tbuckley/cabot-search huggingface dataset:
691. A Light-Inducible CRISPR/Cas9 System for Control of Endogenous Gene Activation
Authors: Lauren P. Toth, Charles A. Gersbach
Journal: Molecular Therapy (2015)

Optogenetic systems enable precise spatiotemporal control of gene expression and cellular behavior that is easily reversible and repeatable. Optogenetic technologies that facilitate dynamic control of endogenous genes would facilitate the systematic analysis of the relationship of gene expression patterns and cell differentiation or disease treatment. We engineered a light-activated CRISPR/Cas9 effector (LACE) system that induces transcription of endogenous genes in the presence of gene-specific guide RNAs (gRNAs) and blue light. This system is based on the plant proteins CRY2 and CIB1 from Arabidopsis thaliana that heterodimerize in response to blue light. The full-length CRY2 was fused to the N-terminus of the transcriptional activator VP64 (CRY2FL-VP64), and an N-terminal fragment of CIB1 was fused to the N- and C-terminus of the catalytically inactive form of Cas9 (CIBN-dCas9-CIBN). When these fusion proteins are expressed with a gRNA, CIBN-dCas9-CIBN localizes to the gRNA target. In the presence of blue light, CRY2FL binds to CIBN, which translocates CRY2FL-VP64 to the gene target and activates transcription. Unlike other optogenetic systems, the LACE system can be targeted to new endogenous loci by solely manipulating the specificity of the gRNA without having to re-engineer the light-inducible proteins. Light-dependent activation of the IL1RN, HBG, or ASCL1 genes was achieved by delivery of the LACE system and four gene-specific gRNAs per promoter region. Illuminated cells in which IL1RN or HBG1/2 was targeted demonstrated significantly greater mRNA levels in the light compared to the dark (p<0.0001 and 0.005, respectively), as well as equivalent activation levels to dCas9-VP64 (p=0.17 and 0.35, respectively). Significant light-dependent activation was also observed when the ASCL1 locus was targeted with the LACE system (p<0.0001). However, in this case mRNA levels were not activated to the same extent as cells that received dCas9-VP64 and the same four ASCL1-targeting gRNAs. In all instances, transfected cells incubated in the dark maintained levels of the targeted gene that did not significantly differ from mock-transfected cells. Importantly, the fusion of CIBN to both N- and C-termini of dCas9 yielded 10- to 100-fold greater gene activation than when CIBN was fused to only one terminus. Endogenous gene expression could be controlled in a reversible and repeatable fashion by modulating the duration of blue light exposure. Spatial patterning of gene expression was also achieved using an eGFP reporter. Cells illuminated though a photomask containing slits of varying width resulted in a corresponding pattern of eGFP-expressing cells. This versatile LACE system is a valuable tool for studying dynamic gene expression patterns in synthetic biology, basic science, and tissue engineering.
400. Improved Transduction of Canine X-Linked Muscular Dystrophy With rAAV9-Microdystrophin By Using MSCs Pretreatment
Authors: Hiromi Hayashita-Kinoh, Hironori Okada, Yuko Nitahara-Kasahara, Tomoko Chiyo, Kiwamu Imagawa, Katsuhiko Tachibana, Shin'ichi Takeda, Takashi Okada
Journal: Molecular Therapy (2015)

Duchenne muscular dystrophy (DMD) is a congenital disease causing progressive deterioration of skeletal and cardiac muscles because of mutations in the dystrophin gene. Supplementation of dystrophin using rAAV is effective to improve pathogenesis of animal models of DMD. However, we have previously reported that local injection of rAAV2 or rAAV8 to canine skeletal muscles without immunosuppression resulted in insufficient transgene expression with potent immune responses. Mesenchymal stem cells (MSCs) regulate graft-versus-host disease (GVHD) by virtue of their immunosuppressive effects. Short-term exposure of MSCs with vector administration efficiently inhibited immune responses to the rAAV capsid and/or transgene products. Human bone-marrow derived MSCs and rAAV9-Luciferase were intravenously injected into the normal dog at 8 weeks old. Seven days after injection, hMSCs were systemically injected again. At 8 days after 1st injection, rAAV9-Luciferase was intramuscularly injected into the tibialis anterior muscle of the same dog. To examine the immune response against rAAV, purified canine peripheral leukocytes were exposed to rAAV9 for 4 hours, and then IFN-γ expression was analyzed using qRT-PCR. Skeletal muscles of the rAAV-Luc injected animals were sampled by biopsy for expression analysis at 4 weeks after intramuscular injection. Intramuscular injection of rAAV-Luc following hMSCs treatment, resulted in higher expression of Luciferase at the injected muscle, compared to the rAAV-Luc transduction alone. Expression of IFN-γ in the purified peripheral blood leukocytes after the rAAV exposure were not induced in the rAAV-Luc with MSCs, suggesting the immune suppressive effects of the MSCs. Our results demonstrate that rAAV-Luc injection with MSCs treatment improved expression of rAAV-derived Luciferase in dogs. This strategy would be effective approach to analyze the expression and function of transgene in vivo. These findings also support the future feasibilities of rAAV-mediated protein supplementation strategies.
Two-Track Epigenetic Remodeling and Backtracking to Embryonic Stem Cell Bivalency in B-Cell Acute Lymphoblastic Leukemias
Authors: Seung-Tae Lee, Marcus O. Muench, Marina Fomin, Jianqiao Xiao, Mi Zhou, Adam J. de Smith, José Martín-Subero, Simon Heath, E. Andres Houseman, Ritu Roy, Margaret R. Wrensch, John K. Wiencke, Catherine Metayer, Joseph L. Wiemels
Journal: Blood (2014)
Non-cytotoxic drug therapy for loco-regional edema and serosal effusions of malignancy
Authors: D. D. Nutu, M. Dediu, R. Anghel, D. Spataru, S. Donea, M. Bostina, S. Stan, L. Calustian, A. Constantinescu
Journal: Journal of Clinical Oncology (2004)

8145 Background: Based on our previous research we hypothesized that three important mechanisms underlie serosal effusions and extensive loco-regional edema of locally advanced malignancies: active vasodilation gradually evolving from local to regional aspects, and increased secretion of autacoids and aldosteron. Hence, vasoconstrictor drugs, autacoid antagonists, and spironolactone should oppose the process. Method: 25 patients (pts) with both tumors of various types and locations, and serosal effusions or locoregional edema for which antitumor treatments were inapplicable or had previously failed, were treated for 3–27 weeks with ergotamine 0.5 mg once daily and t.i.d. propranolol 20–40 mg, ibuprofen 200 mg, cyproheptadine 2 mg, ranitidine 150 mg, and spironolactone 25–50 mg. Pts with pain or on vasodilating drugs were excluded while dexametasone for anorexia and local wound care were allowed. Regression with =50% of either serosal effusions (i.e. at least doubling the interval of instrumental draining), regional edema, or the ulceration size /oozing was the criterion of efficacy. Results: The treatment was effective in 9 of 11 (81.8%) pts with serosal effusions (with renewed response of 4 re-treated recurrences after treatment discontinuation in 3 pts), and in 10 of 14 (71.4%) pts with local ulceration /oozing or edema extending to a whole limb. Complete remission was achieved in 9 pts, in three only after the addition of dexametasone 8 mg twice a week. No significant adverse effects occurred. Conclusions: This drug combination provides substantial palliation for loco-regional edema and serosal effusions related of malignancy. The additive effect of dexametasone deserves further exploration. No significant financial relationships to disclose.
Abstract 1007: Elucidating a role for the translation initiation factor, eIF4G1, in resistance to therapy in inflammatory breast cancer (IBC)
Authors: Arianna Price, Courtney M. Edwards, Myron K. Evans, Gayathri Devi
Journal: Cancer Research (2015)

Abstract Inflammatory breast cancer (IBC) is a highly invasive and aggressive breast cancer with relatively low survival rates compared with other locally advanced breast cancers. IBC is characterized by its rapid development of resistance to radio- and chemotherapies, which may in part be due to altered translational control driven by overexpression of the translation initiation factor, eIF4G1. This protein has been shown to enhance translation of eukaryotic mRNAs with internal ribosome entry sites (IRES) and enhanced translation in IBC may play a role in the survival of tumor cells during cellular stress, when translation would normally be suppressed. Some inhibitors of apoptosis proteins have 5′-untranslated regions (UTRs) that allow for IRES-mediated translation, and inhibition of apoptosis is one mechanism by which IBC cells have previously been shown to become resistant to therapy. Translational up-regulation of an inhibitor of apoptosis protein in therapy-resistant clones of IBC cell lines has been observed, which is of importance as the 5′UTR of such proteins contain highly active IRES elements. Regulation of such protein expression by eIF4G1-mediated IRES translation may be one reason for this observed therapeutic resistance. We knocked down eIF4G1 in a resistant IBC cell line (rSUM149) and assessed cellular viability after treatment with a powerful apoptosis inducer, TRAIL to evaluate the effect of targeted eIF4G1 knockdown on drug resistance in IBC. We observed that targeted knockdown of eIF4G1 sensitizes rSUM149 cells to TRAIL, significantly decreasing cellular viability. Additionally, using a novel culture system we show that inhibition of eIF4G1 expression decreases both the number of size of tumor emboli, a characteristic feature of IBC progression. These findings demonstrate a vital role for eIF4G1 for the development of therapeutic resistance and ultimately the aggressive pathobiology of IBC. Citation Format: Arianna Price, Courtney M. Edwards, Myron K. Evans, Gayathri Devi. Elucidating a role for the translation initiation factor, eIF4G1, in resistance to therapy in inflammatory breast cancer (IBC). [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 1007. doi:10.1158/1538-7445.AM2015-1007
Telomere Shortening, Regenerative Capacity, and Cardiovascular Outcomes
Authors: Muhammad Hammadah, Ibhar Al Mheid, Kobina Wilmot, Ronnie Ramadan, Naser Abdelhadi, Ayman Alkhoder, Malik Obideen, Pratik M. Pimple, Oleksiy Levantsevych, Heval M. Kelli, Amit Shah, Yan V. Sun, Brad Pearce, Michael Kutner, Qi Long, Laura Ward, Yi-An Ko, Kareem Hosny Mohammed, Jue Lin, Jinying Zhao, J. Douglas Bremner, Jinhee Kim, Edmund K. Waller, Paolo Raggi, David Sheps, Arshed A. Quyyumi, Viola Vaccarino
Journal: Circulation Research (2016)

Leukocyte telomere length (LTL) is a biological marker of aging, and shorter LTL is associated with adverse cardiovascular outcomes. Reduced regenerative capacity has been proposed as a mechanism. Bone marrow-derived circulating progenitor cells are involved in tissue repair and regeneration.Main objective of this study was to examine the relationship between LTL and progenitor cells and their impact on adverse cardiovascular outcomes.We measured LTL by quantitative polymerase chain reaction in 566 outpatients (age: 63±9 years; 76% men) with coronary artery disease. Circulating progenitor cells were enumerated by flow cytometry. After adjustment for age, sex, race, body mass index, smoking status, and previous myocardial infarction, a shorter LTL was associated with a lower CD34+ cell count: for each 10% shorter LTL, CD34+ levels were 5.2% lower (P<0.001). After adjustment for the aforementioned factors, both short LTL (<Q1) and low CD34+ levels (<Q1) predicted adverse cardiovascular outcomes (death, myocardial infarction, coronary revascularization, or cerebrovascular events) independently of each other, with a hazard ratio of 1.8 and 95% confidence interval of 1.1 to 2.0, and a hazard ratio of 2.1 and 95% confidence interval of 1.3 to 3.0, respectively, comparing Q1 to Q2-4. Patients who had both short LTL (<Q1) and low CD34+ cell count (<Q1) had the greatest risk of adverse outcomes (hazard ratio =3.5; 95% confidence interval, 1.7-7.1).Although shorter LTL is associated with decreased regenerative capacity, both LTL and circulating progenitor cell levels are independent and additive predictors of adverse cardiovascular outcomes in coronary artery disease patients. Our results suggest that both biological aging and reduced regenerative capacity contribute to cardiovascular events, independent of conventional risk factors.
NK-Cell Recovery and Immune Reconstitution after Haploidentical Hematopoietic Cell Transplantation Using Either CD34 Selected Grafts and Adoptive NK-Cell Transfer or CD3/CD19 Depleted Grafts: Comparison of Two Strategies for NK Cell Based Immunotherapy.
Authors: Chiara Gentilini, Matthias Haegele, Arne Muessig, Axel Nogai, Constanze Kliem, Kristina Bartsch, Nadezda Bazara, Wichard Vogel, Christoph Faul, Lothar Kanz, Eckhard Thiel, Dietger W. Niederwieser, Lutz Uharek, Wolfgang A. Bethge
Journal: Blood (2007)
Resetting microbiota by <i>Lactobacillus reuteri</i> inhibits T reg deficiency–induced autoimmunity via adenosine A2A receptors
Authors: Baokun He, Thomas K. Hoang, Ting Wang, Michael Ferris, Christopher M. Taylor, Xiangjun Tian, Meng Luo, Dat Q. Tran, Jain Zhou, Nina Tatevian, Fayong Luo, Jose G. Molina, Michael R. Blackburn, Thomas H. Gomez, Stefan Roos, J. Marc Rhoads, Yuying Liu
Journal: The Journal of Experimental Medicine (2016)

Regulatory T (T reg) cell deficiency causes lethal, CD4+ T cell–driven autoimmune diseases. Stem cell transplantation is used to treat these diseases, but this procedure is limited by the availability of a suitable donor. The intestinal microbiota drives host immune homeostasis by regulating the differentiation and expansion of T reg, Th1, and Th2 cells. It is currently unclear if T reg cell deficiency–mediated autoimmune disorders can be treated by targeting the enteric microbiota. Here, we demonstrate that Foxp3+ T reg cell deficiency results in gut microbial dysbiosis and autoimmunity over the lifespan of scurfy (SF) mouse. Remodeling microbiota with Lactobacillus reuteri prolonged survival and reduced multiorgan inflammation in SF mice. L. reuteri changed the metabolomic profile disrupted by T reg cell deficiency, and a major effect was to restore levels of the purine metabolite inosine. Feeding inosine itself prolonged life and inhibited multiorgan inflammation by reducing Th1/Th2 cells and their associated cytokines. Mechanistically, the inhibition of inosine on the differentiation of Th1 and Th2 cells in vitro depended on adenosine A2A receptors, which were also required for the efficacy of inosine and of L. reuteri in vivo. These results reveal that the microbiota–inosine–A2A receptor axis might represent a potential avenue for combatting autoimmune diseases mediated by T reg cell dysfunction.
Abstract 4149: Epidermal growth factor-induced cyclooxygenase-2 enhances head and neck squamous cell carcinoma metastasis through fibronectin up-regulation
Authors: Jinn-Yuan Hsu, Kwang-Yu Chang, Shang-Hung Chen, Chung-Ta Lee, Sheng-Tsung Chang, Hung-Chi Cheng, Wen-Chang Chang, Ben-Kuen Chen
Journal: Cancer Research (2015)

Abstract Epidermal growth factor receptor (EGFR) activation is a major cause of cell metastasis in many cancers, such as head and neck squamous cell carcinoma (HNSCC). However, whether the induction of cyclooxygenase-2 (COX-2) correlates with EGF-enhanced HNSCC metastasis remains unclear. Interestingly, we found that EGF induced COX-2 expression mainly in HNSCC. The tumor cell transformation induced by EGF was repressed with COX-2 knockdown, and this repression was reversed by simultaneously treating the cells with EGF and prostaglandin E2 (PGE2). The down-regulation of COX-2 expression or inhibition of COX-2 activity significantly blocked EGF-enhanced cell migration and invasion, but the addition of PGE2 compensated for this blockage in COX-2-knockdown cells. Interestingly, COX-2 depletion inhibited EGF-induced matrix metalloproteinase-1 (MMP-1), MMP3 and fibronectin expression and Rac1/cdc42 activation; this reduction in MMPs and the fibronectin/Rac1/cdc42 axis by the depletion of COX-2 was also rescued when the cells were treated with PGE2. Furthermore, the depletion of fibronectin impeded the COX-2-enhanced binding of HNSCC cells to endothelial cells and tumor cells metastatic seeding of the lungs. These results provide new insight that EGF-induced COX-2 enhances HNSCC metastasis via activation of the fibronectin signaling pathway. The inhibition of COX-2 expression and activation suggests a potential strategy for the treatment of EGFR-mediated HNSCC metastasis. Citation Format: Jinn-Yuan Hsu, Kwang-Yu Chang, Shang-Hung Chen, Chung-Ta Lee, Sheng-Tsung Chang, Hung-Chi Cheng, Wen-Chang Chang, Ben-Kuen Chen. Epidermal growth factor-induced cyclooxygenase-2 enhances head and neck squamous cell carcinoma metastasis through fibronectin up-regulation. [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 4149. doi:10.1158/1538-7445.AM2015-4149
Genome-Wide Crispr-Cas9 Screen Identifies Functionally Relevant Micro-RNAs in FLT3-ITD+ AML
Authors: Jared Wallace, Dominique Kagele, Ruozhen Hu, Marah Runtsch, Margaret Alexander, Thomas Huffaker, Timothy Mosbruger, Timothy Dahlem, W. Zac Stephens, Dinesh S. Rao, Rodney R. Miles, June Round, Ryan M. O'Connell
Journal: Blood (2015)
Patterns of Immune Reconstitution in Patients with Acute Promyelocytic Leukemia Treated with Single Agent Arsenic Trioxide and Its Impact On Time to Molecular Remission
Authors: Ansu Abu Alex, Saravanan Ganesan, Poonkuzhali Balasubramanian, Rayaz Ahmed, Aby Abraham, Auro Viswabandya, Biju George, Ezhilarasi Chendamarai, Alok Srivastava, Vikram Mathews
Journal: Blood (2012)
The Role of Behavioral Interventions in Buprenorphine Maintenance Treatment: A Review
Authors: Kathleen M. Carroll, Roger D. Weiss
Journal: American Journal of Psychiatry (2016)

Although counseling is a required part of office-based buprenorphine treatment of opioid use disorders, the nature of what constitutes appropriate counseling is unclear and controversial. The authors review the literature on the role, nature, and intensity of behavioral interventions in office-based buprenorphine treatment.
Abstract 253: Synergistic antitumor activity of PD-1 signaling blockade and CD27 costimulation correlates with enhanced ratio of effector to regulatory T cells at the tumor site
Authors: Lawrence J. Thomas, Li-Zhen He, Anna Wasiuk, Lauren E. Gergel, James M. Boyer, Sarah M. Round, Henry C. Marsh, Tibor Keler
Journal: Cancer Research (2015)

Abstract Varlilumab (CDX-1127) is a fully human agonist antibody to CD27, a TNF receptor superfamily member expressed on the majority of T cells and subsets of NK cells and B cells. As such, varlilumab is an immunotherapy designed to harness the body's natural immune response by enhancing the activation of T cells that can specifically recognize and kill cancer cells. In an ongoing Phase 1 clinical trial, varlilumab has been very well tolerated and has demonstrated clear biologic activity and promising signs of clinical activity in advanced, treatment-refractory patient populations. This clinical data provides the rationale for combination studies with other immunomodulatory therapies, including inhibitors of PD-1 signaling. In this study we investigated the activity of varlilumab in combination with anti-PD-L1 using a disseminated murine B cell tumor, BCL1 in human CD27 transgenic mice. In this model of i.v. inoculated BCL1 cells, we have previously shown that varlilumab can delay tumor growth, but rarely results in cures. Despite expression of PD-L1, the BCL1 tumors were completely resistant to anti-PD-L1 monotherapy. However, the combination of varlilumab and anti-PD-L1 resulted in a significant improvement in survival and approximately 50% long-term survivors. To investigate the mechanism resulting in the synergistic activity, we collected spleens (the major site of tumor growth) from tumor bearing animals treated with individual or combined regimens and processed them for flow cytometry analysis. We found that animals treated with the combination of varlilumab and anti-PD-L1 had a higher ratio of infiltrating immune cells to tumor cells relative to monotherapy or saline controls. In particular, the ratio of effector (CD8+) T cells to Treg and the number of IFNγ+ CD4 and CD8 T cells were highly increased with the combination treatment. These data demonstrate that CD27 costimulation combined with checkpoint inhibition with anti-PD-L1 can result in improved immune profile in the tumor and synergistic survival benefit. This study, along with the good safety profile of varlilumab reported in the Phase 1 study, support the design of future combination studies in patients with cancer. Citation Format: Lawrence J. Thomas, Li-Zhen He, Anna Wasiuk, Lauren E. Gergel, James M. Boyer, Sarah M. Round, Henry C. Marsh, Tibor Keler. Synergistic antitumor activity of PD-1 signaling blockade and CD27 costimulation correlates with enhanced ratio of effector to regulatory T cells at the tumor site. [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 253. doi:10.1158/1538-7445.AM2015-253
All in a Day’s Work — Equity vs. Equality at a Public ICU in Brazil
Authors: Flávia R. Machado
Journal: New England Journal of Medicine (2016)

Every day, Brazilian intensivists face cruel choices about which patients will be given the care they need. Should a 32-year-old woman with respiratory failure receive extracorporeal membrane oxygenation? Or should the money be spent instead on 1250 doses of antibiotics?
Retrospective analysis of emergency hospital admission due to chemoradiotherapy for head and neck cancer patients
Authors: Hayato Kamata, Shinya Suzuki, Kiwako Ikegawa, Hisanaga Nomura, Tomohiro Enokida, Tetsuro Wakasugi, Tomoko Yamazaki, Makoto Tahara, Shinichiro Saito
Journal: Annals of Oncology (2015)