tbuckley/cabot-search huggingface dataset:
234. Efficient Generation of Stable Genetically Modified Human iPSC-Derived Macrophages for Innovative Gene and Cell Therapeutic Strategies
Authors: Mania Ackermann, Uta Müller-Kuller, Sylvia Merkert, Ulrich Martin, Manuel Grez, Axel Schambach, Thomas Moritz, Nico Lachmann
Journal: Molecular Therapy (2016)

Hematopoietic differentiation of human induced pluripotent stem cells (iPSC) holds great promise for disease modeling, drug testing, and in particular for the development of novel gene and cell therapeutic strategies. In the past, interest has been directed primarily at the generation of reconstituting hematopoietic stem cells, a cell type as of yet problematic to obtain from hPSCs. Recently however, also long-lived mature myeloid cells such as tissue resident macrophages have been described and further characterized. Therefore, transplantation of macrophages may serve as an innovative treatment approach in several diseases such as pulmonary alveolar proteinosis (PAP) and adenosine deaminase deficiency (ADA). In this line, we have established an efficient, embryoid body-based human iPSC-differentiation protocol employing IL-3 in combination with M-SCF to continuously (up to 6 months) generate large numbers (5-10×106/week/6 well plate) of >95% pure monocyte/macrophages (iPSC-MΦ) via an intermediate "myeloid cell forming complex". The iPSC-MΦ revealed typical morphology, surface phenotype and functionality comparable to their in vivo derived counterparts. In order to evaluate the suitability of this protocol also for gene therapy, we transduced human iPSC with 3rd generation SIN lentiviral vectors employing a minimal and safety optimized ubiquitous chromatin opening element (CBX3-UCOE) to prevent differentiation-associated epigenetic silencing. Here, GFP reporter gene expression driven by the elongation factor 1α short (EFS) promoter resulted in >90% GFP+ iPSC-MΦ after myeloid differentiation. In contrast, the EFS promoter without the CBX3-UCOE was subject to massive epigenetic silencing. Moreover, also a myeloid-biased myeloid-related protein 8 (Mrp8) promoter was efficiently protected from silencing, whilst tissue specificity was not affected by the CBX3-UCOE. Of note, the protective activity of the CBX3-UCOE was associated with decreased levels of repressive histone marks (H3K27me3, H3K9me3) and increased levels of active marks (phosphorylated polymerase). Alternatively, we also targeted an expression cassette to the AAVS1 locus employing zinc finger nuclease mediated homologous recombination. Selected clones showed targeted and robust CMV early enhancer/chicken β actin (CAG) promoter-driven fluorochrome expression in the pluripotent status as well as in terminally differentiated iPSC-MΦ. In summary, we here describe silencing resistance lentiviral vectors and genome editing strategies as well as a differentiation protocol allowing for the efficient and long-term generation of stable genetically modified iPSC-MΦ, thus paving the way for novel gene and cell based therapies.
Phase II Study of the JAK2 Inhibitor, INCB018424, In Patients with Refractory Leukemias Including Post-Myeloproliferative Disorder (MPD) Acute Myeloid Leukemia (sAML)
Authors: Alireza Eghtedar, Srdan Verstovsek, Jorge E. Cortes, Zeev Estrov, Jan A. Burger, Carol Bivins, Stefan Faderl, Alessandra Ferrajoli, Gautam Borthakur, Solly George, Robert Newton, Hagop Kantarjian, Farhad Ravandi
Journal: Blood (2010)
Abstract 1547: The clinical implication of MMP-1 and TIMP-1 on prognosis in patients with non-small cell lung cancer
Authors: Ho Jung An, Jin-Hyoung Kang, Yoon-Jin Lee, Soon Uk Hong, Kyo Young Lee, Jeong-Oh Kim
Journal: Cancer Research (2015)

Abstract Introduction: The interaction between tumor cells and tumor micro-environment (TME) plays important role in tumor initiation, progression, treatment resistance, and prognosis in solid tumors including non-small cell lung cancer (NSCLC). The aim of this study is to evaluate the prognostic role of TME-related molecules in NSCLC patients who received surgical resection. To find out a possibility of a potential serum biomarker, protein expression in tissue from surgical specimen and pre-operative serum were investigated together. Methods: For 85 NSCLC patients who underwent surgical resection from August 2006 to September 2010 and had pre-operative serum samplings, we screened the serum levels of 60 TME-related molecules with angiogenesis array kit for 8 patients who had recurrences and 4 patients who did not. For matrix metalloproteinase (MMP)-1 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 in which significant difference was observed between recurrence and recurrence-free groups, protein expressions were investigated using ELISA in serum and immunohistochemistry in tumor tissue. Results: High serum MMP-1 was related to carcinoembryonic antigen (CEA) level≥5 at diagnosis (P = 0.034), higher frequency of the presence of vascular invasion (P = 0.041). High expression of MMP-1 in tumor cells (intensity score≥2) was related to tumor differentiation (P = 0.057), whereas high MMP-1 expression in stroma was significantly associated with lymph node metastasis and TNM stage (P = 0.021 and P = 0.022, respectively). In the case of TIMP-1, expression in tumor cells was higher for adenocarcinoma but higher in stromal cells for squamous cell carcinoma (P = 0.046). There was no correlation between the tissue and serum levels of MMP-1 and TIMP-1. High serum MMP-1 showed a trend for short overall survival (P = 0.080). When serum MMP-1 was combined with CEA level or presence of vascular invasion, the prognostic implication for overall survival was statistically significant. Neither tumor nor stromal positivity for MMP-1 and TIMP-1 was associated with survival outcome; however, a tumor/stroma TIMP-1 intensity ratio≥1 was strongly associated with early recurrence in multivariate analysis (HR = 13.1, P = 0.008). Conclusion: In surgically resected NSCLC patients, serum MMP-1 showed a possibility as a prognostic biomarker. In addition, the tumor/stroma TIMP-1 ratio (≥1) in tissue may be a useful to predict tumor recurrence. Citation Format: Ho Jung An, Jin-Hyoung Kang, Yoon-Jin Lee, Soon Uk Hong, Kyo Young Lee, Jeong-Oh Kim. The clinical implication of MMP-1 and TIMP-1 on prognosis in patients with non-small cell lung cancer. [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 1547. doi:10.1158/1538-7445.AM2015-1547
Abstract 3594: P-glycoprotein attenuates Src activation and DNA repair activity via increased C-terminal Src kinase-binding protein, a negative regulator of Src, in multidrug-resistant cells
Authors: Li-Fang Lin, Ming-Hsi Wu, Tsann-Long Su, Te-Chang Lee
Journal: Cancer Research (2015)

Abstract Multidrug resistance (MDR) remains a significant obstacle to the success of cancer chemotherapy. MDR is often associated with increased expression of ATP-binding cassette transporter family members, which extrude anticancer drugs out of cells. The MDR1 gene product, P-glycoprotein (P-gp) is one of the most well-known ABC transporters and expels a broad range of anticancer drugs, such as vinblastine, vincristine, doxorubicin and paclitaxel. Overexpression of P-gp in tumor tissues is thus a prognostic indicator associated with poor response to chemotherapy and poor clinical outcome. Our previous study has shown that P-gp overexpression attenuates the repair activity of DNA interstrand crosslinks (ICLs) and hence contributes the susceptibility of MDR cells to DNA ICL agents [1]. In the present study, we explored the molecular mechanisms underlying which P-gp interferes with Src-activated DNA repair activity. We first confirmed that ectopic expression of P-gp attenuates DNA ICL repair activity and becomes more susceptible to DNA ICL agents than vector control cells. Our present results showed that the C-terminal Src kinase-binding protein (Cbp), a negative regulator of Src, is significantly enhanced in MDR cells. Cbp silencing by siRNA technique apparently enhances cisplatin-induced Src activation and DNA repair activity, and hence increases the resistance of MDR cells to ICL agents. By aid of immuneprecipitation and immunofluorescence staining techniques, we demonstrated that P-gp not only interacts with Cbp and Src but also enhances the formation of inhibitory Csk-Cbp complex that reduces phosphorylation of Src activation residue Tyr416 and increases phosphorylation of Src negative regulatory Tyr527 residue. Taken together, the current results indicate that P-gp overexpression in MDR cells attenuates DNA repair activity by mediating Cbp dependent mechanism to interfere with Src activation. These results implicate that increasing the expression of Cbp is potentially a therapeutic strategy to increase the susceptibility of cancer cells to chemotherapeutic agents. 1. Lee PC, Lee HJ, Kakadiya R, Sanjiv K, Su TL, Lee TC: Multidrug-resistant cells overexpressing P-glycoprotein are susceptible to DNA crosslinking agents due to attenuated Src/nuclear EGFR cascade-activated DNA repair activity. Oncogene 2013, 32:1144-1154. Citation Format: Li-Fang Lin, Ming-Hsi Wu, Tsann-Long Su, Te-Chang Lee. P-glycoprotein attenuates Src activation and DNA repair activity via increased C-terminal Src kinase-binding protein, a negative regulator of Src, in multidrug-resistant 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 3594. doi:10.1158/1538-7445.AM2015-3594
The Neuronal Protein GAP-43 Associates with Microtubules and Lipid Rafts in Platelets and Megakaryocytes and Is Involved in Platelet Genesis
Authors: Antonija Jurak Begonja, Lilly Nguyen, Joseph E. Italiano, Herve Falet, Karina F. Meiri, John H. Hartwig
Journal: Blood (2011)
411. CD5 CAR for the Treatment of T-Cell Malignancies
Authors: Maksim Mamonkin, Malcolm K. Brenner
Journal: Molecular Therapy (2015)

Chimeric antigen receptors (CARs) have emerged as a powerful therapeutic tool by redirecting patients' own T cells to tackle hematologic malignancies – in particular, those of B cell origin. However, T cell malignancies remain a more challenging task for CAR T cells owing to the shared expression of targetable tumor-associated antigens between normal and cancerous T cells, potentially invoking fratricide of CAR T cells and/or profound immunodeficiency. We designed a novel CAR targeting CD5, a common marker expressed in ~80% of T-cell leukemia/lymphoma blasts as well as in normal T cells. Upon transduction with CD5 CAR, more than 95% of T cells expressed the CAR while producing limited and transient fratricide. Fratricide of CD5 CAR T cells was mainly directed against naïve phenotype and central memory cells, eliminating more than 50% of cells in these subsets and resulting in preferential expansion of effector memory and effector T cells. CD5 CAR T cells acquired resistance to self-killing by 3 days post-transduction and could then expand normally, up to 100,000-fold. Expansion of CD5 CAR T cells coincided with downregulation of CD5 protein from the cell surface while preserving CAR expression. CD5 CAR T cells mounted an effective anti-tumor response, selectively killing five CD5-positive T-ALL and T lymphoma cell lines in 4-hour chromium release assays while displaying no activity against CD5-negative cell lines. Upon longer-term coculture, CD5 CAR T cells eliminated >95% of leukemia cells from 3 T-ALL lines within 48h and 100% by day 7. We also observed the ability of CD5 CAR T cells to eliminate leukemia cells in sequential killing assays where we recurrently replenished fresh target cells for at least 4 iterations. Lack of functional exhaustion in sequential killing assays supports the fitness of CD5 CAR T cells for eradicating large numbers of tumor cells in vivo. CD5 CAR T cells dramatically suppressed systemic in vivo disease progression in 3 different xenograft mouse models, doubling median survival. Importantly, CD5 CAR T cells demonstrated significant cytokine production and cytotoxicity against primary T-ALL blasts (n=6), highlighting the therapeutic potential of CD5 CAR for patients with T cell malignancies. Overall, we demonstrated for the first time that CD5 CAR redirects T cells to eliminate CD5-positive malignant T cells in vitro and in vivo while producing only limited fratricide of the normal T cell population.
92. Evaluation of Producer Cell Line Platform for Production of Oversized AAV-FVIII Vectors
Authors: Bindu Nambiar, Cathleen Sookdeo, Patricia Berthelette, Robert Jackson, Brenda Burnham, Shelley Nass, David Souza, Catherine R. O'Riordan, Karen Vincent, Seng H. Cheng, Donna Armentano, Sirkka Kyostio-Moore
Journal: Molecular Therapy (2016)

Recombinant adeno-associated virus (rAAV) vectors are being evaluated as gene delivery vehicles in several clinical trials. The 4.7 kb wild-type (WT) size genome of AAV presents a challenge for incorporating larger transgenes with incomplete vector genome (vg) packaging being a frequent outcome. To test the feasibility of producing oversized rAAV production using the producer cell line (PCL) method, we generated slightly oversized rAAV vectors (harboring 5.1 or 5.4 kb sized vgs) containing a liver-restricted promoter (mTTR) and a codon-optimized cDNA encoding human B-domain deleted FVIII (FVIIIco). Genomes were packaged into the AAVrh8R serotype vector using the PCL process and compared to matched vectors generated via the triple transfection (TXN) method. Vectors were then characterized for production yields, integrity of packaged genomes and homogeneity. The data showed that the PCL platform was able to produce oversized AAV vectors at levels that were 10- to 100-fold higher than the TXN process with yields greater than 100,000 vg/cell. The PCLs were stable with consistent production maintained up to 60 passages. Southern and dot blot analyses of the packaged genomes demonstrated encapsidation of genomes larger than 4.7 kb in the PCL generated vector while the majority of genomes packaged via the TXN method were 4.7 kb in size. Furthermore, the PCL process generated more vector DNA-containing particles and less packaging of non-vector DNA. Testing the PCL generated vectors in the hemophilia A knock-out (KO) mouse model showed a 2-fold higher plasma FVIII activity (Coatest) and vg copies in the livers than obtained with the vectors made by the TXN process. In summary, the PCL production process generated higher yields of oversized rAAV/FVIIIco vectors as well as higher quality vectors than the TXN method. Hence, the PCL platform may be used for producing greater quality oversized rAAV vectors at levels that can meet the needs for clinical studies.
A High Incidence of Patients with MTHFR (methylenetetrahydrofolate reductase) Gene Mutations Noted in An Appalachian Population Seen for Thrombophilia at the West Virginia University Outpatient Hematology Clinic, 2006–2008
Authors: Mudussara Asad Khan, Vijaya k Gadiyaram, Thomas Hogan, Ramin Altaha, Edward Crowell, Peter Perrotta, Gerald Hobbs
Journal: Blood (2008)
Abstract 5193: Ataxia telangiectasia mutated relates to epithelial-mesenchymal transition in colorectal cancer
Authors: Hidena Takahashi, Masashi Tsuruta, Hirotoshi Hasegawa, Koji Okabayashi, Ryo Seishima, Shimpei Matsui, Toru Yamada, Takayuki Kondo, Takehiro Shimada, Mutsuhito Matsuda, Masashi Yahagi, Yusuke Yoshikawa, Yusuke Asada, Kiyoaki Sugiura, Yoshiyuki Suzuki, Yuki Tajima, Junpei Nakadai, Yuko Kitagawa
Journal: Cancer Research (2015)

Abstract Background: The Ataxia telangiectasia mutated (ATM) is a serine/threonine protein kinase that is recruited and activated by DNA double-strand breaks. Recently, ATM was reported to correlate with breast cancer metastasis by regulating phosphorylated Snail which was known as a marker of epithelial-mesenchymal transition (EMT), however, it remains unclear in colorectal cancer (CRC). In this study we attempted to clarify the impact of ATM on the mechanism of EMT in CRC. Methods: Potential ATM levels were evaluated in human CRC cells (HT29 and SW480) by qPCR and western blot analysis. Cells were induced EMT by treated with TGF-b, were confirmed by the change of EMT markers (i.e. N-cadherin, E-cadherin, Snail and twist). Next, ATM levels were down-regulated by siRNA and the impact on EMT were evaluated by EMT markers. Invasive potential was evaluated by scratch assay. Results: HT29 showed significantly higher levels of ATM comparing to SW480 (p = 0.042). N-cadherin (p = 0.065), Snail (p = 0.034) and twist (p = 0.052) were highly observed in HT-29, while similar level of E-cadherin in SW480 (p = 0.692). Scratch assay revealed that approximately 4 times high migration was observed in HT29. TGF-b induced EMT such as elevation of N-cadherin and Snail (E-cadherin was not affected), leading to promotion of cell migration in SW480. ATM was successfully 48.4% down-regulated (p = 0.001) in SW480, which resulted in reducing cell migration (p = 0.033). Conclusions: ATM might be a critical regulator of EMT in colorectal cancer invasion. Citation Format: Hidena Takahashi, Masashi Tsuruta, Hirotoshi Hasegawa, Koji Okabayashi, Ryo Seishima, Shimpei Matsui, Toru Yamada, Takayuki Kondo, Takehiro Shimada, Mutsuhito Matsuda, Masashi Yahagi, Yusuke Yoshikawa, Yusuke Asada, Kiyoaki Sugiura, Yoshiyuki Suzuki, Yuki Tajima, Junpei Nakadai, Yuko Kitagawa. Ataxia telangiectasia mutated relates to epithelial-mesenchymal transition in colorectal cancer. [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 5193. doi:10.1158/1538-7445.AM2015-5193
Abstract A2-01: SHP-1 is a negative regulator of epithelial-mesenchymal transition in hepatocellular carcinoma
Authors: Li-Ching Fan, Chung-Wai Shiau, Wei-Tien Tai, Man-Hsin Hung, Pei-Yi Chu, Hang Lin, Hui-Chuan Yu, Kuen-Feng Chen
Journal: Cancer Research (2015)

Abstract Epithelial-to-mesenchymal transition (EMT) is well known to involve in tumor invasion and metastasis. SH2 domain-containing phosphatase 1 (SHP-1) functions as a potent tumor suppressor and also acts as a negative regulator of p-STAT3 (Tyr705) oncogenic signaling. However, little is known about the molecular mechanism(s) through which SHP-1 regulates EMT during hepatocellular carcinoma (HCC) progression. Here, we firstly reported that endogenous SHP-1 protein levels were significantly down-regulated in cells with mesenchymal characteristics and negatively correlated with p-STAT3 (Tyr705) and vimentin but positively correlated with E-cadherin. SHP-1 overexpression abolished TGF-beta-induced p-STAT3 (Tyr705) and EMT, as well inhibited migration and invasion but further rescued by STAT3 overexpression. Constitutively overexpressed SHP-1 tyrosine phosphatase activity by D61A mutated SHP-1 markedly reduced TGF-beta-induced p-STAT3 (Tyr705) and EMT features, but not altered by C453S catalytic dead mutant SHP-1. Consequently, SHP-1 acted as a powerful suppressor in preventing EMT by exerting its activity directly targeted p-STAT3 (Tyr705). Most notably, we discovered a novel SHP-1 agonist SC-43 better than sorafenib to exert more potent anti-EMT effects in vitro as well anti-metastatic growth in vivo. In conclusion, SHP-1 is a potent suppressor of HCC EMT and metastasis, thus, highlighting that SHP-1 may serve as a potential therapeutic target for HCC EMT and metastasis. Note: This abstract was not presented at the conference. Citation Format: Li-Ching Fan, Chung-Wai Shiau, Wei-Tien Tai, Man-Hsin Hung, Pei-Yi Chu, Hang Lin, Hui-Chuan Yu, Kuen-Feng Chen. SHP-1 is a negative regulator of epithelial-mesenchymal transition in hepatocellular carcinoma. [abstract]. In: Proceedings of the AACR Special Conference on Translation of the Cancer Genome; Feb 7-9, 2015; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(22 Suppl 1):Abstract nr A2-01.
Ex-Vivo Reduction of Allograft T Cell Dose Does Not Prevent Acute Graft-vs-Host Disease after Reduced-Intensity Hematopoietic Stem Cell Transplantation.
Authors: Nancy M. Hardy, Frances Hakim, Seth Steinberg, Michael Krumlauf, Rebecca Babb, Jeanne Odom, Daniel Fowler, Ronald Gress, Michael R. Bishop
Journal: Blood (2006)
Selective Blockade of the Ubiquitous Checkpoint Receptor CD47 Is Enabled by Dual-Targeting Bispecific Antibodies
Authors: Elie Dheilly, Valéry Moine, Lucile Broyer, Susana Salgado-Pires, Zoë Johnson, Anne Papaioannou, Laura Cons, Sébastien Calloud, Stefano Majocchi, Robert Nelson, François Rousseau, Walter Ferlin, Marie Kosco-Vilbois, Nicolas Fischer, Krzysztof Masternak
Journal: Molecular Therapy (2017)

CD47 is a ubiquitously expressed immune checkpoint receptor that is often upregulated in cancer. CD47 interacts with its counter-receptor SIRPα on macrophages and other myeloid cells to inhibit cancer cell phagocytosis and drive immune evasion. To overcome tolerability and “antigen sink” issues arising from widespread CD47 expression, we generated dual-targeting bispecific antibodies that selectively block the CD47-SIRPα interaction on malignant cells expressing a specific tumor-associated antigen; e.g., CD19 or mesothelin. These bispecific κλ bodies are fully human, native IgG1 molecules, combining tumor targeting and selective CD47 blockade with immune activating mechanisms mediated by the Fc portion of the antibody. CD47-neutralizing κλ bodies efficiently kill cancer cells in vitro and in vivo but interact only weakly with healthy cells expressing physiological levels of CD47. Accordingly, a κλ body administered to non-human primates showed a typical IgG pharmacokinetic profile and was well tolerated. Importantly, κλ bodies preserve their tumoricidal capabilities in the presence of a CD47 antigen sink. Thus, dual-targeting κλ bodies allow for efficacious yet safe targeting of CD47 in cancer. Such a bispecific design could be applied to limit the extent of neutralization of other ubiquitously expressed therapeutic targets. CD47 is a ubiquitously expressed immune checkpoint receptor that is often upregulated in cancer. CD47 interacts with its counter-receptor SIRPα on macrophages and other myeloid cells to inhibit cancer cell phagocytosis and drive immune evasion. To overcome tolerability and “antigen sink” issues arising from widespread CD47 expression, we generated dual-targeting bispecific antibodies that selectively block the CD47-SIRPα interaction on malignant cells expressing a specific tumor-associated antigen; e.g., CD19 or mesothelin. These bispecific κλ bodies are fully human, native IgG1 molecules, combining tumor targeting and selective CD47 blockade with immune activating mechanisms mediated by the Fc portion of the antibody. CD47-neutralizing κλ bodies efficiently kill cancer cells in vitro and in vivo but interact only weakly with healthy cells expressing physiological levels of CD47. Accordingly, a κλ body administered to non-human primates showed a typical IgG pharmacokinetic profile and was well tolerated. Importantly, κλ bodies preserve their tumoricidal capabilities in the presence of a CD47 antigen sink. Thus, dual-targeting κλ bodies allow for efficacious yet safe targeting of CD47 in cancer. Such a bispecific design could be applied to limit the extent of neutralization of other ubiquitously expressed therapeutic targets.