tbuckley/cabot-search huggingface dataset:
Abstract LB-060: A phosphoproteomic comparison of BRAF(V600E) and MKK1/2 inhibition in melanoma
Authors: Scott Stuart, Natalie Ahn, Stephane Houel, William Old
Journal: Cancer Research (2015)

Abstract Inhibitors of oncogenic B-RAF(V600E) and MKK1/2 have yielded remarkable responses in B-RAF(V600E)-positive melanoma patients. However, the efficacy of these inhibitors is limited by the inevitable onset of resistance. Despite the fact that these inhibitors target the same pathway, combination treatment with B-RAF(V600E) and MKK1/2 inhibitors has been shown to improve both response rates and progression-free survival in B-RAF(V600E) melanoma patients. To provide insight into the molecular nature of the combinatorial response, we used quantitative mass spectrometry to characterize the inhibitor-dependent phosphoproteome of human melanoma cells treated with the B-RAF(V600E) inhibitor PLX4032 (vemurafenib) or the MKK1/2 inhibitor AZD6244 (selumetinib). In three replicate experiments, we quantified changes at a total of 23,986 phosphosites on 4,722 proteins. This included 1,317 phosphosites that reproducibly decreased in response to at least one inhibitor. Phosphosites that responded to both inhibitors grouped into networks that included the nuclear pore complex, growth factor signaling, and transcriptional regulators. While the majority of phosphosites were responsive to both inhibitors, we identified 16 sites that decreased only in response to PLX4032, suggesting rare instances where oncogenic B-RAF signaling occurs in an MKK1/2-independent manner. Only two phosphosites were identified that appeared to be uniquely responsive to AZD6244. When cells were treated with the combination of AZD6244 and PLX4032 at subsaturating concentrations (30 nM), responses at nearly all phosphosites were additive. We conclude that AZD6244 does not substantially widen the range of phosphosites inhibited by PLX4032 and that the benefit of the drug combination is best explained by their additive effects on suppressing ERK1/2 signaling. Comparison of our results to another recent ERK1/2 phosphoproteomics study revealed a surprising degree of variability in the sensitivity of phosphosites to MKK1/2 inhibitors in human cell lines, revealing unexpected cell specificity in the molecular responses to pathway activation. Citation Format: Scott Stuart, Natalie Ahn, Stephane Houel, William Old. A phosphoproteomic comparison of BRAF(V600E) and MKK1/2 inhibition in melanoma. [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 LB-060. doi:10.1158/1538-7445.AM2015-LB-060
Low Morbidity but Prolonged Viral Shedding Characterize Parainfluenza Virus 3 Infections in Allogeneic Peripheral Stem Cell Transplant Recipients.
Authors: Zakari Y. Aliyu, Mary-Ann Bordner, David K. Henderson, Elaine M. Sloand, Neal S. Young, Richard Childs, John Bennett, John Barrett
Journal: Blood (2004)
459. Evaluation of Miltenyi ExpAct and TransAct CD3/28 Beads for CAR-T Cell Manufacturing
Authors: Xiuyan Wang, Jinrong Qu, Jolanta Stefanski, Oriana Borquez-Ojeda, Anniesha Hack, Qing He, Teresa Wasielewska, Fang Du, Michel Sadelain, Isabelle Rivière
Journal: Molecular Therapy (2016)

Adoptive transfer of chimeric antigen receptor (CAR) engineered T cells is a promising emerging strategy to treat cancer patients. Large-scale manufacturing of cGMP-grade CAR T cells using patient T cells selected and activated by CTS™ Dynabeads® CD3/CD28 (Dynabeads) followed by transduction with retroviral vectors is being used in the context of many clinical trials by our laboratory and others. Although we have established a robust protocol using Dynabeads, it is important to explore alternative sources to pre-empt supply chain limitations of this critical reagent. To this end, we evaluated T cell activation with either Miltenyi TransAct CD3/28 (TransAct) beads or Miltenyi ExpAct Treg (ExpAct) beads. In small-scale experiments, PBMCs were directly activated with TransAct or ExpAct beads and compared with our standard T cell selection and activation using Dynabeads. Overall, the transduction efficiency and expansion of T cells were comparable upon activation with all three reagents. The TransAct bead-stimulated cells exhibited comparable effector memory (EM)/central memory (CM) phenotype to that of the Dynabeads stimulated cells. In line with the EM/CM phenotype, CAR T cells stimulated with either TransAct or Dynabeads and tranduced with CD19-targeted CAR demonstrated robust and comparable antitumor activity in a systemic NSG/CD19+ NALM6 tumor mouse model. We further tested the efficacy of TransAct beads using positively or negatively selected T cells in a large-scale cGMP grade CAR-T cell manufacturing setting. Both the transduction efficiency and expansion of selected CD3+ cells activated with TransAct beads and Dynabeads were comparable. CD19-targeted CAR T cells activated by either TransAct or Dynabead were subjected to an in vivo stress test by using decreasing amount of CAR-T cells to treat systemic CD19+NALM6 tumors in NSG mice. In this experimental setting, T cells stimulated with TransAct beads demonstrated equivalent if not better anti-tumor activity than T cells stimulated with Dynabeads. In conclusion, our pre-clinical results suggest that TransAct beads support efficient transduction and expansion of CAR T cells. TransAct activated T cells exhibit antitumor activity equivalent to Dynabeads activated T cells in our NSG/CD19+NAML6 stress test. Therefore, Miltenyi TransAct beads can be used as an alternative to Dynabeads to stimulate T cells in clinical trials aiming at evaluating CAR T cell safety and antitumor activity.
Differentiation between the EGFR antibodies necitumumab (Neci), cetuximab (Cetux), and panitumumab (Pan): Glycosylation and IgE reactivity.
Authors: D. Yu, J. Wuertz, A. Taqui, Z. Li, F. E. Fox, J. Qian, T. Liu, M. Li, M. Hsieh, Q. Zhou
Journal: Journal of Clinical Oncology (2011)

e13023 Background: Cetux, Pan and Neci are monoclonal antibodies (mAb) against the human EGFR. Hypersensitivity reactions (HSR) to Cetux have been linked to preexisting anti-α-Gal IgE, suggesting that IgE reactivity to biologics may predict HSR. This study evaluated the levels of α-Gal and N-glycolyl neuraminic acid (NGNA) expressed by Cetux, Pan and Neci, and assessed the reactivity of the three mAb with normal human serum IgE. Methods: To evaluate the levels of α-Gal and NGNA expressed by Cetux, Pan and Neci, the N-linked oligosaccharides were released by PNGase F from the mAb and analyzed by NP-HPLC and MALDI-TOF mass spectrometry. A sensitive electrochemiluminescent assay was developed to detect drug-specific IgE in human serum. Results: Cetux has the highest expression of α-Gal and NGNA. Neci has at least 10 fold less α-Gal or NGNA and Pan has no detectable amount. Sixty lots of serum from individual normal donors from Tennessee were screened for IgE reactivity with mAb either expressing α-Gal and NGNA (Cetux and mab1) or not (mab2). Fifteen serum lots exhibited high IgE reactivity with Cetux and mab1 but minimal reactivity with mab2, and were designated high IgE. The rest of serum lots showed minimal reactivity with the three mAb and were designated low IgE. Fifteen serum lots from each group were used to assess the IgE reactivity to the mAb. Cetux exhibited high reactivity with each lot of the high IgE sera, while Pan and Neci had minimal reactivity. All three mAb did not exhibit any significant reactivity with the low IgE sera. An α-Gal compound, GN334, abolished Cetux-mediated IgE signal completely, indicating that the IgE is specific for α-Gal. GN334 had little or no effect on Pan- and Neci-mediated IgE signal. In comparison, GN334 did not have effect in the low IgE sera across mab tested. Conclusions: High expression of α-Gal and NGNA was detected on Cetux compared to low or no expression by Neci and Pan respectively. IgE reactivity data correlated with the glycosylation expression levels and suggest that Neci and Pan have no or little reactivity against human serum IgE in comparison to Cetux. Further studies are warranted to determine if this leads to a lower incidence of HSR.
Abstract LB-030: Hepatocellular carcinoma-derived exosomes promote motility of immortalized hepatocyte through transfer of oncogenic proteins and RNAs
Authors: Nathalie Wong, Mian He
Journal: Cancer Research (2015)

Abstract Exosomes are increasingly recognized as important mediators of cell-cell communication in cancer progression through the horizontal transfer of RNAs and proteins to neighboring or distant cells. Hepatocellular carcinoma (HCC) is a highly malignant cancer, whose metastasis is largely influenced by the tumor microenvironment. The possible role of exosomes in the interactions between HCC tumor cell and its surrounding hepatic milieu are however largely unknown. In this study, we comprehensively characterized the exosomal RNA and proteome contents derived from three HCC cell lines (HKCI-C3, HKCI-8 and MHCC97L) and an immortalized hepatocyte line (MIHA) using Ion Torrent sequencing and mass spectrometry, respectively. RNA deep sequencing and proteomic analysis revealed exosomes derived from metastatic HCC cell lines carried a large number of pro-tumorigenic RNAs and proteins, such as MET proto-oncogene, S100 family members and the caveolins. Of interest, we found exosomes from motile HCC cell lines could significantly enhance the migratory and invasive abilities of non-motile MIHA cell. We further demonstrated that uptake of these shuttled molecules could trigger PI3K/AKT and MAPK signaling pathways in MIHA with increased secretion of active MMP-2 and MMP-9. Our study showed for the first time that HCC-derived exosomes could mobilize normal hepatocyte, which may have implication in facilitating the protrusive activity of HCC cells through liver parenchyma during the process of metastasis. Citation Format: Nathalie Wong, Mian He. Hepatocellular carcinoma-derived exosomes promote motility of immortalized hepatocyte through transfer of oncogenic proteins and RNAs. [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 LB-030. doi:10.1158/1538-7445.AM2015-LB-030
Abstract 633: Amanitin-based ADCs with an improved therapeutic index
Authors: Torsten Hechler, Christoph Müller, Andreas Pahl, Jan Anderl
Journal: Cancer Research (2015)

Abstract Antitumoral activity of monoclonal antibodies can be dramatically enhanced by conjugation to toxic small molecules. Beside the recent approval of Kadcyla (T-DM1) and Adcetris (SGN-35) more than 30 antibody-drug conjugates (ADC) have entered clinical trials, promising to strengthen the therapeutic capabilities for cancer treatment in the next decade. Surprisingly most ADCs are based on one of few toxic compounds only and on an even smaller number of toxicity mechanisms: Most antibodies are coupled to the microtubuli-targeting auristatins and maytansins. Toxins that operate through such a mechanism could suffer from limited activity in different cancer indications and in cells expressing resistance mechanisms. Accordingly the use of new drugs that function via alternative toxicity mechanisms could enhance the therapeutic potential of ADCs. In the present study we evaluated the antitumoral potency of monoclonal antibodies conjugated to small molecules from the amatoxin family. Amanitin, the most well-known toxin of the amatoxin family, binds to the eukaryotic RNA pol II and thereby inhibits the cellular transcription at very low concentrations. In our experiments, we tested random- and site-specific strategies to covalently conjugate amanitin to antibodies and generated conjugates with low aggregation and high affinity for the target antigen. We compared the cytotoxic activity of stable and cleavable linker ADCs and the stability of such constructs in plasma. Overall we observed high stability in plasma after incubation for four days. Moreover we demonstrated superior efficacy of amanitin-based ADCs in cancer xenograft models compared to tubulin-inhibiting toxins. For site-specific conjugation antibodies were engineered at different locations to incorporate cysteines (Thiomabs). Thiomabs with different positions of the engineered cysteine were compared in vitro and in vivo to identify the best position for amanitin-thiomab-conjugates. Safety profiling of the two best Thiomabs in cynomolgous monkey revealed a substantial in improvement in tolerability and of the therapeutic index by using site-specific engineered amanitin-ADCs. The data support the development of amanitin-based ADCs by using a toxin with a new mode of action and with a favorable therapeutic index. Citation Format: Torsten Hechler, Christoph Müller, Andreas Pahl, Jan Anderl. Amanitin-based ADCs with an improved therapeutic index. [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 633. doi:10.1158/1538-7445.AM2015-633
A High Through-Put Screen Identifies MCP-1 (CCL2) As a Novel Regulator of Iron Homeostasis and a Modifier of Hereditary Hemochromatosis Disease Severity
Authors: Martina U. Muckenthaler, Maja Vujic Spasic, Katarzyna Mleczko-Sanecka, Mingang Zhu, Rainer Pepperkok, Heiko Runz, Ulrich Maus, Joana Gomes, Jorge P Pinto, Matthew Lawless, Andreas E Kulozik, Graf¸ca Porto
Journal: Blood (2011)
TEMPI: A Reversible Syndrome Following Treatment with Bortezomib
Authors: Wilfried A. Schroyens, Casey L. O'Connell, Martha Q Lacy, Eyal C. Attar, Noopur Raje, Bradley G. Somer, Flavia G Rosado, James Hoyer, Donald I. Feinstein, David B. Sykes
Journal: Blood (2012)
637. Treatment of Human B Lymphoma By a Minicircle DNA Vector Expressing the CD3xCD20 Bispecific Antibody in a Xenograft Mouse Model
Authors: Fei Ma, Jing Zhang, Yee Hou, Yu-Jiang Zhong, Xiao-Hu Hou, Cheng-Yi He, Zhi-Ying Chen
Journal: Molecular Therapy (2015)

Breakthroughs have been achieved in cancer immunotherapies recently. Some patients experienced a durable remission or even cure by the treatments activating patients' own T cells. However, these single target technologies have failed to result in any therapeutic effect in a large percent of patients, and evidence suggests that further advances depend on the effective strategy for coping with cancer heterogeneity and dynamics. We hypothesize that minicircles, a class of optimized DNA vectors, are able to cope with these problems; that is to apply minicircles to express a panel of engineered antibodies capable killing all cell lineages in a cancer by inducing antibody-dependent cellular cytotoxicity or retargeting CTLs to become anticancer T cells. To prove this concept, we constructed a minicircle to express a bispecific antibody (BsAB) capable of binding CD3 on T cell and CD20 on B-lymphoma cell simultaneously, and tested if it could function like its BsAb counterpart blinotumomab in treating B-lymphoma. Six to eight week-old female SCID/Beige mice were inoculated with 2×5e5 Raji cells through tail vein injection, and 4 μg of the minicircle was injected into the mouse liver via hydrodynamic technique 24 hours later. Selected mice received 3 intravenous doses of human DC-CIK cells, 5e6 each, every other day starting 2 days after Raji cell inoculation. All the 14 Raji cell-bearing mice receiving no further treatment died in 3 weeks, the 10 mice that received DC-CIDK as well died in 7 weeks, while the10 mice received both MC. CD3xCD20 and DC-CIK survived for additional 5 weeks. Histology examination demonstrated human CD3-positive T cells and CD20-positive B-lymphoma cells in spleens. In another fine-tuning experiment, we imaged the mice receiving the same number of Raji cell expressing luciferase gene when they demonstrated paralyzed rear legs, the typical syndrome indicating the late stage of this disease. We observed strong luciferase signal along the hips and spinal of the mice. Taken together, these data demonstrated that we have successfully established the mouse model of human B-lymphoma, and that minicrcle was able to generate therapeutic level of BsAb in vivo. Further experiments are undergoing to characterize the minicircle-BsAb anti-cancer system. As compared to the virus vectors, minicircle is safer, more convenient to make, easier to target multiple cancer cell lineages simultaneously, our efficacy data support the notion that minicircles have the potential to serve as powerful immunotherapy vectors in fighting human cancer.
Acute Rheumatic Fever with Erythema Marginatum
Authors: Makoto Saito, Shuji Hatakeyama
Journal: New England Journal of Medicine (2016)

A 36-year-old man presented with fever and pain in both shoulders and knees, which had been preceded by a sore throat. His symptoms abated after administration of an NSAID. One week later, painless, nonpruritic, red annular macules appeared on the upper limbs and abdomen.
ITD-Flt3 Enhances Migration of Hematopoietic Cells through Aberrant CXCR4 Signaling Pathways Overlapping but Functionally Distinct From SDF1/CXCR4 Signaling Axis in Normal Hematopoietic Cells.
Authors: Seiji Fukuda, Satomi Mori, Mariko Abe, Seiji Yamaguchi, Louis M. Pelus
Journal: Blood (2009)
INI1 and mib-1 expression in childhood chordomas.
Authors: H. Yin, K. M. O. Dorris, L. M. Wagner, M. H. Collins, A. Towbin, R. Nagarajan
Journal: Journal of Clinical Oncology (2011)

9556 Background: Children and adolescents comprise a small but important subset of patients with chordoma who may have distinct pathologic features and clinical behaviors. Previous reports suggest pediatric patients with chordoma are more likely than adults to have clival tumors and atypical/anaplastic histology, and the latter has been associated with a greater likelihood of recurrence, metastases, and death. A small series has also suggested that aggressive chordomas may be characterized by loss of INI1 expression. Methods: To further investigate factors associated with poor outcome, we performed a retrospective CCHMC institutional review of all cases of chordoma. Pathology was reviewed, as were demographic data and details of therapy. Immunohistochemistry for proliferative index (mib-1) and INI1 expression was performed on available samples. Results: Eight cases (5 female) of pediatric chordoma were identified with a mean age of 9.2 yrs (0.8 -19yrs). Clival location was most common (n=5), and no patient had metastatic disease at diagnosis. Three of 8 had a gross total resection, 4 had a subtotal resection and 1 was unknown. Five received radiation, 2 did not and 1 was unknown. Two received chemotherapy (1 initially and 1 at relapse). Anaplastic histology was noted in three patients (2 with rhabdoid features), who all died from disease progression at a mean of 1.4 yrs. The remaining patients had classic histology, and 4 of 5 remain alive and disease-free with mean follow-up of 6.4 yrs. Immunostaining was performed on 6 available cases, and all 3 tested patients with anaplastic histology had mib-1staining of at least 40% and absent INI1 protein expression. These findings contrast with surviving patients, who had intact INI1 expression and mib-1 staining of 1% or less. Patients with absent INI1 expression and anaplastic histology had a mean age of 3.4 yrs (0.8-8.4 yrs) at diagnosis, compared to 12.6 yrs (6.4-13.0 yrs) for patients with classic histology. Conclusions: Chordoma is an uncommon pediatric tumor that may have aggressive behavior and early age at presentation associated with loss of INI1 protein expression. Immunohistochemistry to assess for mib-1 and INI1 may help identify higher risk patients who could be considered for intensive multiagent therapy.
Abstract 1830: Untargeted metabolomics reveals smokers’ characteristic profiles
Authors: Ping-Ching Hsu, Renny S. Lan, Theodore Brasky, Catalin Marian, Amrita K. Cheema, Habtom W. Ressom, Christopher A. Loffredo, Wallace Pickworth, Peter G. Shields
Journal: Cancer Research (2015)

Abstract Introduction. Smoking-related biomarkers for lung cancer and other disease risks are needed to enhance early detection strategies and to provide a science base for FDA decision making for the regulation of tobacco products. Methods. An untargeted metabolomics approach was used in a cross-sectional study of smokers (n = 112) where subjects smoked 2 cigarettes one hour apart, and blood was collected immediately before and after each cigarette. The global metabolomic profiles from the two blood samples were determined using UPLC-Q-TOF/MS (n = 448 assays). The study design allows for the direct detection of smoking-related effects since each subject served as his/her own control. Results were analyzed by using a false discovery rate q value &amp;lt;0.05. Results. Pair-wise comparisons of pre- and post-cigarettes showed 24 features (metabolites) significantly changed due to smoking. The analysis identified important changes in menthol-glucuronide (p = 6.99E-10) and endogenously occurring metabolites including glutamate and glycerophospholipids. Menthol-glucuronide was positively correlated with glutamate, and negatively correlated with oleamide, lysoPC(16:1(9Z)), and an unknown metabolite with m/z of 517.31+ (p &amp;lt; 0.01), implying a menthol specific effect. Some shared and different metabolites were found by race and gender. Pathway analysis revealed smoking-related effects on carbohydrate metabolism, lipid metabolism, and small molecule biochemistry. Network analysis revealed eleven molecules involved in cancer, notably inhibition of cAMP. Conclusions. This first time identification of a menthol metabolite in blood samples from smokers serves as proof-of-principle for using metabolomics to identify new tobacco-exposure biomarkers. This study also identified several endogenous non-smoke metabolites not previously related to smoking carcinogenesis that may serve as biomarkers of exposure and/or effect (e.g., glutamate), and provides new biological insights for the effects of tobacco smoke on cellular functions. Citation Format: Ping-Ching Hsu, Renny S. Lan, Theodore Brasky, Catalin Marian, Amrita K. Cheema, Habtom W. Ressom, Christopher A. Loffredo, Wallace Pickworth, Peter G. Shields. Untargeted metabolomics reveals smokers’ characteristic profiles. [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 1830. doi:10.1158/1538-7445.AM2015-1830
Characterization of Early Natural Killer Cell Reconstitution Following Autologous Transplantation in Multiple Myeloma.
Authors: Don Benson, Courtney E Bakan, Lana Alghothani, Craig C. Hofmeister, Yvonne A Efebera, Megan K Smith, Carli N Greenfield, Pascale Andre, Patrick Squiban, Francois Romagne, Michael A. Caligiuri, Steven M Devine
Journal: Blood (2009)