tbuckley/cabot-search huggingface dataset:
The ICOS:ICOSL Costimulatory Pathway Plays an Important Role in GVHD and Bone Marrow (BM) Graft Rejection.
Authors: Patricia Taylor, Angela Panoskaltsis-Mortari, Gordon Freeman, Arlene Sharpe, Randolph Noelle, Alexander Rudensky, Tak Mak, Jonathan Serody, Bruce Blazar
Journal: Blood (2004)
700. Successful Repeated Hepatic Gene Delivery in Mice and Non-Human Primates Achieved by Sequential Administration of AAV5 and AAV1 Vector Serotypes
Authors: Anna Majowicz, David Salas, Nerea Zabaleta, Estefania Rodríguez-Garcia, Gloria Gonzalez Aseguinolaza, Harald Petry, Valerie Ferreira
Journal: Molecular Therapy (2016)

The major challenge in AAV-based gene therapy is the presence of circulating neutralizing antibodies (NAB) against AAV vector capsids. NAB can be present in patient's blood prior to AAV treatment due to naturally acquired infections with the wild type AAV virus (pre-existing NAB). Anti-AAV NABs are also raised after first administration of AAV in the course of gene therapy treatment. There is a need to develop strategies that would permit a repeated AAV gene delivery not only to be able to treat the patients that have pre-existing NAB but also for the patients previously treated with recombinant AAV, that might experience overtime a decrease in therapeutic protein expression due to the natural turnover of transduced cells. To address those issues, we explored the feasibility of using the AAV5 and AAV1 serotypes for repeated, liver-targeted gene delivery in non-human primates (NHPs). Sequential AAV-based gene delivery with AAV5 and AAV1 proved to be successful as an expression of the two reporter transgenes used in the study was observed (hSEAP and hFIX). In contrast, the re-administration of the same serotype (AAV5-hSEAP followed by AAV5-hFIX) was unsuccessful due to the total inhibition of secondary AAV5 transduction by anti-AAV5 NAB. In order to determine the long-term stability of transgenes activity/expression, the NHP cross administration experiment was continued up to 1 year after primary injection (AAV5- hSEAP or PBS). All NHPs cross administered with AAV5-hSEAP followed by AAV1-hFIX showed hSEAP and hFIX protein expression in plasma. In a long-term follow-up, decreases of transgenes activity/expression were observed in 5 out of the 12 NHPs injected. The reductions of transgenes activity/expression were associated with specific immune responses directed against hSEAP or hFIX. Interestingly, DNA and mRNA levels in the animals presenting transgene directed immunity were comparable to those observed in the other animals that did not lose the transgene activity/expression. Additionally, fluorescent in situ hybridization (FISH) was performed on the liver sections of the animals to localize AAV vector DNA and transgene mRNA. In summary, our data demonstrates that a successful re-administration can be achieved in AAV-based gene therapy when combining AAV5 and AAV1 in NHPs.
Involvement of MAPK pathway in hypoxia-induced up-regulation of urokinase plasminogen activator receptor in a human prostatic cancer cell line, PC3MLN4
Authors: K. H. Lee, E. Y. Choi, J.-L. Lee, M.-S. Hyun, J. R. Kim, J.-O. Park-Hah
Journal: Journal of Clinical Oncology (2004)

9684 Background: Hypoxia has been known to be involved in many pathological processes including tumor formation, where it has been associated with resistance to treatment and malignant progression. Clinical studies have shown that tumor hypoxia is associated with invasive growth and metastasis and may be an important prognostic factor adversely influencing survival in patients with tumors. Methods: To investigate the mechanisms involved in hypoxia-induced invasive growth and metastasis, hypoxia-mediated uPAR expression, cellular invasiveness, and mitogen activated protein kinase (MAPK) activation were measured in a prostate cancer cell line, PC3MLN4. In order to investigate whether exposure to hypoxia increases the expression of uPAR in PC3MLN4 cells, the cells were incubated under either hypoxia or normoxia for 24 hrs and then the level of uPAR was measured using a flow-cytometry. Results: Hypoxia induced a 2.7 fold increase in the level of cell surface uPAR expression compared to normoxia. And also found hypoxia-induced up-regulation of uPAR mRNA . Hypoxic cells exhibited an invasion rate of 3-fold higher than normoxic cells in standard two-chamber invasion assay. Such a hypoxia-induced cellular invasiveness was blocked by an anti-uPAR monoclonal antibody. Phosphorylations of ERK and p38 kinase were also more extensive in hypoxic cells than in normoxic cells. To test whether ERK and p38 MAP kinase involved in the hypoxia-induced uPAR up-regulation, cells were pretreated with PD 98059 or SB 203580 and the uPAR level was measured by Western blotting. Hypoxia-induced uPAR up-regulation was inhibited by pretreatments with PD 98059 and SB 203580. Cell growth also increased in hypoxic cells. Conclusions: Hypoxia increased tumor cell invasion by up-regulating uPAR expression, which might be mediated through ERK and p38 kinase signaling pathway in the prostate cancer cell line and uPAR may be a good novel therapeutic target. No significant financial relationships to disclose.
Abstract 1751: Y15 and Pyridoxal 5′ Phosphate enhances the cytotoxic profile of Cisplatin and Paclitaxel in platinum resistant ovarian cancer cells
Authors: Monique P. Reboe, Terry-ann Waite, Arkene S. Levy, Sivanesan Dhandayuthapani, Appu Rathinavelu
Journal: Cancer Research (2015)

Abstract Overexpression of the Tyrosine Kinase, Focal Adhesion Kinase (FAK) has been implicated in the development of resistance to front line agents, cisplatin and paclitaxel, in the treatment of ovarian cancer. Moreover vitamins such as Pyridoxal 5′ phosphate (PLP) has been shown to induce cytotoxicity in cancers. Therefore, the aim of this study was to assess the cytotoxic efficacy of FAK inhibition using 1,2,4,5-Benzenetetraamine tetrahydrochloride (Y15) and PLP combined with cisplatin and paclitaxel. Platinum resistant OVCAR-3 cells were treated with varying concentrations of Y15 (10μM, 15 μM, 30 μM, 100 μM), and PLP (0.25mM, 1mM, 2Mm). Cells were also treated with Y15 + PLP, Y15 + Cisplatin + Paclitaxel, PLP + cisplatin + paclitaxel and Cisplatin + paclitaxel for 24 hrs in triplicate and control cells with 2N HCl or media only. The cytotoxic profile of each treatment was assessed using the automated trypan blue assay and DNA fragmentation assay was performed to evaluate the mechanism of cell death. Dose dependent and statistically significant cytotoxicities were seen with Y15 and PLP compared to the controls with IC 50 values 80 μM (p &amp;lt;0.05) and 2 mM (p&amp;lt;0.05) respectively. The use of Y15 in combination with PLP showed enhanced cytotoxic efficacy in OVCAR-3 cell (p&amp;lt;0.0001). Similarly, Y15 significantly increased percentage cell death when combined with cisplatin + paclitaxel (62%, p&amp;lt;0.0001) in comparison to cisplatin + paclitaxel (35%). The use of PLP with cisplatin + paclitaxel also showed a greater cytotoxic profile of 55% (p&amp;lt;0.002). Cell death by apoptosis was not confirmed by DNA fragments in Y15 and PLP positive treatment groups. Y15 and PLP significantly enhances the cytotoxic profile of cisplatin, paclitaxel in resistant OVCAR-3 cells suggesting that targeting FAK activity through the inhibition of Y397 site and the use of PLP augments resistance to paclitaxel and cisplatin. However further studies are needed to determine the mechanistic pathway of Y15 and Pyridoxal 5′Phosphate induce cytotoxicity. Citation Format: Monique P. Reboe, Terry-ann Waite, Arkene S. Levy, Sivanesan Dhandayuthapani, Appu Rathinavelu. Y15 and Pyridoxal 5′ Phosphate enhances the cytotoxic profile of Cisplatin and Paclitaxel in platinum resistant ovarian 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 1751. doi:10.1158/1538-7445.AM2015-1751
555. Development of a Post-Exposure Treatment for Ebola Virus Infections Based on AAV Vectors and Zmapp Antibody Cocktail
Authors: Marc-Andre Robert, Amine Kamen, Gary Kobinger, Rénald Gilbert, Bruno Gaillet
Journal: Molecular Therapy (2016)

The recent Ebola outbreak in West Africa has been the deadliest in the history. To prevent future recurrence of such outbreak, better treatments and effective vaccines against Ebola virus are desirable. Among such promising treatments, the Zmapp cocktail containing neutralizing antibodies (13C6, 2G4 and 4G7) has successfully treated some patients. However, the feasibility of using it on large populations especially in developing countries is questionable. To address this potential issue, we propose to employ recombinant vectors derived from adeno-associated virus (rAAV). There are several advantages of using rAAV: because of 1) their safety profile; 2) only one injection (or a few) would be required; 3) the high stability of lyophilized rAAVs at ambient temperature and; 4) the panel of available serotypes. Because of these interesting features, we are currently developing a treatment based on three rAAVs to deliver the genes for the Zmapp cocktail of antibodies. We have already produced at small scale a rAAV expressing the 2G4 antibody. The DNA sequences for the heavy chain and light chains were codon-optimized for better expression in humans and were designed to be expressed from the same gene. A strong promoter (CAG) resistant to silencing in vivo was chosen to drive gene expression of the antibody. The rAAV were produced by transfection using our patented cGMP compatible HEK293 cell line. The production was performed in suspension culture in the absence of serum. Secretion of 2G4 antibody by rAAV transduced cells (HEK293 and CHO cells) was confirmed. The results demonstrated that rAAV-CAG-2G4 was functional and allowed for the correct assembly of the heavy and light chains of 2G4. Purification of 200 mL of rAAV-CAG-2G4 production was performed by ultracentrifugation on an iodixanol density-step gradient. Two other rAAVs coding 13C6 and 4G7 antibodies are in the processed of being constructed and produced in a similar manner. We are also in the process of comparing the efficacy of two serotypes of AAV (9 and DJ) in mice by intranasal delivery. Using the best serotype, the rAAVs will be produced and purified from a starting suspension culture of 20 L. Their efficacy for treating Ebola infections will then be evaluated in a mouse model infected by the virus.
A Case Report of Aggressive Adult Neuroblastoma Mimicking Acute Leukemia with Fulminant Course and Fatal Outcome
Authors: Irina Panovska, Martin Ivanovski, Sanja Trajkova, Aleksandra Pivkova, Sonja Genadieva-Stavric, Liljana Hadzi-Pecova, Arif Latifi, Dejan Trajkov, Mirko Spiroski, Lidija Cevreska
Journal: Blood (2008)
Abstract 2442: XPO1 is selinexor prime target: validation by mutating cysteine 528 on both XPO1 alleles using CRISPR/Cas9 genome editing
Authors: Jasper E. Neggers, Thomas Vercruysse, Maarten Jacquemyn, Els Vanstreels, Erkan Baloglu, Sharon Shacham, Marsha Crochiere, William Senapedis, Yosef Landesman, Dirk Daelemans
Journal: Cancer Research (2015)

Abstract Human Exportin-1 (XPO1) is the key nuclear-cytoplasmic transport protein that exports a variety of different cargo proteins out of the cell's nucleus. Inhibition of XPO1-mediated protein export results in nuclear retention of cargo proteins including tumour suppressor proteins and is a novel therapeutic strategy for cancer. Selinexor, an orally bioavailable XPO1 inhibitor, effectively demonstrates potent activity against multiple types of cancer. Here we directly demonstrate the high selectivity of selinexor for XPO1 inside living cells. Selinexor binds to the cargo-binding pocket of XPO1 and inhibits the formation of XPO1-cargo complex. The binding of selinexor to XPO1 involves a slowly reversible Michael addition type interaction of its acrylate moiety with cysteine 528 residue of XPO1. Using CRISPR/Cas9 genome editing we have now created a homozygous Jurkat T-ALL cell line carrying a Cys528Ser mutation in XPO1. This cell line was used to study the specificity of selinexor for XPO1 inside the cancer cell. Selinexor was highly toxic to wild-type Jurkat cells in the low nanomolar range while the mutant cells were resistant up to micromolar concentrations. These results were confirmed by resistance to induction of caspase dependent apoptosis as well as cell cycle arrest in mutant cells. The single Cys528Ser mutation thus confers resistance to selinexor. In addition, selinexor failed to accumulate cargo proteins in the nucleus of mutant cells. Moreover, the Cys528Ser substitution conferred resistance to the direct interaction of this drug to XPO1 in cells. In conclusion, these results validate XPO1 as the selinexor's prime target and pinpoint its unique selectivity to the Cys528 residue of XPO1. Our findings also demonstrate the general utility of CRISPR/Cas9 genome editing for introducing point mutations in the genome of cancer cells for drug-target specificity validation. Citation Format: Jasper E. Neggers, Thomas Vercruysse, Maarten Jacquemyn, Els Vanstreels, Erkan Baloglu, Sharon Shacham, Marsha Crochiere, William Senapedis, Yosef Landesman, Dirk Daelemans. XPO1 is selinexor prime target: validation by mutating cysteine 528 on both XPO1 alleles using CRISPR/Cas9 genome editing. [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 2442. doi:10.1158/1538-7445.AM2015-2442
360. Magnetofection: A Versatile Approach for Messenger RNA Delivery
Authors: Zohreh Sadat Badieyan, Manish K. Aneja, Christian Plank
Journal: Molecular Therapy (2015)

Transcript therapy, using stabilized non-immunogenic messenger RNA (SNIM RNA), overcomes the potential risks of mutagenicity and immunogenicity, normally associated with DNA based gene delivery and recombinant protein therapy, respectively. Magnetofection, as a delivery system, has been shown to enhance DNA based viral and non-viral gene delivery. In this method, viral or non-viral gene carriers containing nucleic acids are associated with magnetic nanoparticles and a gradient magnetic field is applied to pull them on to the target cell surface. In this study, we established a magnetofection based protocol for enhanced mRNA delivery, using non-viral carriers. Firstly, we compared magnetofection, using different magnetic nanoparticles, to lipofection and polyfection, to find the best protocol for in vitro mRNA delivery. Then, the established protocol was generalized for co-transfection of up to three reporter SNIM RNAs (Luc, eGFP and RFP) in mono-culture of Primary Mouse Embryonic Fibroblasts (PMEF), or co-culture of PMEF and Porcine Fetal Fibroblast (PFF). In parallel, expression kinetics post magnetofection of d2eGFP SNIM RNA were determined. Magnetofection significantly improved transfection efficiency in mono culture as well as co-culture of applied cells. Using this protocol, co-transfection of different SNIM RNAs did not inhibit the transfection of every single SNIM RNA into PMEF or PFF cells. Moreover, rapid and higher peak of expression was observed with using magnetic nanoparticles. To conclude, magnetofection of mRNA presents itself as an efficient mRNA delivery system, thereby bringing transcript therapies a step closer to their clinical uses.
Dendritic Cell Vaccination Enhances Immune Responses and Induces Regression of HER2pos DCIS Independent of Route: Results of Randomized Selection Design Trial
Authors: Lea Lowenfeld, Rosemarie Mick, Jashodeep Datta, Shuwen Xu, Elizabeth Fitzpatrick, Carla S. Fisher, Kevin R. Fox, Angela DeMichele, Paul J. Zhang, Susan P. Weinstein, Robert E. Roses, Brian J. Czerniecki
Journal: Clinical Cancer Research (2016)

Abstract Purpose: Vaccination with HER2 peptide-pulsed DC1s stimulates a HER2-specific T-cell response. This randomized trial aimed to establish safety and evaluate immune and clinical responses to vaccination via intralesional (IL), intranodal (IN), or both intralesional and intranodal (ILN) injection. Experimental Design: Fifty-four HER2pos patients [42 pure ductal carcinoma in situ (DCIS), 12 early invasive breast cancer (IBC)] were enrolled in a neoadjuvant HER2 peptide-pulsed DC1 vaccine trial. Patients were randomized to IL (n = 19), IN (n = 19), or ILN (n = 16) injection. Immune responses were measured in peripheral blood and sentinel lymph nodes by ELISPOT or in vitro sensitization assay. Pathologic response was assessed in resected surgical specimens. Results: Vaccination by all injection routes was well tolerated. There was no significant difference in immune response rates by vaccination route (IL 84.2% vs. IN 89.5% vs. ILN 66.7%; P = 0.30). The pathologic complete response (pCR) rate was higher in DCIS patients compared with IBC patients (28.6% vs. 8.3%). DCIS patients who achieved pCR (n = 12) and who did not achieve pCR (n = 30) had similar peripheral blood anti-HER2 immune responses. All patients who achieved pCR had an anti-HER2 CD4 immune response in the sentinel lymph node, and the quantified response was higher by response repertoire (P = 0.03) and cumulative response (P = 0.04). Conclusions: Anti-HER2 DC1 vaccination is a safe and immunogenic treatment to induce tumor-specific T-cell responses in HER2pos patients; immune and clinical responses were similar independent of vaccination route. The immune response in the sentinel lymph nodes, rather than in the peripheral blood, may serve as an endpoint more reflective of antitumor activity. Clin Cancer Res; 23(12); 2961–71. ©2016 AACR.
Updated clinical and biomarker results from a phase I study of vandetanib with radiation therapy (RT) with or without cisplatin in locally advanced head and neck squamous cell carcinoma (HNSCC).
Authors: V. Papadimitrakopoulou, J. Heymach, S. J. Frank, J. Myers, H. Lin, H. T. Tran, C. Chen, F. R. Hirsch, P. B. Langmuir, J. R. Vasselli, S. M. Lippman, D. Raben
Journal: Journal of Clinical Oncology (2011)

5510 Background: Vandetanib (V) is an oral inhibitor of VEGFR, EGFR and RET signaling. Methods: Eligible patients (pts) with previously untreated, unresected HNSCC received once-daily V (starting dose 100 mg) for 14 days (d) followed by V + RT (2.2 Gy/d, 5 d/wk; total 66 Gy) for 6 wks (regimen 1) or V + RT (2 Gy/d, 5 d/wk; total 70 Gy) + cisplatin (30 mg/m2 weekly) for 7 wks (regimen 2). The primary objective was to determine the safety, tolerability and maximum tolerated dose (MTD) of V. We used multianalyte biomarker profiling (Luminex Corp) and ELISA to assess 57 plasma markers before and after 14 d of V and at completion of RT (d50), and EGFR, p16, E-cadherin and vimentin immunohistochemistry in tumor tissue. Clinical results were correlated with biomarkers. Results: Of 33 pts (median age 53 yrs; 28 male; all M0) who received treatment, 30 completed therapy (regimen 1, n=12; regimen 2, n=18). In regimen 1, there were no DLTs in the 100 mg cohort and 1 DLT in the 200 mg cohort (each n=6). In regimen 2, there were no DLTs in the 100 mg cohort and 3 DLTs in the 200 mg cohort (each n=6); 100 mg was defined as the MTD and the cohort expanded to 15 pts. The most common grade ≥3 AEs were dysphagia (27%), stomatitis (24%) and mucosal inflammation (21%); 4 pts discontinued V due to AEs. All 29 pts evaluable for efficacy (RECIST) achieved a complete response. At data cut-off, 11/29 had completed 2-yr follow-up without recurrence and 6 had recurrence. Steady-state exposure to V was unaffected by RT/cisplatin. Blood biomarkers were analyzed in 31 (baseline), 30 (d15) and 26 (d50) pts; 27 pts had tissue analyses. High levels of baseline plasma HGF, IL-8, MIF and eotaxin were associated with shorter progression-free survival (PFS; P<0.05). Significant increases in six plasma factors (HGF, OPN, IL-6, soluble IL-2R, IP-10 and MCP-1) were observed after RT, but not V alone. Tumor p16 expression was associated with superior PFS (P=0.01). Conclusions: The combination of V with RT ± cisplatin is tolerable with encouraging efficacy. Baseline HGF, IL-8, MIF and eotaxin are potentially useful markers of adverse outcome. Inflammatory and hypoxia-regulated cytokines appear to be upregulated after RT.
Phase I Study of Non-Engrafting Allogeneic, Mismatched, Unmanipulated Peripheral Blood Mononuclear Cell Infusions to Treat Poor-Prognosis Acute Myeloid Leukemia
Authors: Elizabeth F Krakow, Julie Bergeron, Silvy Lachance, Denis-Claude Roy, Jean-Sebastien Delisle
Journal: Blood (2015)
Abstract 2193: Hypoxia-induced G9a histone methyltransferase is crucial for blunting RUNX3 tumor suppressor function
Authors: You Mie Lee
Journal: Cancer Research (2015)

Abstract Recent our work shed light on the critical roles of RUNX3 in early tumorigenesis. Hypoxia has been reported to be an early tumorigenic insult. Previously, we showed that RUNX3 is silenced by G9a HMT under hypoxia. However, the possible direct modification of RUNX3 by G9a induced by hypoxia and its outcome are not yet investigated. Here we show that RUNX3 physically interacts with G9a. Under hypoxic conditions, G9a directly methylates lysines 129 and 171 of RUNX3. G9a-methylated RUNX3 is localized to the cytoplasm and degrade through Smurf-mediated ubiquitination pathway. Hypoxia and G9a overexpression decreased RUNX3-responsive genes by DNA microarray analysis. Consistently, tumor suppressive role of RUNX3 was decreased by G9a-mediated methylation in gastric cancer xenograft model. Taken together, our results demonstrate that methylation by G9a is a key chemical modification for the regulation of RUNX3 tumor suppressor function in hypoxic conditions and in early tumorigenic stage. Citation Format: You Mie Lee. Hypoxia-induced G9a histone methyltransferase is crucial for blunting RUNX3 tumor suppressor function. [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 2193. doi:10.1158/1538-7445.AM2015-2193