tbuckley/cabot-search huggingface dataset:
284. Long-Term Therapeutic Immune Reconstitution in XSCID Canine Model via In Vivo Foamy Virus Delivery of Common Gamma Chain
Authors: Frieda Chan, Olivier Humbert, Christopher Burtner, Daniel Humphrys, Jennifer Adair, Grant Trobridge, Patricia O'Donnell, Nicholas Hubbard, Troy Torgerson, Andrew Scharenberg, David Rawlings, Peter J. Felsburg, Hans-Peter Kiem
Journal: Molecular Therapy (2016)

X-linked combined immunodeficiency disease (XSCID) is caused by mutation in the common gamma chain, γC (interleukin-2 receptor subunit gamma, IL2RG) in both humans and canines. It is characterized by the inability of T-cell development leading to absence of T-cells in peripheral blood, lack of T-cell mediated immune response, low IgA and IgG levels, and early infant mortality. In the 1990s, human XSCID clinical trials utilizing gamma-retroviral vectors to deliver the IL2RG gene caused leukemia in 5 out of 20 patients due to vector integration in or near proto-oncogenes. Recent studies showed Foamy virus based vectors as an excellent alternative for in vivo gene-therapy because it is non-pathogenic in humans while exhibiting increased serum stability and favorable integration pattern. Previously, we have demonstrated CD3+ T-cell reconstitution in the canine model via intravenous injection of foamy virus expressing human elongation factor-1 alpha promoter (Ef1α)-yC. Unfortunately, the treated animals contained a low number of gene corrected progenitors at a sub-therapeutic level. Here, we achieved long-term therapeutic immune-reconstitution by intravenous delivery of a human phosphoglycerate kinase promoter (Pgk)-mediated γC foamy viral vector into XSCID neonatal canines. Long-term (2 years) post-injection follow-up demonstrated therapeutic levels of CD3+ T-cell expansion. Within the T-cell population, gene correction with Pgk-γC stabilized at ~80%. We validated T-cell functionality by using spectratyping analysis, which exhibited a diverse repertoire of receptor gene rearrangement. Retroviral integration site analysis (RIS) indicated polyclonal contribution to the reconstituted T-cells. Immunoglobulin ELISA assays showed that IgA and IgG levels in peripheral blood are comparable to normal healthy controls. We immunized the gene-corrected canine recipients with bacteriophage ϕx174 and confirmed production of specific IgG antibodies, showing the ability for isotype switching in B-lymphocytes. Currently, the gene-corrected canines exhibit comparable health and physical attributes to normal controls. Furthermore, semen from the gene-corrected male canine was used via artificial insemination to produce a litter of viable offsprings. In summary, our data demonstrate that Pgk-γC foamy viral vector delivered long-term therapeutic gene correction in a large-animal model for XSCID gene therapy. Most importantly, these results indicate that in vivo Pgk-γC foamy vector administration is a viable option for long-term immune reconstitution in future XSCID human clinical trials.
118. Inhibition of Hepatitis B Virus Replication In Vivo Following Delivery of Antiviral TALENs With Recombinant Adeno-Associated Viral Vectors
Authors: Samantha Nicholson, Buhle Moyo, Kristie Bloom, Claudio Mussolino, Toni Cathomen, Koichi Watashi, Abdullah Ely, Patrick Arbuthnot
Journal: Molecular Therapy (2015)

Hepatitis B virus (HBV) is hyperendemic to sub-Saharan Africa where there are approximately 600 000 deaths per year associated with chronic HBV infection. HBV may have long periods of dormancy in carriers of the virus, which is dependent on persistence of the nuclear replication intermediate covalently closed DNA (cccDNA). Current HBV therapies, including nucleot(s)ide analogs, target the actively transcribed virus. The cccDNA is not cleared and may act as a reservoir for re-initiation of viral replication following treatment withdrawal. This poses a significant challenge for the successful treatment of chronic HBV sufferers, and makes the cccDNA intermediate an important target for novel HBV treatments. Employing gene editing technology to disable cccDNA has emerged as a promising novel therapeutic strategy. TALENs targeting the surface and the core viral open reading frames, previously described by our group, demonstrated efficacy against HBV targets. To advance this technology to therapeutic application, efficient delivery and evaluation of the TALENs' safety and target specificity in vivo and in vitro need to be established. This study aimed to evaluate the safety and efficacy of TALENs delivered by adeno-associated viral (AAV) vectors in vivo and in vitro. The HBV-infectable HepG2-hNTCP-C4 cell line was used to establish that the TALENs could disable cccDNA. Capacity for capsid modification, low immunogenicity and good hepatotropic transduction efficiency are favorable features that were considered in selecting these vectors. However, the delivery of TALENs by AAVs is technically complicated, as sequences encoding the two subunits of the complete TALENs exceed the capacity of single stranded AAVs. Consequently, we generated AAVs that encode each of the TALEN subunits. To constitute the complete TALENs, pairs of vectors were thus administered to mice or cultured cells. In vitro testing of AAV-TALENs was undertaken using the HepG2-hNTCP-C4 cell line. These cells present a novel tool for testing anti-HBV therapeutics as they overexpress the human sodium taurocholate co-transporting polypeptide (hNTCP) gene allowing them to be infected by HBV. The viral life-cycle is accurately recapitulated and enables assessment of effects of therapeutic agents on cccDNA. Since viral gene expression is dependent on transcription from the cccDNA intermediate in these cells, measurement of markers of HBV replication may be used as an indicator of cccDNA function. This study evaluated the potential of AAV delivered TALENs in vivo and in vitro by testing their activity and evaluating off-target activity, immunogenicity and liver toxicity. The results provide evidence for the utility of applying AAVs to the delivery of anti-HBV TALENs and offers further support for the feasibility of employing TALENs to treat chronic HBV infection.
Two Single Nucleotide Polymorphisms of the ETS2 Transcriptional Factor Gene Predispose Individuals to High-Risk Acute Myelogenous Leukemia (AML).
Authors: Il-Kwon Lee, Jeong-Hwa Choi, Yeo-Kyeoung Kim, Hee Nam Kim, Kyeong-Soo Park, Je-Jung Lee, Myung Geun Shin, Chan Choi, Hoon Kook, Tae Ju Hwang, Moo Rim Park, Deog Yeon Jo, Jong Ho Won, Hyeoung-Joon Kim
Journal: Blood (2005)
Safety data and patterns of progression in met diagnostic subgroups in OAM4558g; A phase II trial evaluating MetMAb in combination with erlotinib in advanced NSCLC.
Authors: A. Vashishtha, P. H. Patel, W. Yu, J. G. Bothos, J. Simpson, T. Maneatis, L. Doessegger, A. C. Peterson, C. Clement-Duchene, G. Robinet, M. Krzakowski, G. R. Blumenschein, J. H. Goldschmidt, D. B. Daniel, D. R. Spigel
Journal: Journal of Clinical Oncology (2011)

7604 Background: MetMAb (M) is a monovalent monoclonal antibody that blocks HGF-induced Met signaling. A Phase II double-blinded, randomized trial of MetMAb+erlotinib (M+E) or placebo+erlotinib (P+E) in 2nd- and /3rd- line NSCLC was recently completed. The M+E arm demonstrated both PFS and OS advantages in the c-Met IHC expression positive 2+ /3+ (Met Dx+) patients only. Methods: Adverse events (AEs) were assessed for MetMAb related toxicities. In addition to AEs, tumor growth rate and onset of new lesions were evaluated to understand their potential contribution to the clinical outcomes. Results: In the M+E arm, median cycles of M were higher in Met Dx+ vs. Met Dx- pts (4 vs. 2) as were median E doses (61 vs. 42 respectively). Overall pattern of AEs were similar in the two treatment arms, with the exception of peripheral edema which was higher on M+E; regardless of c-Met expression levels. Time to onset of any Grade ≥3 AE in M+E treated patients was also similar across groups. In the Met Dx- group, Grade 3 AEs on M+E were higher than on P+E (56% vs. 35%), but the rate was similar to that observed for Met Dx+ pts on M+E (57%) or P+E (53%). AEs leading to death were highest in Met Dx- pts on M+E (n=5), whereas the number of AEs leading to treatment discontinuation was highest in Met Dx+ pts on M+E (n=6): no consistent patterns were observed for either. The rate of tumor growth was not significantly different between treatment arms or Met sub groups. 55% of Met Dx+ pts progressed with new lesions vs 43% of Met Dx- (P+E), but the addition of M had no effect on this rate. The power to detect a difference is limited by the number of subjects in the study, ie., 65 with Met Dx+ (35 on M+E and 30 on P+E) and 56 with Met Dx- (27 on M+E and 29 on P+E). Conclusions: The combination of M+E did not substantially alter the safety profile of erlotinib and differences in AEs do not clearly explain the PFS and OS outcomes observed in Met Dx subgroups on M+E. Furthermore, the addition of M to E in Met Dx- patients did not appear to alter tumor growth rates or patterns of disease progression.
Abstract 5097: APE1/Ref-1 promotes cell adhesion and migration in cervical cancer cells
Authors: Mihwa Kim, Ho Jin You, Dae Joon Kim
Journal: Cancer Research (2015)

Abstract Apurinic and apyrimidinic endonuclease 1 (APE1)/redox effector factor-1 (Ref-1) is known to be involved in DNA base excision repair pathway and redox-dependent regulation of a number of transcription factors, such as AP1, p53, and Pax-8. It is expressed in numerous human cancers such as pancreatic cancer, prostate cancer, breast cancer, cervical cancer, and colon cancer. Our previous studies suggested APE1/Ref-1 can promote cancer progression through its involvement in cell proliferation, metastasis and angiogenesis; however, the molecular mechanism by which APE1/Ref-1 functions in these processes has not been characterized. In this study, we are the first to report that APE1/Ref-1 activates cell adhesion-related proteins to trigger cell migration. To investigate whether APE1/Ref-1 is involved in cell adhesion in normal fibroblasts, an adhesion assay was performed using GM00637 human fibroblast cell lines stably overexpressing APE1/Ref-1. Cell adhesion to vitronectin, a known ligand of the adhesion molecules integrin ανβ3 and integrin ανβ5, was significantly increased in APE1/Ref-1-overexpressing GM00637 cells compared to control fibroblast cells. FACS analysis showed APE1/Ref-1 induces the expression and activation of adhesion molecules including integrin αν and β3 in APE/Ref-1-overexpressing GM00637 cells. To evaluate whether APE1/Ref-1-mediated regulation of integrin ανβ3 is involved in cell migration in cervical cancer cells, a migration assay was performed using APE1/Ref-1-specific siRNA and an integrin ανβ3-specific antibody. Cell migration was significantly decreased in the absence of APE1/Ref-1 expression in cervical cancer cells, which was comparable with inhibition of integrin ανβ3. Consistent with these results, APE1/Ref-1 increased the activation and expression of integrin ανβ3 in both cervical cancer cells and cervical cancer patient tissue. Combined, these studies suggest that APE/Ref-1 can promote cell adhesion and migration through integrin ανβ3 signaling and thereby this signaling mechanism could contribute to cervical cancer progression. Citation Format: Mihwa Kim, Ho Jin You, Dae Joon Kim. APE1/Ref-1 promotes cell adhesion and migration in cervical 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 5097. doi:10.1158/1538-7445.AM2015-5097
Developing a decision support tool for responding to patients’ reported levels of information needs, family anxiety, depression, and breathlessness.
Authors: Liesbeth van Vliet, Richard Harding, Claudia Bausewein, Sheila Payne, Irene J. Higginson
Journal: Journal of Clinical Oncology (2014)

173 Background: Routine clinical use of Patient Reported Outcome Measures (PROMs) such as the Palliative Care Outcome Scale (POS) may be prevented by a lack of guidance on how to respond to reported symptoms. When using POS in clinical care, clinicians encounter the most difficulties with responding to information needs, depression and family anxiety while breathlessness remains a difficult to treat symptom. We aimed to create a Decision Support Tool (DST) on how to respond to different levels of these patient-reported symptoms. Methods: A systematic search for guidelines and systematic reviews on these topics was conducted (in Pubmed, Cochrane and York DARE databases, Googlescholar, NICE, National Guideline Clearinghouse, Canadian Medical Association, Google.com). In a two-round online Delphi study purposefully sampled international experts (clinicians, researchers, patient representatives) judged the appropriateness (1-9 scale + do not know option) of drafted recommendations for each POS answer category (0-4) and provided qualitative remarks. Recommendations with a median of 7-9 and &lt;30% of scores between 1-3 and 7-9 were included in the DST. Quality was assessed using an adapted GRADE approach. Results: Twenty-five out of 38 (66%) experts participated in round 1, 23 out of 37 (62%) in round 2. Higher POS scores were related to more included recommendations. The DST consists of both a manual and flow-charts of included recommendations for each topic. Overall, psychosocial interventions were recommended for lower levels of depression and breathlessness than drug interventions (e.g., goal-setting/coping versus morphine for breathlessness). Good communication and emotional support were recommended for low family anxiety levels, but a social needs assessment only for higher levels. For information needs recommendations were least discriminative; almost all recommendations (e.g., assess patients’ understanding of information, show empathy) seemed always relevant. Conclusions: The developed DST can assist clinical responses to patient-reported symptoms in palliative care. Future work is needed to test the effect of using the DST on patients’ outcomes.
Effect of pathohistologic complete remission (pCR) rate on the prediction of long-term prognosis following chemoradiotherapy (XRT) with cetuximab (Cet), capecitabine (CAP), and oxaliplatin (OX; Cet-CAPOX) in patients (pts) with locally advanced rectal cancer (LARC).
Authors: T. Liersch, C. Weiss, K. Dellas, M. Hipp, R. Sauer, C. Rödel, D. Arnold
Journal: Journal of Clinical Oncology (2011)

e14035 Background: Preoperative XRT with Cet, CAP and OX (Cet-CAPOX) simultaneously to 50.4 Gy in fractions of 1.8 Gy has shown to be feasible, with no unexpected acute toxicity or increased perioperative morbidity, in LARC (Rödel et al., Int J Radiat Oncol Biol Phys., 2008). However, this regimen failed to show an increase in pathohistological response, incl. pCR rate, compared to reported data of 5-FU based XRT. Here, long-term results are reported. Methods: 58 pts with LARC were included in the phase I/II trial, with 40 pts evaluable for long-term efficacy (phase II). Pts characteristics: median age 63 (37-83) yrs, ECOG PS 0/1 80%/20%, at pre-therapeutic staging: cT2/3/4 3%/85%/12%, cN0/+ 20%/80%. Results: Following XRT, quality assessed TME surgery was done in all pts: low anterior resection in 70%, APR in 23%, Hartmann`s procedure in 7%. Histopathologically confirmed R0 rate was 95%. Pathology reports: ypT0N0 10%, ypT1-2N0 30%, ypT1-2N+ 5%, ypT3-4N0 25%, ypT3-4N+ 30%. Pathological tumor regression grading (TRG) according to Dworak et al.: TRG 4 (pCR) 10%, TRG 3 (>50% of tumor mass eradicated) 40%, TRG 2 (25-50%) 30%, TRG 0/1 20%. Adjuvant chemotherapy (not protocol defined) was given in 70%: 5FU/FA or CAP alone 30%, with OX 40%. With a median follow-up of 40 (3-49) months, 8 of 40 pts. (20%) have died: 1 with local R2 status and occult peritoneal metastases at staging died 5 weeks after surgery due to peritoneal tumor progression. No further locoregional failure occurred in 39 pts up to now. 3 pts. (8%) died unrelated to the malignant disease. 4 pts. (10%) developed distant metastases (liver: 1; lung: 1, both: 1, peritoneum: 1). Disease-free survival at 4 years was 77%. Conclusions: Notwithstanding the limited number of pts and follow-up time, the disappointingly poor results in terms of pathohistological response (incl. pCR) did not translate into poor long-term results. Taken into account the results of the EXPERT-C trial (Dewdney et al., ASCO GI 2011), pCR and TRG are likely insufficient parameters to assess the benefit of a cetuximab-containing perioperative XRT regimen.
Abstract 1363: VEGF reduction facilitates angiogenesis by CXCR2
Authors: Mary A. Kosir, Donghong Ju
Journal: Cancer Research (2015)

Abstract PURPOSE: With multiple pathways for angiogeneis, the purpose of the study is to quantify the expression of angiogenic tyrosine kinase receptors during endothelial tubule formation while inhibiting a G-protein linked CXCR2 receptor that binds angiogenic CXC chemokines. METHODS: Using endothelial tubule formation assays on Matrigel and Growth Factor Reduced (GFR) Matrigel, and without added VEGF, human umbilical vascular endothelial cells (HUVEC) were treated with a selective small molecule non-peptide inhibitor of CXCR2 called SB225002 (N-(2-hydroxy-4-nitrophenyl)-N9-(2-bromophenyl)urea compared with control. Previous dose-inhibition curves established optimal dosing of SB225002. After 6 hours of incubation, endothelial tubule formation was quantified by counting tubule length in four random microscopic fields using the microscope camera software NIS-Elements AR 3.10 in triplicate and compared to control. In parallel, endothelial cells were collected from the Matrigel under the same conditions for quantification of angiogenesis receptors and growth factors by RT-PCR in triplicate. RESULTS: There was significant inhibition of endothelial tubule formation on Matrigel (and GFR Matrigel) by the addition of SB225002 (40-58% of inhibition) under conditions of no added VEGF when compared to control. The expression of mRNA in HUVEC cells harvested from Matrigel was not different for VEGFR1-3, FGFR1-3, and PDGFRA,B with or without the inhibitor. Under conditions of no added VEGF, the expression of CXCR2 mRNA was the same with or without the inhibitor, but the endothelial tubule formation was significantly inhibited. CONCLUSION: Under growth factor reduced conditions, including no added VEGF, the endothelial tubule formation and angiogenenic tyrosine kinase receptor expression was similar to standard matrigel. However, when a selective non-peptide inhibitor of CXCR2 (SB225002) was added, endothelial tubule formation was inhibited without change in expression of other angiogenesis receptors from HUVEC cells retrieved from the matrigel. Therefore, tubule formation continues with the chemokine/CXCR2 axis and may be an additional pathway that circumvents other attempts at angiogenesis inhibition targeting tyrosine kinase inhibitors. Citation Format: Mary A. Kosir, Donghong Ju. VEGF reduction facilitates angiogenesis by CXCR2. [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 1363. doi:10.1158/1538-7445.AM2015-1363
The E3 Ubiquitin Ligase TRIM36, a Transcriptional Target of Tob, Is Expressed in Anergic T Cells and Mediates Unresponsiveness through Proteolysis of Signaling Proteins PLC- γ1 and PKC-𝛉.
Authors: Dimitrios Tzachanis, Alla Berezovskaya, Esther M. Lafuente, Lequn Li, Gordon J. Freeman, Vassiliki A. Boussiotis
Journal: Blood (2004)
587. Genetic Delivery of a miRNA Cluster with Polycistronic siRNAs Reduces Expression of Epidermal Growth Receptor in Human Glioblastoma Cells
Authors: Martin J. Hicks, Imari Patel
Journal: Molecular Therapy (2016)

Glioblastoma multiforme (GBM), the most common malignancy of the brain, is a grade IV astrocytoma. It is one of the most rapidly growing and invasive types of glial tumors. The standard therapy includes surgical removal, radiation and chemotherapy with a median survival of about 14 months. In addition, systemic therapies are limited by the blood-brain barrier. To bypass the barrier, we are developing a gene delivery strategy to inhibit the expression of tyrosine kinase receptors (TKR), which are commonly upregulated in GBM. One TKR, epidermal growth factor receptor (EGFR), is overexpressed in GBM leading to uncontrolled growth and proliferation. Our approach is to recruit the RNA interference pathway. Although small interfering RNAs (siRNAs) are often utilized to silence gene expression, exogenously expressed siRNAs are not an effective strategy to treat human disease due to both extracellular and intracellular nucleases as well as activation of cellular immunity against foreign nucleic acids. To bypass these degradatory mechanisms, we use a natural miRNA cluster genetic background to effectively deliver the DNA encoding multiple anti-EGFR siRNAs by inserting them into the tertiary structure of the miRNA cluster, miR-17-92. The anti-EGFR polycistronic miRNA cluster (pAAV-miR-IP1) expresses six siRNAs directed against EGFR, specifically targeting the extracellular ligand binding domain, transmembrane domain, intracellular tyrosine kinase domain and 3' untranslated region of the EGFR transcript. The vector, pAAV-miR-IP1, was transfected into the human GBM cell lines, A172 and U87MG. Results demonstrate that pAAV-miR-IP1 was expressed at high levels with a subsequent reduction in EGFR mRNA expression. Additional strategies include using the polycistronic delivery mechanism to target multiple TKRs in addition to EGFR.
Biomarkers of Bone Remodeling in Multiple Myeloma Patients to Tailor Bisphosphonate Therapy
Authors: Chirayu Patel, Andrew Yee, Tyler A. Scullen, Neeharika Nemani, Loredana Santo, Anuj Mahindra, Paul G. Richardson, Jacob P. Laubach, Irene M. Ghobrial, Robert Schlossman, Kenneth C. Anderson, Nikhil C Munshi, Noopur Raje
Journal: Blood (2012)
Phase I Trial of the Combination of Lenalidomide, Thalidomide and Dexamethasone In Relapsed/Refractory Multiple Myeloma
Authors: Jatin J Shah, Robert Z. Orlowski, Raymond Alexanian, Michael Wang, Sheeba Thomas, Muzaffar H. Qazilbash, Simrit Parmar, Nina Shah, Qaiser Bashir, Uday Popat, Donna Weber
Journal: Blood (2010)