tbuckley/cabot-search huggingface dataset:
618. bFGF or VEGF Gene Transfer Via AAV2 Vectors To the Tendon Activates Multiple Genes Critical To Tendon Growth and Regeneration: An In Vivo Study
Authors: Xiao Tian Wang, Ya Fang Wu, You Lang Zhou, Jin Bo Tang, Paul Y. Liu
Journal: Molecular Therapy (2015)

Purpose: Tendon healing is innately weak and transfer of growth factor gene therapy may activate multiple genes necessary or specific for tendon growth and regeneration. In this study we transferred bFGF or VEGF gene by AAV2 vectors to the healing tendon in a chicken flexor tendon model. We investigated how endogenous tendon healing related genes were activated after the delivery of AAV2-bFGF or AAV2-VEGF and the differences between epitenon and endotenon using a laser capture microdissection ((LCM). Methods: Eighteen digital flexor tendons from the big toes of 9 chickens were used. The tendons were completely transected and divided into three groups: AAV-bFGF injection, AAV-VEGF injection and no-injection control. After AAV injection, the tendons were surgically repaired. At 4 weeks after surgery, the tendons were harvested. The formalin-fixed, paraffin-embedded tendons were sectioned. LCM was used to dissect epitenon and endotenon separately. The tissue was subsequently analyzed for gene expression of collagen type I alpha 1 (COL1A1), collagen type III alpha 1 (COL3A1), proliferating cell nuclear antigen (PCNA), fibronectin 1 (FN1), scleraxis (SCX) and tenascin C (TNC) using RT-qPCR analysis. Results: In AAV-VEGF and AAV-bFGF injected tendons, we found significant increases in expression of FN1, PCNA, SCX, COL1A1 and COL3A1 genes (p < 0.05 or p<0.01) compared with no-injection control. In the AAV-bFGF injected tendon, SCX was expressed 5-fold higher in the endotenon than in the epitenon. The increases in TNC gene expression did not reach statistical significance, but interestingly, TNC gene was expressed highly in the endotenon (10-fold greater than in the epitenon) in AAV-VEGF injected tendon. The TNC in the endotenon of AAV-VEGF-injected tendon was raised by 55-fold compared with that of the non-injected tendon. Discussion: Our results demonstrated a drastic increase in expression of studied genes specific to tendon growth and regeneration after AAV2-bFGF or AAV2-VEGF therapy. We also found precisely the epitenon and endotenon are equally responsive to the gene therapy. This study provides evidence at genetic levels to support the effective changes of the gene expression by the gene therapy, which may be key mechanistic findings to support the use of these gene therapies.
Abstract 4198: mTORC2 directs breast morphogenesis through Rictor-dependent PKCα/Rac1 signaling independent of Akt
Authors: Dana M. Brantley-Sieders, Meghan M. Morrison, Donna J. Hicks, Rebecca S. Cook
Journal: Cancer Research (2015)

Abstract Akt phosphorylation is a major driver of cell survival, motility, and proliferation in development and disease, causing increased interest in upstream regulators of Akt like mTOR complex 2 (mTORC2). We used genetic disruption of Rictor to impair mTORC2 activity in mouse mammary epithelia, which decreased Akt phosphorylation, ductal length, secondary branching, cell motility, and cell survival. These effects were recapitulated with a pharmacological dual inhibitor of mTORC1/mTORC2, but not upon genetic disruption of mTORC1 function via Raptor knock-out. Surprisingly, Akt re-activation was insufficient to rescue survival, branching, or motility of mTORC2-impaired mammary epithelial cells (MECs). However, activation of the mTORC2 substrate protein kinase C (PKC)-α fully rescued branching, invasion, and survival of MECs after genetic or pharmacological mTORC2 inhibition. PKCα-mediated signaling through the small GTPase Rac1 was necessary for mTORC2-dependent mammary morphogenesis, revealing a novel role for Rictor/mTORC2 in survival and motility of untransformed MECs through an Akt-independent, PKCα/Rac1-dependent mechanism. Ongoing analyses of mammary specific Rictor-deletion in genetically engineered mouse models of breast cancer support a key role for mTORC2 in tumor progression and metastasis. Citation Format: Dana M. Brantley-Sieders, Meghan M. Morrison, Donna J. Hicks, Rebecca S. Cook. mTORC2 directs breast morphogenesis through Rictor-dependent PKCα/Rac1 signaling independent of Akt. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4198. doi:10.1158/1538-7445.AM2015-4198
Abstract LB-043: Highly specific and sensitive nanoimmunoassay of PARP1 as cell death measurement toolkit
Authors: Michael A. Pontikos, Amos Zimmermann, Camilo Moncada, Karin V. Abarca Heidemann, Jordan B. Miller, Jonathan Birabaharan, Michael Boyer, Robin M. Zuck, Philip L. Lorenzi, Carl Ascoli
Journal: Cancer Research (2015)

Abstract We developed and validated a highly specific toolkit to analyze PARP1 in a panel of control and siRNA knockdown cell lysates by multiple immunoassays. We can specifically and quantitatively distinguish PARP1 from 17 other PARP family members using highly specific antibodies and optimized methods. Poly (ADP-ribose) polymerase-1 (PARP1) is a chromatin associated, ADP-ribosylating enzyme essential for multiple cellular functions, including cardiac remodeling, vasoconstriction, regulation of astrocyte and microglial function, long term memory, aging, transcription regulation, and DNA repair. More recently, it has been implicated in a new form of cell death termed parthanatos. PARP1 can also promote tissue survival by shifting the balance of cell death programs between autophagy and necrosis. Since PARP1 can promote tumorigenicity, it has gained traction as a therapeutic target in cancer. In that regard, clinical studies have shown vulnerability to PARP inhibitors in DNA repair defective cancers. One of the difficulties in analyzing PARP1 activity is the promiscuity of the reagents toward other PARP family members. Furthermore, the ability to examine multiple tissue samples in parallel has been limited. In this study we developed highly sensitive and specific antibodies against PARP1 fragments. Using siRNAs against 18 PARP family members, we validated antibody specificity by western blotting and nanoimmunoassay (NIA). NIA is a highly sensitive platform capable of screening up to 96 cancer samples at sub-microliter volume. Moreover, this platform permits quantitative analysis of non-modified PARP1 as well as post-translational modifications such as PARylation and phosphorylation using a single antibody for the measurement of all species. We anticipate this workflow will be amenable to a wide range of protein targets, ushering in a new frontier in diagnostic analysis. Citation Format: Michael A. Pontikos, Amos Zimmermann, Camilo Moncada, Karin V. Abarca Heidemann, Jordan B. Miller, Jonathan Birabaharan, Michael Boyer, Robin M. Zuck, Philip L. Lorenzi, Carl Ascoli. Highly specific and sensitive nanoimmunoassay of PARP1 as cell death measurement toolkit. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-043. doi:10.1158/1538-7445.AM2015-LB-043
38. Pharmacological Regulation of Vesicular Trafficking as a Strategy to Enhance Recombinant AAV Transduction
Authors: Garrett E. Berry, Aravind Asokan
Journal: Molecular Therapy (2015)

Vector dose-related toxicity has been highlighted as a potential safety concern in recent preclinical studies and clinical trials with recombinant AAV vectors. Thus, investigation of strategies to enhance AAV transduction may allow for administration of lower vector doses, thereby mitigating toxicity. Several host cell factors have been shown to restrict viral infection during post-entry trafficking steps through a variety of mechanisms. In the case of AAV, the 26S proteasome is thought to reduce transduction efficiency by restricting nuclear entry via ubiquitin-dependent capsid degradation. The AAA+ ATPase p97/valosin-containing protein (VCP) is a segregase and unfoldase which is closely associated with the proteasomal degradation pathway and disassembles multimeric protein complexes for subsequent processing. VCP modulates numerous cellular processes ranging from endosomal sorting to autophagy and ER-associated degradation. In the current study, we sought to determine whether VCP plays a role as a restriction factor in the AAV infectious pathway. Chemical inhibition of VCP by Eeyarestatin I (EerI) increased AAV transduction by nearly an order of magnitude in a cell type and serotype independent manner. This effect was associated with increased recovery of viral genomes and capsid accumulation within the nucleus, but not cell surface binding or uptake. Interestingly, VCP appears to restrict AAV infection by regulating vesicular trafficking of viral particles. In particular, VCP inhibition redistributes AAV capsids to Rab7 and LAMP1 positive vesicles, while simultaneously disrupting capsid association with syntaxin-5 positive vesicles recently shown to be essential for Golgi transport. These results support the notion that AAV vesicular transport can be redirected to enhance transduction efficiency. In summary, our results identify VCP as a novel host restriction factor that regulates the post-entry trafficking of recombinant AAV vectors, and highlight VCP as a potential target for modulating AAV transduction in the clinic.
Long Non-Coding RNA MALAT1 Is Associated with the Progression of Multiple Myeloma and Induced By Cellular Stress
Authors: Yuko Kuroda, Kei Kimura, Yuta Masuda, Arito Yamane, Hikaru Hattori, Kenichi Tahara, Ayaka Kaneko, Itsumi Suda, Noriyuki Takahashi, Gotoh Nanami, Kie Nagai, Tomomi Nagashima, Makiko Takizawa, Hiromi Koiso, Takuma Ishizaki, Takeki Mitsui, Hirono Iriuchishima, Takayuki Saitoh, Hiroaki Shimizu, Akihiko Yokohama, Norifumi Tsukamoto, Tetsuhiro Kasamatsu, Yusuke Minato, Yoshihisa Nojima, Hirokazu Murakami, Hiroshi Handa
Journal: Blood (2015)
367. In Vitro Electrotransfer of Small and Large Plasmids in Mesenchymal Stem Cells: Calcium, Temperature and PH Impact
Authors: Hugues O. Gontier, Léa L. Lesueur, Lluis M. Mir, Franck M. Andre
Journal: Molecular Therapy (2015)

Nucleic acids electrotransfer has been shown to be a safe and efficient non-viral technique in a wide variety of cells (including primary cells) even with large nucleic acids. Using a small 3.5 kbp GFP reporter plasmid we previously obtained electrotransfer efficiency of up to 90%, with around 70% cell viability, in Mesenchymal Stem Cells (MSCs). However with larger plasmids (about 15 kbp), we observed highly decreased cell viability and electrotransfer efficacy in MSCs. Our results suggested that this could be the consequence of the increased time necessary for larger plasmids to cross the plasma membrane. Here we further studied the mechanisms of small and large plasmid electrotransfer. We investigated the roles of calcium, temperature and pH. Our results shed light on some of the mechanisms involved in gene electrotransfer as well as provide means to enhance electrotransfer efficacy and cell viability for small and large plasmids.
Abstract 9478: GRK2-Mediated Phosphorylative Desensitization of AdipoR1 in the Failing Heart
Authors: Yajing Wang, Erhe Gao, Wayne Bond Lau, Yuexing Yuan, Theodore Christopher, Bernard Lopez, Walter Koch, Xin Ma
Journal: Circulation (2014)

Adiponectin (APN) is a novel adipokine whose cardioprotective actions in acute myocardial infarction (MI) are well-defined. However, the role of APN in chronic heart failure (HF) is controversial. Experimental studies demonstrate APN deficiency exacerbates HF progression. Paradoxically, clinical observations report APN increases in relation to worsening HF. Here, we determine whether APN receptors 1 and 2 (AdipoR1/AdipoR2, the 7-transmembrane domain-containing receptors mediating APN actions) are phosphorylatively modified post-MI, similar to G-protein coupled receptors (GPCR), impeding APN signaling. Receptor phosphorylation, kinase expression, and APN function were determined via in vivo, ex vivo, and in vitro models. Neither AdipoR1 nor AdipoR2 were phosphorylated in the normal heart. However, AdipoR1 was phosphorylated post-MI, peaking at 7 days, and remained significantly phosphorylated thereafter. The extent of post-MI AdipoR1 phosphorylation positively correlated with the expression level of GPCR kinease-2 (GRK2). Cardiac-specific GRK2 knockdown virtually abolished post-MI AdipoR1 phosphorylation, whereas virus-mediated GRK2 overexpression significantly phosphorylated AdipoR1 and blocked APN metabolic-regulatory/anti-inflammatory signaling. Ex vivo experiments demonstrated AMPK activation and anti-TNFα effect of APN were significantly inhibited in cardiomyocytes isolated from non-ischemic cardiac regions 7 days post-MI. In vivo experiments demonstrated that acute APN administration induced cardiac GLUT4 translocation, and eNOS phosphorylation was blunted 7 days post-MI. Continuous APN administration beginning 7 days post-MI failed to protect against adverse cardiac remodeling and HF progression in WT mice, which was restored by cardiac-specific GRK2 knockdown. These results demonstrate for the first time that the cardiometabolic-regulatory, anti-inflammatory, and cardioprotective functions of APN are significantly impaired by GRK2-mediated AdipoR1 phosphorylative desensitization during post-MI HF development, suggesting that inhibiting GRK2-mediated AdipoR1 phosphorylation may be a novel therapeutic target, restoring APN signaling and mitigating post-MI remodeling and HF.
Blockage of SDF-1-CXCR4 Axis by AMD 3100 Can Be a Novel Therapy for Acute Lymphoblastic Leukemia by Targeting the Extramedullary Sites of Leukemic Cells.
Authors: Itaru Kato, Akira Niwa, Hisanori Fujino, Katsutsugu Umeda, Satoshi Saida, Souichi Adachi, Mamoru Ito, Toshio Heike, Tatsutoshi Nakahata
Journal: Blood (2009)
Screening of JAK2 V617F Mutation in 84 Patients with Thrombocythemia and Erythrocytosis. Is Necesary for Diagnosis of a Chronic Myeloproliferative Disorders?.
Authors: Marta A. Sobas, M. Martinez, M. Perez-Encinas, T. Gonzalez, C. Quinteiro, F. Barros, A. Bendana, E. Ansoar, M. Rabunal, S. Gonzalez, N. Alonso, J. Diaz, J. Bello
Journal: Blood (2006)
Rapid Mobilization of Murine Hematopoietic Stem and Progenitor Cells with TG-0054, a Novel CXCR4 Antagonist.
Authors: Ying-Huey Huang, Yi-Chun Liu, Chi-Feng Yen, Hua-Chien Chen, Shu-Jen Chen, Chi-Hsin R. King, Ming-Chu Hsu
Journal: Blood (2009)
Abstract 5103: Loss of p27 upregulates MnSOD in a STAT3-dependent manner, disrupts intracellular redox activity and enhances cell migration
Authors: Jing An, Dongyun Zhang, Chuanshu Huang
Journal: Cancer Research (2015)

Abstract Cell migration is a dynamic process that is central to a variety of physiological functions as well as disease pathogenesis. The modulation of cell migration by p27 (officially known as CDKN1B) has been reported, but the exact mechanism(s) whereby p27 interacts with downstream effectors that control cell migration have not been elucidated. By systematically comparing p27+/+ mouse embryonic fibroblasts (MEFs) with genetically ablated p272/2 MEFs using wound-healing, transwell and time-lapse microscopic analyses, we provide direct evidence that p27 inhibits both directional and random cell migration. Identical results were obtained with normal and cancer epithelial cells using complementary knockdown and overexpression approaches. Additional studies revealed that overexpression of manganese superoxide dismutase (MnSOD, officially known as SOD2) and reduced intracellular oxidation played a key role in increased cell migration in p27-deficient cells. Furthermore, we identified signal transducer and activator of transcription 3 (STAT3) as the transcription factor responsible for p27-regulated MnSOD expression, which was further mediated by ERK- and ATF1-dependent transactivation of the cAMP response element (CRE) within the Stat3 promoter. Collectively, our data strongly indicate that p27 plays a crucial negative role in cell migration by inhibiting MnSOD expression in a STAT3-dependent manner. Citation Format: Jing An, Dongyun Zhang, Chuanshu Huang. Loss of p27 upregulates MnSOD in a STAT3-dependent manner, disrupts intracellular redox activity and enhances cell migration. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5103. doi:10.1158/1538-7445.AM2015-5103
Association of Weight Loss Maintenance and Weight Regain on 4-Year Changes in CVD Risk Factors: the Action for Health in Diabetes (Look AHEAD) Clinical Trial
Journal: Diabetes Care (2016)

Short-term weight loss improves cardiovascular disease (CVD) risk factors. We sought to determine the longer-term effects of maintaining weight loss or, conversely, regaining weight.We used data from Action for Health in Diabetes (Look AHEAD), a randomized trial of intensive lifestyle intervention (ILI) compared to a control condition in overweight/obese individuals with type 2 diabetes. ILI participants were grouped according to weight change patterns, as follows: 1) no weight loss (±3% at years 1 and 4); 2) moderate weight loss (3-8% at years 1 and 4); 3) large weight loss (8-20% at years 1 and 4); 4) moderate loss/full regain (3-8% at year 1/±3% at year 4); 5) large loss/full regain (8-20% at year 1/± 3% year 4); and 6) large loss/partial regain (8-20% at year 1/3-8% at year 4) and changes in CVD risk factors were compared.Adjusting for baseline differences and medication use, larger weight losses produced greater improvements in HbA1c, systolic blood pressure, HDL cholesterol, and triglycerides at years 1 and 4 (all P ≤ 0.02). Despite maintenance of weight loss, HbA1c levels worsened between years 1 and 4, and remained below baseline only in those with large weight losses. We found no negative associations of losing and regaining weight relative to not having lost weight. Moreover, those who had large initial weight loss but full regain of weight had greater improvements in HbA1c levels at year 4 than those with smaller or no initial weight loss.Larger initial weight loss should be encouraged in individuals with type 2 diabetes, despite the possibility of regain.
Abstract 1067: Characterization of DNA hypermethylation in two cases of peritoneal mesothelioma
Authors: Ritsuko Oikawa, Yoshiyuki Watanabe, Ryota Hama, Hiroyuki Yamamoto, Fumio Itoh
Journal: Cancer Research (2015)

Abstract Background: Malignant mesothelioma (MM) is a rare disease with a poor prognosis. Pleural mesothelioma, which is the most common type of MM, is considered to be caused by asbestos exposure and is increasing in incidence, with about 15,000 new cases diagnosed worldwide annually. On the other hand, peritoneal mesothelioma is a very rare type of MM; thus its pathogenesis is even less understood than pleural mesothelioma. Recent research on the pathogenesis of malignant pleural mesothelioma has indicated that both epigenetic and genetic alterations contribute to tumorigenesis. Here, we hypothesize that peritoneal mesothelioma also has an epigenetic alteration in the same genes (KAZALD1, TMEM30B and MAPK13). Our goal is to identify DNA methylation of these three candidate genes in two peritoneal mesothelioma cases. Methods: Laser capture microdissection (LCM) was used to separate diseased sections of formalin-fixed paraffin-embedded (FFPE) samples from one surgically resected tissue (epithelial type) and one autopsy tissue (sarcomatous type). Genomic DNA was subsequently extracted by the standard phenol chloroform method. The DNA was then treated with sodium bisulfite, and pyrosequencing analysis was used to quantitatively analyze the methylation of candidate genes reported to be hypermethylated in malignant pleural mesothelioma (KAZALD1, TMEM30B and MAPK13). Results: TMEM30B and MAPK13 were not methylated in either case. However, KAZALD1 was highly methylated in sarcomatoid type peritoneal mesothelioma. Discussion: We first report that the KAZALD1 gene was hypermethylated in sarcomatoid type malignant peritoneal mesothelioma. Citation Format: Ritsuko Oikawa, Yoshiyuki Watanabe, Ryota Hama, Hiroyuki Yamamoto, Fumio Itoh. Characterization of DNA hypermethylation in two cases of peritoneal mesothelioma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1067. doi:10.1158/1538-7445.AM2015-1067
PTH-079 Direct-to-test endoscopic ultrasound
Authors: P Lochhead, P S Phull
Journal: Gut (2010)

<h3>Introduction</h3> Endoscopic ultrasound (EUS) has become an invaluable investigation in the assessment of pancreatobiliary disease. EUS is superior to abdominal ultrasound in pancreatobiliary imaging having a sensitivity of &gt;90% for choledocholithiasis and affording better views of the pancreas. Traditionally, patients tend to undergo EUS following specialist review. We are not aware of any reports of direct-to-test EUS for patients who have been assessed only by a general practitioner (GP). We propose that, in selected patients with suspected choledocholithiasis, direct-to-test EUS is safe and avoids specialist clinic review and the attendant delay, expense and inconvenience. Here we present our initial data on direct-to-test EUS in a teaching hospital setting. <h3>Methods</h3> Twenty-four patients who had been referred to the gastroenterology department at our institution by their general practitioners between February 2005 and July 2009 were selected to be offered direct-to-test EUS. Selected patients had liver chemistry and/or symptomatology compatible with biliary pathology; all but one patient had had an abdominal ultrasound scan before referral. A patient information leaflet was sent in the post along with the appointment letter. <h3>Results</h3> All patients accepted direct-to-test EUS appointment; the average time from referral to procedure was 6 weeks. The average patient age was 59 years (range 21–83); 17 out of 24 patients were female (71%). The clinical details were: biliary-type pain with abnormal liver chemistry but normal ducts on ultrasound (n=14); abnormal liver chemistry with duct dilatation but no stones identified on ultrasound (n=4); biliary-type pain, normal chemistry, with duct dilatation but no stones on ultrasound (n=3); progressive cholestatic liver chemistry post cholecystectomy with no duct dilatation on ultrasound (n=2); unexplained recent pancreatitis with a normal ultrasound (n=1). The EUS findings for the 24 patients were: normal examination (n=13), choledocholithiasis (n=3), gallstones (n=5), hepatic steatosis (n=2), periampullary tumour (n=1). Three of the cases of gallstones had not been previously identified on abdominal ultrasound scanning. The overall yield for pathology was 46%. There were no noted complications in any of the 24 cases. On the basis of the EUS examination, three patients were referred for ERCP, two for cholecystectomy, and one for a Whipple9s procedure. This equates to a 25% onward referral rate for definitive treatment based on the direct-to-test EUS findings. <h3>Conclusion</h3> Our initial data support the hypothesis that direct-to-test EUS for selected patients with suspected biliary disease is a safe investigation, with a high yield of significant pathology.
Ibritumomab Tiuxetan (Zevalin) Followed by High-Dose Chemotherapy and Autologous Stem-Cell Transplantation Results in Improved Survival of Patients with Chemo-Refractory Non-Hodgkin’s Lymphoma Expected To Have Poor Outcome with Standard Pre-Transplant Conditioning.
Authors: Avichai Shimoni, Tsila Zwass, Yaacov Oksman, Izhar Hardan, Noga Shem-Tov, Ronit Yerushalmi, Abraham Avigdor, Isaac Ben-Bassat, Arnon Nagler
Journal: Blood (2006)
A Unique Leukemia Mouse Model Established From AML Patient with IDH2 R140Q and FLT3-ITD Mutations Among Other Common AML Mutations
Authors: Jinping Liu, Ran Wu, Na Wang, Di Wang, Liang Huang, Jie Cai, Hong Wang, Taiping Chen, Jean-Pierre Wery, Yiyou Chen, Henry Qixiang Li
Journal: Blood (2012)