tbuckley/cabot-search huggingface dataset:
PTH-009 Colorectal cancer diagnoses in non-responders to Bowel Cancer Screening Programme and faecal occult blood negative responders
Authors: H Dixon, L Dodd, J James, K Matthewson
Journal: Gut (2010)

<h3>Introduction</h3> All 60–69-year-olds in the North of Tyne (NOT) Bowel cancer screening (BCS) catchment population have been offered faecal occult blood (FOB) testing since February 2008. All FOB positive patients are offered colonoscopy. We have studied the frequency of colorectal cancer (CRC) diagnoses in patients who declined FOB testing and those who accepted but were FOB negative. <h3>Methods</h3> All patients aged 60–69 who were referred to the three colorectal multidisciplinary teams (MDT) serving the NOT BCS catchment population between February 2008 and September 2009 wih CRC were identified. These patients were cross-referenced with the BCS database and their FOB testing status ascertained as positive, negative, no response, opt out or known previous diagnosis of CRC. <h3>Results</h3> Total number of patients with CRC=507 Number of patients with pre-existing colorectal cancer diagnosis excluded from analysis=206 Number of CRC patients potentially diagnosable through BCS=301 Number diagnosed through BCS=80 (26.5%) Number with negative FOB=42 (14.0%) Number of non-responders=133 (44.1%) Number of opt outs=46 (15.2%) Of all patients with CRC who had FOB testing, 65.6% were positive and 34.4% were negative <h3>Conclusion</h3> 26.5% of 60–69-year-olds in the NOT BCS area with CRC are being diagnosed by the programme. FOB testing misses 34.4% of cases. 179 (59.3%) of patients with CRC were non-responders or opt outs, and of these 65.6% might have been diagnosed earlier had they agreed to participate.
A phase II study of lonafarnib (LF) in patients with locally advanced and metastatic breast cancer (MBC): Hoosier Oncology Group BRE07-126.
Authors: B. Leyland-Jones, K. Miller, P. Silverman, C. Shen, C. E. Williams, T. Breen, G. W. Sledge
Journal: Journal of Clinical Oncology (2011)

598 Background: LF is a potent, specific, reversible farnesyl transferase inhibitor (FTI). FT is an enzyme that catalyzes the covalent attachment of a 15-carbon farnesyl group to conserved amino acid residues at the carboxy terminus of certain proteins. FTIs were initially developed to target the ras family of oncoproteins (H-ras, K-ras, and N-ras), G-proteins that exist in a mutated (oncogenic) form in about 30% of human cancers. Oral administration of LF blocked tumor growth of a variety of human xenografts (glioblastoma, lung, pancreatic, prostate, colon, bladder, and CML) in nude mice. LF inhibits growth both ER+ and ER−, as well as HER2+ and HER2− breast cancer. We conducted a nonrandomized, open label study of LF in pts with advanced and MBC. Methods: Primary objective was to determine progression-free survival (PFS) of LF in pts with locally advanced and MBC, unlimited prior treatment, and ECOG PS 0-1. Secondary objectives included the evaluation of clinical benefit response rate (CR+PR+SD > 180 day duration), overall response rate (ORR), and toxicity. Pts 1- 4 were treated with 200mg PO BID daily on days 1-21 of every 21 day cycle until PD or unacceptable toxicity. However, due to early onset of intolerable diarrhea, the study design was modified to a dose escalation model. Pts were pre-treated with Loperamide and started on LF 200mg total daily dose (TDD). If LF was tolerated for 7 days, pts were escalated to 300mg TDD and then 400mg TDD. Results: 29 eligible pts were treated with a median of 2 cycles (range 1-7). Median age was 56 yrs (range 41-76 years), and 100% were female and ECOG PS 1. Median # of prior regimens was 8 (range 0-20). 15/24 (63%), 11/24 (46%) and 3/20 (15%) were ER, PR and HER2 positive, respectively. The most frequent grade 3/4 toxicities comprised diarrhea (23%) and dehydration (8%). Two grade 5 toxicities: PD NOS and encephalopathy. The best response data for the 17 evaluable pts are 5 (29%) stable disease (SD) and 12 (71%) progressive disease (PD). Median PFS was 9 wks. Conclusions: Results based on the first 17 pts does not support improved PFS or ORR compared to historical controls. Further follow up and biomarker correlations are ongoing and will be presented at the meeting.
585. Multiplexed Gene Transfer To Human T-Cells By Combining Sleeping Beauty Transposon System With Methotrexate Selection
Authors: Nataly A. Kcherovsky, Gary W. Liu, Lisa F. Horowitz, Michael C. Jensen, Suzie H. Pun
Journal: Molecular Therapy (2015)

Engineered human T-cells are a promising therapeutic modality for cancer immunotherapy. T-cells expressing chimeric antigen receptors combined with additional genes to enhance T-cell proliferation, survival, or tumor targeting may further improve efficacy but require multiple stable gene transfer events. Methods are therefore needed to increase production efficiency for multiplexed engineered cells. In this work, we demonstrate multiplexed, non-viral gene transfer to a human T-cell line with efficient selection (≈50%) of cells expressing up to three recombinant open reading frames. The efficient introduction of multiple genes to T-cells was achieved using the Sleeping Beauty transposon system delivered in minicircles by nucleofection. We demonstrate rapid selection for engineered cells using methotrexate (MTX) and a mutant human dihydrofolate reductase resistant to methotrexate-induced metabolic inhibition. Preferential amplification of cells expressing multiple transgenes was achieved by two successive rounds of increasing MTX concentration. We are currently applying this non-viral gene transfer method with MTX selection to engineer primary human T-cells with multiple genetic modifications.
496. Elucidating Design Rules Governing Extracellular Vesicle-Mediated Therapeutic Protein Delivery
Authors: Michelle E. Hung, Joshua N. Leonard
Journal: Molecular Therapy (2016)

Extracellular vesicles (EVs) are secreted biological nanoparticles that have great potential as therapeutic delivery vehicles - they are well-tolerated in vivo and naturally capable of transferring RNA and proteins between cells. Our ability to engineer EVs as therapeutic delivery vehicles is limited by an incomplete understanding of how EVs load biomolecular cargo and deliver it to recipient cells. In particular, the biophysical rules governing mRNA and protein delivery by EVs have not been elucidated. Open questions include: Does size limit mRNA loading efficiency into EVs? To what extent is EV mRNA cargo translated in recipient cells? What factors impact the degree to which RNA and protein cargo are delivered to the cytoplasm of recipient cells? To quantitatively investigate the above questions, we leveraged our Targeted and Modular EV Loading (TAMEL) platform, which enables active loading of specific cargo RNA into EVs. TAMEL can enrich cargo mRNA loading into EVs up to 40-fold relative to passive loading. By directly comparing active loading efficiencies between mRNAs of different lengths, we characterized what type of RNAs can be loaded into EVs. While active loading of mRNA-length (> 1.5 kb) cargo molecules was significant, active loading was much more efficient for smaller (~0.5 kb) RNA molecules, providing the first direct evidence for the impact of cargo RNA size on loading into EVs. We next leveraged the TAMEL platform to elucidate the limiting steps in EV-mediated delivery of mRNA and protein to prostate cancer cells, as a therapeutically relevant model system. In this model system, we did not observe translation of EV-delivered mRNA in recipient cells, indicating this is a limiting step in functional delivery of EV cargo. In contrast, we observed robust EV-mediated delivery of dTomato reporter protein, and thus further explored EVs as therapeutic protein delivery vehicles. To probe the efficacy of EV-mediated therapeutic protein delivery, we investigated using EVs to deliver the prodrug converting enzyme cytosine deaminase fused to uracil phosphoribosyl transferase (CD-UPRT), which converts the prodrug 5-FC to the toxic 5-FU. Importantly, CD-UPRT can function without without requiring endosomal escape because both 5-FC and 5-FU are membrane permeable. We also explored strategies for EV-mediated delivery of Cas9 nuclease, which must overcome the EV loading barrier imposed by its NLS sequence as well as escape the endosome in recipient cells. To address the loading challenge, we investigated a strategy for conditional NLS reconstitution to allow enhanced loading of Cas9 into EVs. We then used Cas9 delivery to assess the degree to which EV-delivered proteins can escape the endosomal/lysosomal pathways and traffic to other subcellular locations. Altogether, our investigations elucidated key design rules and central limiting steps that may guide the further development and utilization of EVs as therapeutic biomolecule delivery vehicles.
Leukemia-Reactive Virus-Specific T Cells Generated by T Cell Receptor (TCR) Transfer Preserve Their Dual Specificity upon Repetitive Stimulation of the Endogenous TCR.
Authors: M.M. van Loenen, R.S. Hagedoorn, M. Hoogeboom, M.G.D. Kester, Roelof Willemze, J.H. Frederik Falkenburg, M.H.M. Heemskerk
Journal: Blood (2006)
Regulational effects of breast cancer stromal cells and normal breast stromal cells on MCF-7 mammosphere formation
Authors: F. Zhang, Y. Xu, C. Song, Y. Ma, F. Nan, H. Wang, S. Lin, M. Huang, Y. Zhang
Journal: Journal of Clinical Oncology (2009)

1056 Background: It is well known that microenvironment plays an important role in tumor progression so we investigated the regulatory effects of breast cancer stromal cells (BCSCs) and normal breast stromal cells (NBSCs) as microenvironment on MCF-7 mammosphere formation. Methods: MCF-7 cells were cultured in suspension to generate mammospheres. The proportion of CD44+CD24- cells was assessed by flow cytometry and the expression of Wint1, notch1, β-catenin, CXCR4, SOX2, and ALDH3A1 was detected by real-time PCR. The stromal cells were purified and identified by immumohistochemistry. BCSCs or NBSCs and MCF-7 cells were co-cultured via Transwell system, the volumes and numbers of mammospheres and the mammosphere-forming efficiency (MFE) were calculated and the expression of Wnt1, β-catenin, Notch1 were detected. Results: The proportion of CD44+CD24- cells in mammospheres and MCF-7 cells was 10.4% and 2.1% (p &lt; 0.05), respectively. Real-time PCR analysis suggested that Wint1, notch1, β-catenin, CXCR4, SOX2, and ALDH3A1 genes in the mammosphere cells were with higher levels than MCF-7 cells by about 2.25, 2.45, 1.72, 4.68, 4.25, 5.38 fold, respectively (p &lt; 0.01). The stromal cells purified were identified as fibroblasts by α-SMA,Vimentin and fibroblast special protein antibody via immumohistochemistry. The time of mammosphere's formation was earlier, the volumes of mammospheres were bigger, and the MFE was higher than control group. The expressions of Wnt1 in co-culture group were significantly upregulated 1.27, 3.18 folds than control group, respectively, while the β-catenin was 1.22, 1.75 folds; Notch1 was 1.31, 2.09 folds; and CXCR4 was 1.73, 2.77 folds, respectively. Conclusions: Mammosphere cells contained higher propotion of breast cancer stem cells and expressed higher levels of cancer stem cell related genes. BCSCs can promote the mammosphere-forming efficiency and upregulate the expression of cancer stem cell related genes. No significant financial relationships to disclose.
Total Genomic Loss Detected by High-Density Single Nucleotide Polymorphism Array Is Predictive of Azacitidine Response in Very Poor IPSS-Revised MDS or AML Patients
Authors: Thomas Cluzeau, Chimène Moreilhon, Nicolas Mounier, Jean-Michel Karsenti, Lionel Mannone, Faezeh Legrand, Valentine Richez, Lauris Gastaud, Antoine Thyss, Georges Garnier, Daniel Re, Nathalie Montagne, Jean Gutnecht, Patrick Auberger, Jill-Patrice Cassuto, Sophie Raynaud
Journal: Blood (2012)
Abstract 2110: Modulating NRAS mRNA translation by nucleic acid clamp-mediated stabilization of the 5′- UTR G-quadruplex
Authors: Taisen Hao, Tracy A. Brooks
Journal: Cancer Research (2015)

Abstract NRAS, a proto-oncogene belonging to the RAS oncogene superfamily, is a intracellular signal cascade mediator, with intrinsic GTPase activity, functioning to initiate the MAPK cell signaling pathway. Among the RAS oncogene superfamily that is mutated in approximately 30% of all cancers, the NRAS mutation accounts for about 15% of RAS related human malignancies, particularly myeloid leukemias and cutaneous melanomas. Recently, a stable G-quadruplex (G4) was previously identified in the 5′-untranslated region (UTR) of NRAS mRNA. This G4 functioned as a translational silencer, downregulating NRAS protein and leading to the hypothesis that a small nucleic acid clamp could be optimized to facilitate the formation of specifically this G4, decrease NRAS expression, and facilitate cell death in melanoma cells. In the present study, a set of clamps were designed and examined for their ability to facilitate the formation of a G4 in the 5′-UTR sequence of NRAS. These clamps are comprised of ribonucleotides complementary to the RNA 5′- and 3′-regions flanking the G4-forming sequence held together by a small linker element. Based on binding studies consisting of electromobility shift assays and circular dichroism, an optimal flank length and linker length was chosen for cellular studies. Effects on cellular viability and NRAS expression are being examined in leukemia and melanoma cells, and RNA ChIP-Seq is being used to localize the intracellular activity of the clamps to the 5′-UTR. These nucleic acid clamps offer a high degree of selectivity for G4s over linear RNA facilitated by the small linker region; moreover, the approach of complementary nucleotides, versus small molecule-mediated G4 stabilization, allows for specificity for just one G4 out of all of the intracellular structures. This approach not only provides a solid base for developing the clamp into a non-small molecule cancer therapy, but also opens a new window for modulating oncogenic gene expression at the transcriptional or translational levels. The current study, in particular, offers a novel approach to anti-NRAS-mediated therapies for the treatment of myeloid leukemias and cutaneous melanomas. Citation Format: Taisen Hao, Tracy A. Brooks. Modulating NRAS mRNA translation by nucleic acid clamp-mediated stabilization of the 5′- UTR G-quadruplex. [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 2110. doi:10.1158/1538-7445.AM2015-2110
Dissecting the Molecular Mechanisms of Aneuploidy in Acute Myeloid Leukemia By Next Generation Sequencing
Authors: Giorgia Simonetti, Antonella Padella, Anna Ferrari, Viviana Guadagnuolo, Elisa Zago, Francesca Griggio, Marianna Garonzi, Simona Bernardi, Cristina Papayannidis, Stefania Paolini, Francesca Volpato, Emanuela Ottaviani, Margherita Bodini, Laura Riva, Serena Formica, Annalisa Astolfi, Carmen Baldazzi, Elisa Dan, Barbara Sinigaglia, Ilaria Iacobucci, Nicoletta Testoni, Daniel Remondini, Pier Giuseppe Pelicci, Raffaele Calogero, Alberto Ferrarini, Massimo Delledonne, Giovanni Martinelli
Journal: Blood (2014)
531. Intravenous Delivery of Toca 511 Gene Therapy in Combination with 5-Fluorocytosine for Intratumoral Production of 5-Fluorouracil in a Colon Cancer Metastasis Model
Authors: Kader Yagiz, Maria E. Rodriguez-Aguirre, Fernando L. Espinoza, Bryan Martin, Tiffany T. Huang, Carlos Ibanez, Derek Ostertag, Noriyuki Kasahara, Harry E. Gruber, Douglas J. Jolly, Joan M. Robbins
Journal: Molecular Therapy (2015)

Despite advances in screening, colorectal cancer (CRC) remains the fourth most commonly diagnosed cancer and the second leading cause of cancer death for both men and women in the US. Approximately one half of patients with CRC develop liver metastases (mCRC). The standard treatment for mCRC is 5-fluorouracil (5-FU) based combination chemotherapy. 5-FU combination chemotherapy has extended the median survival of these patients from 6 to >20 months. We are pursuing a unique investigational approach to treat cancer via in situ production of 5-FU. Toca 511 (vocimagene amiretrorepvec), a retroviral replicating vector (RRV), selectively replicates and spreads in malignant cells and encodes an optimized yeast cytosine deaminase (CD) gene. Within infected cells, the CD enzyme is expressed and converts 5-FC (flucytosine, an orally available anti-fungal drug) to the anti-cancer drug 5-FU. Both a direct cytotoxic effect and an extended immunotherapeutic effect have been reported using this approach. Toca 511, in conjunction with subsequent oral extended-release 5-fluorocytosine (Toca FC), is currently under investigation in patients with recurrent high grade glioma. In these studies, Toca 511 is delivered either intratumorally (NCT01156584), by injection into the surgical resection bed (NCT01470794), or intravenously (NCT01985256) in subjects scheduled for subsequent resection. We tested the suitability of the intravenous approach for the treatment of mCRC in a mouse syngeneic liver metastasis model. CT-26-luciferase colon carcinoma cells were delivered via intrasplenic injection producing multiple tumor foci within the liver. Intravenous delivery of RRV resulted in expression of the vector encoded transgene in tumor foci but not in adjacent normal liver tissue. Intravenous delivery of Toca 511 followed by courses of 5-FC resulted in shrinkage or elimination of tumor foci and improved survival in this model of mCRC. The data is supportive of future clinical trials of intravenous Toca 511 followed by cycles of Toca FC in metastatic CRC.
Gsα deficiency in the dorsomedial hypothalamus underlies obesity associated with Gsα mutations
Authors: Min Chen, Yogendra B. Shrestha, Brandon Podyma, Zhenzhong Cui, Benedetta Naglieri, Hui Sun, Thuy Ho, Eric A. Wilson, Yong-Qi Li, Oksana Gavrilova, Lee S. Weinstein
Journal: Journal of Clinical Investigation (2016)

Gsα, encoded by Gnas, mediates hormone and neurotransmitter receptor–stimulated cAMP generation. Heterozygous Gsα-inactivating mutations lead to obesity in Albright hereditary osteodystrophy (AHO) patients, but only when the mutations occur on the maternal allele. This parent-of-origin effect is due to Gsα imprinting in the CNS, although the relevant CNS regions are unknown. We have now shown that mice with a Gnas gene deletion disrupting Gsα expression on the maternal allele, but not the paternal allele, in the dorsomedial nucleus of the hypothalamus (DMH) developed obesity and reduced energy expenditure without hyperphagia. Although maternal Gnas deletion impaired activation of brown adipose tissue (BAT) in mice, their responses to cold environment remained intact. Similar findings were observed in mice with DMH-specific deficiency of melanocortin MC4R receptors, which are known to activate Gsα. Our results show that Gsα imprinting in the DMH underlies the parent-of-origin metabolic phenotype that results from Gsα mutations and that DMH MC4R/Gsα signaling is important for regulation of energy expenditure and BAT activation, but not the metabolic response to cold.
Abstract SY01-04: Chloroquine derivatives as anticancer agents
Authors: Ravi K. Amaravadi
Journal: Cancer Research (2015)

Abstract Mouse models of cancer have demonstrated that therapy-induced autophagy in advanced cancer can promote tumor cell survival and resistance to therapy. Chloroquine derivatives impair autophagy by impairing lysosomal function. Preclinical studies in many, but not all, cancer models have shown that combining chloroquine derivatives with other anticancer agents produces autophagic vesicle accumulation followed by enhanced cancer cell death. Based on these results, clinical trials involving the hydroxychloroquine (HCQ) have been launched for patients with refractory malignancies. The results of the first 6 HCQ clinical trials (5 in humans, 1 in dogs) represent the first deliberate attempt to modulate autophagy therapeutically in patients. These studies indicate that autophagy can be modulated therapeutically with HCQ in patients. While the majority of patients did not experience unexpected toxicity, specific HCQ- chemotherapy combinations did produce enhanced myelosuppression, indicating treatment with HCQ and other chloroquine derivatives should continue to only be considered in the context of clinical trials. Encouraging antitumor activity of HCQ based combinations was observed in patients with melanoma, colon cancer, renal cell carcinoma, and in pet dogs, but randomized studies will be needed to determine if the activity can be attributed to the addition of HCQ. Preliminary results of currently enrolling or more recently completed clinical trials involving hydroxychloroquine will be highlighted. A serum-based approach based on secretome analysis of melanoma cells grown in three dimensional culture has identified potential quantitative biomarkers of low and high tumor cell autophagy. This approach could potentially lead to predictive and pharmacodynamic assays of response to autophagy modulation. To improve the potency of chloroquine derivatives against cancer we have previously reported a dimeric chloroquine Lys05, which is being evaluated for clinical development. Based on analysis of 70 new derivatives of Lys05, compounds which localize to the lysosome, inhibit autophagy and exhibit nanomolar cytotoxicity profiles have been identified. Finally we will discuss ongoing efforts to develop a more detailed understanding of the mechanism of action chloroquine as an anticancer agent. Citation Format: Ravi K. Amaravadi. Chloroquine derivatives as anticancer agents. [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 SY01-04. doi:10.1158/1538-7445.AM2015-SY01-04