tbuckley/cabot-search huggingface dataset:
113. HSV Vector Development for Targeted Gene Delivery
Authors: Bonnie L. Reinhart, Marco Marzulli, Daniela Leronni, Yoshitaka M. Miyagawa, Justus B. Cohen, Joseph C. Glorioso
Journal: Molecular Therapy (2015)

Gene therapy often requires selective gene delivery to specific cellular subpopulations to provide the most effective therapeutic outcome and avoid off-target effects. This can be accomplished by viral vector targeting involving replacement of the natural virus-cell interactions that trigger virus entry, with novel cell-specific interactions, thereby permitting delivery of therapeutic transgenes to specific cell types. Strategies for full retargeting of HSV require (i) mutagenesis-mediated virus detargeting from its cognate receptors (HVEM and nectin1) recognized by the virus attachment/entry component glycoprotein D (gD) and (ii) the introduction of new ligands into gD that allow entry through recognition of cognate cellular receptors. To target an HSV vector for entry exclusively into cells expressing the receptors GFRa1 or TrkA, we employed the ligands GDNF and NGF, respectively. We replaced the signal peptide and HVEM binding domain of gD with pre-pro-(pp)GDNF to create a GFRa1 targeting protein, gD(Y38)_GDNF, that can still bind nectin1. Virus expressing gD(Y38)_GDNF was propagated on cells expressing nectin1 and purified virus was shown to enter nectin1-deficient cells in a GFRa1-dependent manner. Likewise, TrkA-dependent virus entry was observed using ppNGF as a targeting ligand. Although propagation of completely retargeted viruses (mutationally inactivated for both HVEM and nectin1 binding) can be achieved in complementing cell lines that express the target receptor, we observed differences among cell lines in retargeted virus spread. HSV receptor-deficient J1.1-2 and B78H1 cells engineered to express the target receptors allowed virus entry, but only B78H1 based cells displayed observable spread. U2OS cells engineered to express GFRa1 demonstrated the most robust virus entry and spread, but these cells express abundant nectin1, providing strong selective pressure for reversion of the nectin1 binding defect of fully retargeted viruses during propagation. In one approach to overcome this problem, we developed genetic selection methods to isolate retargeted virus variants that both entered and spread on J1.1-2 cells transduced with the target receptor. We found that selected variants had acquired mutations in other envelope glycoproteins, including one glycoprotein involved in envelope-cell fusion events (gH) and two that were previously shown to contribute to the spreading process (gE and gI). We expect that the combination of virus mutants and engineered cell lines for efficient production of stably retargeted HSV gene therapy vectors will allow us to achieve HSV retargeting to a broader range of receptors.
447. Lentiviral Gene Therapy in a Preclinical Model of Omenn Syndrome
Authors: Maria Carmina Castiello, Valentina Capo, Elena Draghici, Virginia Maina, Niek P. van Til, Gerard Wagemaker, Marita Bosticardo, Anna Villa
Journal: Molecular Therapy (2015)

Patients with Omenn syndrome (OS), carrying hypomorphic recombination activating genes (RAG) mutations, develop a peculiar severe combined immunodeficiency associated with autoimmune-like manifestations. RAG1 and RAG2 genes play an essential role in the generation of antigen receptors during lymphocyte development. HLA-matched bone marrow transplantation is curative, however, hematopoietic stem cell gene therapy (GT) could represent an alternative treatment for patients lacking a suitable donor. The aim of the present study was to investigate the efficacy of GT in a knock-in murine model carrying the Rag2 R229Q mutation that mimics most of the symptoms of human OS. To this aim, we are testing a lentiviral vector encoding for codon optimized human RAG2 (hRAG2co-LV vector) under the control of the ubiquitous chromatin opening element (UCOE). We initially measured the efficiency of transduction of Rag2R229Q/R229Q lineage negative (lin-) cells by testing the presence of the integrated viral vector by real-time PCR in lin- cells transduced at a multiplicity of infection (MOI) 5 and 10 and in vitro cultured for 8 days. We observed that the hRAG2co-LV vector transduced Rag2R229Q/R229Q lin- cells very efficiently at MOI 5. Then, we tested the in vivo efficacy of hRAG2co-LV-mediated GT infusing transduced lin- BM cells isolated from male Rag2R229Q/R229Q mice in lethally irradiated females. Control Rag2R229Q/R229Q mice received wild-type (BMT-WT) or untransduced (BMT-OS) lin- BM cells. Analysis of immune cells in peripheral blood was evaluated overtime starting from six weeks after the treatment. GT-treated mice showed the appearance and progressive increase of B cells and amelioration of T cell frequency and absolute count as compared to BMT-OS mice. In parallel, the frequency of myeloid cells started to decrease 13 weeks after GT indicating a normalization of peripheral immune cell distribution. Our preliminary data demonstrate the feasibility of GT in the preclinical model of OS. Additional experiments aimed at evaluating long-term efficacy and safety are ongoing.
Exosomes derived from miR-140-5p-overexpressing human synovial mesenchymal stem cells enhance cartilage tissue regeneration and prevent osteoarthritis of the knee in a rat model
Authors: Shi-Cong Tao, Ting Yuan, Yue-Lei Zhang, Wen-Jing Yin, Shang-Chun Guo, Chang-Qing Zhang
Journal: Theranostics (2016)

Osteoarthritis (OA) is the most common joint disease throughout the world. Exosomes derived from miR-140-5p-overexpressing synovial mesenchymal stem cells (SMSC-140s) may be effective in treating OA. We hypothesized that exosomes derived from SMSC-140 (SMSC-140-Exos) would enhance the proliferation and migration abilities of articular chondrocytes (ACs) without harming extracellular matrix (ECM) secretion.SMSCs were transfected with or without miR-140-5p. Exosomes derived from SMSCs or SMSC-140s (SMSC-Exos or SMSC-140-Exos) were isolated and identified. Proliferation, migration and ECM secretion were measured in vitro and compared between groups. The mechanism involving alternative Wnt signalling and activation of Yes-associated protein (YAP) was investigated using lentivirus, oligonucleotides or chemical drugs. The preventative effect of exosomes in vivo was measured using Safranin-O and Fast green staining and immunohistochemical staining.Wnt5a and Wnt5b carried by exosomes activated YAP via the alternative Wnt signalling pathway and enhanced proliferation and migration of chondrocytes with the side-effect of significantly decreasing ECM secretion. Highly-expressed miR-140-5p blocked this side-effect via RalA. SMSC-140-Exos enhanced the proliferation and migration of ACs without damaging ECM secretion in vitro, while in vivo, SMSC-140-Exos successfully prevented OA in a rat model.These findings highlight the promising potential of SMSC-140-Exos in preventing OA. We first found a potential source of exosomes and studied their merits and shortcomings. Based on our understanding of the molecular mechanism, we overcame the shortcomings by modifying the exosomes. Such exosomes derived from modified cells hold potential as future therapeutic strategies.
PTH-021 A survey of endoscopy workload in northern England: lower gastrointestinal endoscopy workload exceeds British Society of Gastroenterology projections
Authors: T J W Lee, M Rutter, C Zwart, C Rees
Journal: Gut (2010)

<h3>Introduction</h3> The BSG working party report on the Provision of Endoscopy Related Services in District General Hospitals (2001) made clear projections of the estimated endoscopic workload in an average district general hospital.1 The Northern Region Endoscopy Group (NREG) is a network connecting all endoscopy units in the North East of England with the aim of promoting collaboration and endoscopy based research. The aim of this study was to measure current endoscopic workload in the Northern Region Endoscopy Goup and compare it to the BSG recommendations. <h3>Methods</h3> Questionnaires were sent to each of the 16 endoscopy units within the Northern Region of England. Completed questionnaires were received from all 16 endoscopy units in nine trusts. <h3>Results</h3> The total population served by endoscopy units within NREG is 3.5 million people. Ninety-five thousand endoscopic procedures were performed across the region last year, 45 194 (47%) were upper GI, 27 541 (29%) colonoscopies, 17 912 (19%) flexible sigmoidoscopies and 2688 (3%) were ERCPs. Across the region 13 gastroscopies per 1000 population, 7.9 colonoscopies, 5.1 flexible sigmoidoscopies and 0.77 ERCPs were performed per 1000 population per annum. <h3>Conclusion</h3> Current endoscopic workload figures for upper GI endoscopy and ERCP are in line with the BSG working party9s estimations of 2001. The burden of lower GI work within the Northern Region is greater than the BSG predictions with nearly twice as many procedures as predicted being performed. Contributing factors include changing referral practices, the Bowel Cancer Screening Programme, and more widespread open access services. Current lower GI endoscopy workload figures are considerably higher than those quoted in the NICE document “Improving Outcomes on Colorectal Cancers” (2004)2 and the Royal College of Physicians publication “Consultant Physicians Working with Patients” (2004).3 Current workload is in line with Healthcare commission9s 2006 survey.4 The implications of these data include: Financial and manpower planning needs to take into account the increased burden on endoscopy services. The BSG estimates for lower GI endoscopy workload should be revised. A similar evaluation of changes in endoscopy workload should be performed at a national level. Abstract submitted on behalf on the Northern Region Endoscopy Group (NREG).
Efficacy in selected tumor types in a phase I study of the c-MET inhibitor ARQ 197 in combination with sorafenib.
Authors: A. A. Adjei, J. A. Sosman, R. E. Martell, G. K. Dy, L. W. Goff, W. W. Ma, L. Horn, G. J. Fetterly, S. A. Michael, J. A. Means, F. Chai, M. Lamar, G. M. Strauss, W. Chiang, J. Jarboe, B. E. Schwartz, I. Puzanov
Journal: Journal of Clinical Oncology (2011)

3034 Background: ARQ 197 (A) is a selective, oral, non-ATP competitive inhibitor of c-MET, a RTK implicated in tumor cell proliferation, invasion and metastasis, currently in phase II/III clinical trials. Based on preclinical synergy of A and sorafenib (S) in multiple cell lines, a phase I study was undertaken to define safety and RP2D of the A+S combination. Methods: A traditional 3+3 dose escalation schema was employed. Two dose levels were evaluated, dose level (DL) 1 (A 360 mg PO BID + S 200 mg PO BID) and DL2 (A 360 mg BID + S 400 mg BID), which was deemed the RP2D, since it represents full single-agent doses of both drugs. Extension cohorts have been opened at this dose in patients with five tumor types: renal cell carcinoma (RCC), hepatocellular carcinoma (HCC), breast cancer (BC), non-small cell lung cancer (NSCLC) or melanoma, based on the preclinical and clinical data. Results: To date, 54 pts (17 F/37 M; mean age: 61.4 yrs; 13 RCC, 8 HCC, 12 NSCLC, 8 melanoma, 7 BC, 6 others) have been treated, 5 at DL1 w/o DLT, 9 at DL2 with 2 DLTs in 1 pt (a Grade 3 fatigue and a Grade 3 dyspnoea) and 40 at the RP2D (extension cohort). Adverse events (AEs) considered related to one or both study drugs occurred in 45 (83.3%) pts with the most common ones (≥10%) being fatigue (35.2%), rash (31.5%), diarrhea (29.6%), anorexia (29.6%), nausea (18.5%), stomatitis (18.5%), alopecia (18.5%), hypophosphataemia (18.5%), vomiting (13.0%), pruritus (13.0%), hand-foot syndrome (11.1%) and lymphopenia (11.1%). Among 48 pts on extension cohorts, 37 (77.1%) were evaluable (defined as having at least one post-treatment CT scan) for efficacy. Their data are summarized in the table. Conclusions: The A+S combination therapy is well tolerated at full single-agent doses and RP2D has been identified. Preliminary evidence of anti-cancer activity has been observed, indicating that the combined inhibition of c-MET and angiogenic signalling demonstrates therapeutic potential. RCC (n=12) HCC (n=5) Melanoma (n=6) NSCLC (n=9) BC (n=5) PR 1 (8.3%) 0 3 (50.0%) 0 0 SD 11 (91.7%) 4 (80.0%) 0 4 (44.4%) 3 (60.0%) Median weeks (range) on treatment 38.1 (15.0-60.4) 19.7 (8.0-61.0) 9.0 (3.9-25.1) 9.1 (2.7-16.0) 12.3 (2.0-33.9)
Abstract 2173: Robust estimation of mutation burden
Authors: Oscar Westesson, Rasmus Nielsen, John St John, Aleah Caulin, Nicholas Hahner, Stewart Stewart, Catherine Foo, Kimberly Lung, Jeff Catalano, Mandy Lee, Petros Giannikopoulos, Will Polkinghorn, Jonathan Wiessman, Aviv Regev, Trever Bivona
Journal: Cancer Research (2015)

Abstract Developing a more robust approach to measure mutational burden is of central importance to improving the characterization of the molecular profile of tumor and may improve our ability to predict tumor progression or response to therapy in patients. Mutational burden is typically calculated as a direct enumeration of called somatic mutations per megabase covered. However, there is a growing appreciation that tumor purity, variable sequencing coverage, and copy number alterations can substantially impact the accurate identification any specific somatic mutation. Furthermore, population genetic theory and empirical data indicate that in many cases the vast majority of somatic mutations appear in only a small subpopulation of tumor cells, a context in which there is a high likelihood that an individual subclonal mutation may not be identified by conventional analysis. This tendency to miss low frequency mutations is highly variable and dependent, in part, upon sample purity and results in a strong source of bias not addressed in existing methods to measure variant allele frequencies. We present a novel computational method that incorporates these sources of bias in a coherent probabilistic framework that enables maximum-likelihood inference of relevant population parameters such as mutation burden. We apply our method to simulated data as well as patient tumor samples diluted with varying known proportions of normal DNA. We show that our approach allows us to generate estimates of mutation burden that are robust to the substantial variations in purity and sequencing coverage that are frequently encountered in patient tumor analysis. Hence, our novel method may improve the accurate detection and quantification of variant alleles in patient tumors to better understand their genetic landscape and guide clinical management. Citation Format: Oscar Westesson, Rasmus Nielsen, John St John, Aleah Caulin, Nicholas Hahner, Stewart Stewart, Catherine Foo, Kimberly Lung, Jeff Catalano, Mandy Lee, Petros Giannikopoulos, Will Polkinghorn, Jonathan Wiessman, Aviv Regev, Trever Bivona. Robust estimation of mutation burden. [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 2173. doi:10.1158/1538-7445.AM2015-2173
PTU-080 Accuracy of 13C mixed triglyceride breath testing for investigation of exocrine pancreatic insufficiency: an audit: Abstract PTU-080
Authors: V Porter, D A Mcnamara, J M C Donald, B Mackie, S Shirron
Journal: Gut (2010)

<h3>Introduction</h3> The two main indirect non-invasive clinical investigations for determination of exocrine pancreatic function are faecal and breath tests. The 13C mixed triglyceride (MTG) breath test is simple, inexpensive and readily available and has a reported sensitivity and specificity of 81% and 85%, respectively. <h3>Methods</h3> 13C MTG breath tests performed over a sixteen month period in a single university affiliated hospital were identified from a database. A retrospective review was performed. Each patient had undergone the same test protocol: Two fasting end-expiratory breath samples were taken prior to the patient being given a test meal. The test meal consisted of 100 g of toast coated with butter (0.25 g×patient weight (kg). The butter was laced with labelled MTG (4 mg×patient9s body weight (kg)). Test breath samples were taken every 30 min for 6 h. A cumulative percent 13C-dose recovery (CDPR) of &lt;23% indicated abnormal exocrine pancreatic function. Samples were run using HeliFANas software. All samples were run at least two times and the results of each of each run were compared to assess validity. <h3>Results</h3> In all, twenty eight patients were identified. Patients were made up of two groups: patients with known chronic pancreatitis and those with symptoms suggestive of pancreatic disease. 46% (n=13) of patients were male and the mean age was 57 (±16 years). Analysis revealed an 18% (5/28) variance between test runs 1 and 2 in this population. Concordance between all runs (1–4) were analysed and results are demonstrated below in Abstract 080. <h3>Conclusion</h3> This study raises concerns regarding the reliability of the 13C MTG breath test. In our population, there was an 18% (5/28) variance between two separate breath analysis. Therefore, five patients would have received an inaccurate diagnosis based on the MTG test alone. However, accuracy can be improved by running samples more than once.
Prospective Evaluation of the International Prognostic Score (IPS) in All Stages of Hodgkin’s Lymphoma Treated with ABVD Plus Involved-Field Radiotherapy (IFRT).
Authors: Santiago Pavlovsky, Claudia Corrado, Miguel A Pavlovsky, Virginia Prates, Lucia Zoppegno, Mario Giunta, Ider Cerutti, Elsa Palomino, Graciela Avila, Francisco Lastiri
Journal: Blood (2008)
Efficacy and thyroidal effects of sunitinib in Japanese patients with metastatic renal cell carcinoma: Hypothyroidism and thyroid atrophy as potential biomarkers for sunitinib?
Authors: N. Shinohara, M. Takahashi, T. Kamishima, H. Ikushima, A. Sazawa, H. Kanayama, K. Nonomura
Journal: Journal of Clinical Oncology (2009)

e16097 Background: Although hypothyroidism is a well-known adverse effect of sunitinib in western patients (pts) with metastatic renal cell carcinoma (MRCC), the effects on thyroid gland of sunitinib in Japanese pts still remain unclear. We therefore evaluated thyroid dysfunction and thyroid atrophy in Japanese RCC pts who received sunitinib. Methods: Fourteen (8 males, 6 females) of 51 pts who were treated in a phase II trial of sunitinib in Japanese pts with MRCC were included in this retrospective study. All 14 pts were euthyroid at baseline. The measurement of serum thyroid function test (fT3, fT4, TSH) was performed at the beginning of each sunitinib treatment cycle. A TSH concentration greater than 10 mU/L was considered as hypothyroidism regardless of subclinical or clinical. CT volumetry of the thyroid gland was performed utilizing the data obtained for tumor assessment in a phase II trial. Tumor response was evaluated based on the RECIST criteria. Results: 9 (64%) of 14 pts achieved RECIST-defined objective response (1 CR, 8 PRs). With regard to thyroid function, hypothyroidism was experienced by 11 (79%) pts. Three of 11 pts had Sunitinib-induced thyrotoxicosis before experiencing hypothyroidism. The mean time to development of hypothyroidism was 55 days (range 13–668). Six pts received typical doses of L-thyroxine. The response rate was 73% (8/11) in pts with hypothyroidism and 33% (1/3) in pts without. With regard to the volume of thyroid gland in 13 pts, 8 pts had more than 50% reduction comparing the volume at the baseline. Median reduction rate in volume of 13 pts was 57% (range 0–95%). Hypothyroidism was experienced by all 8 pts with more than 50% reduction in volume and 2 of 5 (40%) without. Furthermore, the response rate was 88% (7/8) in former pts and 40% (2/5) in latter pts. Conclusions: In addition to high anti-tumor efficacy, hypothyroidism and thyroid atrophy were commonly observed in Japanese MRCC pts who received sunitinib. Although further study should be required, these abnormal findings in thyroid gland following treatment with sunitinib may be potential biomarkers for tumor response to sunitinib. [Table: see text]
Abstract 2563: Modulation of DNA damage responses and cytotoxicity of bleomycin by vitamin C
Authors: Blazej Rubis, Anatoly Zhitkovich
Journal: Cancer Research (2015)

Abstract Bleomycin (BL) is an iron-chelating chemical that is used for treatment of several types of cancer. The administration of this drug to patients is also associated with significant side effects, including life-threatening lung fibrosis. Biological factors affecting chemotherapeutic effectiveness of BL and the severity of its major side effects remain poorly understood. Cytotoxic effects of BL are attributed to its ability to cause DNA breaks via oxidative reactions by chelated iron. Ascorbate is a main cellular antioxidant, which can also promote redox cycling of iron by reducing its ferric form. Standard cultures of human cells contain less than 1% of physiological amounts of vitamin C, raising concerns whether common in vitro models adequately recapitulate major mechanistic aspects of BL toxicity in vivo. In this work, we investigated the effects of intracellular ascorbate on cell killing and DNA damage-activated stress responses by BL. We found that replenishment of human lung carcinoma cells with ascorbate increased cytotoxicity and clonogenic lethality of BL. Examination of DNA damage responses in BL-treated cells found that cellular ascorbate enhanced some signaling branches while suppressing others. Ascorbate-restored cells also displayed an increased dependence on nonhomologous end-joining DNA repair for their long-term survival. Overall, our results show that restoration of ascorbate levels increases sensitivity of cancer cells to killing by BL and alters their DNA damage responses. Low ascorbate levels in hypoxic tumors can contribute to their resistance to BL-based therapy. Citation Format: Blazej Rubis, Anatoly Zhitkovich. Modulation of DNA damage responses and cytotoxicity of bleomycin by vitamin C. [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 2563. doi:10.1158/1538-7445.AM2015-2563