tbuckley/cabot-search huggingface dataset:
75. Development of an LMP-Specific T Cell Bank for Third Party Use as a Curative Strategy Post-Transplant Lymphoproliferative Disease After Solid Organ Transplant
Authors: Lauren McLaughlin, Lauren Roesch, E. Kathy Mintz, Sarah McCormick, Cecilia Barese, Elizabeth Williams, Neha Joshi, Fahmida Hoq, Michael Keller, C. Russell Cruz, Mitchell S. Cairo, Jessica Hochberg, Patrick J. Hanley, Catherine M. Bollard
Journal: Molecular Therapy (2016)

EBV-associated tumors in the immune deficient host express type II and III latency antigens including latent membrane protein 1 (LMP1) and LMP2, which can serve as potential targets for immunotherapy. Several studies have documented the safety and efficacy of LMP-specific T cells for patients with EBV-associated malignancies, but clinical applications may be limited by the time to generate LMP-specific T cell products as well as the availability of an appropriate donor source. We hypothesize that the administration of "off the shelf" third party LMP-specific cytotoxic lymphocytes (LMP-CTLs) will rapidly restore EBV-specific T-cell immunity and prevent relapse in patients with post-transplant lymphoproliferative disease PTLD post solid organ transplant (SOT). To develop the third party T cell bank, we manufactured healthy donor-derived LMP-specific T cells from eligible donors with a wide range of HLA types in our good manufacturing practices (GMP) facility. This T-cell bank is for several clinical trials including a proposed multicenter Children's Oncology Group (COG) trial (ANHL1522) for patients with PTLD after SOT. Currently, 15 LMP-specific T-cell products have been manufactured from healthy donors and released for third-party use using autologous monocytes and lymphoblastoid cell lines (LCL), transduced with an adenoviral vector expressing ΔLMP1 and LMP2. T-cell products were active against LMP2 (mean:172 SFU/1×10^5 cells; range:13-655), LMP1 (33; 1-322), and LCL (87; 0-424) as determined by IFN-γ ELISPOT assay. Epitope mapping of LMP-specific T cells using IFN-γ ELISPOT assay demonstrates that these products recognize a broad epitope repertoire within LMP1 and LMP2. At the time of cryopreservation, the T-cell products comprised a mean of 45% CD8+ T-cells, 35% CD4+ T-cells, and 9% NK cells. No B cells or monocytes were detected in the final products. Thus far, one patient with NK/T cell non-Hodgkin Lymphoma received third party LMP-specific T cells achieving a very good partial response. No infusion-related toxicities were observed, and LMP-specific T cells were detectable post-infusion. Thus, third party LMP-specific T cells appear to be a safe and promising therapeutic modality for patients with EBV-associated lymphomas, and a third party bank will make this therapeutic more readily available to patients with PTLD post-SOT.
Novel Leukemia Stem Cell-Targeted Therapy for Acute Myeloid Leukemia Based on Dual Inhibition of Ezh1/Ezh2
Authors: Shuhei Fujita, Daisuke Honma, Nobuaki Adachi, Kazushi Araki, Emi Takamatsu, Kazumasa Aoyama, Atsushi Iwama, Issay Kitabayashi
Journal: Blood (2015)
Predictors of treatment outcomes in early stage hepatocellular carcinoma (HCC) detected in a surveillance program
Authors: S. L. Chan, F. K. Mo, J. Koh, E. P. Hui, S. C. Yu, P. B. Lai, H. L. Chan, V. T. Chan, A. T. Chan, W. Yeo, T. Mok
Journal: Journal of Clinical Oncology (2009)

4584 Background: Surveillance for HCC in hepatitis B virus (HBV) carriers aims to improve survival by detection of early resectable tumor. In 2005, our group reported low surgical resection rates despite detection of small sized tumors in a prospective surveillance cohort of 1018 HBV carriers (Mok TS et al. J Clin Oncol 05). In current study, we aim to identify predictive factors for treatment outcomes in HCC detected from our surveillance program. Methods: The prospective cohort was recruited at the Prince of Wales Hospital between Oct 1997 and Nov 2000. We updated the database in Dec 2008 and performed univariate and multivariate analysis on clinical (age, sex, cirrhosis, ascites, anti-viral therapy, bilirubin, ALT, albumin, INR), tumorous (size, number, resection) and virologic factors (HBV DNA, Genotype) for prediction of outcome in HCC patients (pts). Results: In the prospective cohort, total 923 HBV carriers were updated (95 lost to follow-up). After median follow-up of 9.95 years, we confirmed diagnosis of 105 HCC. Median age = 51 (range: 40–69); M:F = 82:23; Child's A and B cirrhosis = 38:67. Fifty seven pts (54.3%) had solitary tumor but only 34 (32.4%) are amenable to resection. Absence of cirrhosis (p=0.0072) and normal albumin level (p=0.0379) are predictors of surgical resection while tumor size and number are not. The median survival of all HCC pts was 2.26 years. Anti-viral therapy during the surveillance period is a strong predictor of survival (3.74 vs. 1.63 years; p=0.0115). In multivariate analysis, both anti- viral therapy (HR=0.35; 95%CI: 0.19–0.73; p=0.0041) and normal bilirubin level (HR=0.30; 95% CI: 0.25–0.80; p=0.0069) were predictive of improved survival in pts with HCC. The benefits of anti-viral therapy applied to both surgical and non-surgical candidates (p=0.17). Conclusions: Anti-viral therapy is a potent predictor of survival of HBV related HCC. Liver function is more important than tumor characteristics in determining the outcome of HCC detected in the surveillance program. No significant financial relationships to disclose.
Abstract 4595: Intra-arrest Hypothermia Preserves Heart Akt Phosphorylation And Cardiac Function Following Cardiac Arrest In Wild-type But Not Akt(+/−) Mice
Authors: David G Beiser, Kimberly R Wojcik, Danhong Zhao, Kimm J Hamann, Terry L Vanden Hoek
Journal: Circulation (2008)

OBJECTIVE: To establish the role of Akt signaling in hypothermia protection following cardiac arrest. METHODS: Cardiac arrest was induced in anesthetized C57B6 wild-type (WT) and Akt1(+/−) heterozygous mice with an intravenous bolus of KCl. Animals were randomized to normothermia ( NT , 37°C) or hypothermia ( HT , 30°C) induced after 6 minutes of arrest and continued for 1 hour following return of spontaneous circulation (ROSC). After 8 minutes, resuscitation was attempted with chest compressions, mechanical ventilation, and fluid administration. Following ROSC, resuscitated animals were monitored for up to 4 hours. Total Akt and phosphorylated Akt (pAkt at Ser473 and Thr308) levels were measured by Western blot analysis in whole heart lysates from 4-hour survivors and shams (n=4 each group). RESULTS: In WT animals, 4-hour survival (HT = 7 of 8, NT = 3 of 8, p &lt; 0.04), cardiac output (p&lt;0.001) and left-ventricular dP/dt max (p&lt;0.001) were significantly improved in HT versus NT animals. In addition, hearts from WT animals randomized to HT displayed increased pAkt (Ser473) levels and over 8-fold higher pAkt(Thr308) levels compared to NT and sham groups (p&lt;0.001). These pAkt changes occurred despite total Akt levels remaining unchanged. Baseline hemodynamics prior to arrest were similar between WT and Akt(+/−) animals, while heart Akt1levels were decreased by approximately 40% in Akt(+/−) heterozygous mice. In contrast to WT mice, HT compared to NT had no effect on Akt(+/−) mouse 4-hour survival rates and left-ventricular hemodynamic measures. Compared to HT treated WT mice, Akt(+/−) animals treated with HT displayed significantly lower maximum left-ventricular pressure, stroke volume, and dP/dt max by 4 hours post-ROSC (p&lt;0.05). CONCLUSIONS: HT improves short-term cardiovascular outcomes and induces signficant heart Akt(Thr308) phosphorylation during the immediate post-ROSC period. The therapeutic benefit of intra-arrest HT on post-ROSC hemodynamics is greatly attenuated in Akt(+/−) animals, suggesting that Akt signaling may play a critical role in mediating HT protection.
307. Rational Development of 12 Different AAV Serotypes as Scaffolds for Peptide Display
Authors: Kathleen Boerner, Eike Kienle, Anna Sacher, Lin-Ya Huang, Marina Bechtle, Ellen Wiedtke, Philipp Bayer, Hans-Georg Kräusslich, Mavis Agbandje-McKenna, Dirk Grimm
Journal: Molecular Therapy (2015)

Recombinant AAV vectors are some of the most attractive vehicles for therapeutic gene delivery available today, for many reasons. One is that AAV capsids are highly amenable to genetic engineering, permitting the creation and identification of vectors with novel properties including target cell specificities. One potent strategy is to insert a randomized peptide library into an exposed AAV2 capsid region, followed by iterative selection on desired cells to enrich single peptides binding to cell-surface receptors and hence mediating AAV transduction. However, several problems hamper the use of AAV2 as scaffold for peptide display: (i) neutralizing anti-AAV2 antibodies that are prevalent in the human population may interfere with transduction of AAV2 peptide mutants, and (ii) if not fully ablated, the AAV2 tropism for its genuine receptors will dampen retargeting efficiencies. Here, we aimed to overcome both problems by developing 11 other AAV serotypes as templates for peptide display, AAV1, 3-9, rh.10, po.1 and 12. We first engineered their capsid genes to contain unique restriction sites and then exploited these for insertion of 18 different oligonucleotides encoding peptides which were pre-selected in AAV2 on various cells. The resulting >200 AAV variants were subsequently produced as YFP-expressing vectors and screened in over 80 human or murine cells, including primary hepatocytes or keratinocytes, as well as T-cells and stem/iPS cells. Strikingly, many capsid-peptide combinations vastly outperformed the AAV2 counterparts, especially those based on AAV serotypes 1, 7-9 and rh.10, and those carrying 7mer peptides with an NxxRxxx motif (x = any amino acid). In addition, we found that disruption of residue R585 in AAV2 boosted the potency of several peptide display mutants by up to two orders of magnitude, whereas the same mutation had no effect in the chimeric capsid AAV-DJ, despite a common region surrounding R585. Another puzzling notion was that some serotypes were inactivated by peptide display, tempting us to model all 12 capsids after insertion of a prototype peptide. This revealed that in contrast to our prediction from linear sequence alignments, the exact position of the displayed peptide differed by up to four residues between the 12 capsids. Indeed, once adjusted accordingly, AAV3 and AAV6 were also markedly enhanced upon peptide insertion. Collectively, our data imply an enormous potential of alternative AAV serotypes as scaffolds for peptide display and provide a set of vital guidelines for their further development as gene therapy vectors. To aid in this process, we also present a protocol for AAV spotting and drying in 96/384-well plates which permits their long-term storage and shipping, and thus facilitates their evaluation in other labs.
Effectiveness of first- or second-line bevacizumab (BV) treatment (tx) in elderly patients (pts) with metastatic colorectal cancer (mCRC) in ARIES, an observational cohort study (OCS).
Authors: M. Kozloff, T. S. Bekaii-Saab, J. C. Bendell, A. L. Cohn, H. Hurwitz, N. Roach, H. Tezcan, S. Fish, E. D. Flick, Y. Mun, D. Dalal, A. Grothey
Journal: Journal of Clinical Oncology (2011)

3625 Background: BV (Avastin) + chemotherapy (CT) prolongs survival for 1st- or 2nd-line tx of mCRC. Prior trials for BV included low proportions of elderly pts, for whom there is little information about effectiveness of BV tx, particularly in 2nd-line. Methods: We assessed progression-free (PFS) and overall survival (OS) for pts <70 and ≥70 yrs receiving 1stor 2nd-line BV + CT in the ARIES OCS, using Kaplan-Meier methods. A Cox proportional hazards regression model was also used with multivariate adjustment for baseline factors: sex, race, ECOG PS, albumin, alkaline phosphatase, hypertension, diabetes mellitus, site of and surgical resection of 1o tumor, prior adjuvant tx. Results: As of 9/20/10, 1550 1st-line and 482 2nd-line pts from 248 US sites have enrolled. BL characteristics, OS and PFS for pts < or ≥70 yrs are shown in the table. Survival characteristics were similar among men and women. The incidences of BV-targeted adverse events were similar for pts <70 and ≥70 yrs within 1st and 2nd-line cohorts. Conclusions: This is the first study to describe effects of 2nd-line BV tx for mCRC in a large population of elderly pts. Within the 2nd-line cohort there were no significant differences between median OS and PFS in pts <70 and ≥70 yrs. Within the 1st-line cohort, median OS in pts ≥70 yrs was significantly lower than in pts <70 yrs. First-line Second-line <70 yrs (n=1,126) ≥70 yrs(n=424) <70 yrs(n=336) ≥70 yrs(n=146) Median age (range) 58.0 (18-69) 75.0 (70-92) 58.0 (24-69) 76.0 (70-96) Sex, male, % 56.7 57.3 54.8 61.0 Race, % White Black Other . 79.3 13.6 7.1 . 84.4 10.8 4.8 . 82.1 11.6 6.3 . 83.6 8.9 7.5 Median follow-up (range) 22.2 (1.1-48.2) 18.7 (0.3-48.1) 17.2 (1.2-45.5) 15.8 (0.6-45.5) Survival characteristics (95% CI) Median PFS 10.3 (9.8-10.9) 9.9 (8.9-10.4) 7.9 (7.2-8.3) 7.9 (6.7-9.2) Hazard ratio (HR) Univariate Multivariate* . 1 (reference) 1 . 1.11 (0.99-1.25) 1.11 (0.98-1.25) . 1 1 . 0.94 (0.77-1.15) 0.96 (0.78-1.19) Median OS 25.1 (23.1-26.9) 19.6 (18.1-21.6) 18.7 (17.0-21.4) 17.0 (13.4-21.8) HR Univariate Multivariate* . 1 1 . 1.29 (1.13-1.48) 1.23 (1.06-1.42) . 1 1 . 1.10 (0.88-1.37) 1.17 (0.93-1.48) * Covariates are described in methods section.
A novel NFIA-NFκB feed-forward loop contributes to glioblastoma cell survival
Authors: JunSung Lee, Edlira Hoxha, Hae-Ri Song
Journal: Neuro-Oncology (2016)

The nuclear factor I-A (NFIA) transcription factor promotes glioma growth and inhibits apoptosis in glioblastoma (GBM) cells. Here we report that the NFIA pro-survival effect in GBM is mediated in part via a novel NFIA-nuclear factor-kappaB (NFκB) p65 feed-forward loop.We examined effects of gain- and loss-of-function manipulations of NFIA and NFκB p65 on each other's transcription, cell growth, apoptosis and sensitivity to chemotherapy in patient-derived GBM cells and established GBM cell lines.NFIA enhanced apoptosis evasion by activating NFκB p65 and its downstream anti-apoptotic factors tumor necrosis factor receptor-associated factor 1 (TRAF1) and cellular inhibitor of apoptosis proteins (cIAPs). Induction of NFκB by NFIA was required to protect cells from apoptosis, and inhibition of NFκB effectively reversed the NFIA anti-apoptotic effect. Conversely, NFIA knockdown decreased expression of NFκB and anti-apoptotic genes TRAF1 and cIAPs, and increased baseline apoptosis. NFIA positively regulated NFκB transcription and NFκB protein level. Interestingly, NFκB also activated the NFIA promoter and increased NFIA level, and knockdown of NFIA was sufficient to attenuate the NFκB pro-survival effect, suggesting a reciprocal regulation between NFIA and NFκB in governing GBM cell survival. Supporting this, NFIA and NFκB expression levels were highly correlated in human GBM and patient-derived GBM cells.These data define a previously unknown NFIA-NFκB feed-forward regulation that may contribute to GBM cell survival.
PTU-099 A Large Prospective Audit of Morbidity and Mortality in Patients with Feeding Gastrostomies Placed for Head and Neck Cancer: Abstract PTU-099 Table 1
Authors: K Keshk, S Lewis, N Pitts
Journal: Gut (2016)

<h3>Introduction</h3> Feeding patients undergoing radical treatment for head and neck cancer via a percutaneous endoscopically or radiologically placed/inserted gastrostomy (PEG, RIG) is common practice. PEG/RIG insertion is not entirely safe and complications may occur with long-term usage. We did a prospective audit of PEG/RIG related complications in patients having them placed prior to surgery for head and neck cancer. <h3>Methods</h3> Community dietitians reviewed all patients with a gastrostomy placed prior to surgery or radiotherapy for head and neck cancer. They prospectively recorded morbidity and mortality between 2008–2014. In addition hospital databases and case notes were examined. Recorded morbidity included insertion site infection, leakage, over granulation, haemorrhage and buried bumper. <h3>Results</h3> In the 206 patients there was one death within 30 days from placement, no deaths attributable to gastrostomy placement. One patient underwent a laparotomy for peritonitis. 95 minor complications occurred in 72 (35%) patients or 1:887 days of having a gastrostomy. The commonest complication was peristomal site infection occurring in 55 (27%) patients or 1:1531 days of having a gastrostomy, all of which settled with antibiotics. There was no buried bumpers or tumour implantation at the insertion site reported in the study. There was no procedure related 30 day mortality. <h3>Conclusion</h3> We have shown a low rate of major complications (0.5%) and a relatively high minor complications rate (35%) of PEGs and RIGs in patients with head and neck cancer. Wound infection can be reduced by careful sterile technique, preoperative antibiotics, a 1 cm abdominal incision and avoiding tightness of the crossbar. We conclude that the use of PEG tubes is a safe method of delivering non-oral nutritional support for patients undergoing treatment for head and neck cancer. <h3>References</h3> 1 Gauderer MW, Ponsky JL, Izant RJ, Jr. Gastrostomy without laparotomy: A percutaneous endoscopic technique. <i>J Pediatr Surg</i> 1980;<b>15</b>:872–5. 2 Mansoor H, Masood MA, Yusuf MA. Complications of percutaneous endoscopic gastrostomy Tube insertion in Cancer patients: A retrospective study. <i>J Gastrintest Cane</i> 2014;<b>45</b>:452–459. 3 Burney RE, Bryner BS. Safety and long-term outcomes of percutaneous endoscopic gastrostomy in patients with head and neck cancer. <i>Surg Endosc</i> 2015;<b>29</b>(12):3685–9. 4 Hunter JG, Lauretano L, Shellito PC. Percutaneous endoscopic gastrostomy in head and neck Cancer patients. <i>Ann Surg</i> 1989;<b>210</b>:42–46. <h3>Disclosure of Interest</h3> None Declared
Abstract 2447: Development of Hsp90 paralog specific inhibitors with mitochondrial drug delivery system
Authors: Byoung Heon Kang
Journal: Cancer Research (2015)

Abstract The mitochondrial pool of Hsp90 and its mitochondrial paralog, TRAP1, suppresses cell death and reprograms energy metabolism in cancer cells; therefore, Hsp90 and TRAP1 have been suggested as target proteins for anticancer drug development. Here, we report that the actual target protein in cancer cell mitochondria is TRAP1, not Hsp90, and current Hsp90 inhibitors cannot effectively inactivate TRAP1 due to insufficient accumulation in the mitochondria. To develop mitochondrial TRAP1 inhibitors, we determined the crystal structures of human TRAP1 complexed with Hsp90 inhibitors. The isopropyl amine of the Hsp90 inhibitor PU-H71 was replaced with the mitochondria-targeting moiety triphenylphosphonium to produce SMTIN-P01. SMTIN-P01 showed a different mode of action from the non-targeted PU-H71, as well as much improved cytotoxicity to cancer cells. In addition, we determined the structure of a TRAP1-adenylyl-imidodiphosphate (AMP-PNP) complex. Based on comparative analysis of TRAP1 structures, we propose a molecular mechanism of ATP hydrolysis that is crucial for chaperone function. Citation Format: Byoung Heon Kang. Development of Hsp90 paralog specific inhibitors with mitochondrial drug delivery system. [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 2447. doi:10.1158/1538-7445.AM2015-2447
309. Optimization of Production of AAVrh.10 Viral Vectors
Authors: Hyunmi Lee, Lucy Quach, Fan Fan, Sara Cram, Dolan Sondhi, Ronald Crystal, Stephen Kaminsky
Journal: Molecular Therapy (2016)

With the increasing interest in the use of adeno-associated virus (AAV) vectors to treat human disorders, there is increasing focus on optimizing AAV production to improve yield, purity and potency. The production of AAV vectors can be conceptualized in two stages: upstream cell culture and downstream purification and formulation. As an example of how AAV production can be optimized, we assessed upstream (cell density at transfection, plasmid and transfection reagent concentration, cell culture nutrients and harvest timing) and downstream (virus recovery, purification methods, potency) steps in the production of AAVrh. 10, a nonhuman primate vector, using adherent 293T cells and a 2 plasmid transfection system using polyethylenimine (PEI). Upstream, vector yield, as measured by quantitative PCR in the cell harvest from transfections at each of several different cell densities peaked at 8 × 104 cell/cm2. Total vector yield as a function of time after transfection peaked at day 3 post-transfection with diminishing returns and greater partitioning into the cell supernatant at longer times. Higher vector yield correlated with a greater total quantity of plasmid and PEI, but changing the PEI:plasmid ratio from 1:1 diminished productivity. Fetal bovine serum concentration had minimal effect. Downstream, freeze-thaw methods were not reproducible. Physically lysing the cells was not as effective for AAV recovery as the detergents, triton or tween, which were equally effective, with the lowest effective detergent concentration important to minimize the residual detergent in the final product. While iodixanol gradients are the traditional AAV purification systems, depth filtration followed by tangential flow filtration worked well for small scale production (>90%), but for large scale was less efficient (>70%). Affinity chromatography using AVB (GE Healthcare Life Sciences), provided an effective purification with maximum vector loads of up to 3 × 1012 vector genomes per ml of packed column. Finally, concentration of the final bulk AAV product was critically dependent on a formulation that stabilizes the solubility of the AAV monomer, with 0.01% pluronic acid providing AAV recovery of nearly 80%. Together, the optimized methods for the best yield, purity and potency relied on transfection with equal amounts of plasmid and PEI and a cell density of 8 × 104 cells/cm2 combined with the use of detergent to disrupt and recover AAV and sequential purification by depth filtration, tangential flow filtration and affinity chromatography. This process improved the yield from harvest to final product from 9% to 40% and achieved final purified AAV at 7 × 109 vector genomes/cm2 of adherent cell culture. These systematic refinements to the individual steps of AAV production demonstrate improved yield and facilitates transfer of a method for GMP production for clinical use.