patient confirmed to have disease, novel variants in some genes and regions are empirically shown to have a sufficiently high probability of pathogenicity to support a "likely pathogenic" classification, even without additional segregation or functional data. This could increase the yield of high confidence actionable variants, consistent with the framework and recommendations of current guidelines. The techniques outlined offer a consistent and unbiased approach to variant interpretation for Mendelian disease genetic testing. We propose adaptations to ACMG/AMP guidelines to incorporate such evidence in a quantitative and transparent manner.