tbuckley/cabot-search huggingface dataset:
Abstract 3117: Full spectrum of in vitro disease models with optimized CRISPR-Cas9 gene editing in difficult-to-transfect blood-lineage cells
Authors: Huanyu Jin, Zhongsheng Yu, Vladimir Pak, Yin Zhang, Padmaja Tummala, Pavithra Rajeswaran, Diana Nguyen, Monika Maleszewska, Jinling Li, Charles Cao, Ruby Yanru Chen-Tsai
Journal: Cancer Research (2018)

Abstract CRISPR/Cas9 technology is a powerful tool for gene editing in mammalian cells. However, the efficiency of gene editing in certain types of cells, especially suspension cells such as several blood-lineage cell lines, is extremely low. This is due to a variety of factors including low transfection efficiency of CRISPR/Cas9 reagents, cytotoxicity, and low or undesired gRNA and Cas9 activities in the cell lines. By optimizing the protocol and transfection conditions, we were able to successfully generate a spectrum of gene modifications in blood-lineage cells. Here, we describe techniques optimized for genome editing in some difficult-to-transfect blood-lineage cell lines and the types of modifications achieved: (1) For leukemia cell lines such as KG-1 and MOLM-13 cells, we compared DNA-based versus RNP-based Cas9 delivery efficiency, and found that plasmid-based Cas9 transfection resulted in cytotoxicity. In contrast, the RNP-based route demonstrated significantly lower cytotoxicity and higher efficiency of gene editing. Furthermore, we found that the application of a DNA-dependent protein kinase inhibitor dramatically enhanced the homology directed repair (HDR) in these cell lines; (2) For T lymphocyte cell lines such as Jurkat cells, we could efficiently delete a targeted 5kb regulatory region of a critical gene. We were also able to sequentially knockout (KO) two genes (double KO) in Jurkat cells using a Cas9-plasmid protocol; (3) In the case of the lymphoblastic cell line T2, we successfully created a large fragment knock-in, as well as a gene knockout cell line model, through DNA-based CRISPR/Cas9 delivery. Our studies demonstrate that for CRISPR/Cas9 genome editing in blood-lineage cells, different cell lines may require different approaches or modified protocols to deliver the CRISPR/Cas9 components for efficient and successful modifications of the targeted genes. Citation Format: Huanyu Jin, Zhongsheng Yu, Vladimir Pak, Yin Zhang, Padmaja Tummala, Pavithra Rajeswaran, Diana Nguyen, Monika Maleszewska, Jinling Li, Charles Cao, Ruby Yanru Chen-Tsai. Full spectrum of in vitro disease models with optimized CRISPR-Cas9 gene editing in difficult-to-transfect blood-lineage 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 3117.
Abstract 13013: Associations Between Unplanned Cardiac Reinterventions and Early Outcomes After Pediatric Cardiac Surgery: Findings From the Society of Thoracic Surgeons Congenital Heart Surgery Database
Authors: John M Costello, Michael C Monge, Kevin D Hill, Sung Hee Kim, Sara K Pasquali, Babatunde Yerokun, Jeffrey P Jacobs, Carl L Backer, Mjaye L Mazwi, Marshall L Jacobs
Journal: Circulation (2016)

Introduction: In children undergoing cardiac surgery, we sought to determine the incidence of unplanned cardiac reinterventions (surgery or therapeutic catheterization) during the same hospitalization as the index operation, identify risk factors for reinterventions, and explore associations between unplanned reinterventions and adverse outcomes. Hypothesis: We hypothesized that younger patients undergoing more complex operations would be at greater risk for unplanned cardiac reinterventions, and that operative mortality and postoperative length of stay (LOS) would be greater in patients who undergo reintervention when compared to those who do not. Methods: Patients ≤18 years of age in the STS Congenital Heart Surgery Database from January 2010-June 2015 were included. Multivariable logistic regression was used to evaluate risk factors for unplanned reintervention, and the impact of reintervention on outcomes including operative mortality and post-operative LOS. Covariates were included to adjust for important patient characteristics (e.g., weight, age, and comorbidities), case complexity, and center effects. Results: Included were 84,404 patients from 117 centers, of whom 21% were neonates and 36% infants. An unplanned cardiac reintervention was performed in 5.4% of patients, including 11.8% of neonates, 5.2% of infants, and 2.8% of children. Independent risk factors for an unplanned reintervention included presence of non-cardiac anomalies and genetic syndromes, non-Caucasian race, younger age, lower weight among neonates and infants, prior cardiothoracic operations, preoperative mechanical ventilation, preoperative risk factors, and higher STAT Mortality Category (adjusted p&lt;0.001 for all). The performance of an unplanned reintervention was an independent risk factor for operative mortality (adjusted OR, 5.34, 95% CI, 4.83-5.91, p&lt;0.001) and longer post-operative LOS (adjusted RR=2.31, 95% CI, 2.20-2.42, p&lt;0.001). Conclusions: Unplanned cardiac reinterventions are not rare, particularly in neonates, and are independently associated with mortality and increased post-operative LOS. Patients at greater risk may be readily identified preoperatively, presenting opportunities for quality improvement.
Effects of Different Atrial Pacing Modes on Atrial Electrophysiology
Authors: Wen-ChungYu, Shih-AnnChen, Ching-TaiTai, An-NingFeng, Mau-SongChang
Journal: Circulation (1997)

Background Multiple-site atrial pacing has been shown to prevent recurrence of atrial fibrillation. However, information about the mechanisms of different atrial pacing modes in prevention of atrial fibrillation was not clear. Methods and Results Forty-two patients without structural heart disease were classified into group 1 and group 2 according to absence or presence of clinical atrial fibrillation, respectively. Atrial conduction time and electrogram width of the right posterior interatrial septum (RPS) were measured during drive-train stimulation (S1) and early extrastimulation (S2). The locations of S1 were the high right atrium (HRA), distal coronary sinus (DCS), or both sites simultaneously. Effective refractory periods (ERPs) of the HRA and DCS were also determined during S1 stimulation at each site and during biatrial pacing. The ERPs were not different between single-site atrial pacing and biatrial pacing. In contrast, early S2 stimulation at the HRA caused greater atrial conduction delay and g...
Abstract 5525: <i>Rgs8</i> and <i>Rgs16</i> protect against pancreatitis and PDA progression
Authors: Shreoshi Pal Choudhuri, Yalda Zolghadri, Luke Mascarenhas, Thomas Wilkie
Journal: Cancer Research (2018)

Abstract Pancreatic ductal adenocarcinoma (PDA) has the highest death rate among major cancers, and new treatments are desperately needed. The mice we developed in this study, KCR8-16, are an excellent mouse model for identification, characterization, and in vivo validation of novel PDA therapeutics. Kras oncogenic mutations (e.g., KrasG12D) are found in over 90% of human PDA. Kras can be activated by protein kinase and G-protein coupled receptor (GPCR) signaling. Regulator of G-protein signaling (Rgs) proteins regulate GPCR signaling by accelerating the GTPase activity of Gq- and Gi class alpha subunits. Activating alleles of Gq that are resistant to Rgs inhibition are found in benign precursors of PDA in humans. We previously reported that Rgs8 and Rgs16 are in vivo reporters of Kras activity in pancreatic intraepithelial neoplasia (PanIN), intraductal papillary mucinous neoplasm (IPMN), and PDA progression in KIC;Rgs16::GFP mice (p48::Cre/+; KrasG12D/+; Cdkn2af/f; Rgs16GFP ) (DMM 8, 2015). To identify the role of Rgs 8 and 16 in PDA, we crossed the Rgs8-16 double knockout into pancreas-specific KrasG12D (KC) mutant mice (termed KCR8-16). We found that deletion of Rgs8 and Rgs16 in KC accelerated PDA progression. Additional pancreatic stress evoked by caerulein treatment caused immediate and pancreas-wide progression to PDA in KCR8-16 mice. Our study suggests that Rgs8 and Rgs16 act as tumor-suppressor genes in PDA initiation and progression. Moreover, KCR8-16 and KIC;Rgs16::GFP mice can be used as an excellent model for identification and rapid in vivo validation of PDA therapeutics. Citation Format: Shreoshi Pal Choudhuri, Yalda Zolghadri, Luke Mascarenhas, Thomas Wilkie. Rgs8 and Rgs16 protect against pancreatitis and PDA progression [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 5525.
The Distribution of Refractory Periods Influences the Dynamics of Ventricular Fibrillation
Authors: Bum-RakChoi, TongLiu, GuySalama
Journal: Circulation Research (2001)

Abstract—The spatial and dynamic properties of ventricular fibrillation (VF) may be random or related to cellular electrical properties of the normal heart. Local activation intervals (AIs) in VF may depend on the local refractory period (RP), and sustained VF may require a steep action potential (AP) restitution curve. In guinea pig hearts, AP durations (APDs) and RPs on the epicardium are shorter at the apex and progressively longer toward the base, producing gradients of RPs that may influence the spatial organization of VF. In the present study, the influence of APDs on VF dynamics is investigated in perfused guinea pig hearts stained with a voltage-sensitive dye by comparing APD gradients to the dynamics of VF elicited by burst pacing. In VF, AIs had no clear periodicity, but average AIs were shorter at the apex (57.5±8.1 ms) than the base (76.1±1.5 ms, n=6, P<0.05) and had gradients similar to APD gradients (correlation coefficient 0.71±0.04). Analysis of local velocity vectors showed no preferentia...
Abstract 4410: miRNA-34a sensitizes triple-negative breast cancer cells to paclitaxel reducing cancer stem cell population and lung colonization
Authors: Montserrat Climent, Paola Bonetti, Francesco Nicassio
Journal: Cancer Research (2018)

Abstract Background: Cancer stem cells (CSCs) have inherent or developed drug resistance for which chemotherapy fails to eliminate all tumor cells, a property that is critically contributing to recurrence and metastasis. Here, we combine miR-34a, a well-known tumor suppressor miRNA that controls proliferation, apoptosis and self-renewal, with Paclitaxel (PTX), a commonly used first-line chemotherapy drug. Based on our preliminary observations (Bonetti et al., manuscript under revision), we hypothesized a combinatorial approach, coupling miRNA- and chemo- treatment, aimed at reducing CSCs in vitro and recurrence in vivo using triple-negative breast cancer (TNBC) as a model. Methodology: As a model, we exploited SUM159pt cells (TNBC mesenchymal-like, with CSC features) to mimic aggressive breast cancer, which harbor a vector for pLUC, which is induced by luciferin in vivo, to monitor tumor growth in real time. Furthermore, we used i) pSlik34a, with tunable expression of miR-34a by doxycycline; ii) treatment with PTX to select for resistant cells (RC); and combined the two, by treating RC with doxycycline. CSC content was evaluated by ALDH (Aldehyde Dehydrogenase) activity, sphere-forming efficiency (SFE) and qRT-PCR of known markers. A similar approach was followed for in vivo studies, with NSG female injected through tail vein injection with 500,000 SUM159/pLUC/pSlik34a cells, and separated into three groups: not treated (NT), PTX alone, and PTX+34a. Results: As observed for aggressive breast cancer, treatment of SUM159 with PTX generated resistant cells (RC) with increased CSC content (ALDH activity and spheres forming ability). When miR-34a was induced in the RC, cells changed morphology, slowed proliferation rate, and decreased CSC content (ALDH activity and spheres). Moreover, a second PTX treatment almost completely abolished the ALDH activity and SFE. The combined effect was evaluated in vivo, following by luminescence the growth of metastases after lung colonization, with PTX+34a mice reducing lung colonization than the PTX treatment or miR-34a treatment alone. Conclusions: Overall, these results indicate that miR-34a sensitize PTX-resistant cells reducing cancer stem-cell capacities, and in this preclinical setting limited the aggressive phenotype of breast cancer, including proliferation, drug resistance, relapse and metastasis causing in delay of lung colonization in vivo. Citation Format: Montserrat Climent, Paola Bonetti, Francesco Nicassio. miRNA-34a sensitizes triple-negative breast cancer cells to paclitaxel reducing cancer stem cell population and lung colonization [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 4410.
Involvement of Rho GTPases in the Transcriptional Inhibition of Preproendothelin-1 Gene Expression by Simvastatin in Vascular Endothelial Cells
Authors: OctavioHernández-Perera, DoloresPérez-Sala, EstrellaSoria, SantiagoLamas
Journal: Circulation Research (2000)

Abstract—Endothelial dysfunction is characterized by an impaired vasodilatory response to endothelial agonists as well as by alterations in adhesion and coagulation processes. 3-Hydroxy-3-methylglutaryl-CoA reductase inhibitors (statins) have been shown to be useful in the reversal of endothelial dysfunction, an effect that may be independent of the reduction in cholesterol levels. Both the l-arginine–nitric oxide–cGMP and endothelin pathways are involved in the regulation of vascular tone. Here, we show that the basal transcription rate of the preproendothelin-1 gene was decreased by simvastatin (10 μmol/L) in bovine aortic endothelial cells. Transfection studies with the preproendothelin-1 gene promoter showed that mevalonate (100 μmol/L) was able to prevent the inhibitory effect mediated by simvastatin. Protein geranylgeranylation, but not farnesylation, proved to be crucial for a correct expression of the preproendothelin-1 gene. The C3 exotoxin from Clostridium botulinum that selectively inactivates ...
Thioredoxin Promotes ASK1 Ubiquitination and Degradation to Inhibit ASK1-Mediated Apoptosis in a Redox Activity-Independent Manner
Authors: YingmeiLiu, WangMin
Journal: Circulation Research (2002)

It has been shown that thioredoxin (Trx) in a reduced form binds to and inhibits apoptosis signal-regulating kinase 1 (ASK1). Apoptotic stimuli such as tumor necrosis factor (TNF) and reactive oxygen species (ROS) activate ASK1 in part by oxidizing Trx (forming intramolecular disulfide between C32 and C35) to release Trx from ASK1. In the present study, we examined if Trx affects ASK1 protein stability and whether the redox activity of Trx is critical in regulating ASK1 activity. First, we showed that overexpression of the wild-type Trx (Trx-WT) in endothelial cells induced ASK1 ubiquitination and degradation. Trx-induced ASK1 ubiquitination/degradation could be blocked by ASK1 activators TNF and TRAF2. We then tested the single-mutation of Trx at the catalytic site C32 or C35 (Trx-C32S or Trx-C35S) and the double-mutation (Trx-CS). The results showed that the single mutants (but not Trx-CS) retained the binding activity for ASK1 and the ability to induce ASK1 ubiquitination/degradation. Unlike Trx-WT, Tr...
Ionic Current Basis of Electrocardiographic Waveforms
Authors: KazutakaGima, YoramRudy
Journal: Circulation Research (2002)

Body surface electrocardiograms and electrograms recorded from the surfaces of the heart are the basis for diagnosis and treatment of cardiac electrophysiological disorders and arrhythmias. Given recent advances in understanding the molecular mechanisms of arrhythmia, it is important to relate these electrocardiographic waveforms to cellular electrophysiological processes. This modeling study establishes the following principles: (1) voltage gradients created by heterogeneities of the slow-delayed rectifier (IKs) and transient outward (Ito) potassium current inscribe the T wave and J wave, respectively; T-wave polarity and width are strongly influenced by the degree of intercellular coupling through gap-junctions. (2) Changes in [K+]o modulate the T wave through their effect on the rapid-delayed rectifier, IKr. (3) Alterations of IKs, IKr, and INa (fast sodium current) in long-QT syndrome (LQT1, LQT2, and LQT3, respectively) are reflected in characteristic QT-interval and T-wave changes; LQT1 prolongs QT ...
Abstract A65: Combination of a thioxodihydroquinazolinone compound with cisplatin eliminates ovarian cancer stem cell-like cells (CSC-LCs) and shows preclinical potential
Authors: Jing Ma, Joseph Salamoun, Peter Wipf, Robert Edwards, Bennett Van Houten, Wei Qian
Journal: Clinical Cancer Research (2018)

Abstract Cancer stem cell-like cells (CSC-LCs) contribute to drug resistance and recurrence of ovarian cancer. Strategies that can eradicate CSC-LCs are expected to substantially improve the outcome of ovarian cancer treatments. We have previously identified a class of thioxodihydroquinazolinone small molecules, which have strong synergistic antitumor activity with platinum drugs, the standard chemotherapeutic agents for ovarian cancer treatment. In the current study, using the activity of aldehyde dehydrogenase (ALDH) as a marker of CSC-LCs, we demonstrated that the combination of thioxodihydroquinazolinone compound 19 with cisplatin is able to deplete ALDH-high CSC-LC population in both established platinum-resistant ovarian cancer cell lines and primary ovarian cancer cells from metastatic ascites of a cisplatin-resistant patient. Compound 19 enhanced the accumulation of intracellular cisplatin in ALDH-high ovarian cancer cells. The combination of compound 19 with cisplatin was also able to reduce the sphere-forming capability of cisplatin-resistant ovarian cancer cells. Using a spheroid-based in vitro metastasis model of ovarian cancer, we demonstrated that the combination of compound 19 with cisplatin prevents spheroid ovarian cancer cells from attaching to substratum and spreading. In vivo, the treatment by the combination of compound 19 with cisplatin significantly reduced tumor burden in a cisplatin-resistant intraperitoneal ovarian cancer xenograft mouse model, as compared to cisplatin-alone treatment. Taken together, our study demonstrated that the thioxodihydroquinazolinone compounds are a new class of agents that in combination with cisplatin are capable of eliminating CSC-LCs in ovarian cancer. Further development of thioxodihydroquinazolinone small molecules may yield a more effective treatment for cisplatin-resistant metastatic ovarian cancer. Citation Format: Jing Ma, Joseph Salamoun, Peter Wipf, Robert Edwards, Bennett Van Houten, Wei Qian. Combination of a thioxodihydroquinazolinone compound with cisplatin eliminates ovarian cancer stem cell-like cells (CSC-LCs) and shows preclinical potential. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr A65.
Biphasic Actions of Prostaglandin E2 on the Renal Afferent Arteriole
Authors: LilongTang, KathyLoutzenhiser, RodgerLoutzenhiser
Journal: Circulation Research (2000)

Abstract—Prostaglandin (PG) E2 is an important modulator of the actions of angiotensin (Ang) II. In the present study, we investigated the renal microvascular actions of PGE2 and the EP receptor subtypes involved. Ibuprofen potentiated Ang II–induced vasoconstriction in in vitro perfused normal rat kidneys and augmented afferent arteriolar, but not efferent arteriolar, responses in the hydronephrotic rat kidney model. This preglomerular effect of endogenous prostanoids was mimicked by exogenous PGE2, which reversed Ang II–induced afferent arteriolar vasoconstriction at concentrations of 0.1 to 10 nmol/L without affecting the efferent arteriole. The PGE2-induced vasodilation was potentiated by the phosphodiesterase inhibitor Ro 20-1724 and was mimicked by 11-deoxy-PGE1 (0.01 to 1 nmol/L). Butaprost, which acts preferentially at EP2 receptors, was relatively ineffective. Whereas 0.1 to 10 nmol/L PGE2 elicited vasodilation, higher concentrations (1 to 10 μmol/L) restored Ang II–induced afferent arteriolar va...
Abstract 3887: Whole exome sequencing to identify genetic markers for trastuzumab-induced cardiotoxicity
Authors: Chihiro Udagawa, Hiromi Nakamura, Hiroshi Ohnishi, Kenji Tamura, Tatsunori Shimoi, Masayuki Yoshida, Teruhiko Yoshida, Hiroshi Okamura, Yasushi Totoki, Tatsuhiro Shibata, Hitoshi Zembutsu
Journal: Cancer Research (2018)

Abstract Although trastuzumab-induced cardiotoxicity is an important determinant to limit the use of this drug, the molecular mechanism of risk for this toxicity is not well understood. To identify a genetic marker(s) determining the risk of trastuzumab-induced cardiotoxicity, we performed whole exome sequencing of germline DNA samples from 9 patients with trastuzumab-induced cardiotoxicity, and conducted a case-control association study of 2,258 genetic variants between 9 cases (with trastuzumab-induced cardiotoxicity) and general Japanese population controls registered in Human Genetic Variation Database (HGVD). To further validate the result of the screening study, we carried out a replication study of 10 variants showing Pmin &amp;lt; 0.001 in the screening study using 234 independent patients treated with trastuzumab, including 10 cases and 224 controls (without trastuzumab-induced cardiotoxicity). In the replication study, we observed that three variants had effect in the same direction as in the screening study (SNV1 in exon 2 of HERADR1, SNV2 in exon 2 of HERADR2 and SNV3 in exon 44 of HERADR3). A combined result of the screening and the replication studies suggested an association of a locus on chromosome 6q12 with trastuzumab-induced cardiotoxicity (SNV3 in HERADR3, combined-Pmin = 0.00056, OR = 13.73). This finding provides new insights into personalized trastuzumab therapy for the patients with HER2 positive cancer. Citation Format: Chihiro Udagawa, Hiromi Nakamura, Hiroshi Ohnishi, Kenji Tamura, Tatsunori Shimoi, Masayuki Yoshida, Teruhiko Yoshida, Hiroshi Okamura, Yasushi Totoki, Tatsuhiro Shibata, Hitoshi Zembutsu. Whole exome sequencing to identify genetic markers for trastuzumab-induced cardiotoxicity [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 3887.
Adrenergic Neurohumoral Influences on Circulation and Lipolysis in Canine Omental Adipose Tissue
Authors: Kathrynballard, Sunerosell
Journal: Circulation Research (1971)

Vascular resistance, capillary filtration coefficient (CFC) and changes in blood volume were determined in canine omental adipose tissue. Release of glycerol and free fatty acids (FFA) were measured. Basal values in acutely denervated tissue for blood flow and CFC were 11 ml/min/100 g (range 4 to 26, N = 36) and 0.05 ml/min/100 g/mm Hg (range 0.018 to 0.090, N = 38), respectively. Stimulation of the sympathetic nerves (1 to 9 Hz) caused initial vasoconstriction, maintained for 1 to 3 minutes. At lower frequencies (1 to 3Hz), the blood flow increased gradually after the initial decrease. At higher frequencies, the constriction often reverted to vasodilatation. The CFC was not changed or increased initially. The clearance rate of locally injected 125I decreased in spite of a constant blood flow and blood volume was reduced initially. After α-receptor blockade, nerve stimulation caused vasodilatation, which in turn was inhibited by β-receptor blockade. Infusion of norepinephrine produced a pattern of vascula...