tbuckley/cabot-search huggingface dataset:
676. An Engineered CAR T Cell Platform for Allogeneic Combination Immunotherapy
Authors: Philippe Duchateau, Julien Valton, Alexandre Juillerat, Laurent Poirot
Journal: Molecular Therapy (2016)

The adoptive transfer of CAR T cell represents a highly promising strategy to fight against multiple cancers. The clinical outcome of such therapies is intimately linked to the ability of effector cells to engraft, proliferate and specifically kill tumor cells within patients. When allogeneic CAR T cell infusion is considered, host versus graft and graft versus host reactions must be avoided to prevent rejection of adoptively transferred cells, host tissue damages and to elicit significant antitumoral outcome. This work proposes to address these three requirements through the development of multidrug resistant TCR-deficient CAR T cells. We demonstrate in vitro and in an in vivo xenograft mice model, that these engineered T cells displayed efficient antitumor activity and proliferated in the presence of single or muliple nucleotide analogues, currently used in clinic as preconditioning lymphodepleting regimens and antineoplastic agents. The absence of TCR at their cell surface along with their nucleotide analogues-resistance properties could prevent their alloreactivity and enable them to resist to lymphodepleting regimens that may be required to avoid their ablation via HvG reaction. By providing a basic frame work to develop a universal T cell compatible with allogeneic adoptive transfer, this work is laying the foundation stone of the large scale utilization of CAR T cell immunotherapies.
In Vitro and In Vivo Mesenchymal Stem Cells Neurotrophin Production but Limited Neuronal Differentiation.
Authors: Patrizia Bossolasco, Davide Soligo, Yvan Torrente, Federica Pisati, Mirella Meregalli, Lidia Cova, Cinzia Calzarossa, Marzia Belicchi, Manuela Gavina, Chiara Marchesi, Vincenzo Silani, Nereo Bresolin, Giorgio Lambertenghi Deliliers, Elio Polli
Journal: Blood (2005)
Anti-Tumor Cytotoxicity of Blood γδ T Cells Expanded from Leukemia Patients Against Autologous Leukemia Cells—Enhancement of the Anti-Tumor Cytotoxicity by Type I IFN.
Authors: Anri Saito, Miwako Narita, Norihiro Watanabe, Ayumi Yokoyama, Asuka Sekiguchi, Nozomi Tochiki, Louie Alldawi, Jun Takizawa, Shigeo Hashimoto, Tatsuo Furukawa, Ken Toba, Yoshifusa Aizawa, Masuhiro Takahashi
Journal: Blood (2005)
Mice That Lack RhoA In Their Platelets Have Normal Actin Dynamics, but Have Macrothrombocytopenia
Authors: Aae Suzuki, Lurong Lian, Liang Zhao, Sang H. Min, Yuhuan Wang, Mortimer Poncz, John K Choi, Christopher L. Carpenter, Charles S. Abrams
Journal: Blood (2010)
Abstract 2433: A novel MDM2 inhibitor suppresses breast cancer growth and metastasis
Authors: Jiang-Jiang Qin, Wei Wang, Sukesh Voruganti, Ruiwen Zhang
Journal: Cancer Research (2015)

Abstract The MDM2 oncogene is amplified and/or overexpressed in many human cancers, including breast cancer, and it has been suggested as a molecular target for treating human cancers. MDM2 is a major negative regulator of tumor suppressor p53. Most MDM2 inhibitors under development are targeting the MDM2-p53 binding, and have little or no effects on cancers without functional p53, such as advanced breast cancer. Therefore, new strategies to target MDM2 are desirable. The present study was designed to develop a new class of MDM2 inhibitors that exert anti-breast cancer activity, regardless of p53 status in the cancers. The selective MDM2 inhibitors were discovered by a computational structure-based screening, yielding a lead compound, termed JapA. Fluorescence polarization (FP)-based binding assay and cellular thermal shift assay were performed to determine the binding between MDM2 and JapA. Its cytotoxicity was initially evaluated in breast cancer cell lines (MCF-7, MCF-7/p53−/−, MDA-MB-231, and MDA-MB-468) and normal human breast cells (MCF-10A). Its anti-breast cancer activities were further assessed in in vitro and in vivo MCF-7 and MDA-MB-231 models. The molecular mechanisms for its targeting effects on MDM2 were elucidated in various cell models in vitro and in vivo. We further found that JapA selectively inhibited breast cancer cell growth, decreased cell proliferation, and induced G2/M phase arrest and apoptosis in breast cancer cells through an MDM2-dependent mechanism, regardless of p53 status. In MCF-7 and MDA-MB-231 xenograft models, JapA inhibited MDM2 and suppressed tumor growth and lung metastasis in a dose-dependent manner, without host toxicity. Furthermore, JapA directly bound to MDM2 protein and reduced MDM2 levels in cancer cells in vitro and in vivo by promoting MDM2 protein degradation and inhibiting MDM2 transcription, which is distinct from the existing MDM2 inhibitors. In conclusion, JapA represents a new class of MDM2 inhibitor that exerts its anticancer activity through directly down-regulating MDM2, and might be developed as a novel cancer therapeutic agent. (Supported by the NIH R01 CA112029, R01 CA121211, and R01 CA186662) Citation Format: Jiang-Jiang Qin, Wei Wang, Sukesh Voruganti, Ruiwen Zhang. A novel MDM2 inhibitor suppresses breast cancer growth and metastasis. [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 2433. doi:10.1158/1538-7445.AM2015-2433
761. Targeted Killing of HIV Infected Cells Using CCR5-Disrupted Anti-HIV-CAR T Cells
Authors: Guillermo S. Romano Ibarra, Malika Hale, Karen Sommer, Iram Khan, Jaya Sahni, Christopher Peterson, Hans-Peter Kiem, Jordan Jarjour, Alexander Astrakhan, Andrew M. Scharenberg, David J. Rawlings, Thor A. Wagner
Journal: Molecular Therapy (2016)

A cure for HIV remains an important treatment goal for 30 million HIV-infected individuals worldwide. Long term control of HIV following a single treatment will require a mechanism to eradicate the latent reservoir of HIV infected cells that are capable of reactivating. A previous phase II clinical trial using an anti-HIV chimeric antigen receptor (αHIV-CAR) targeting the CD4-binding site on HIV envelope was partially effective. To optimize this approach we have developed a series of CARs based on the scFV of broadly neutralizing HIV antibodies targeting four different structural regions of the HIV envelope: the V1/V2 loop, the V3 loop, the CD4 binding site, and the membrane-proximal external region. αHIV-CAR T cells targeting different epitopes were compared and were able to kill > 80% of HIV-infected cells grown in the presence of ART. However, a limitation of αHIV-CAR T cells is that the αHIV-CAR also serves as a receptor for HIV and allows HIV infection of αHIV-CAR T cells. Therefore we disrupted the major co-receptor required for HIV cell entry, CCR5, using a megaTAL nuclease, as a means of protecting the CAR-expressing cells from HIV infection. We used two strategies for achieving these dual modifications in primary human T cells. For both strategies, the CCR5 megaTAL nuclease was delivered by mRNA electroporation, which has previously been shown to induce a high rate of bi-allelic NHEJ-mediated gene disruption. αHIV-CAR expression cassettes were delivered into the host genome via lentiviral vectors (LV), or were targeted to the megaTAL nuclease cleavage site in CCR5 using an adeno-associated virus (AAV) that included CCR5 homology arms. Both strategies resulted in stable expression of the CAR construct and specific activation of CAR+ T cells in the presence of an HIV+ cell line. In the presence of actively replicating virus, CCR5-megaTAL treated CAR+ T cells out-performed CAR+ T cells generated by LV delivery alone as measured by reduction in HIV capsid protein. To enable testing in non-human primates (NHP), we re-optimized the cell-editing protocol with NHP lymphocytes. Primary T cells from pigtail macaques were successfully transfected with CCR5 megaTAL mRNA and achieved a CCR5 disruption rate of 50%. Cells were subsequently transduced with αHIV-CAR LV resulting in ~70% of manipulated cells with αHIV-CAR. Expansion of the cells in vitro resulted in 60-fold expansion over 8 days. CAR+ NHP cells specifically killed HIV infected human cells demonstrating that NHP-derived αHIV-CAR+ T cells retained killing function. In conclusion, it is feasible to construct αHIV-CAR+ T cells that are protected from HIV infection in human and NHP cells, and warrants further study in vivo.
Abstract 510: A novel role for Olig2 in the regulation of glioma invasion
Authors: Shiv K. Singh, Nicole Giannonatti, Robert Kupp, Costanza Lo Cascio, Shwetal Mehta
Journal: Cancer Research (2015)

Abstract Glioblastoma multiforme (GBM) are highly infiltrative, aggressive and lethal primary brain tumors in adults that are resistant to conventional treatments. A major clinical obstacle in the treatment of GBMs is the ability of glioma cells to invade the surrounding brain parenchyma, preventing their complete surgical resection. Several studies have shown that invasive glioma stem cells (GSCs) extend beyond the surgically resected regions. These invasive cells are the cause for tumor recurrence and poor prognosis in GBM patients. The molecular and cellular mechanism involved in the regulation of the switch from proliferation to invasion/migration in GBM is poorly understood. We investigated the role of Olig2, a Central Nervous System (CNS) specific transcription factor, in promoting the switch from proliferating to invading glioma cell. Genetically relevant murine glioma model as well as patient-derived glioma stem cells (GSCs) were utilized to identify the effect of Olig2 on glioma invasion. Functionally, Olig2-mediated invasion was assessed by 3D-transwell invasion assays, organotypic slice cultures as well as PDX tumors in immunocompromised mice. In addition, murine and patient-derived GSCs were utilized to identify direct genetic targets of Olig2 involved in glioma invasion by ChIP-seq and RNA-seq. Our data demonstrate that Olig2-expressing murine glioma stem/progenitor-like cells are highly invasive as compared to Olig2 knockout cells. We further corroborated the requirement of Olig2 for glioma invasion in patient-derived GSCs by utilizing shRNA-mediated knockdown of Olig2. Olig2 depletion resulted in reduced glioma cell invasion in at least four independent human GSCs. Recent studies have shown that activation of transcription factors such as Zeb1 and Twist1 occurs in invasive high grade gliomas. We show that knockdown of Olig2 in murine as well as human GSCs leads to a significant decrease in the expression of invasion genes such as Zeb1, Vimentin, Twist1, and Stat3 as compared to control cells. In conclusion, our studies highlight a novel role for the Olig2 signalling network in promoting the switch from proliferation to invasion in GBM, and provide a plausible target for inhibiting glioma invasion. Citation Format: Shiv K. Singh, Nicole Giannonatti, Robert Kupp, Costanza Lo Cascio, Shwetal Mehta. A novel role for Olig2 in the regulation of glioma invasion. [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 510. doi:10.1158/1538-7445.AM2015-510
Abstract 440: Nicotinamide phosphoribosyltransferase (NAMPT): a new cytokine in tumor progression
Authors: Ambra Grolla, Simone Torretta, Angela Amoruso, Giuseppe Orsomando, Armando Genazzani
Journal: Cancer Research (2015)

Abstract An important role has been shown for Nicotinamide phosphoribosyltransferase (NAMPT; also known as PBEF or visfatin) in cancer, inflammation and metabolic diseases. Alongside maintaining intracellular NAD levels constant by contributing to the salvage pathway of NAD production, it is now clear that NAMPT is also a pleiotropic player in cell signalling: (i) as a key regulator of NAD consuming-enzymes, including sirtuins and PARPs; and (ii) as an extracellular cytokine that binds to a yet unknown extracellular receptor. It has been shown that a regulated positive secretory process exists, although the exact mechanism of release is at present under investigation. A mounting amount of cell types have been shown to release eNAMPT, including adipocytes, hepatocytes, cardiomyocytes and activated immune cells. In my work, I verified that eNAMPT is released by different types of tumour cells, specifically by melanoma, neuroblastoma and glioblastoma, mesotelioma, and prostate cancer cells in vitro. In particular, we analysed the mechanisms of its release and we found both classical and non-classical pathway involvement. eNAMPT release could be modulated by tumoural microenvironment conditions and once released by tumour cells, in our hands, eNAMPT has paracrine and autocrine effects: it activates MAPK and Akt pathways in melanoma cells and induces M1 polarization in human monocytes. Mainly, I demonstrated for the first time that eNAMPT levels in plasma of tumour-bearing mice are increased because part of this circulating protein derives directly from tumour itself. This parallels observations that eNAMPT is increased in cancer patients, and indeed this increase can be, at least, in part attributed to the tumour itself. All these data suggested that the tumour microenvironment might be enriched of this cytokine-like protein, which could have a role in tumour progression and severity. Citation Format: Ambra Grolla, Simone Torretta, Angela Amoruso, Giuseppe Orsomando, Armando Genazzani. Nicotinamide phosphoribosyltransferase (NAMPT): a new cytokine in tumor progression. [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 440. doi:10.1158/1538-7445.AM2015-440
Abstract 2104: Evidence for E2F/EWS-FLI1 oncoprotein synergism using systems biology
Authors: Raphaela Schwentner, Theodore Papamarkou, Maximilian Kauer, Vassilios Stathopoulos, Fan Yang, Sven Bilke, Paul S. Meltzer, Mark Girolami, Heinrich Kovar
Journal: Cancer Research (2015)

Abstract Cell cycle deregulation and enhanced proliferation of cells is one of the hallmarks of oncogenesis. In Ewing Sarcoma, a highly aggressive pediatric cancer, the chimeric transcription factor EWS-FLI1 deregulates cell cycle by targeting several cell cycle regulators including the E2F family of transcription factors. ChIP-seq studies showed a significant overlap of EWS-FLI1 and E2F3/4 binding in Ewing sarcoma cells. We show that EWS-FLI1 is able to directly activate E2F3, followed by the combinatorial binding of EWS-FLI1 and E2F3 on their target genes synergistically activating their transcription. Furthermore we propose a model where EWS-FLI1 directly exchanges repressive E2F4/p130 by E2F3/pRB thereby driving cells into enhanced cell proliferation. However, so far we were unable to experimentally demonstrate a physical interaction of EWS-FLI1 with any E2F or pocket protein. As an alternative approach to the study of functional synergy between EWS-FLI1 and E2F3, we used time resolved RNA and protein expression data as a basis for mathematical modeling of EWS-FLI1 dependent E2F target gene regulation. By Bayesian model selection, we were able to postulate the formation of a complex between EWS-FLI1 and E2F3 as the most likely explanation for the observed ETS/E2F synergy. This study therefore provides an example of how computational systems approaches can complement experimental data in the discovery of disease mechanisms. This study was supported in part by the Austrian Science Fund (FWF), [grant 22328-B09], and by the 7th framework program of the European Commission, [grant 259348] (‘ASSET’). Citation Format: Raphaela Schwentner, Theodore Papamarkou, Maximilian Kauer, Vassilios Stathopoulos, Fan Yang, Sven Bilke, Paul S. Meltzer, Mark Girolami, Heinrich Kovar. Evidence for E2F/EWS-FLI1 oncoprotein synergism using systems biology. [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 2104. doi:10.1158/1538-7445.AM2015-2104
Abstract P176: Resumption of Cardiopulmonary Resuscitation after Defibrillation Induces Recurrence of Ventricular Fibrillation
Authors: Jocelyn Berdowski, Liesbeth van Zyll de Jong, Rudolph W Koster
Journal: Circulation (2008)

Introduction- Resuscitation guidelines 2005 advise to resume cardiopulmonary resuscitation (CPR) immediately after defibrillation. We hypothesized that this promotes recurrence of ventricular fibrillation (VF). Methods- This was a prospective per-patient randomized controlled trial. We randomized Automated External Defibrillators (AEDs) used by first responders to either a) perform post-shock analysis and prompt rescuers to a pulse check - guidelines 2000 - or b) to resume CPR immediately after defibrillation - guidelines 2005. Between June 2006 and April 2008, we collected continuous ECG and impedance signal of all patients with a non-traumatic out-of-hospital cardiac arrest to whom a study AED was applied. We annotated and calculated time intervals between the first effective shock, moment of CPR resumption, moment of refibrillation, and subsequent defibrillation shock. Intervals are expressed as median and inter quartile range. Results- We collected 120 ECGs of patients with a shockable rhythm: 61 were randomized to guidelines 2000 and 59 patients to guidelines 2005. In respectively 54% (33/61) and 56% (33/59) of the patients VF recurred after the first effective shock. Rescuers resumed CPR after 32 (24–45) and 8 (7–9) seconds (P&lt;0.01), VF recurred at 35 (21–57) and 13 (8–25) seconds after the first effective shock (figure 1 ; P=0.02), the time interval between resumption of CPR and refibrillation was 4 (−8–20) and 6 (1–22) seconds (P=0.59), and VF sustained for 98 (63–204) and 150 (98–261) seconds (P=0.15). Conclusion- CPR induces recurrent VF. Immediate resumption of CPR after effective defibrillation is associated with early and longer lasting recurrence of VF.
Abstract 4432: A-803467, a tetrodotoxin-resistant sodium channel blocker, modulates ABCG2-mediated MDR in vitro and in vivo
Authors: Nagaraju Anreddy, Atish Patel, Yun-Kai Zhang, Yi-Jun Wang, Suneet Shukla, Rishil J. Kathawala, Priyank Kumar, Pranav Gupta, Suresh V. Ambudkar, John ND Wurpel, Zhe-Sheng Chen
Journal: Cancer Research (2015)

Abstract ABCG2 is a member of the ABC transporter superfamily, which has been implicated in the development of multidrug resistance (MDR) in cancer. Its diverse range of substrates includes many antineoplastic agents such as topotecan, doxorubicin and mitoxantrone. ABCG2 expression has been significantly increased in some solid tumors and hematologic malignancies, which is correlated to poorer clinical outcomes. In addition, ABCG2 expression is a distinguishing feature of cancer stem cells, whereby this membrane transporter impacts resistance to the chemotherapeutic drugs. To enhance the chemosensitivity of cancer cells, attention has been focused on MDR modulators. In this study, we investigated the ability of a tetrodotoxin-resistant sodium channel blocker, A-803467 to reverse ABCG2-mediated MDR. We found that A-803467 at non-toxic concentration could significantly increase the cellular sensitivity to ABCG2 substrates in drug-resistant cells overexpressing either wild-type or mutant ABCG2. Mechanistic studies indicated that A-803467 (7.5 μM) significantly increased the intracellular accumulation of mitoxantrone by inhibiting the transport activity of ABCG2, without altering its expression level. In addition, A-803467 stimulated the ATPase activity of ABCG2 in a concentration-dependent manner, indicating that A-803467 might be a substrate of ABCG2. Binding interactions of A-803467 were found to be in transmembrane region of homology modeled human ABCG2. Interactions of A-803467 with ABCG2 were relatively stronger when compared to the interactions of topotecan with ABCG2. Furthermore, A-803467 (30 mg/kg) with topotecan (3 mg/kg) significantly decreased the growth rate and tumor size of ABCG2 overexpressing tumors in a xenograft nude mouse model. Our findings indicate that A-803467 has the potential to be used in combination with ABCG2 chemotherapeutic substrates to improve the response in drug resistant cancers. Citation Format: Nagaraju Anreddy, Atish Patel, Yun-Kai Zhang, Yi-Jun Wang, Suneet Shukla, Rishil J. Kathawala, Priyank Kumar, Pranav Gupta, Suresh V. Ambudkar, John ND Wurpel, Zhe-Sheng Chen. A-803467, a tetrodotoxin-resistant sodium channel blocker, modulates ABCG2-mediated MDR in vitro and in vivo. [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 4432. doi:10.1158/1538-7445.AM2015-4432
Abstract 5089: Desmoplastic stroma affects growth and invasion of progressively mutated human pancreatic cancer cells <i>in vitro</i>
Authors: Ruchi Malik, Tiffany Luong, Seda Karamil, Peter Lelkes, Edna Cukierman
Journal: Cancer Research (2015)

Abstract Desmoplastic (desmo) stroma is a prognostic marker for pancreatic cancer, yet mechanisms behind its tumorigenic effects remain poorly understood. Tumor stroma, known to act permissive to naïve cells, may exacerbate tumor progression in mutated cells, whereas normal stroma acts restrictive to even aggressively mutated cells. The mechanistic details of these responses remain unclear. Previous studies suggested that a 3D microenvironment (i.e. Matrigel) is necessary for the manifestation of the metastatic phenotype in vitro in aggressively K-Ras mutated pancreatic cancer cells. Here we describe a new 3D in vitro model to study how human pancreatic stromal extracellular matrices (ECMs) interact with pancreatic epithelial cells, how these interactions activate specific integrins and downstream signaling pathways such as PI3K and ERK1/2, and how these signaling pathways may synergize with intrinsic mutational cues to enhance progression of pancreatic cancer. Our model is comprised of physiologically relevant desmo and normal 3D cell-derived matrices as well as polyacrylamide gel-based bioengineered ECMs. The influences of cell-derived and bioengineered ECMs were tested using a series of isogenic pancreatic ductal epithelial cells (PDEC) displaying different aggressive behavior. In preliminary studies, we established that physiological desmo ECMs are tumor permissive (i.e. induce growth and/or invasion) for all PDEC cells (especially kRas mutated cells) when compared with matched restrictive/normal ECMs. In addition, K-Ras cells cultured on stromal activated (i.e. desmo) myofibroblast-derived 3D ECMs resulted in enhanced ERK1/2 activity, similar to what had been previously observed with Matrigel based system. In order to discern distinct cellular and signal responses mediated by cell-derived ECM in the various PDEC cells, we independently modulated microenvironmental cues such as stiffness, topography, and biochemical composition in the bioengineered ECMs. Our initial data suggests that the various PDEC cells differentially respond to alterations in microenvironmental cues. We anticipate that our studies will lead to better understanding of ECM-PDEC interactions in more detail, and help in identification of novel biomarker for early detection or prevention of pancreatic cancer. Citation Format: Ruchi Malik, Tiffany Luong, Seda Karamil, Peter Lelkes, Edna Cukierman. Desmoplastic stroma affects growth and invasion of progressively mutated human pancreatic cancer cells in vitro. [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 5089. doi:10.1158/1538-7445.AM2015-5089