tbuckley/cabot-search huggingface dataset:
165. Liver-Directed Gene Therapy for Murine Glycogen Storage Disease Type IB
Authors: Joonhyun Kwon, Jun-Ho Cho, Young Mok Lee, Goo-Young Kim, Javier Anduaga, Janice Chou
Journal: Molecular Therapy (2016)

Glycogen storage disease type Ib (GSD-Ib) deficient in the glucose-6-phosphate transporter (G6PT or SLC37A4) is characterized impaired glucose homeostasis, myeloid dysfunction, and long-term complication of hepatocellular adenoma (HCA). We have shown that gene therapy mediated by a recombinant (r) AAV8 vector expressing G6PT directed by the chicken β-actin promoter/CMV enhancer enabled the G6pt-/- mice lived to over 51 weeks but all 5 transduced G6pt-/- mice expressed only low levels of hepatic G6PT activity and two developed multiple HCAs with one undergoing malignant transformation. We now examined the safety and efficacy of rAAV8-GPE-G6PT, a rAAV8 vector expressing G6PT directed by the gluconeogenic tissue-specific human G6PC promoter/enhancer (GPE). Of the fifteen rAAV8-GPE-G6PT-treated G6pt-/- mice that lived over age 60 weeks expressed 2-62% of wild-type hepatic G6PT activity with only one developed HCA. The treated mice, including the HCA-bearing mouse exhibit a leaner phenotype along with normal blood metabolite, display normal glucose tolerance profiles, maintain normoglycemia over a 24-hour fast, and retain insulin sensitivity. We further show that activation of hepatic ChREBP signaling that improves glucose tolerance and insulin sensitivity is one mechanism that protects the rAAV-GPE-G6PT-treated G6pt-/- mice against age-related obesity and insulin resistance.
163. Hepatocyte Targeted Endosomal Escape Agents
Authors: Christopher W. White, Nicholas J. Baumhover, Kevin G. Rice
Journal: Molecular Therapy (2015)

The successful development of potent non-viral DNA and mRNA delivery agents to transfect liver hepatocytes in vivo requires the use of an endosomal escape molecule. Two major strategies have emerged, one that utilizes membrane lytic peptides, and a second that uses proton sponge polymers. Both approaches would require administration of excess endosomal release agent with a smaller dose of targeted polyplex. The present study compared these approaches by conducting 384-well in vitro transfections on primary hepatocytes, as previously described1. An N-glycan targeted melittin was designed to target hepatocytes via the asialoglycoprotein receptor. The synthesis used a purified triantennary N-glycan from bovine fetuin as an potent high affinity asialoglycoprotein receptor targeting ligand that was conjugated by disulfide bound to an N-terminal Cys on melittin, a 26 amino acid membrane lytic peptide. Following targeted entry into hepatocytes via receptor mediated endocytosis, melittin is bioactivated by reduction of the disulfide bond to release the N-glycan, leading to endosomal lysis. RBC hemolysis assays established the membrane lytic activity of triantennary-melittin was muted prior to reduction, which allowed it to be safely i.v. dosed in mice. A second strategy involved the modification of PEI by reductive amination with lactose followed by acetylation, resulting in an asialoglycoprotein receptor targeted proton sponge polymer. The resulting LacAcyl-PEI was purified by RP-HPLC and characterized by NMR. The i.v. dosing of LacAcyl PEI in mice demonstrated that it possessed a much greater safety margin than unmodified PEI. These novel endosomal escape agents are being compared for their ability to mediate in vitro gene transfer of glycan targeted DNA and mRNA in miniaturized primary hepatocyte transfection assays.
701. Long-Term Complications of Glycogen Storage Disease Type IA in the Dog Model Treated With Gene Replacement Therapy
Authors: Elizabeth D. Brooks, Dustin J. Landau, Talmage T. Brown, Yohannes G. Asfaw, Dwight D. Koeberl
Journal: Molecular Therapy (2015)

Glycogen storage disease type Ia (GSD-Ia) in dogs closely resembles human GSD-Ia where untreated patients develop complications associated with glucose-6-phosphatase (G6Pase) deficiency (hypoglycemia, hyperlipidemia, growth retardation, and early death). For the past 3 decades survival of human patients that are placed under intensive nutritional management with uncooked corn starch has improved; however, long-term complications persist including renal failure, nephrolithiasis, hepatic adenomas, and a high risk for hepatocellular carcinoma. Affected dogs fail to thrive with dietary therapy alone, but treatment with gene replacement therapy using adeno-associated viral vectors (AAV) expressing G6Pase has greatly prolonged life and prevented hypoglycemia. In this study, 7 GSD-Ia affected dogs treated with AAV-G6Pase were followed up to 8 years of life to describe long-term complications. All required readministration of AAV vector(s) pseudotyped as a new serotype to avoid anti-AAV antibodies, due to decreased ability to maintain normoglycemia during fasting. Two dogs developed chronic renal disease as denoted by polyuria, polydipsia, and azotemia at 6 and 8 years of age. Calcium oxalate and phosphate urolithiasis was detected in 2 dogs; 1 dog had evidence of polycystic ovarian disease. Four of the 7 dogs were euthanized due to reaching humane endpoints related to liver and/or kidney disease, at 3 to 8 years of life, and 1 dog succumbed to acute hypoglycemia. Necropsy revealed several focal hepatic lesions in all deceased dogs; 2 dogs had lesions confirmed histologically as hepatocellular carcinoma, the largest tumor reached 8 cm in diameter (Fig. 1). One dog demonstrated lesions consistent with hepatocellular adenoma and in the other 2, there was hepatocellular hyperplasia. There was no significant difference in amount of vector DNA in hepatic tumors compared with normal tissue (n=4 dogs), suggesting that insertional mutagenesis by the AAV vector was not the mechanism for tumor formation (Fig. 2). Glomerular sclerosis, fibrosis of Bowman's capsules and focal interstitial nephritis were found in the 2 dogs with clinical renal disease. Two dogs, both female, remain in good health with no evidence of liver masses appreciable on ultrasound examination; they are 4 and 5 years of age. Hepatocellular carcinoma, kidney disease, polycystic ovaries and urolithiasis are common findings among GSD-Ia patients. Here we show that the canine GSD-Ia model demonstrates similar long-term complications in spite of AAV-mediated gene replacement therapy. Further development of gene therapy is needed to prevent long-term complications of GSD-Ia.
Effects of eltrombopag on human cord blood hematopoietic stem cell/primitive progenitor cell expansion.
Authors: H. Sun, Y. Tsai, J. Liesveld, K. L. Dawson, C. L. Erickson-Miller, Y. Chen
Journal: Journal of Clinical Oncology (2011)

6632 Background: Umbilical cord blood transplantation has emerged as a promising therapy but is low in stem cells. Agents enhancing expansion of CB hematopoietic stem cell (HSC)/hematopoietic progenitor cell (HPC) are highly desirable. Eltrombopag (epag) is a non-peptide, c-MPL agonist that activates c-MPL of human and chimps. We investigate effects of epag on expansion of HSC/HPC of human CB. Methods: (1) In vitro: CB CD34+ cells were treated with rhTPO or epag in presence of rhSCF and rhFL for 7 days. After staining with anti-CD41, anti-CD34 and anti-CD38, cells were analyzed by Flow. (2) Intracellular signaling: serum-starved cells expanded from CB CD34+ were treated with rhTPO or epag. Phosphorylation of STAT3, STAT5 and AKT in c-MPL signal pathway was analyzed by Flow after staining with anti-phospho-STAT3, anti-phospho-STAT5, anti-phospho-AKT anti-CD41, anti-CD34, and anti-TPO-receptor. (3) In vivo: Sublethally irradiated NOD/SCID mice were transplanted with CB CD34+ and gavaged daily with epag or water (28 d) and sacrificed between 1-9 wks. Blood count and bone marrow (BM) extracts were analyzed by Flow after staining with anti-human-CD41a-PE, anti-mouse-CD61-FITC, anti-human-CD45-FITC, anti-mouse CD45-PE, anti-human-CD34-FITC and anti-human-CD41a-PE antibodies. Results: (1) Epag increased CD34+ by 26%, CD34+CD38- by 42%, and CD41+ by 400% in culture, compared with130%,160% and 900%, respectively by rhTPO. The % CD34+, CD34+CD38- and CD41+ were similar comparing rhTPO with epag. (2) Both rhTPO and epag induced phosphorylation of STAT5 in CD34+CD41-, CD34-CD41+ and CD34-CD41- cells. rhTPO-induced pSTAT5 was cell-type dependent, while epag-induced pSTAT5 was not. rhTPO-induced the phosphorylation of STAT3 and AKT in CD34-CD41+ cells, while pSTAT3 and pAKT were not detectable in epag treated CD34-CD41+ cells. (3) Epag led to an increase of circulating human platelets and WBCs without affecting murine platelets or WBCs in NOD/SCID. Epag augmented expansion of human CD45+, CD34+ and CD41+ cells in BM without effects on mouse BM cells. Conclusions: Eltrombopag expanded HSC/HPC of CB in vitro, promoted human platelet and WBC proliferation/differentiation in NOD/SCID mice, and human HSC/HPC in mouse BM.
Comparative Single-Institute Analysis of Cord Blood Transplantation from Unrelated Donors with Unrelated Bone Marrow or Related Peripheral Blood Stem-Cell Transplants in Elderly Patients with Hematologic Diseases after Reduced Intensity-Conditioning Regimen.
Authors: Naoyuki Uchida, Atsushi Wake, Kazuhiro Masuoka, Kazuya Ishiwata, Masanori Tsuji, Hisashi Yamamoto, Shinsuke Takagi, Daisuke Kato, Sachiko Seo, Naofumi Matsuno, Shigesaburo Miyakoshi, Akiko Yoneyama, Shuichi Taniguchi
Journal: Blood (2007)
Preventing Platelet Alloimmunization for Patients with Hematologic Malignancies: Rapid HLA-class I and HPA Immunogenetic Profiling to Enable Selection of Optimally Matched Platelets
Authors: Ghazala Hashmi, Dipika Patel, Yi Zhang, Michael Seul, Fleur M. Aung, Pedro Cano, Emil J. Freireich, Benjamin Lichtiger
Journal: Blood (2012)
Association of the expression of the glioma-associated oncogene homolog (GLI) 1 with nuclear expression of NF-κB and unfavorable overall survival of patients with pancreatic cancer.
Authors: S. Yang, S. H. Kuo, C. Hsu, J. Lee, Y. Tien, C. Hsu, A. Cheng
Journal: Journal of Clinical Oncology (2011)

4112 Background: Hedgehog signaling pathway is involved in the moleculopathogenesis of pancreatic cancer. In pancreatic cancer, NF-κB up-regulates sonic hedgehog (Shh) expression and further contributes to the activation of hedgehog (Hh) signaling pathway. In this study, we examined the association of Hh signaling pathway with NF-κB activation and clinical outcome of pancreatic cancer patients. Methods: We reviewed the medical records and pathologic specimens of 87 patients with histologically proven primary pancreatic cancer diagnosed at our institutions from January 1, 1995 through December 30, 2007. Of the pancreatic cancer patients, 74 with available tumor specimens for further evaluated for the expression of NF-κB (RelA/p65), sonic hedgehog (Shh), and Gli1 by immunohistochemistry. The expression of NF-κB activation and Hh signaling pathway correlated with the clinicopathologic features and overall survival (OS) of these patients. Results: The frequencies of expression of Shh, and nuclear expression of Gli1 and NF-κB were found in 68 (92%) of 74 cases, 29 (39%) of 74 cases, and 23 (31%) of 74 cases, respectively. The expression of Gli1 was closely associated with nuclear expression of NF-κB (P <0.001). Clinically, the initial stage (P = 0.006), the location of primary tumor (P = 0.001), and the response to chemotherapy (P = 0.003) were significantly associated with OS. Patients with nuclear expression of Gli1 had significant worse prognosis than those without (median survival [MS], 7.9 months versus 13.9 months, P = 0.009). Similarly, patients with nuclear expression of NF-κB had a shorter OS than those with negative or cytoplasmic expression of NF-κB (MS, 6.3 months versus 13.9 months, P <0.001). However, Shh expression lost of its prognostic significance of OS. Conclusions: Our results indicate that nuclear expression of Gli1 or NF-κB is a strong predictor of poor prognosis in pancreatic cancer. Additional investigation of the close interaction of Gli1 and NF-κB and its biologic significance may help us improve the treatment efficacy of this group of tumors.
Abstract A2-41: The Amplinome™ Test – A droplet digital PCR (ddPCR) based pan-cancer test to assess gene amplification status in solid tumors
Authors: Amy Wong, Jennifer Pecson, Girish Putcha, Lincoln Nadauld, Austin P. So
Journal: Cancer Research (2015)

Abstract Gene amplifications constitute &amp;gt;30% of all actionable mutation types present in cancer. However, despite their prevalence and the availability of targeted drug therapies, the development of molecular assays to detect gene amplifications has been limited by technical challenges such as limited DNA quantity and tumor heterogeneity. To address this, we describe the Amplinome™ test, a test that leverages the high precision and accuracy of droplet digital PCR (ddPCR) to determine the amplification status of 12 commonly amplified genes targeted by FDA approved drugs. Twelve taqman assays were designed to target AURKA, BRAF, CDK4, CDK6, EGFR, ERBB2/HER2, ERBB3/HER3, FGFR1, JAK2, MET, SRC, and VEGFA, and duplexed with the reference gene RNaseP (Epoch, Bothell, WA). Each duplexed assay was then assembled with 2 μL of purified DNA and analyzed via ddPCR (Bio-rad QX-200 ddPCR Platform, Hercules CA). Genes were identified as “amplified” if the observed target:reference ratio was &amp;gt;1.48 (p &amp;lt; 0.005), which adjusts the ASCO-CAP threshold of 2.2 recommended for HER2 amplification for a minimum tumor content of 40%. The Amplinome test identified 29 of 49 patients having actionable amplifications in at least one of the 12 assayed genes. In contrast, only 6 of 49 patients were identified as having amplifications in these genes through a commercially available sequencing test. An overall concordance rate of 90% (532/588 calls) between the two tests was observed. Discordant calls (56/588) arose from amplifications identified by the Amplinome test but not sequencing (55/56). One discordant call (HER3) was identified as amplified via sequencing, but confirmed to be unamplified via FISH. The observed 10-fold increase in sensitivity versus a sequencing-based clinical test suggests that the prevalence of actionable amplifications may be significantly underestimated in cancer. This sensitivity, combined with its ability to provide results within an actionable timeframe (&amp;lt;1 week), the underlines the potential value of Amplinome™ test in providing treatment options for oncologists and their patients. Citation Format: Amy Wong, Jennifer Pecson, Girish Putcha, Lincoln Nadauld, Austin P. So. The Amplinome™ Test – A droplet digital PCR (ddPCR) based pan-cancer test to assess gene amplification status in solid tumors. [abstract]. In: Proceedings of the AACR Special Conference on Translation of the Cancer Genome; Feb 7-9, 2015; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(22 Suppl 1):Abstract nr A2-41.
The prognostic role of serum telomerase activity in advanced breast cancer receiving anthracyclines
Authors: H. Akbulut, S. Temizkan, A. Arican, N. Zengin, B. Yalcin, F. Icli
Journal: Journal of Clinical Oncology (2004)

861 Background: Breast cancer is the second leading cause of cancer deaths throughout the world. Cytotoxic chemotherapy including anthracyclines is the main stay of the treatment of the advanced breast cancer. However, the resistance is an important problem for the most of the patients. Various prognostic factors have been studied to determine the anthracycline resistance and prognosis. In this preliminary study, we aimed to assess the relation of serum telomerase activity with the response to anthracyclines and role of the serum enzyme activity as a prognostic factor in patients with advanced breast cancer. Methods: Nineteen chemo naive patients with advanced breast cancer were included. The first line chemotherapy regimens included the standard doses of doxorubicine plus cyclophosphamide in 14 patients, cyclophosphamide, fluorouracil and doxorubicine in 4 and cyclophosphamide, fluorouracil and epirubicine in 1 patient. Serum telomerase activity was measured before chemotherapy and every 2 cycles thereafter by using a PCR-ELISA trap assay. ER/PR, c-erbB2 expression of tumor tissues were determined by immunohistochemistry. Results: The median age was 47 years. The overall response rate was 57.9%. Half of the patients have ER/PR and 44.4% have erb-B2 positive tumors. The rate of serum telomerase positivity was 39.5%. Though not significant the patients without erb-B2 expression or non-detectable serum telomerase activity has tended to have better response rates when compared to positive ones (83.3%, P=0.177 and 83.3%, P=0.206, respectively). Though not significant, the patients without serum telomerase have better 1-year survival rate when compared to positive ones (62.5% vs. 47.6%, P=0.350). Conclusions: In conclusion, the results of the c current preliminary study shows that serum telomerase activity could be detected in the sera of the cancer patients and detectable serum levels could be a poor prognostic factor for response to the anthracycline containing regimens and prognosis in patients with advanced breast cancer. No significant financial relationships to disclose.
Abstract 950: Antitumor activity of a novel phosphodiesterase 10 inhibitor in an orthotopic mouse model of lung cancer
Authors: Veronica Ramirez-Alcantara, Michele Schuler, Bing Zhu, Nan Li, Evrim Gurpinar, Dennis Otali, Joshua Canzoneri, Adam Keeton, Bernard Gary, Suzanne Russo, Lori Coward, Gregory Gorman, William Grizzle, Xi Chen, Michael Boyd, Gary Piazza
Journal: Cancer Research (2015)

Abstract BACKGROUND: We recently reported that phosphodiesterase 10A (PDE10) is elevated in certain tumor types and essential for tumor cell growth (Li et al., Oncogene 2014). Here we evaluate anti-tumor efficacy, pharmacokinetics, tissue distribution, and tolerance of a novel PDE10 inhibitor, ADT-048, in mice. METHODS: ADT-048 was orally administrated to C57BL/6 mice and free levels quantified in plasma and tissues by LC-MS. Anti-tumor activity was evaluated in athymic nude-Foxn1nu mice after inoculating the left lung with 1×106 A549 lung tumor cells transfected with luciferase. Tumor growth was monitored by in situ bioluminescence using the In Vivo Imaging System and efficacy was evaluated by necropsy grading. RESULTS: ADT-048 inhibited A549 lung tumor cell growth with an IC50 value of 0.7 μM, while appreciably higher concentrations were required to inhibit the growth of normal airway epithelial cells (IC50 = 35 μM). Pharmacokinetic studies demonstrated a half-life suitable for once a day dosing, while tissue distribution studies demonstrated a unique capacity to achieve high concentrations in lungs relative to plasma and other tissues. ADT-048, at a dose of 100 mg/Kg, displayed significant anti-tumor activity as evident by reduced luminescence and tumor grading following 28 days of treatment. CONCLUSIONS: A novel PDE10 inhibitor, ADT-048, was found to display highly potent and selective tumor cell growth inhibitory activity in vitro. ADT-048 was well tolerated in mice and had attractive pharmaceutical properties with a unique capacity to achieve high lung concentrations following oral administration. Efficacy studies demonstrate strong anti-tumor activity of ADT-048, which represents a candidate for clinical development. Supported by NIH grants 1R01CA155638 and 1R01CA131378 (Piazza). Citation Format: Veronica Ramirez-Alcantara, Michele Schuler, Bing Zhu, Nan Li, Evrim Gurpinar, Dennis Otali, Joshua Canzoneri, Adam Keeton, Bernard Gary, Suzanne Russo, Lori Coward, Gregory Gorman, William Grizzle, Xi Chen, Michael Boyd, Gary Piazza. Antitumor activity of a novel phosphodiesterase 10 inhibitor in an orthotopic mouse model of lung cancer. [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 950. doi:10.1158/1538-7445.AM2015-950
Abstract 2648: A novel dual inhibitor for AKT and EGFR signaling inhibits the growth of lung cancer cells
Authors: Arokya Mary Sashi Papu John, Murali Ankem, Chendil Damodaran
Journal: Cancer Research (2015)

Abstract Lung cancer is the leading cause of deaths worldwide due to cancer, of which non-small cell lung cancer is more common making up about 80 percent of lung cancer cases. Although early detection of lung cancer is feasible, targeted therapies importance are growing for advanced lung cancer. EGFR inhibitor is one of the potent options for non-small cell lung cancer (NSCLC), either alone or in combinations with chemotherapy drugs, erlotinib, gefitinib and afatinib; however the complete remission is not yet achieved. Hence, our goal is to dissect the role behind EGFR resistance in NSCLC. Our results suggested a concomitant activation of AKT which was also evident in EGFR expressed of NSCLC. So, we hypothesized that inhibiting both AKT as well as EGFR signaling, could be a promising strategy for treatment of lung cancer. Recently we identified a small molecule, 6-dimethyl, dihydro pyranone (6ddp), that inhibits both AKT (pAKTser473) and EGFR signaling in NSCLC, which resulted growth inhibition of A549 and H460 lung cancer cell lines. Interestingly, the IC50 concentrations were at 1.5nm and 0.3nm in both the lung cancer cell lines. Inhibition of both pro-survival signaling by a small molecule, activated induction of apoptosis by up regulating caspase mediated pro-apoptotic signaling (cleaved caspase) in NSCLC cells. Interestingly, the potent molecule inhibits epithelial-mesenchymal transition (EMT) in A549 and H460 cells by modulating the kinetics of E-Cadherin, β-Catenin expression. Currently, we are planning to validate these interesting findings in animal models, which is essential to translate into clinics. Citation Format: Arokya Mary Sashi Papu John, Murali Ankem, Chendil Damodaran. A novel dual inhibitor for AKT and EGFR signaling inhibits the growth of lung cancer cells. [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 2648. doi:10.1158/1538-7445.AM2015-2648
Analysis of the Inflammatory and Hemostatic Changes and Their Relation to Outcomes in an Experimental Model of Heatstroke in Baboons.
Authors: George Roberts, Veronique Olliver, Sylvie Chollet-Martin, Fallah Al-Mohanna, Rafat El-Sayed, Shaheen Nakeeb, Abdelmoneim Eldali, Dominique de Prost, Heidi Davis, Boleslaw Lach, Abderrezak Bouchama
Journal: Blood (2004)
Differential in Vitro Drug Resistance Profile Between First and Second Relapsed Acute Lymphoblastic Leukemia and Acute Myeloblastic Leukemia in Children
Authors: Jan Styczynski, Beata Kurylo-Rafinska, Malgorzata Kubicka, Beata Kolodziej, Monika Pogorzala, Krzysztof Czyzewski, Robert Debski, Magdalena Wisniewska, Mariusz Wysocki, Michal Matysiak, Iwona Malinowska, Walentyna Balwierz, Edyta Juraszewska, Jacek Wachowiak, Benigna Konatkowska, Mariola Woszczyk, Igor Olejnik, Maryna Krawczuk-Rybak, Marta Kuzmicz, Jerzy Kowalczyk, Maria Jolanta Stefaniak, Wanda Badowska, Tomasz Szczepanski, Renata Tomaszewska, Elzbieta Adamkiewicz-Drozynska, Lucyna Maciejka-Kemblowska, Grazyna Sobol, Agnieszka Mizia-Malarz, Joanna Szczepanek
Journal: Blood (2015)