bluuebunny/crossref_metadata_2025 huggingface dataset:
The Biochemistry and Physiology of Mitochondrial Fatty Acid β-Oxidation and Its Genetic Disorders
Authors: Houten Sander M. , Violante Sara , Ventura Fatima V. , Wanders Ronald J.A.
Published: October 2015

Mitochondrial fatty acid β-oxidation (FAO) is the major pathway for the degradation of fatty acids and is essential for maintaining energy homeostasis in the human body. Fatty acids are a crucial energy source in the postabsorptive and fasted states when glucose supply is limiting. But even when glucose is abundantly available, FAO is a main energy source for the heart, skeletal muscle, and kidney. A series of enzymes, transporters, and other facilitating proteins are involved in FAO. Recessively inherited defects are known for most of the genes encoding these proteins. The clinical presentation of these disorders may include hypoketotic hypoglycemia, (cardio)myopathy, arrhythmia, and rhabdomyolysis and illustrates the importance of FAO during fasting and in hepatic and (cardio)muscular function. In this review, we present the current state of knowledge on the biochemistry and physiological functions of FAO and discuss the pathophysiological processes associated with FAO disorders.
Control and Functions of Fixational Eye Movements
Authors: Rucci Michele , Poletti Martina
Published: October 2015

Humans and other species explore a visual scene by making rapid eye movements (saccades) two to three times every second. Although the eyes may appear immobile in the brief intervals between saccades, microscopic (fixational) eye movements are always present, even when an observer is attending to a single point. These movements occur during the very periods in which visual information is acquired and processed, and their functions have long been debated. Recent technical advances in controlling retinal stimulation during normal oculomotor activity have shed new light on the visual contributions of fixational eye movements and the degree to which these movements can be controlled. The emerging body of evidence, reviewed in this article, indicates that fixational eye movements are important components of the strategy by which the visual system processes fine spatial details; they enable both precise positioning of the stimulus on the retina and encoding of spatial information into the joint space–time domain.
Hepatocyte X-box binding protein 1 deficiency increases liver injury in mice fed a high-fat/sugar diet
Authors: Liu Xiaoying , Henkel Anne S. , LeCuyer Brian E. , Schipma Matthew J. , Anderson Kristy A. , Green Richard M.
Published: October 2015

Fatty liver is associated with endoplasmic reticulum stress and activation of the hepatic unfolded protein response (UPR). Reduced hepatic expression of the UPR regulator X-box binding protein 1 spliced (XBP1s) is associated with human nonalcoholic steatohepatitis (NASH), and feeding mice a high-fat diet with fructose/sucrose causes progressive, fibrosing steatohepatitis. This study examines the role of XBP1 in nonalcoholic fatty liver injury and fatty acid-induced cell injury. Hepatocyte-specific Xbp1-deficient ( Xbp1−/−) mice were fed a high-fat/sugar (HFS) diet for up to 16 wk. HFS-fed Xbp1−/−mice exhibited higher serum alanine aminotransferase levels compared with Xbp1fl/flcontrols. RNA sequencing and Gene Ontogeny pathway analysis of hepatic mRNA revealed that apoptotic process, inflammatory response, and extracellular matrix structural constituent pathways had enhanced activation in HFS-fed Xbp1−/−mice. Liver histology demonstrated enhanced injury and fibrosis but less steatosis in the HFS-fed Xbp1−/−mice. Hepatic Col1a1 and Tgfβ1 gene expression, as well as Chop and phosphorylated JNK (p-JNK), were increased in Xbp1−/−compared with Xbp1fl/flmice after HFS feeding. In vitro, stable XBP1-knockdown Huh7 cells (Huh7-KD) and scramble control cells (Huh7-SCR) were generated and treated with palmitic acid (PA) for 24 h. PA-treated Huh7-KD cells had increased cytotoxicity measured by lactate dehydrogenase release, apoptotic nuclei, and caspase3/7 activity assays compared with Huh7-SCR cells. CHOP and p-JNK expression was also increased in Huh7-KD cells following PA treatment. In conclusion, loss of XBP1 enhances injury in both in vivo and in vitro models of fatty liver injury. We speculate that hepatic XBP1 plays an important protective role in pathogenesis of NASH.
Phase synchrony reveals organization in human atrial fibrillation
Authors: Vidmar David , Narayan Sanjiv M. , Rappel Wouter-Jan
Published: October 2015

It remains unclear if human atrial fibrillation (AF) is spatially nonhierarchical or exhibits a hierarchy of organization sustained by sources. We utilize activation times obtained at discrete locations during AF to compute the phase synchrony between tissue regions, to examine underlying spatial dynamics throughout both atria. We construct a binary synchronization network and show that this network can accurately define regions of coherence in coarse-grained in silico data. Specifically, domains controlled by spiral waves exhibit regions of high phase synchrony. We then apply this analysis to clinical data from patients experiencing cardiac arrhythmias using multielectrode catheters to simultaneously record from a majority of both atria. We show that pharmaceutical intervention with ibutilide organizes activation by increasing the size of the synchronized domain in AF and quantify the increase in temporal organization when arrhythmia changes from fibrillation to tachycardia. Finally, in recordings from 24 patients in AF we show that the level of synchrony is spatially broad with some patients showing large spatially contiguous regions of synchronization, while in others synchrony is localized to small pockets. Using computer simulations, we show that this distribution is inconsistent with distributions obtained from simulations that mimic multiwavelet reentry but is consistent with mechanisms in which one or more spatially conserved spiral waves is surrounded by tissue in which activation is disorganized.
Myosin MgADP release rate decreases at longer sarcomere length to prolong myosin attachment time in skinned rat myocardium
Authors: Tanner Bertrand C. W. , Breithaupt Jason J. , Awinda Peter O.
Published: October 2015

Cardiac contractility increases as sarcomere length increases, suggesting that intrinsic molecular mechanisms underlie the Frank-Starling relationship to confer increased cardiac output with greater ventricular filling. The capacity of myosin to bind with actin and generate force in a muscle cell is Ca2+ regulated by thin-filament proteins and spatially regulated by sarcomere length as thick-to-thin filament overlap varies. One mechanism underlying greater cardiac contractility as sarcomere length increases could involve longer myosin attachment time ( t on) due to slowed myosin kinetics at longer sarcomere length. To test this idea, we used stochastic length-perturbation analysis in skinned rat papillary muscle strips to measure t on as [MgATP] varied (0.05–5 mM) at 1.9 and 2.2 μm sarcomere lengths. From this t on-MgATP relationship, we calculated cross-bridge MgADP release rate and MgATP binding rates. As MgATP increased, t on decreased for both sarcomere lengths, but t on was roughly 70% longer for 2.2 vs. 1.9 μm sarcomere length at maximally activated conditions. These t on differences were driven by a slower MgADP release rate at 2.2 μm sarcomere length (41 ± 3 vs. 74 ± 7 s−1), since MgATP binding rate was not different between the two sarcomere lengths. At submaximal activation levels near the pCa50 value of the tension-pCa relationship for each sarcomere length, length-dependent increases in t on were roughly 15% longer for 2.2 vs. 1.9 μm sarcomere length. These changes in cross-bridge kinetics could amplify cooperative cross-bridge contributions to force production and thin-filament activation at longer sarcomere length and suggest that length-dependent changes in myosin MgADP release rate may contribute to the Frank-Starling relationship in the heart.
Lipopolysaccharide-induced pulmonary endothelial barrier disruption and lung edema: critical role for bicarbonate stimulation of AC10
Authors: Nickols Jordan , Obiako Boniface , Ramila K. C. , Putinta Kevin , Schilling Sarah , Sayner Sarah L.
Published: October 2015

Bacteria-induced sepsis is a common cause of pulmonary endothelial barrier dysfunction and can progress toward acute respiratory distress syndrome. Elevations in intracellular cAMP tightly regulate pulmonary endothelial barrier integrity; however, cAMP signals are highly compartmentalized: whether cAMP is barrier-protective or -disruptive depends on the compartment (plasma membrane or cytosol, respectively) in which the signal is generated. The mammalian soluble adenylyl cyclase isoform 10 (AC10) is uniquely stimulated by bicarbonate and is expressed in pulmonary microvascular endothelial cells (PMVECs). Elevated extracellular bicarbonate increases cAMP in PMVECs to disrupt the endothelial barrier and increase the filtration coefficient ( Kf) in the isolated lung. We tested the hypothesis that sepsis-induced endothelial barrier disruption and increased permeability are dependent on extracellular bicarbonate and activation of AC10. Our findings reveal that LPS-induced endothelial barrier disruption is dependent on extracellular bicarbonate: LPS-induced barrier failure and increased permeability are exacerbated in elevated bicarbonate compared with low extracellular bicarbonate. The AC10 inhibitor KH7 attenuated the bicarbonate-dependent LPS-induced barrier disruption. In the isolated lung, LPS failed to increase Kf in the presence of minimal perfusate bicarbonate. An increase in perfusate bicarbonate to the physiological range (24 mM) revealed the LPS-induced increase in Kf, which was attenuated by KH7. Furthermore, in PMVECs treated with LPS for 6 h, there was a dose-dependent increase in AC10 expression. Thus these findings reveal that LPS-induced pulmonary endothelial barrier failure requires bicarbonate activation of AC10.
High-intensity interval training evokes larger serum BDNF levels compared with intense continuous exercise
Authors: Saucedo Marquez Cinthia Maria , Vanaudenaerde Bart , Troosters Thierry , Wenderoth Nicole
Published: October 2015

Exercise can have a positive effect on the brain by activating brain-derived neurotrophic factor (BDNF)-related processes. In healthy humans there appears to be a linear relationship between exercise intensity and the positive short-term effect of acute exercise on BDNF levels (i.e., the highest BDNF levels are reported after high-intensity exercise protocols). Here we performed two experiments to test the effectiveness of two high-intensity exercise protocols, both known to improve cardiovascular health, to determine whether they have a similar efficacy in affecting BDNF levels. Participants performed a continuous exercise (CON) protocol at 70% of maximal work rate and a high-intensity interval-training (HIT) protocol at 90% of maximal work rate for periods of 1 min alternating with 1 min of rest (both protocols lasted 20 min). We observed similar BDNF kinetics in both protocols, with maximal BDNF concentrations being reached toward the end of training ( experiment 1). We then showed that both exercise protocols significantly increase BDNF levels compared with a rest condition (CON P = 0.04; HIT P < 0.001), with HIT reaching higher BDNF levels than CON ( P = 0.035) ( experiment 2). These results suggest that shorter bouts of high intensity exercise are slightly more effective than continuous high-intensity exercise for elevating serum BDNF. Additionally, 73% of the participants preferred the HIT protocol ( P = 0.02). Therefore, we suggest that the HIT protocol might represent an effective and preferred intervention for elevating BDNF levels and potentially promoting brain health.
The effect of leptin replacement on sleep-disordered breathing in the leptin-deficientob/obmouse
Authors: Pho H. , Hernandez A. B. , Arias R. S. , Leitner E. B. , Van Kooten S. , Kirkness J. P. , Schneider H. , Smith P. L. , Polotsky V. Y. , Schwartz A. R.
Published: October 2015

Obese leptin-deficient ( ob/ob) mice demonstrate defects in upper airway structural and neuromuscular control. We hypothesized that these defects predispose to upper airway obstruction during sleep, and improve with leptin administration. High-fidelity polysomnographic recordings were conducted to characterize sleep and breathing patterns in conscious, unrestrained ob/ob mice (23 wk, 67.2 ± 4.1 g, n = 13). In a parallel-arm crossover study, we compared responses to subcutaneous leptin (1 μg/h) vs. vehicle on respiratory parameters during NREM and REM sleep. Upper airway obstruction was defined by the presence of inspiratory airflow limitation (IFL), as characterized by an early inspiratory plateau in airflow at a maximum level (V̇imax) with increasing effort. The severity of upper airway obstruction (V̇imax) was assessed along with minute ventilation (V̇e), tidal volume (VT), respiratory rate (RR), inspiratory duty cycle, and mean inspiratory flow at each time point. IFL occurred more frequently in REM sleep (37.6 ± 0.2% vs. 1.1 ± 0.0% in NREM sleep, P < 0.001), and leptin did not alter its frequency. V̇imax (3.7 ± 1.1 vs. 2.7 ± 0.8 ml/s, P < 0.001) and V̇e increased (55.4 ± 22.0 vs. 39.8 ± 16.4 ml/min, P < 0.001) with leptin vs. vehicle administration. The increase in V̇e was due to a significant increase in VT(0.20 ± 0.06 vs. 0.16 ± 0.05 ml, P < 0.01) rather than RR. Increases in V̇e were attributable to increases in mean inspiratory flow (2.5 ± 0.8 vs. 1.8 ± 0.6 ml/s, P < 0.001) rather than inspiratory duty cycle. Similar increases in V̇e and its components were observed in non-flow-limited breaths during NREM and REM sleep. These responses suggest that leptin stabilized pharyngeal patency and increased drive to both the upper airway and diaphragm during sleep.
Epigenetics and cytoprotection with heat acclimation
Authors: Horowitz Michal
Published: October 2015

Studying “phenotypic plasticity” involves comparison of traits expressed in response to environmental fluctuations and aims to understand tolerance and survival in new settings. Reversible phenotypic changes that enable individuals to match their phenotype to environmental demands throughout life can be artificially induced, i.e., acclimation or occur naturally, i.e., acclimatization. The onset and achievement of acclimatory homeostasis are determined by molecular programs that induce the acclimated transcriptome. In heat acclimation, much evidence suggests that epigenetic mechanisms are powerful players in these processes. Epigenetic mechanisms affect the accessibility of the DNA to transcription factors, thereby regulating gene expression and controlling the phenotype. The heat-acclimated phenotype confers cytoprotection against novel stressors via cross-tolerance mechanisms, by attenuation of the initial damage and/or by accelerating spontaneous recovery through the release of help signals. This indispensable acclimatory feature has a memory and can be rapidly reestablished after the loss of acclimation and the return to the physiological preacclimated phenotype. The transcriptional landscape of the deacclimated phenotype includes constitutive transcriptional activation of epigenetic bookmarks. Heat shock protein (HSP) 70/HSP90/heat shock factor 1 memory protocol demonstrated constitutive histone H4 acetylation on hsp70 and hsp90 promotors. Novel players in the heat acclimation setup are poly(ADP-ribose)ribose polymerase 1 affecting chromatin condensation, DNA linker histones from the histone H1 cluster, and transcription factors associated with the P38 pathway. We suggest that these orchestrated responses maintain euchromatin and proteostasis during deacclimation and predispose to rapid reacclimation and cytoprotection. These mechanisms represent within-life epigenetic adaptations and cytoprotective memory.
Combined, but not individual, blockade of ASIC3, P2X, and EP4 receptors attenuates the exercise pressor reflex in rats with freely perfused hindlimb muscles
Authors: Stone Audrey J. , Copp Steven W. , Kim Joyce S. , Kaufman Marc P.
Published: October 2015

In healthy humans, tests of the hypothesis that lactic acid, PGE2, or ATP plays a role in evoking the exercise pressor reflex proved controversial. The findings in humans resembled ours in decerebrate rats that individual blockade of the receptors to lactic acid, PGE2, and ATP had only small effects on the exercise pressor reflex provided that the muscles were freely perfused. This similarity between humans and rats prompted us to test the hypothesis that in rats with freely perfused muscles combined receptor blockade is required to attenuate the exercise pressor reflex. We first compared the reflex before and after injecting either PPADS (10 mg/kg), a P2X receptor antagonist, APETx2 (100 μg/kg), an activating acid-sensing ion channel 3 (ASIC) channel antagonist, or L161982 (2 μg/kg), an EP4 receptor antagonist, into the arterial supply of the hindlimb of decerebrated rats. We then examined the effects of combined blockade of P2X receptors, ASIC3 channels, and EP4 receptors on the exercise pressor reflex using the same doses, intra-arterial route, and time course of antagonist injections as those used for individual blockade. We found that neither PPADS ( n = 5), APETx2 ( n = 6), nor L161982 ( n = 6) attenuated the reflex. In contrast, combined blockade of these receptors ( n = 7) attenuated the peak (↓27%, P < 0.019) and integrated (↓48%, P < 0.004) pressor components of the reflex. Combined blockade injected intravenously had no effect on the reflex. We conclude that combined blockade of P2X receptors, ASIC3 channels, and EP4 receptors on the endings of thin fiber muscle afferents is required to attenuate the exercise pressor reflex in rats with freely perfused hindlimbs.
Influence of statins on distinct circulating microRNAs during prolonged aerobic exercise
Authors: Min Pil-Ki , Park Joseph , Isaacs Stephanie , Taylor Beth A. , Thompson Paul D. , Troyanos Chris , D'Hemecourt Pierre , Dyer Sophia , Chan Stephen Y. , Baggish Aaron L.
Published: October 2015

Statins exacerbate exercise-induced skeletal muscle injury. Muscle-specific microRNAs (myomiRs) increase in plasma after prolonged exercise, but the patterns of myomiRs release after statin-associated muscle injury have not been examined. We examined the relationships between statin exposure, in vitro and in vivo muscle contraction, and expression of candidate circulating myomiRs. We measured plasma levels of myomiRs, circulating microRNA-1 (c-miR-1), c-miR-133a, c-miR-206, and c-miR-499-5p from 28 statin-using and 28 nonstatin-using runners before (PRE), immediately after (FINISH), and 24 h after they ran a 42-km footrace (the 2011 Boston marathon) (POST-24). To examine these cellular-regulation myomiRs, we used contracting mouse C2C12 myotubes in culture with and without statin exposure to compare intracellular and extracellular expression of these molecules. In marathoners, c-miR-1, c-miR-133a, and c-miR-206 increased at FINISH, returned to baseline at POST-24, and were unaffected by statin use. In contrast, c-miR-499-5p was unchanged at FINISH but increased at POST-24 among statin users compared with PRE and runners who did not take statins. In cultured C2C12 myotubes, extracellular c-miR-1, c-miR-133a, and c-miR-206 were significantly increased by muscle contraction regardless of statin use. In contrast, extracellular miR-499-5p was unaffected by either isolated statin exposure or isolated carbachol exposure but it was increased when muscle contraction was combined with statin exposure. In summary, we found that statin-potentiated muscle injury during exercise is accompanied by augmented extracellular release of miR-499-5p. Thus c-miR-499-5p may serve as a biomarker of statin-potentiated muscle damage.
Role of miR206 in genistein-induced rescue of pulmonary hypertension in monocrotaline model
Authors: Sharma Salil , Umar Soban , Centala Alexander , Eghbali Mansoureh
Published: October 2015

Pulmonary hypertension (PH) is a progressive lung disease associated with proliferation of smooth muscle cells and constriction of lung microvasculature, leading to increased pulmonary arterial pressure, right ventricular failure, and death. We have previously shown that genistein rescues preexisting established PH by significantly improving lung and heart function. (Matori H, Umar S, Nadadur RD, Sharma S, Partow-Navid R, Afkhami M, Amjedi M, Eghbali M. Hypertension 60: 425–430, 2012). Here, we have examined the role of microRNAs (miRs) in the rescue action of genistein in monocrotaline (MCT)-induced PH in rats. Our miR microarray analysis on the lung samples from control, PH, and genistein-rescue group revealed that miR206, which was robustly upregulated to ∼11-fold by PH, was completely normalized to control levels by genistein treatment. Next, we examined whether knockdown of miR206 could reverse preexisting established PH. PH was induced in male rats by 60 mg/kg of MCT, and rats received three intratracheal doses of either miR206 antagomir (10 mg/kg body wt) or scrambled miR control at days 17, 21, and 26. Knockdown of miR206 resulted in significant improvement in the cardiopulmonary function, as right ventricular pressure was significantly reduced to 38.6 ± 3.61 mmHg from 61.2 ± 5.4 mmHg in PH, and right ventricular hypertrophy index was decreased to 0.35 ± 0.04 from 0.59 ± 0.037 in PH. Knockdown of miR206 reversed PH-induced pulmonary vascular remodeling in vivo and was associated with restoration of PH-induced loss of capillaries in the lungs and induction of vascular endothelial growth factor A expression. In conclusion, miR206 antagomir therapy improves cardiopulmonary function and structure and rescues preexisting severe PH in MCT rat model possibly by stimulating angiogenesis in the lung.
Macrophage Blockade Using CSF1R Inhibitors Reverses the Vascular Leakage Underlying Malignant Ascites in Late-Stage Epithelial Ovarian Cancer
Authors: Moughon Diana L. , He Huanhuan , Schokrpur Shiruyeh , Jiang Ziyue Karen , Yaqoob Madeeha , David John , Lin Crystal , Iruela-Arispe M. Luisa , Dorigo Oliver , Wu Lily
Published: October 2015

Malignant ascites is a common complication in the late stages of epithelial ovarian cancer (EOC) that greatly diminishes the quality of life of patients. Malignant ascites is a known consequence of vascular dysfunction, but current approved treatments are not effective in preventing fluid accumulation. In this study, we investigated an alternative strategy of targeting macrophage functions to reverse the vascular pathology of malignant ascites using fluid from human patients and an immunocompetent murine model (ID8) of EOC that mirrors human disease by developing progressive vascular disorganization and leakiness culminating in massive ascites. We demonstrate that the macrophage content in ascites fluid from human patients and the ID8 model directly correlates with vascular permeability. To further substantiate macrophages' role in the pathogenesis of malignant ascites, we blocked macrophage function in ID8 mice using a colony-stimulating factor 1 receptor kinase inhibitor (GW2580). Administration of GW2580 in the late stages of disease resulted in reduced infiltration of protumorigenic (M2) macrophages and dramatically decreased ascites volume. Moreover, the disorganized peritoneal vasculature became normalized and sera from GW2580-treated ascites protected against endothelial permeability. Therefore, our findings suggest that macrophage-targeted treatment may be a promising strategy toward a safe and effective means to control malignant ascites of EOC. Cancer Res; 75(22); 4742–52. ©2015 AACR.
ATDC/TRIM29 Drives Invasive Bladder Cancer Formation through miRNA-Mediated and Epigenetic Mechanisms
Authors: Palmbos Phillip L. , Wang Lidong , Yang Huibin , Wang Yin , Leflein Jacob , Ahmet McKenzie L. , Wilkinson John E. , Kumar-Sinha Chandan , Ney Gina M. , Tomlins Scott A. , Daignault Stephanie , Kunju Lakshmi. P. , Wu Xue-Ru , Lotan Yair , Liebert Monica , Ljungman Mats E. , Simeone Diane M.
Published: October 2015

Bladder cancer is a common and deadly malignancy but its treatment has advanced little due to poor understanding of the factors and pathways that promote disease. ATDC/TRIM29 is a highly expressed gene in several lethal tumor types, including bladder tumors, but its role as a pathogenic driver has not been established. Here we show that overexpression of ATDC in vivo is sufficient to drive both noninvasive and invasive bladder carcinoma development in transgenic mice. ATDC-driven bladder tumors were indistinguishable from human bladder cancers, which displayed similar gene expression signatures. Clinically, ATDC was highly expressed in bladder tumors in a manner associated with invasive growth behaviors. Mechanistically, ATDC exerted its oncogenic effects by suppressing miR-29 and subsequent upregulation of DNMT3A, leading to DNA methylation and silencing of the tumor suppressor PTEN. Taken together, our findings established a role for ATDC as a robust pathogenic driver of bladder cancer development, identified downstream effector pathways, and implicated ATDC as a candidate biomarker and therapeutic target. Cancer Res; 75(23); 5155–66. ©2015 AACR.
Identification of Cancer-Associated Fibroblasts in Circulating Blood from Patients with Metastatic Breast Cancer
Authors: Ao Zheng , Shah Sanket H. , Machlin Leah M. , Parajuli Ritesh , Miller Philip C. , Rawal Siddarth , Williams Anthony J. , Cote Richard J. , Lippman Marc E. , Datar Ram H. , El-Ashry Dorraya
Published: October 2015

Metastasis is facilitated by cancer-associated fibroblasts (CAF) in the tumor microenvironment through mechanisms yet to be elucidated. In this study, we used a size-based microfilter technology developed by our group to examine whether circulating CAF identified by FAP and α-SMA co-expression (cCAF) could be distinguished in the peripheral blood of patients with metastatic breast cancer. In a pilot study of patients with breast cancer, we detected the presence of cCAFs in 30/34 (88%) patients with metastatic disease (MET group) and in 3/13 (23%) patients with localized breast cancer (LOC group) with long-term disease-free survival. No cCAFs as defined were detected in healthy donors. Further, both cCAF and circulating tumor cells (CTC) were significantly greater in the MET group compared with the LOC group. Thus, the presence of cCAF was associated with clinical metastasis, suggesting that cCAF may complement CTC as a clinically relevant biomarker in metastatic breast cancer. Cancer Res; 75(22); 4681–7. ©2015 AACR.
Phase Ib Randomized, Double-Blinded, Placebo-Controlled, Dose Escalation Study of Polyphenon E in Patients with Barrett's Esophagus
Authors: Joe Andrew K. , Schnoll-Sussman Felice , Bresalier Robert S. , Abrams Julian A. , Hibshoosh Hanina , Cheung Ken , Friedman Richard A. , Yang Chung S. , Milne Ginger L. , Liu Diane D. , Lee J. Jack , Abdul Kazeem , Bigg Michelle , Foreman Jessica , Su Tao , Wang Xiaomei , Ahmed Aqeel , Neugut Alfred I. , Akpa Esther , Lippman Scott M. , Perloff Marjorie , Brown Powel H. , Lightdale Charles J.
Published: October 2015

This study was conducted to determine the safety and efficacy of the green tea–derived Polyphenon E (Poly E) in patients with Barrett's Esophagus (BE). Subjects were randomized to a 6-month, twice daily (BID) oral treatment of placebo or Poly E (200, 400, or 600 mg). Endoscopic evaluation, including biopsies, was performed before and after treatment. The primary objective was to demonstrate safety; secondary objectives investigated catechin accumulation and effects in clinical specimens. Of the 44 enrolled subjects, 11 received placebo, and 33 received Poly E. No dose-limiting toxicities were encountered, and a maximum tolerated dose (MTD) was not reached. The recommended phase II dose was 600 mg twice daily. The most common treatment-related adverse events (AE) in Poly E–treated subjects were grade I and II nausea, grade I belching, and grade I lactate dehydrogenase (LDH) elevation. No treatment-related AEs were reported in placebo-treated subjects, aside from grade I laboratory abnormalities. Pill counts and subject diaries were not consistently collected, and compliance was difficult to determine. However, on the basis of an intention-to-treat analysis, there was a significant relationship between Poly E dose and esophageal EGCG level—mean changes (pmol/g) of 0.79 (placebo), 6.06 (200 mg), 35.67 (400 mg), and 34.95 (600 mg); P = 0.005. There was a possible relationship between Poly E dose and urine PGE-M concentration. In conclusion, Poly E was well-tolerated, and treatment with Poly E (400 and 600 mg) but not Poly E (200 mg) or placebo resulted in clinically relevant and detectable EGCG accumulation in the target organ, esophageal mucosa. Cancer Prev Res; 8(12); 1131–7. ©2015 AACR.
Abstract A14: Overcoming therapeutic MAb resistance in agressive HER2 positive breast carcinomas by adoptive immunotherapy using optizimed effectors cells
Authors: Valsesia-Wittmann Sandrine , Clemenceau Beatrice , Jallas Anne-Catherine , Blay Jean-Yves , Marabelle Aurelien , Caux Christophe , Vie Henri
Published: October 2015

Trastuzumab, a monoclonal antibody targeting HER2 has been demonstrated to improve survival of HER2 overexpressing metastatic breast cancer. However, half of patients who initially respond to Trastuzumab develop resistance within one year of treatment initiation. Alteration of Antibodies Dependant Cellular Cytotoxicity (ADCC) mechanisms is one of the rational mechanisms for resistance to therapeutic Mab.

The goal of this project is to overcome this resistance by adoptive transfer using genetically engineered optimized NK cells. We thus compare efficacy of two different approaches: (i) NK cells armed with a high affinity Fc domain (FcγRIIIα/CD16) linked to its transduction chain FCεRIγ to generate NK-CD16/γ cells that allow recognition and interaction with Mab, inducing ADCC and (ii) NK cells expressing Trastuzumab sequence fused to the transduction chain FCεRIγ to generate NK-Her/γ that directly kill the HER2 positively cells.

In vitro, both strategies demonstrated high cytotoxic efficacy against HER2 positive cells, with direct killing systematically more efficient (reaching up to 95% with ratio E:T 5:1). Interestingly, two steps killing with NK-CD16/γ enabled a very specific cytotoxicity only in the presence of specific antibody. We demonstrated that efficacy of lysis is linked (i) to effector dose, (ii) levels of CD16/γ expression on effectors cells, (iii) HER2 expression but not level on target cells

In vivo, in Trastuzumab resistant HER2+ xenograft immunodeficient NSG mice model, complete regression was obtained only when using NK-CD16/γ in the presence of Trastuzumab. Even if the NK-Her/γ was found by IF or FACS in the tumor after IV or IP injection, they could not induce tumor regression. Analyses of NK dysfunction will be presented.

Restoration or improvement of effectors cells might be considerate as an issue in therapeutic Mab resistance treatment. Successful immune-based therapies will likely ultimately integrate strategies that combine immunotherapy approaches and immune-modulating drugs, in order to maximize their antitumor activity.

Citation Format: Sandrine Valsesia-Wittmann, Beatrice Clemenceau, Anne-Catherine Jallas, Jean-Yves Blay, Aurelien Marabelle, Christophe Caux, Henri Vie. Overcoming therapeutic MAb resistance in agressive HER2 positive breast carcinomas by adoptive immunotherapy using optizimed effectors cells. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A14.
Abstract A23: Potentiating immunological memory in mice using aptamer targeted siRNA delivery to inhibit mediators of effector differentiation in CD8+ cytotoxic T lymphocytes
Authors: Berezhnoy Alexey , Anugraha Rajagopalan , Malek Thomas , Gilboa Eli
Published: October 2015

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR11) of the Conference Proceedings.

Citation Format: Alexey Berezhnoy, Rajagopalan Anugraha, Thomas Malek, Eli Gilboa. Potentiating immunological memory in mice using aptamer targeted siRNA delivery to inhibit mediators of effector differentiation in CD8+ cytotoxic T lymphocytes. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A23.
Abstract A33: Preparation of local anti-wart and anti-inflammatory rub of polyglycidol based saturated lipid (DDP-PG) with HPMC containing curcumin
Authors: Shaheen Sharif Mohammad , Bakardzhiev Pavel , Momekova Denitsa , Koseva Neli S. , Rangelov Stanislav
Published: October 2015

Curcumin is absorbed poorly and has a limited bioavailability to work promptly as an anti-cancer drug or as an anti-inflammatory agent. If curcumin could be delivered directly into the cell, it might be a potential therapeutic against the inflammation and thereby cancer signaling cascades. In order to reach this goal here a topical medicament (rub) containing curcumin has been developed as an alternative therapeutic especially for skin inflammation and cancer thereby. We synthesized polyglycidol based block copolymers DDP-PG (MW 2700), which was incorporated into HPMC (7%) to prepare the rub containing curcumin. One more formulation containing monoolein rac glycerol (MO) with DDP-PG was incorporated. The diffusion of curcumin was evaluated in vitro using a biomembrane (fish swim bladder). The diffusion of curcumin form DDP-PG based formulation did not differ from that of MO, which is the indication of their suitability as biomaterials for the preparation of a transdermal penetrating vehicle. Subsequently, curcumin containing DDP-PG was evaluated directly for therapeutic purposes treating local apoptotic skin, inflammatory sites and benign tumor mass (wart). It showed skin cured very promisingly from apoptosis, which might be worst if not treated in time. It cleared the inflammation perfectly and reduced dramatically the size of the wart (a benign tumor) as well.

Citation Format: Sharif Mohammad Shaheen, Pavel Bakardzhiev, Denitsa Momekova, Neli S. Koseva, Stanislav Rangelov. Preparation of local anti-wart and anti-inflammatory rub of polyglycidol based saturated lipid (DDP-PG) with HPMC containing curcumin. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A33.
Abstract A41: Galectin-9 has tumor promoting properties that may be targetable in metastatic melanoma
Authors: Enninga Elizabeth Ann L. , Nevala Wendy K. , Markovic Svetomir N.
Published: October 2015

Introduction: Galectin-9 (Gal-9) is a carbohydrate-binding protein that interacts with T-cell immunoglobulin mucin 3 (TIM3) to modulate immune homeostasis. The Gal-9/TIM3 interaction down regulates activated T cells which promotes tumor growth, much like program death 1 (PD-1) and its ligand (PD-L1). Still, the effect Gal-9 has on the immune environment of melanoma patients is weakly understood. This study focuses on the role of Gal-9 in immunoregulation of healthy persons and metastatic melanoma patients.

Methods: Soluble Gal-9 levels were determined from plasma collected from patients and healthy persons using standard ELISA methods. Proliferation and apoptosis of cells in the presence and absence of Gal-9 was determined by mixed lymphocyte reactions and Tunnel staining. Supernatants from these reactions were collected and run using a cytokine bead assay to determine changes in immune parameters with and without Gal-9.

Results: Soluble Gal-9 was significantly higher in metastatic melanoma patients compared to healthy controls (3929 vs. 877 pg/mL, p=0.0003 respectively). In the presence of Gal-9, the proliferative effect of an allostimulus could be overridden until the level of responder cells to stimulator cells was 4:1. Using an apoptosis assay, Gal-9 was found to increase the level of cellular apoptosis; however, apoptosis was decreased as the allostimulus increased. Healthy cells incubated with Gal-9 were found to increase secretion of growth factors (EGF and FGF-2), chemokines (CXCL1, CXCL8, CCL2 and CCL3), T helper 1 cytokines (IFN-g and IL-7) which have shown tumor promoting affects and T helper 2 cytokines (IL-13 and IL-4).

Conclusions: Gal-9 may potentially play a role in suppressing anti-tumor immunity. This suggests the use of a Gal-9 neutralizing antibody for therapeutic benefit in melanoma.

Citation Format: Elizabeth Ann L. Enninga, Wendy K. Nevala, Svetomir N. Markovic. Galectin-9 has tumor promoting properties that may be targetable in metastatic melanoma. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A41.
Abstract A55: PD-1 blockade synergizes with cisplatin radiation therapy aiding in clearance of murine HPV+ oropharyngeal carcinoma
Authors: Vermeer Daniel W. , Powell Steven , Spanos William C. , Lee John H.
Published: October 2015

Squamous cell carcinomas of the head and neck (SCCHN) are the 6th most common malignancy world-wide with approximately 650,000 diagnoses per year. Human papillomavirus (HPV) causes up to 80% of oropharyngeal (tonsil/base of tongue) SCCHN. Despite presenting at an advanced stage, multiple studies show that HPV+ SCCHN are more curable relative to their HPV- counterparts. Clearance of SCCHN during treatment with cisplatin radiation therapy (CRT) is not only related to direct cytotoxicity, but is also dependent on immune mediated clearance. This immune clearance is strongly dependent on an intact CD4+ and CD8+ cellular response to tumor cells harboring these viral oncogenes. Furthermore, the immune system recognizes and clears HPV + SCCHN only after the start of CRT. The immunologic tolerance to these cancers prior to treatment could be explained by immune checkpoints aimed at preventing unregulated immune activity. The programmed death 1 receptor (PD-1) and its ligand, PD-L1, are one such checkpoint axis. Recent data suggests that the PD-1:PD-L1 pathway may play an important role in immune resistance in head and neck cancer. We evaluated PD-1 in combination with CRT in an immune competent HPV+ SCCHN mouse model. HPV + mouse SCCHN cells were injected subcutaneously into immune competent mice and tumors were treated after growing to palpable size. Using the mouse analog to the human PD-1 blocker, pembrolizumab, we determined that while single-agent therapy does not have significant activity on primary disease or distant metastasis, PD-1 blockade strongly synergizes with CRT. An overall survival of 58% was evident in mice treated with combination PD-1 and CRT versus 8% in mice treated with isotype control (IgG1) in combination with CRT (p = 0.006). These studies provide the impetus for future work investigating less toxic induction of immune responses in combination with blockade of immune checkpoint pathways.

Citation Format: Daniel W. Vermeer, Steven Powell, William C. Spanos, John H. Lee. PD-1 blockade synergizes with cisplatin radiation therapy aiding in clearance of murine HPV+ oropharyngeal carcinoma. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A55.
Abstract A71: Modulation of macrophages in breast tumors towards an anti-tumor phenotype
Authors: Guerriero Jennifer L. , Letai Anthony
Published: October 2015

While several avenues of manipulating the adaptive immune response have met with striking clinical success in recent years, there is also a growing appreciation of the innate immune system in mediating tumor reduction and elimination. In fact, tumor associated macrophages (TAMs) are the most abundant immune population in a breast tumor. Although M1 macrophages have potent anti-tumor activity, TAMs are phenotypically polarized to M2 macrophages, which support tumors. Clinically, a high tumor density of TAMs has been associated with chemoresistance and a worse clinical outcome. Using a tool, BH3 profiling, that is unique to the Letai Laboratory, we have identified that macrophage phenotype can alter the sensitivity of neighboring tumor cells to apoptotic signaling, and enhance their response to chemotherapy.

The Letai Laboratory has developed a unique and novel assay to predict a cancer cell's response to chemotherapy called BH3 profiling. BH3 profiling is used to determine whether a cell is relatively close to the threshold of apoptosis (“primed” for death), or relatively far from the threshold (“unprimed”). We have found that the priming of cancer cells, as measured by BH3 profiling, is by itself a predictor of clinical response to conventional chemotherapy. Using the BH3 profiling strategy, we show for the first time that M1 macrophages “prime” breast tumor cells for apoptosis, which correlates to their increased chemosensitivity. Interestingly, we have found some breast tumor cells to be resistant to M1 macrophage induced priming. We have performed gene expression analysis on these two phenotypes of breast tumor cells, to uncover why some, but not others, are sensitive to M1 macrophages. We also have identified how M1 macrophages increase the priming of tumor cells, making them more chemosensitive. These results suggest modulating TAMs towards an anti-tumor phenotype may have great promise for cancer therapy. To this end, we are investigating strategies to convert M2 TAMs to a M1 phenotype. We have conducted a high throughput screen (HTS) at the Institute of Chemistry and Cell Biology (ICCB) – Longwood, at Harvard Medical School, to identify small molecules that polarize macrophages to an anti-tumor phenotype. We have identified several interesting small molecules and are working to validate their clinical significance.

Citation Format: Jennifer L. Guerriero, Anthony Letai. Modulation of macrophages in breast tumors towards an anti-tumor phenotype. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A71.
Abstract B20: Enhanced affinity T cell receptor based therapy can surmount immune and physical barriers to treat pancreatic ductal adenocarcinoma
Authors: Stromnes Ingunn M. , Brockenbrough Scott J. , Schmitt Thomas M. , Hingorani Sunil R. , Greenberg Philip D.
Published: October 2015

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR08) of the Conference Proceedings.

Citation Format: Ingunn M. Stromnes, Scott J. Brockenbrough, Thomas M. Schmitt, Sunil R. Hingorani, Philip D. Greenberg. Enhanced affinity T cell receptor based therapy can surmount immune and physical barriers to treat pancreatic ductal adenocarcinoma. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B20.
Abstract B30: Signal peptide –mediated immunization against mouse mammary tumor virus (MMTV)-associated tumors
Authors: Braitbard Ori , Gross Tamar , Roniger Maayan , Bar-Sinai Allan , Hochman Nira , Hochman Jacob
Published: October 2015

Signal peptides (SPs) are N-terminal extensions on nascent secretory and membrane proteins that mediate insertion into, or translocation across the membrane of the endoplasmic reticulum (ER). A growing number of SPs have been shown to carry out post-ER targeting functions. A case in point is the SP of the envelope precursor protein of Mouse Mammary Tumor Virus (MMTV), a beta retrovirus known to cause lymphoma and mammry carcinoma in laboratory mice. This signal peptide (named MMTV-p14 , or p14 for short) is uniquely localized to nucleoli of cells that harbor the virus where it binds to specific protein targets and functions as a nuclear export factor for viral transcripts. P14 is also a phosphoprotein tumor modulator, the phosphorylation status of which determines whether it will function, in vivo, in a tumor promoting or tumor suppressing capacity.

Here we report that p14 is expressed also on the surface of a variety of cells that contain MMTV (murine lymphoma and mammary carcinoma, and human breast cancer cells ectopically expressing p14). Furthermore p14 is immunogenic and vaccination with purified recombinant p14 (using different adjuvants) is sufficient to immunize mice against malignant cells that harbor MMTV. This can be adoptively transferred into naïve mice.

Furthermore, newly derived monoclonal anti-p14 antibodies with different epitope specificity are now available to study their effect on MMTV associated tumors in vivo as well as for targeted drug delivery to these tumors

In view of reports suggesting involvement of MMTV in about 30% of human breast cancers we propose p14 and anti-p14 antibodies as candidates for immunization against MMTV associated malignancies, and as a functional example for other viral signal peptides.

Citation Format: Ori Braitbard, Tamar Gross, Maayan Roniger, Allan Bar-Sinai, Nira Hochman, Jacob Hochman. Signal peptide –mediated immunization against mouse mammary tumor virus (MMTV)-associated tumors. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B30.
Abstract B59: The combined effects of piperlongumine, vitamin C, and vitamin D on Caenorhabditis elegans and Volvox carteri
Authors: Winters Ashley M.
Published: October 2015

Apoptosis, part of the everyday cell growth, essential for killing tumor cells, vital in organism's survival, kills pernicious cells, thus ridding the organism of mutations. The p53 gene is a tumor suppressor that is mutated in over half of all human tumor cells. When activated, it can stimulate DNA repair and apoptosis, making it crucial in the killing of tumor cells. Piperlongumine, vitamin C, and vitamin D have shown to increase p53 activity, thus increasing survival. Caenorhabdits elegans (C.elegans) underwent ultraviolet-C radiation, subjected to these reagents, and were counted daily in an eight day observation period for survival. Western blot analysis was used to compare the p53 gene activity. Similarly, Volvox carteri were observed for population after heat shock and the administration of the reagents. The results showed the combination of all three reagents led to the largest increase in p53 protein activity, lifespan in the nematodes, and population in the green algae. In C. elegans, the combination of vitamin C and D, statistically, led to the same increase in lifespan as piperlongumine alone. It was found that increasing reagent dosage induced a heighten population in Volvox carteri, while the control drastically decreased. These findings provide further evidence that there indeed exists a p53 gene in Volvox carteri.

Citation Format: Ashley M. Winters. The combined effects of piperlongumine, vitamin C, and vitamin D on Caenorhabditis elegans and Volvox carteri [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B59.