tbuckley/cabot-search huggingface dataset:
62. Aptamer-siRNA Conjugate Directed Transcriptional Gene Silencing in HIV-1 Infected T Cells
Authors: Daniel C. Lazar, Jiehau Zhou, Sheena Saayman, John J. Rossi, Kevin V. Morris, Marc S. Weinburg
Journal: Molecular Therapy (2015)

The primary standard of care against human immunodeficiency virus (HIV) -1 is highly active antiretroviral therapy which is plagued by toxicity and patient compliance. As such, alternative therapies are sought to provide sustained viral inhibition with reduced treatment intervention. One new potential approach, termed transcriptional gene silencing (TGS), involves the use of small non-coding RNAs (sncRNAs) to inhibit gene expression via the induction of concomitant silent-state epigenetic marks on histones and DNA. We previously demonstrated the ability to silence HIV-1 via TGS by targeting sites within the 5' LTR promoter [1, 2]. However, the lack of efficient, systemic delivery of these RNAs to infected cells hinders successful clinical progression. To address this hurdle, we describe here the development of chimeric molecules based on the previously reported dual function of the gp120 (A-1) aptamer conjugated to 27-mer Dicer-substrate anti-HIV siRNA (dsiRNA) to deliver these sncRNAs to virally infected cells. The efficacy of such a system had been previously demonstrated both in vitro and in vivo to target selectively gp120-expressing cells and to deliver siRNAs for the inhibition of HIV-1 through post-transcriptional gene silencing (PTGS) [3]. To validate the ability for nuclear delivery of sncRNAs, we demonstrated, by fluorescent confocal microscopy, 15% nuclear delivery for our novel chimera-delivered dsiRNAs in a gp120 expressing CHO cells. Using nuclear-cytoplasmic fractionation of HIV-infected PBMCs, dsiRNAs targeting the susceptible regions within the 5' LTR showed preferential nuclear localization when compared to a scrambled siRNAs. Additionally, two 800 nM doses at day 5 and 7 post-infection (p.i.) induced 10-fold suppression of viral p24 levels as measured at day 12 p.i. in CEM cells. The LTR-362 27-mer aptamer chimera proved more potent than A-1 tat/rev 27mer chimera, a positive control that inhibits HIV via PTGS. To validate the inhibition occurred via TGS, 5'-Azacytidine and trichostatin A were added at day 5 p.i, and resulted in transcriptional derepression by the A-1 LTR-263 27-mer, but not the A-1 Tat/Rev 27mer. Nuclear run-on analysis provided further confirmation of TGS inhibition. This is the first study to demonstrate RNA-induced TGS of an existing viral infection in both human PBMC and CEM cell lines following application of aptamer-siRNA chimeras.
647. Repeated Infusion of AAV2-hAADC Into the Thalamus of Non-Human Primates: Effect of Anti-AAV2 Antibodies on Transgene Expression
Authors: Lluis Samaranch, Piotr Hadaczek, Adrian P. Kells, John Bringas, Diane Stockinger, Waldy San Sebastian, Michael Macayan, Sridhar Samineni, Philip Pivirotto, John Forsayeth, Krystof Bankiewicz
Journal: Molecular Therapy (2015)

AAV-based gene therapy has considerable potential as a therapeutic tool to treat many monogenic diseases by restoring deficient expression of key genes after a single injection of an AAV hosting a functional copy of the gene. Under some circumstances, these diseases may need more than a single administration of the AAV-based therapeutic agent. One of the main concerns related to repeated AAV delivery is inhibition of cell transduction at the second infusion site by neutralizing antibodies (NAb) generated after the first exposure. In the present study, we evaluated the immune response after repeated AAV2 delivery to the brain in AAV2 seronegative Rhesus macaques. We infused AAV2-hAADC into left and right thalamus, with a 91-day delay between vector infusions. We determined the levels of anti-AAV2 NAb in sera weekly to monitor antibody production until necropsy 130 days after the first infusion. To ensure high levels of NAb before second infusion, we boosted the NAb production with 2 intramuscular AAV2-VLP immunizations. AADC expression in the thalamus was analyzed in post-mortem tissue by immunohistochemistry and inhibition of transduction was observed in the second infused hemisphere. Immune markers such as GFAP, Iba1, MHCII, CD8 and CD20 were also analyzed, as well NeuN and H&E, to investigate the extent of the immunologic activation after a second brain delivery. Surprisingly, these markers evinced a more dramatic immune response at the first injection site than at the second, resulting even in neuronal loss at the infusion site. CD20 staining revealed the abundant presence of B-lymphocytes within the infusion site. A20 antibody immunostaining revealed the presence of the AAV2 capsid in both hemispheres suggesting a possible trigger of the immune response. The present work demonstrates that very high levels of NAb generated after a primary injection can impair the transduction levels in the second injection site. Moreover, it shows that infiltration of B-lymphocytes can induce tissue damage under conditions of AAV2 capsid persistence. Further studies will be performed to determine the chronology of the events and then establish optimal design and timeline for serial multiple-administration of the AAV-based therapeutic agents. The data further suggest that use of serotypes, such as AAV2, that clear from tissues slowly may not be suitable for therapies in which readministration is being considered.
PWE-035 Diagnostic yield of capsule endoscopy in small bowel Crohn's disease: an experience from a district general hospital
Authors: V Mitra, A Naqvi, S Soloman, K Kapur
Journal: Gut (2010)

<h3>Introduction</h3> The small bowel is a common site for Crohn9s disease but remains relatively inaccessible endoscopically for the majority of its part. There has been variable success in diagnosing active Crohn9s disease by conventional radiological imaging methods.1 The British Society of Gastroenterology (BSG) guidelines on small bowel capsule endoscopy (CE) recommend that CE should be considered in patients with a high suspicion of small bowel Crohn9s disease undetected by conventional means.2 <h3>Methods</h3> We carried out a retrospective audit of all patients referred for CE in our hospital between 2005 and 2009 with suspected small bowel Crohn9s disease undetected by conventional means to evaluate the diagnostic yield of CE and its impact on management. <h3>Results</h3> Small bowel Crohn9s disease was suspected in patients with symptoms of abdominal pain or diarrhoea and weight loss, positive family history of inflammatory bowel disease, extra-intestinal manifestations or investigational findings including iron deficiency anaemia (IDA), raised CRP/ESR or hypoalbuminaemia. All patients presenting with symptoms of abdominal pain had small bowel radiology before proceeding to CE in line with BSG guidelines.<sup>4</sup> 202 patients underwent CE during the study period of which 34 (16.8%) patients were referred to exclude small bowel Crohn9s disease. 19 (56%) patients were females. A diagnosis of Crohn9s disease was made in 13 (38%) patients on CE based on the findings of erosions, aphthous ulcers etc. 13 (38%) patients had a normal examination, 2 (6%) had NSAID induced ulcers, 1 (3%) was diagnosed with coeliac disease, 1 (3%) had erosive gastropathy, 2 (6%) had discrete aphthous ulcers of query significance and 2 (6%) had limited views due to poor bowel preparation for which a repeat CE examination was requested. CE changed management in 19 (56%) patients and included the following – 12 patients with Crohn9s disease were managed conservatively while one had surgery, NSAIDs were stopped in two, proton pump inhibitor was started in one, gluten free diet was initiated in one and two patients underwent repeat endoscopic examination which excluded Crohn9s disease. <h3>Conclusion</h3> In our selected cohort, small bowel Crohn9s disease was diagnosed in 38% of patients that was undetectable by conventional means. This figure is slightly lower than that mentioned in the literature (43–71%).3 Other significant diagnoses were obtained in 12% of cases (NSAID induced ulcers, coeliac disease and erosive gastropathy) and were managed conservatively with good outcome.
A phase 0 exploratory study to assess the pharmacodynamic effects of single intratumoral dose of a novel bispecific targeting/immune-activating agent on the melanoma tumor microenvironment.
Authors: L. A. Fecher, Y. McGrath, D. D. Williams, N. J. Hassan, M. Kalos, L. M. Schuchter, R. K. Amaravadi, A. Chew, E. A. Veloso, B. J. Czerniecki, G. Karakousis, G. P. Linette, B. K. Jakobsen, C. H. June
Journal: Journal of Clinical Oncology (2011)

TPS160 Background: IMCgp100 is a novel biologic comprising a soluble T cell receptor (TCR) fused to an antibody fragment (scFv) being developed for the treatment of metastatic melanoma. The TCR component binds to the melanoma associated antigen (Ag) gp100 which is overexpressed and presented by HLA A2 on melanoma cells. The scFv component activates T cells in physical contact with the target via CD3. Thus, IMCgp100 is expected to bind melanoma cells and stimulate the immune system to attack the target tissue via cytotoxic lymphocyte killing and via stimulation of accessory immune mechanisms.The purpose of this trial is to assess the pharmacodynamic effects of a single intratumoral dose of IMCgp100 on the tumour microenvironment and to establish what local concentration of drug is required for optimal effect. Such data will aid in future trial design and facilitate clinical development. Methods: This 2 stage exploratory study will assess the effects of 2 different doses of IMCgp100 in subjects with advanced unresectable melanoma not currently requiring systemic treatment and/or in a window between treatments. Eligible subjects must have completed other cancer therapies 14 days prior to injection, ECOG PS ≤ 2, no active, uncontrolled infection, no steroids within 2 weeks of injection. In the first stage, subjects will receive a single intratumoral injection of 0.00017mg of IMCgp100 into a target cutaneous or subcutaneous metastasis, 7-15mm in size. This should generate a local concentration of 1nM; shown to induce maximal tumour cell killing in vitro. One week later, biopsies of the target lesion and a designated reference lesion will be performed. Primary endpoints include: assessment of tumor and reference tissue for lymphocyte infiltration and Ag expression by IHC; molecular analysis of immune cell infiltration and activation status using Q-RT-PCR, flow cytometry, Luminex analysis. Secondary endpoints will measure markers of systemic immune activity and safety. The second stage will assess a 10 fold higher dose. Enrollment for the trial is now open. Up to 20 subjects may be enrolled to yield 6 evaluable, HLA 2+ subjects.
Platinum-Based Compounds Induce Expression of p73 and Restores Drug- Sensitivity in p53 Dysfunctional Chronic Lymphocytic Leukemia (CLL) Cells.
Authors: Sanne H. Tonino, Chantal Mulkens, Yoshinori Nonomura, Guangli Suo, Thomas J. Kipps, Marinus H.J. van Oers, Eric Eldering, Arnon P. Kater
Journal: Blood (2008)
An investigation about the efficacy of measuring serum CA 19-9 in patients with advanced urothelial carcinoma
Authors: Hiroshi Yaegashi, Takahiro Nohara, Kazutaka Narimoto, Kouji Izumi, Satoru Ueno, Yoshifumi Kadono, Yasuhide Kitagawa, Hiroyuki Konaka, Atsushi Mizokami, Mikio Namiki
Journal: Annals of Oncology (2015)
Abstract 391: Utility of circulating tumor cell (CTC) in advanced gastric cancer
Authors: Won Suk Lee, Jeong Min Kim, Woo Sun Kwon, So Jung Lim, Tae Soo Kim, Ga Yun Kim, Sung Ho Choi, Byung Hee Jeon, Min Kyu Jung, Joong Bae Ahn, Hyun Cheol Chung, Sun Young Rha
Journal: Cancer Research (2015)

Abstract Circulating tumor cells (CTCs), known as the “leukemic phase” of solid tumor, have been associated with poor prognosis and clinical outcome in various cancer including breast and colorectal. However, fewer studies reported characteristic and usefulness of CTC as an indicator in gastric cancer (GC). In this study, we evaluated utility of CTC in advanced GC cancer. First, we confirmed detection rate of GC cancer cell lines as a spiking test, and then defined CTC as DAPI+, EpCAM+ and CD45- cells through immunofluorescence stating. To determine a cut-off for CTC of GC, we assessed the number of CTCs in blood of 22 healthy volunteers, and observed 1 (3/22) to 2 (1/22) of CTCs. Based on these results, we determined CTC positivity of GC as more than 3 CTCs. When we evaluated CTC positive rate in Stage IV GC patients, 28 patients were CTC positive (30.1%) with the median number of CTC 5 (range 3 to over 100). Interestingly, 3 out of 28 patients were showed more than 100 CTCs. While, in operable GC patients (Stage I to III), CTC positivity was 18.4% (7/38) and median number of CTC was 4 (range 3 to 9) in CTC positive patients. These results showed that Stage IV GC patients have high number of CTCs than operable GC patients. Tumor histology and Lauren classification did not affect to CTC positivity in Stage IV and operable GC patients. We also observed the potential use of CTC in serial follow-ups and one of the cases shows the decreasing CTCs followed by clinical response. In conclusion, this study demonstrates that detection of CTC may be useful prognostic indicator of GC cancer patients. Citation Format: Won Suk Lee, Jeong Min Kim, Woo Sun Kwon, So Jung Lim, Tae Soo Kim, Ga Yun Kim, Sung Ho Choi, Byung Hee Jeon, Min Kyu Jung, Joong Bae Ahn, Hyun Cheol Chung, Sun Young Rha. Utility of circulating tumor cell (CTC) in advanced gastric 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 391. doi:10.1158/1538-7445.AM2015-391
Abstract 1607: Identification of low number circulating tumor cells (CTCs) for cancer treatment monitoring
Authors: Thomas J. George, Weian Sheng, Jose I. Varillas, Chen Liu, Z. Hugh Fan
Journal: Cancer Research (2015)

Abstract For most solid tumors, the clinical use of CTCs is limited by current instrument restrictions on detection of infrequent CTCs. Based on the FDA-approved criteria used in current CTC detection instruments, we developed a microfluidic device capable of capturing, enumerating and releasing low numbers of CTCs in the peripheral blood of patients with solid tumors. The device is the size of a microscope slide, consisting of an inlet, an outlet, and eight parallel channels connecting them. Micromixers inside the channels enhance interactions between CTCs and capture agents on the channel surfaces. Epithelial cell-adhesion molecule (EpCAM) antibodies are immobilized onto the surfaces via avidin-biotin chemistry. A blood sample from a cancer patient is introduced through the inlet. Cells captured by anti-EpCAM studded channels are verified as CTCs using triple-color IHC definitions (EpCAM+, cytokeratin+, CD45-, and DAPI+). Using pancreatic cancer as a model for a low frequency CTC condition, device capture efficiency and cell purity was tested under various flow rates (shear stresses) using L3.6pl cells spiked into blood. Capture efficiency was defined as the% of target cells isolated relative to target cells present in a sample, representing device sensitivity. Cell purity was defined as the% of target cells isolated relative to all cells captured (both target and non-target cells), representing device specificity. Capture efficiency (90%) and cell purity (84%) were further optimized by changing the flow rate. A higher flow rate resulted in higher cell purity as non-target bound cells are dislodged by shear forces. This capability of parameter adjustment and performance control is enabled by the microfluidics platform used as opposed to CTC isolation using other commercially approved technologies. The device was then clinically tested in detecting CTCs and monitoring the response to anti-cancer drug treatment in patients with advanced pancreatic cancer. We identified CTCs in 113 out of 115 clinical blood samples (98%). CTC enumeration (CTC number/mL of blood) both correlated with and predated changes in the radiographic tumor size over the treatment period, ranging from 10 to 35 cycles (2 weeks per cycle) of therapy. Microfluidic devices possess the capacity to enumerate infrequent CTCs which also appear to correlate with treatment response. Note: This abstract was not presented at the meeting. Citation Format: Thomas J. George, Weian Sheng, Jose I. Varillas, Chen Liu, Z. Hugh Fan. Identification of low number circulating tumor cells (CTCs) for cancer treatment monitoring. [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 1607. doi:10.1158/1538-7445.AM2015-1607
Abstract 1975: Role and activation mechanisms of myeloid zinc finger-1 (MZF1) in ErbB2-induced breast cancer invasion
Authors: Tuula Kallunki, Ditte M. Brix, Knut Kristoffer B. Clemmensen, Siri A. Tvingsholm, Bo Rafn, José Moreira, Irina Gromova, Marja Jäättelä
Journal: Cancer Research (2015)

Abstract ErbB2 overexpression gives rise to a highly invasive, aggressive phenotype associated with an increased activity of lysosomal cysteine cathepsins B and L and with a pericellular accumulation of lysosomes at the close vicinity of the cell membrane, enabling lysosomal exocytosis and secretion of lysosomal contents to the extracellular space. We recently showed that ErbB2 expression level correlates positively with cysteine cathepsin B and L levels in primary breast cancer and that ErbB2 regulates the expression of cathepsins B and L leading to their increased activity in several ErbB2-positive breast cancer cell lines. We found that the increased cysteine cathepsin activity is essential for the invasion of ErbB2 overexpressing breast cancer cells in 3-dimensional (3D) Matrigel cultures and that it is mediated by a novel signaling network that involves TGFβR, ERK2-MAPK, PAK4, PKCα and CDC42bpβ kinases and two transcription factors, Myeloid Zinc Finger 1 (MZF1) and ETS1, which regulate the ErbB2-induced expression of cathepsins B and L. We currently work on understanding the mechanisms that increase the transcriptional activity of MZF1 in breast cancer. Thus far we have identified two basic mechanisms that can increase MZF1 target gene expression: cancer-induced, increased levels of MZF1 and ErbB2 signaling-induced, increased transcriptional activity of MZF1. Our results thus far show that the level of MZF1 can be directly downregulated by specific, non-coding RNAs. The cancer-induced increase in the MZF1 level is most likely due to changes in the expression of these non-coding RNAs. Preliminary data employing two-dimensional SDS-PAGE as well as the factual involvement of several kinases downstream of ErbB2 for its transcriptional activity, suggests that MZF1, a prominent phosphoprotein, is activated by phosphorylation in response to ErbB2 signaling. Towards this end we have set up various experiments to verify the phosphorylation of MZF1 in response to ErbB2 signaling. We now aim at the identification of the ErbB2-regulated phosphorylation sites. With this work we expect to collect more information about the molecular mechanisms underlying ErbB2-induced invasion. Increased knowledge of the mechanisms is expected to enable the development of novel therapies and biomarkers for aggressive, ErbB2-positive breast cancer. Citation Format: Tuula Kallunki, Ditte M. Brix, Knut Kristoffer B. Clemmensen, Siri A. Tvingsholm, Bo Rafn, José Moreira, Irina Gromova, Marja Jäättelä. Role and activation mechanisms of myeloid zinc finger-1 (MZF1) in ErbB2-induced breast cancer invasion. [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 1975. doi:10.1158/1538-7445.AM2015-1975
S14 Patients’ perceptions of an exercise programme delivered following discharge from hospital after critical illness (the revive trial)
Authors: K McDowell, JM Bradley, DF McAuley, B Blackwood, B O’Neill
Journal: Thorax (2016)

<h3>Introduction</h3> The REVIVE RCT investigated the effectiveness of an individually tailored (personalised) exercise programme for patients discharged from hospital after critical illness.<sup>1</sup> By including qualitative methods, we aimed to explore their perceptions of engaging in the 6 week programme to facilitate a better understanding of the intervention and trial outcomes. <h3>Methods</h3> Patients allocated to the exercise group were invited to participate in semi-structured interviews following their final outcome assessment (6 months following randomisation). Interviews were conducted by a trained member of the research team not involved in the intervention. Interviews were audio recorded, transcribed verbatim and content analysis used to explore themes arising from the data. <h3>Results</h3> Of 30 patients allocated to the exercise group 21 completed interviews. Seven core themes were identified (1) sequelae of critical illness and critical care recovery; (2) satisfaction and endorsement of the exercise programme; (3) beneficial impact of the exercise programme on physical and psychological health; (4) facilitators of beneficial impact; (5) barriers to beneficial impact; (6) challenges to continuing exercise; (7) contrasting views on outcome measures. Patients provided insight into the physical and mental sequelae they experienced following critical illness. There was a strong sense of patients’ need for the exercise programme and its importance for their recovery following discharge home. The programme was described as invaluable, and provided feelings of motivation and hope. Key facilitators of beneficial impact included supervision, tailoring of the exercises to personal needs, and the manual. Barriers to the beneficial impact of the programme included poor mental health, existing physical limitations and lack of motivation. Patients’ views of the questionnaires and performance based outcome measures in the REVIVE trial varied. Many patients were unsure about what would be the best way of measuring how the programme affected their health. <h3>Conclusion</h3> The benefits of physical rehabilitation programmes, needs to be counterbalanced against patients’ mental health status post-ICU and any pre-admission limitations, if they are to be successful. Including this qualitative component improved our understanding of the mechanisms underpinning the impact of the programme and how programmes should be evolved for future trials. <h3>Reference</h3> O’Neill B, <i>et al</i>. Effectiveness of a programme of exercise on physical function in survivors of critical illness following discharge from the ICU: study protocol for a randomised controlled trial (REVIVE). <i>Trials</i> 2014;<b>15</b>:146.