tbuckley/cabot-search huggingface dataset:
Abstract 227: Cardiomyocytes from Human Pluripotent Stem Cells Predict Chemotherapy-Induced Cardiotoxicity
Authors: Maxime Mioulane, Gabor Foldes, Sian Harding
Journal: Circulation Research (2012)

The potential of human pluripotent stem cells-derived cardiomyocytes (hPSC-CM) to mimic human cardiac toxicity tests is largely unknown. Here we investigate the interaction between the chemotherapeutic agents Doxorubicin (DOX) and Lapatinib (LAP) and the protective effect of beta2 adrenergic signalling in hPSC-CM and murine myocytes. We hypothesized that death and survival pathways in hPSC-CM differ from those in animal-derived myocytes and thereby may predict better clinical outcome of chemotherapeutics. Human induced pluripotent or embryonic stem cell H7 cell lines were differentiated into cardiomyocytes (hiPSC-CM and hESC-CM, respectively), treated with drugs and stained with fluorescent dyes (Hoechst 33342, mitochondrial probes, effector caspases activation and impermeant nuclear dyes). Cell death profile was generated from high content screening and analysis (Cellomics). In isolated hPSC-CM, the anthracycline DOX induced apoptosis in a concentration-dependent manner (1-100µM), whereas LAP did not produce any toxicity. However, the combination of the two chemotherapeutic agents worsened the toxicity profile compared to DOX alone (p&lt;0.001, n=12), consistent with clinical observations. In contrast, rat neonates were sensitive to LAP and showed a different cell death profile in DOX and drug combination experiments. Activation of apoptotic caspases in hESC-CM clusters was coincident with the dissipation of the mitochondrial membrane potential, while the potential was preserved in isolated cells during this phase. Beta2 adrenergic stimulation did not reverse DOX pro-apoptotic effect but modified toxicity-related nuclear events. In conclusion, we have shown that hPSC-CM reproduce the clinical adverse cardiac effects after treatment with anti-cancer drugs, making these cells a more relevant system for future drug development than animal-derived cells. However, our data suggest that cell death pathways involved may depend on culture condition for cardiomyocytes.
C-Terminal Tails of Sulfonylurea Receptors Control ADP-Induced Activation and Diazoxide Modulation of ATP-Sensitive K+ Channels
Authors: TetsuroMatsuoka, KenjiMatsushita, YusukeKatayama, AkikazuFujita, KiyoshiInageda, MasayukiTanemoto, AtsushiInanobe, ShizuyaYamashita, YujiMatsuzawa, YoshihisaKurachi
Journal: Circulation Research (2000)

Abstract—The ATP-sensitive K+ (KATP) channels are composed of the pore-forming K+ channel Kir6.0 and different sulfonylurea receptors (SURs). SUR1, SUR2A, and SUR2B are sulfonylurea receptors that are characteristic for pancreatic, cardiac, and vascular smooth muscle–type KATP channels, respectively. The structural elements of SURs that are responsible for their different characteristics have not been entirely determined. Here we report that the 42 amino acid segment at the C-terminal tail of SURs plays a critical role in the differential activation of different SUR-KATP channels by ADP and diazoxide. In inside-out patches of human embryonic kidney 293T cells coexpressing distinct SURs and Kir6.2, much higher concentrations of ADP were needed to activate channels that contained SUR2A than SUR1 or SUR2B. In all types of KATP channels, diazoxide increased potency but not efficacy of ADP to evoke channel activation. Replacement of the C-terminal segment of SUR1 with that of SUR2A inhibited ADP-mediated chann...
<i>FHIT</i>, a Novel Modifier Gene in Pulmonary Arterial Hypertension
Authors: Svenja Dannewitz Prosseda, Xuefei Tian, Kazuya Kuramoto, Mario Boehm, Deepti Sudheendra, Kazuya Miyagawa, Fan Zhang, David Solow-Cordero, Joshua C. Saldivar, Eric D. Austin, James E. Loyd, Lisa Wheeler, Adam Andruska, Michele Donato, Lingli Wang, Kay Huebner, Ross J. Metzger, Purvesh Khatri, Edda Spiekerkoetter
Journal: American Journal of Respiratory and Critical Care Medicine (2018)

Pulmonary arterial hypertension (PAH) is characterized by progressive narrowing of pulmonary arteries, resulting in right heart failure and death. BMPR2 (bone morphogenetic protein receptor type 2) mutations account for most familial PAH forms whereas reduced BMPR2 is present in many idiopathic PAH forms, suggesting dysfunctional BMPR2 signaling to be a key feature of PAH. Modulating BMPR2 signaling is therapeutically promising, yet how BMPR2 is downregulated in PAH is unclear.We intended to identify and pharmaceutically target BMPR2 modifier genes to improve PAH.We combined siRNA high-throughput screening of >20,000 genes with a multicohort analysis of publicly available PAH RNA expression data to identify clinically relevant BMPR2 modifiers. After confirming gene dysregulation in tissue from patients with PAH, we determined the functional roles of BMPR2 modifiers in vitro and tested the repurposed drug enzastaurin for its propensity to improve experimental pulmonary hypertension (PH).We discovered FHIT (fragile histidine triad) as a novel BMPR2 modifier. BMPR2 and FHIT expression were reduced in patients with PAH. FHIT reductions were associated with endothelial and smooth muscle cell dysfunction, rescued by enzastaurin through a dual mechanism: upregulation of FHIT as well as miR17-5 repression. Fhit-/- mice had exaggerated hypoxic PH and failed to recover in normoxia. Enzastaurin reversed PH in the Sugen5416/hypoxia/normoxia rat model, by improving right ventricular systolic pressure, right ventricular hypertrophy, cardiac fibrosis, and vascular remodeling.This study highlights the importance of the novel BMPR2 modifier FHIT in PH and the clinical value of the repurposed drug enzastaurin as a potential novel therapeutic strategy to improve PAH.
Abstract LB-342: Targeted gene control with bifunctional molecules to harness endogenous chromatin-modifying activity
Authors: Anna M. Chiarella, Kyle V. Butler, Jian Jin, Nathaniel A. Hathaway
Journal: Cancer Research (2018)

Abstract Epigenetic pathways are commonly found disrupted in many human diseases including cancer, yet much remains unknown about the mechanisms by which dysregulation of chromatin regulatory pathways leads to disease pathology. To investigate and control these complex interactions in live cells, we developed a novel system that will unveil the underlying mechanisms of proteins responsible for modulating chromatin states using a gene-specific manipulation strategy. To explore how dynamic changes in acetylation levels impact chromatin structure we generated an endogenous reporter system at the murine Oct4 locus with a Gal4 DNA binding array upstream of an EGFP reporter allele. In our system, a FKBP protein fused to a Gal4 DNA-binding-domain serves as a protein anchor to the DNA binding array. Next, we synthesized novel cell-permeable, bifunctional molecules that include FK506 (which binds to FKBP) linked to a component that selectively binds to a histone tail chromatin-modifying enzyme. We characterized the effects of the Chemical Epigenetic Modifiers (CEM)s using flow cytometry, high-content microscopy and chromatin immunoprecipitation. We show that our lead repressive CEM can repress GFP expression by 50%. Next, we have made our system more modular by creating a deactivated Cas9 (dCas9)-FKBP fusion protein. The FKBP portion allows us to recruit our CEMs to any region of the genome to which we design a guide RNA. To study the addition of histone acetylation we utilized our new activating, HAT-binding CEMs and designed our dCas9-FKBP to target low-expression reporters. Our lead activating CEM increases expression over 20-fold. We are now using our activating CEM system to target tumor suppressor genes in hard to treat cancers. To show proof-of-principle, we have targeted the tumor suppressor p53 in a colon cancer cell line. We hypothesize that our activating CEM platform will activate the repressed P53 allele leading to slower tumor cell proliferation and apoptosis. This work represents a novel therapeutic strategy that is capable of modulating gene expression in a reversible and dose-dependent manner. Citation Format: Anna M. Chiarella, Kyle V. Butler, Jian Jin, Nathaniel A. Hathaway. Targeted gene control with bifunctional molecules to harness endogenous chromatin-modifying activity [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 LB-342.
Aggregated Low-Density Lipoprotein Uptake Induces Membrane Tissue Factor Procoagulant Activity and Microparticle Release in Human Vascular Smooth Muscle Cells
Authors: VicentaLlorente-Cortés, MartaOtero-Viñas, SandraCamino-López, OriolLlampayas, LinaBadimon
Journal: Circulation (2004)

Background— Tissue factor (TF) is the main initiator of the arterial blood coagulation system, and aggregated LDL (agLDL) are found in the arterial intima. Our hypothesis is that agLDL internalization by vascular smooth muscle cells (VSMCs) may trigger TF-procoagulant activity. Methods and Results— Cultured human VSMCs were obtained from human coronary arteries of explanted hearts during transplant operations. VSMCs were incubated with native LDL (nLDL) or agLDL. TF mRNA was analyzed by real-time polymerase chain reaction, and cellular and released TF protein antigen were analyzed by Western blot. TF microparticle (MP) content was analyzed by flow cytometry and TF activity by a factor Xa generation test. Both nLDL and agLDL strongly and equally increased TF mRNA and cell membrane protein expression, by approximately 5- and 9-fold, respectively. A sustained TF procoagulant activity was induced by agLDL but not by nLDL (agLDL 2.46±0.22 versus nLDL 0.72±0.12 mU/mg protein at 12 hours). AgLDL increased TF ant...
Atrial Reentrant Tachycardia After Surgery for Congenital Heart Disease
Authors: ThomasPaul, BrittaWindhagen-Mahnert, ThomasKriebel, HaraldBertram, RenateKaulitz, ThomasKorte, MichaelNiehaus, JürgenTebbenjohanns
Journal: Circulation (2001)

Background—The purpose of the present study was to determine the role of a novel, noncontact mapping system for assessing a variety of atrial reentrant tachycardias (ART) in patients after the surgical correction of congenital heart disease. Methods and Results—In 14 patients, an electrophysiological study using the Ensite 3000 system was performed to assess ARTs resistant to medical treatment. Sixteen different forms of ART were inducible in the 14 patients studied. The reentrant circuit of all ARTs could be characterized and localized with respect to anatomic landmarks such as atriotomy scars, intraatrial patches/baffles, and cardiac structures. In 15 of the 16 ARTs (in 13 of the 14 patients), a target area of the reentrant circuit for radiofrequency current application (ie, an area of conduction between 2 anatomical obstacles such as surgical barriers and cardiac structures of electrical isolation) could be localized within the systemic venous atrium. Nine patients exhibited macroreentry, and 4 showed ...
Abstract 3775: Use of RLI-15 a clinical grade fusion protein with IL-15 superagonistic activity for the activation of anti-tumor immune response
Authors: Irena Adkins, Lenka Sadilkova, Nada Hradilova, Jakub Tomala, Barbora Tomalova, Marek Kovar, Romana Mikyskova, Milan Reinis, Guy de Martynoff, David Bechard, Ulrich Moebius, Radek Spisek
Journal: Cancer Research (2018)

Abstract RLI-15, a superagonist fusion protein of interleukin (IL)-15 and the IL-15 receptor α (IL-15Rα) sushi+ domain represents a promising candidate for the induction of anti-tumor immunity. RLI-15 was designed to bypass the need of endogenous IL-15Rα, thereby leveraging the activity of IL-15 in vivo on target immune cells. RLI-15 stimulates the proliferation and the cytotoxic activity of natural killer (NK) cells and memory CD8+ T cells with no significant expansion and activation of regulatory T cell compartment. RLI-15 was previously shown to exhibit a potent anti-metastatic activity in B16F10 melanoma and Renca renal cell carcinoma mouse models. RLI-15 also significantly delayed tumor growth and prolonged survival when combined with anti-PD1 therapy in CT26 and MC38 colon carcinoma models. Here, we report that the combination treatment with clinical-grade RLI-15 and an anti-PD1 antibody leads to a significant anti-tumor efficacy in a TRAMP-C2 prostate cancer mouse model with 70 % of mice remaining tumor free after the treatment. We evaluated the optimal schedule of such combination therapy to set the basis for the design of upcoming clinical trials. We further tested how the administration schedule affects the pharmacodynamics properties of clinical-grade RLI-15 and translates into the anti-tumor efficacy in metastatic Renca and CT26 mouse models. In cynomolgous monkeys, various schedules of administration of RLI-15 showed a dose-dependent expansion of peripheral blood lymphocytes, predominantly of NK cell and memory CD8+ T cell compartments. The toxicity in mice and cynomolgous monkeys was evaluated to determine the maximal tolerated dose of RLI-15. Furthermore, the activity of clinical-grade RLI-15 was tested in vitro on human PBMCs and the superiority over IL-2 and IL-15 stimulatory capacity has been confirmed. The complex analysis of RLI-15 behavior and of the induced anti-tumor immune response will be explored in the design of a planned Phase I clinical study in patients with both solid tumors and hematological malignancies. Citation Format: Irena Adkins, Lenka Sadilkova, Nada Hradilova, Jakub Tomala, Barbora Tomalova, Marek Kovar, Romana Mikyskova, Milan Reinis, Guy de Martynoff, David Bechard, Ulrich Moebius, Radek Spisek. Use of RLI-15 a clinical grade fusion protein with IL-15 superagonistic activity for the activation of anti-tumor immune response [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 3775.
Abstract 2769: Targeting malignant mesothelioma with an antibody against the tumor extracellular matrix protein fibulin-3
Authors: Mohan Sobhana Nandhu, Chandra Goparaju, Sharon L. Longo, Harvey I. Pass, Mariano S. Viapiano
Journal: Cancer Research (2018)

Abstract Malignant mesothelioma (MM) is an aggressive tumor of mesothelial tissues that has poor prognosis and limited therapeutic options. Pass et al. (N Engl J Med 2012) identified the extracellular matrix protein fibulin-3 as an accurate biomarker of MM progression, which can be detected in the tumor parenchyma and in pleural effusions. Here, we report a systematic study of the mechanisms of fibulin-3 in MM and its value as a molecular target. Fibulin-3 activated canonical NFkB signaling and regulated the expression of NFkB-downstream genes, including the proteases MMP9 and MMP13, and the proinvasive proteins TNC and MLCK. Fibulin-3 knockdown downregulated NFkB signaling, reduced MM cell viability and potentiated the cytotoxicity of cisplatin in vitro. We next developed a function-blocking antibody ("mAb428.2") against an epitope of fibulin-3 required for NFkB activation (Nandhu et al., Clin Cancer Res 2017). This antibody blocked fibulin-3 signaling in vitro and reduced the expression of this protein, as well as NFkB-regulated MMP9, in subcutaneous (sc) MM tumors implanted in nude mice. Next, we treated mice carrying sc or intrapleural models of MM with mAb428.2 or control IgG1 (8x 30 mg/kg, q24h) delivered IV. Our anti-fibulin-3 antibody caused a significant reduction of tumor burden and extension of overall survival in both models. Overall, downregulation and inhibition of fibulin-3 proved a successful approach to enhance MM therapy. Given the value of this protein as a MM biomarker, we expect that detection of fibulin-3 may serve to identify patients with the highest chance to respond to combination therapies including anti-fibulin-3 approaches, achieving maximum therapeutic against these aggressive tumors. Citation Format: Mohan Sobhana Nandhu, Chandra Goparaju, Sharon L. Longo, Harvey I. Pass, Mariano S. Viapiano. Targeting malignant mesothelioma with an antibody against the tumor extracellular matrix protein fibulin-3 [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 2769.
Abstract 587: Small solute carrier and sirtuin genes are potential prognostic biomarkers for renal cell carcinoma
Authors: Andre R. Jordan, Naureen Mullani, Asif Talukder, Vinata B. Lokeshwar
Journal: Cancer Research (2018)

Abstract Introduction: Five-year survival of metastatic renal cell carcinoma (mRCC) patients is &amp;lt; 10% and African American (AA) males have the highest incidence. In the kidney, Small Solute Carrier (SLC) and Sirtuin (SIRT) gene families regulate redox signaling, oxidative stress responses, transport of glycolysis and Krebs cycle metabolites, etc. We examined the prognostic potential of eleven SLC and SIRT genes to predict clinical outcome in RCC patients and evaluated the biological functions of SLC13A3 in RCC cells. Methods: Expression of SLC (13A3, 22A6, 47A1, 5A10) and SIRT 1-7 transcript levels was evaluated in a TCGA RCC-dataset (n=604 patients). 13A3 expression was measured by quantitative RT-PCR (qPCR) in matched normal and RCC specimens from 53 patients. Biomarker levels were correlated with clinical outcome, using Mann-Whitney test. RCC cells stably-expressing SLC13A3, were examined for proliferation and cell cycle under normoxia and hypoxia. Results: In the TCGA dataset, SIRT4, SIRT5, SLC22A6, SLC5A10, and SLC47A1 downregulation and SIRT2, SIRT6 and SIRT7 upregulation correlated with higher tumor-stage and metastasis (P&amp;lt; 0.001). Kaplan-Meier plots showed that these biomarkers stratified patients into low- or high-risk for survival (P&amp;lt;0.001). qPCR showed 40-fold downregulation of SLC13A3 expression in RCC tissues when compared to normal kidney (P&amp;lt; 0.0001). Downregulation was 40-fold in White/Hispanic patients, but 198-fold in AA patients (P=0.0049) and correlated with tumor stage and metastasis (P=0.009). SLC13A3 expression in RCC cells increased GSH transport by 2.5-fold and inhibited cell proliferation by 3-4-fold (P&amp;lt;0.001), due to cell-cycle arrest at G1-S or G2-M checkpoints. Treatment of RCC cells with DNA-demethylating agent 5-Azacytidine and Trichostatin A, induced SLC13A3 expression by 50-fold (P&amp;lt;0.001). Conclusion: SLC/SIRT family members are potential prognostic biomarkers for RCC. This first study on SLC13A3 reveals that it is a potential tumor suppressor and its downregulation in RCC tissues correlates with metastasis and racial disparity. Citation Format: Andre R. Jordan, Naureen Mullani, Asif Talukder, Vinata B. Lokeshwar. Small solute carrier and sirtuin genes are potential prognostic biomarkers for renal cell carcinoma [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 587.
Aspirin (5 mmol/L) Inhibits Leukocyte Attack and Triggered Reactive Cell Proliferation in a 3D Human Coronary In Vitro Model
Authors: RainerVoisard, RobertFischer, MarkusOßwald, SamiVoglic, RegineBaur, MiloratSusa, WolfgangKoenig, VinzenzHombach
Journal: Circulation (2001)

Background—Leukocyte attack (LA) and the triggered reactive proliferation of smooth muscle cells (SMCs) are key events for the development of early atherosclerosis and restenosis. In the present study, we used a 3D human coronary in vitro model of LA (3DLA model) to examine the effect of high-dose aspirin on the adhesion and chemotaxis of leukocytes and the reactive proliferative response of SMCs. Methods and Results—For dose-finding, the effect of aspirin (1, 2, 5, and 10 mmol/L) on the tumor necrosis factor-α–induced upregulation of intercellular adhesion molecule-1 was analyzed in monocultures of human coronary endothelial cells (HCAEC) and the SMCs of the human coronary media (HCMSMC). In cytoflow and Northern blot experiments, the expression of intercellular adhesion molecule-1 was slightly reduced after incubation with 5 mmol/L aspirin, and strong inhibition was found after incubation with 10 mmol/L. In 3DLA models, HCAECs and HCMSMCs were cultured on both sides of a porous filter. For LA, human mon...
Predictors of symptomatic hyperviscosity in Waldenström macroglobulinemia
Authors: Jithma P. Abeykoon, Saurabh Zanwar, Stephen M. Ansell, Jeffrey Winters, Morie A. Gertz, Rebecca L. King, David Murray, Thomas Habermann, David Dingli, Eli Muchtar, Ronald S. Go, Nelson Leung, David J Inwards, Francis K. Buadi, Angela Dispenzieri, Martha Q. Lacy, Yi Lin, Wilson I. Gonsalves, Taxiarchis Kourelis, Thomas E. Witzig, Carrie Thompson, S. Vincent Rajkumar, Robert A. Kyle, Shaji Kumar, Prashant Kapoor
Journal: American Journal of Hematology (2018)

Symptomatic hyperviscosity is a well-established phenomenon in Waldenström macroglobulinemia (WM). Monoclonal IgM can variably impact intrinsic serum viscosity, leading to widely disparate symptomatic thresholds for development of hyperviscosity-related symptoms. Data regarding the predictors of symptomatic hyperviscosity and outcomes related to this complication remain scarce and a recent study proposed that IgM >6000 mg/dL be considered a new criterion for initiating therapy in otherwise asymptomatic (smoldering) WM to pre-empt hyperviscosity-related injury. Herein, we attempt to identify predictors of the development of symptomatic hyperviscosity and its impact in patients with WM. Of 997 WM patients evaluated from January, 1996 through June, 2017, symptomatic hyperviscosity was observed in 130 (13%) patients. Overall survival (OS) of these 130 patients was similar to that of patients without symptomatic hyperviscosity (median: 11.5 vs 11.6 years; P = 0.63). On multivariate-analysis, only viscosity >1.8 cp (risk ratio: 4.0, P = 0.02) assessed at the time of WM diagnosis was an independent predictor for the development of subsequent symptomatic hyperviscosity. Among patients with smoldering WM and IgM >6000 mg/dL at diagnosis (n = 13) who were managed expectantly, the median time-to-initial therapy was 6.9 years and only 15% developed hyperviscosity-related symptoms subsequently. In summary, the occurrence of symptomatic hyperviscosity does not impact OS. Serum viscosity at diagnosis of WM, and not IgM concentration, represents the single most important independent predictor for development of subsequent hyperviscosity-related symptoms. Patients with smoldering WM and high serum IgM can be safely observed in the absence of any indications per the Consensus recommendations to initiate WM-directed therapy.
The Forces Needed to Evoke Sounds from Cardiac Tissues, and the Attenuation of Heart Sounds
Authors: Williamdock
Journal: Circulation (1959)

There has been no study of the efficiency of any sound-producing system. Here there is a description of a device that records the force of each pull applied to a valve segment or strip of myocardium while the sound evoked is recorded. The relation of force to sound intensity, and the attenuation of sounds in air, blood, and ventricular and chest walls provide some quantitative data on a neglected phase of cardiac physiology.