tbuckley/cabot-search huggingface dataset:
481. CRISPR/Cas9-Mediated In Vivo Genome Editing to Correct the OTC spfash Mutation in Newborn Mice
Authors: Lili Wang, Yang Yang, Peter Bell, Deirdre McMenamin, Zhenning He, John White, Hongwei Yu, Chenyu Xu, Hiroki Morizono, Kiran Musunuru, Mark L. Batshaw, James M. Wilson
Journal: Molecular Therapy (2016)

Many genetic liver diseases, including OTC deficiency, present in newborns with repeated, often lethal, metabolic crises. Adeno-associated virus (AAV) neonatal gene therapy in this setting would require multiple vector administrations to maintain the therapeutic effects because the non-integrating genome is lost as developing hepatocytes proliferate. As such, we reasoned that newborn liver may be an ideal setting for AAV-mediated gene correction using CRISPR/Cas9, a powerful genome-editing tool consisting of the Cas9 nuclease and a single-guide RNA (sgRNA). We developed a strategy using an AAV vector with high liver tropism (AAV8) to correct the point mutation in newborn spfash mice using Cas9 enzyme from Staphylococcus aureus (SaCas9). An animal model of OTC deficiency, the male sparse fur ash (spfash) mouse, has a G-to-A point mutation at the donor splice site at the end of exon 4 of the OTC gene, which leads to abnormal splicing and a 20-fold reduction in OTC mRNA and protein. We developed a two-vector approach to incorporate all 3 components of CRISPR/Cas9 into AAV. Vector 1 expresses the SaCas9 gene from a liver-specific TBG promoter, while vector 2 contains both the sgRNA1 sequence expressed from a U6 promoter and the 1.8 kb donor OTC DNA sequence. Spfash pups were injected intravenously on postnatal day 2 with mixtures of vector 1 and vector 2 and subsequently evaluated for indel (insertion and deletion) formation and functional correction of the spfash mutation. Following gene correction (3 and 8 weeks), indels were detected by deep sequencing in 31% of OTC alleles, and HDR-based correction of the G-to-A mutation was observed in 10% of OTC alleles. Liver sections were analyzed by immunohistochemistry for OTC expression, showing 15% OTC-positive cells at 3 weeks and 13% at 8 weeks. Direct measurements of OTC enzyme activity from liver homogenates and OTC mRNA from total cellular RNA from liver revealed similarly high levels of correction in treated animals. We further assessed the impact of gene correction on the clinical manifestations of OTC deficiency by evaluating the tolerance of spfash mice to a one-week course of high-protein diet. Spfash mice treated with the gene-editing vectors had significantly lower plasma ammonia levels and showed a survival improvement as compared with untreated spfash mice. This study provides convincing evidence for efficacy in an authentic animal model of a lethal human metabolic disease following in vivo genome editing.
Abstract 2798: Quercetin inhibits Cr(VI)-induced malignant transformation of human lung epithelial cells by targeting miR-21-Pdcd4 signaling pathway
Authors: Pratheeshkumar Poyil
Journal: Cancer Research (2015)

Abstract Hexavalent chromium [Cr(VI)] is an important human carcinogen associated with pulmonary diseases and lung cancer. Cancer prevention using natural products has become an integral part of cancer control. Quercetin is one of the most abundant dietary flavonoids widely present in many fruits and vegetables, possesses potent antioxidant activity capable of protecting normal cells from various stimuli-induced oxidative stress and cell death. MicroRNA-21 (miR-21) is a key regulator of oncogenic processes. It is significantly elevated in the majority of human tumors and functionally linked to cellular proliferation, invasion and migration. Studies have shown that miR-21 exerts its oncogenic activity by targeting the tumor suppressor gene programmed cell death 4 (Pdcd4). The present study examined the effect of quercetin on the inhibition of Cr(VI) induced carcinogenesis and the role of miR-21-Pdcd4 signaling involved. Our results showed that quercetin decreased ROS generation induced by Cr(VI) exposure in Beas-2B cells. Chronic Cr(VI) exposure induced malignant cell transformation, increased miR-21 expression and caused inhibition of Pdcd4, which were significantly inhibited by the treatment of quercetin in a dose dependent manner. Stable knockdown of miR-21 or overexpression of pdcd4 in Beas-2B cells significantly reduced the Cr(VI) induced cell transformation. Furthermore, Quercetin also inhibited the Cr(VI) induced beta-catenin/TCF-dependent transcription. Taken together, these results demonstrate that quercetin is able to protect Beas-2B cells from Cr(VI)-induced carcinogenesis by targeting miR-21-Pdcd4 signaling. Keywords: Hexavalent chromium, Quercetin, miR-21-Pdcd4 signaling Citation Format: Pratheeshkumar Poyil. Quercetin inhibits Cr(VI)-induced malignant transformation of human lung epithelial cells by targeting miR-21-Pdcd4 signaling pathway. [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 2798. doi:10.1158/1538-7445.AM2015-2798
TP53 Alterations Including Missense Mutations, Aberrant Exon-Junctions and Internal Intron Retentions Are Frequent and May Contribute to MDM2 Antagonist-Resistance in B-Acute Lymphoblastic leukemia
Authors: Ilaria Iacobucci, Anna Ferrari, Stefania Trino, Annalisa Lonetti, Cristina Papayannidis, Alberto Ferrarini, Luca Venturini, Maria Chiara Abbenante, Sarah Parisi, Federica Cattina, Simona Soverini, Stefania Paolini, Domenico Russo, Marco Vignetti, Fabrizio Pane, Pellegrino Musto, Massimo Delledonne, Michele Baccarani, Giovanni Martinelli
Journal: Blood (2011)
Dynamics Changes in CCL3 and CCL4 Plasma Chemokine Levels in Patients with Chronic Lymhocytic Leukemia (CLL) Managed with Observation
Authors: Mariela Sivina, Lillian Werner, Laura Rassenti, William G. Wierda, Michael J. Keating, Susan O'Brien, Donna Neuberg, Thomas J. Kipps, Jan A. Burger
Journal: Blood (2014)
Triple-negative, low-grade gliomas: A highly agressive tumor with dismal prognosis.
Authors: P. Metellus, B. Coulibaly, C. Colin, A. Maues de Paula, A. Barlier, A. Loundou, S. Fuentes, H. Dufour, M. Barrie, O. L. Chinot, L. Ouafik, D. Figarella-Branger
Journal: Journal of Clinical Oncology (2011)

2023 Background: The phenotypic heterogeneity of low-grade gliomas (LGGs) is still inconsistently explained by genetic alterations characteristic in patients treated according to present standards of care. Here we investigate the combined analysis of three molecular alterations according to clinical and radiologic data in a series of 89 LGGs to provide new insights into LGGs pathogenesis. Methods: IDH1 codon 132 and IDH2 codon 172 sequencing was performed in a series of 89 LGGs and correlated with clinical presentation, MR imaging characteristics, genomic profile and outcome. Also p53 expression and 1p19q deletion status was assessed in all cases. Results: A total of 74 IDH1 mutations at codon 132 and 2 IDH2 mutations at codon 172 were found, including 68 R132H (91.9%), 4 R132C (5.4%), 2 R132S (2.7%) and 2 R172M (5%). The IDH mutations were significantly associated with 1p19q deleted genotype (P =0.027) and p53 expression (P=0.012). The presence (v absence) of IDH mutations was associated with a better outcome (5-year survival rate, 91% v 54%, respectively, P<0.001). After adjustment for age, tumor location and size, radiologic infiltration pattern and extent of surgery, multivariate analysis confirmed that IDH mutations was an independent favorable prognostic factor (hazard ratio=34.6; 95% CI, 2.72 to 329.32, P =0.006). Furthermore, we showed that patients with IDH-nonmutated tumors were significantly older (P <0.05) and that these tumors involved significantly more frequently the insula (P<0.05), were larger in size (>6cm, P<0.05), displayed an infiltrative pattern on MRI (P<0.05) and were almost all p53 negative with no 1p19q deletion (P <10-6). Conclusions: The combined absence of IDH mutations, p53 expression and 1p19 codeletion in LGGs identifies a novel entity coined “Triple negative” tumors with distinctive location, infiltrative behavior, specific molecular alterations and dismal outcome. These findings could significantly modify LGG classification and may represent a new tool to guide patient-tailored therapy.
707. Prolonged Survival and Improved Phenotypes of Lethal Hypophosphatasia Model Mice By Adeno-Associated Virus-Mediated Muscle Transduction of Bone-Targeted Alkaline Phosphatase
Authors: Aki Nakamura, Koichi Miyake, Atsushi Watanabe, Yukihiko Hirai, Noriko Miyake, Osamu Iijima, Kumi Adachi, Hideaki Kinoshita, Taku Noguchi, Shinichi Abe, Takashi Shimada, Takashi Okada
Journal: Molecular Therapy (2015)

Hypophosphatasia (HPP) is a systemic skeletal disease, caused by the deficiency of tissue-nonspecific alkaline phosphatase (TNALP). The TNALP knockout (Akp2 −/-) mice phenotypically mimic the severe infantile form of HPP and usually die by 3 weeks of age suffering from epileptic seizures and apnea. Although considerable amount of attention has been focused on the enzyme replacement therapy as a "Breakthrough therapy", but repeated injection of large amounts of recombinant TNALP is required to achieve clinical benefit. As an alternative approach, we have recently shown that Akp2–/– mice can be treated by a single intravenous injection of recombinant adeno-associated virus (rAAV) vector expressing bone-targeted TNALP with deca-aspartates at the C terminus (TNALP-D10) by the tissue non-specific CAG promoter at the neonatal period. However, the safety of a systemic injection of the rAAV vector into the neonates has not yet been established, because of transduction into non-target tissue. In this study, to exploit a safe and clinically applicable protocol, we constructed a new self-complementary type 8 AAV (scAAV8) vector harboring the muscle creatine kinase (MCK) promoter to express TNALP-D10 and examined the feasibility of muscle transduction for HPP gene therapy. To ascertain whether the MCK promoter retained the ability to muscle-specific expression, scAAV8 expressing enhanced green fluorescence protein driven by the MCK promoter (scAAV8-MCK-EGFP) were generated and injected into the quadriceps femoris muscle (5×1011 vector genome/body (v.g./body)) of newborn C57BL6 mice (n=3). And, to treat Akp2–/– mice, we constructed scAAV8-MCK-TNALP-D10 and injected (2.5×1012 v.g./body) of newborn Akp2–/– mice (n=10). Muscle directed transduction was demonstrated after the injection of scAAV8-MCK-EGFP. The plasma ALP activity was elevated and sustained at therapeutic level, (>1.0 unit/ml) for at least 3 months of scAAV8-MCK-TNALP-D10 treated Akp2–/– mice. The treated Akp2–/– mice grew well and survived over 3 months with healthy appearance (9/10), while untreated Akp2 −/- mice died within 3 weeks (n=10; P<0.001). Improved mineralization of the knee joints was demonstrated by X-ray images and micro CT analysis. Ectopic calcification under this condition was not detected in the treated mice. These results suggest that rAAV-mediated muscle TNALP-D10 transduction strategy would be promising to treat the severe infantile form of HPP.
Abstract 442: Decreased tumor progression by the knockdown of interleukin 17 receoptor A in the B16F10 melanoma model
Authors: Ya-Shan Chen, Chao-Lin Liu, Hui-Shan Chen, Su-Ting Wu, Chia-Rui Shen
Journal: Cancer Research (2015)

Abstract Introduction: Interleukin 17 (IL-17), correlating with advanced stage or poor prognosis of cancer patients, appears to contribute to tumor progression. Here we demonstrated the presence of IL-17 receptor A (IL-17RA) on the tumor cells directly promoting tumor growth, angiogenesis and metastasis. Methods: B16F10 and its derived IL-17RA deficient melanoma cells were stimulated with recombinant IL-17 (γmIL-17) or subcutaneously implanted into C57BL/6 mice. Cell proliferation, cell invasion, tumor growth, metastasis and angiogensis were monitored and anti-tumor immunity was also assessed. Results: As compared with those in the control B16F10 cells, the γmIL-17 inducing cell proliferation, migration and invasion were significantly reduced in the derived IL-17RA-deficient cells. The tumor growth and metastasis were also apparently reduced in the mice bearing the IL-17RA deficient tumors. In fact, not ony the expression of MMP2 and MMP9 but also the production of VEGF and HGF were reduced both in vitro and in vivo. Additionally, the increased levels of IFNγ+ and Granzyme B+ immune cells as well as the reduced numbers of Tregs and MDSCs appeared in the mice bearing the IL-17RA deficient tumors, indicating the lack of IL-17 enhancing the CTL activity and other anti-tumor immunity. Conclusion: These findings support the hypothesis that IL-17/IL-17RA signaling directly promotes tumor growth as well as recruits MDSCs and Treg cells, resulting in tumor progression. It suggests IL-17RA on tumors as a potential therapeutic target for cancer therapy. Citation Format: Ya-Shan Chen, Chao-Lin Liu, Hui-Shan Chen, Su-Ting Wu, Chia-Rui Shen. Decreased tumor progression by the knockdown of interleukin 17 receoptor A in the B16F10 melanoma model. [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 442. doi:10.1158/1538-7445.AM2015-442
Percutaneous Coronary Intervention With Bioresorbable Scaffolds in a Young Child
Authors: Tamim M. Nazif, Sanjog Kalra, Ziad A. Ali, Dimitrios Karmpaliotis, Mariel E. Turner, Thomas J. Starc, Yang Cao, Charles C. Marboe, Michael B. Collins, Martin B. Leon, Ajay J. Kirtane
Journal: JAMA Cardiology (2016)

Although much less frequent than in adults, coronary artery disease requiring revascularization may develop in children because of homozygous familial hypercholesterolemia or other underlying conditions. Percutaneous coronary intervention (PCI) with a bioresorbable scaffold (BRS) may have advantages over metallic coronary stents in this population.To present a case of the successful treatment of unstable, multivessel coronary artery disease in a child with PCI with BRS implantation. This case highlights the limitations of conventional metal stents and the potential benefits of using BRSs in children.This is a case report from an academic tertiary care institution of a 3-year-old boy with homozygous familial hypercholesterolemia and unstable coronary artery disease requiring revascularization. We also briefly review the related literature.Intravascular imaging-guided PCI of the proximal right coronary artery and the left main and proximal left circumflex arteries was performed with BRSs.The primary outcomes were acute procedural success and survival to liver transplant (3 months after PCI).Following BRS implantation, the patient recovered and remained free of cardiovascular symptoms 3 months after PCI. He subsequently underwent an orthotopic liver transplant for definitive treatment of homozygous familial hypercholesterolemia but died of noncardiac complications. A postmortem examination, including a histological assessment, revealed both BRSs to be patent with nonobstructive neointimal hyperplasia.To our knowledge, this is the first report of PCI with BRSs in a child. This represents an application of a BRS with potentially important implications for the future treatment of coronary artery disease in children and warrants further study.
583. Translating RNAi Therapy for Spinocerebellar Ataxia 1 to the Clinic
Authors: Megan S. Keiser
Journal: Molecular Therapy (2016)

Spinocerebellar ataxia 1 (SCA1) is among a group polyglutamine expansion diseases and is characterized by cerebellar ataxia and neuronal degeneration in the cerebellum and brainstem. Currently, there are no effective treatment strategies for this disease. RNA interference (RNAi) is a naturally occurring process that mediates gene silencing and is currently being investigated as a therapy for dominant diseases such as SCA1. Previously, we used AAV vectors to deliver RNAi triggers to transgenic and knock-in mouse models of SCA1 and noted improved neuropathological, motor phenotypes and transcriptional changes. We have also completed studies in non-human primates (NHPs) evaluating the biodistribution, safety and efficacy of vector delivery to the deep cerebellar nuclei (DCN) in NHPs. We next performed dosing studies in pre-and post-symptomatic mice to identify the lowest efficacious dose, the highest tolerated dose. For this, groups of pre-symptomatic mice were given 1 of 4 doses at 5 weeks of age and motor function assayed after symptom onset for untreated mice (34 weeks of age) and immediately sacrificed for post-necropsy analysis. We identified a ceiling dose that conferred toxicity, a low dose that had no effect, and two doses that prevented phenotypic rotarod deficits relative to control injected SCA1 littermates. Concurrently, post-symptomatic mice were injected at 12 weeks of age at 3 escalating doses. Motor function tests at rotarod at 20 weeks of age identified a dose that not only prevented further deficit but significantly improved performance relative to baseline performance. Thus, our AAV-mediated delivery of RNAi to the SCA1 model can reverse motor impairment in mice, and is scalable to nonhuman primates, two important considerations in advancing this therapy to the clinic.
A retrospective analysis of 5-fluorouracil plus cisplatin in metastatic or recurrent esophageal squamous cell carcinoma
Authors: Shuji Hiramoto, Ken Kato, Hirokazu Shoji, Natsuko Okita, Atsuo Takashima, Yoshitaka Honma, Satoru Iwasa, Tetsuya Hmaguchi, Yasuhide Yamada, Yasuhiro Shimada
Journal: Annals of Oncology (2015)
Abstract 2086: Role of the transcription factor NFE2L3 in TNFα signaling pathway in colorectal cancer
Authors: Marina Bury, Volker Blank
Journal: Cancer Research (2015)

Abstract Patients with inflammatory bowel disease have an increased risk of developing colorectal cancer (CRC), which is the third leading cause of cancer related deaths in the world. Our laboratory is studying the transcription factor NFE2L3 [nuclear factor (erythroid-derived 2)-like 3]. NFE2L3, a member of the Cap’n’Collar family, is a basic-region leucine zipper transcription factor that was identified 15 years ago. Contrary to its extensively studied homolog NFE2L2, the role of NFE2L3 in cancer is still unknown. We demonstrated that NFE2L3 functions as heterodimer by complexing with small Maf (musculoaponeurotic fibrosarcoma) proteins for DNA binding. According to the clinical databases, NFE2L3 is up-regulated in CRC compared to normal tissues. Using the colorimetric MTT viability assay, we determined the optimal concentration to treat colon cell lines with the tumor necrosis factor-alpha (TNFα), a pro-inflammatory cytokine associated with CRC tumorigenesis. We showed that protein levels of NFE2L3 and small Maf proteins are up-regulated by TNFα in CRC cells. We used genomic approaches for identifying genes that are modulated by NFE2L3 after TNFα treatments. The identification of the precise role of NFE2L3 in TNFα signaling pathway will lead to a better understanding of colorectal cancers and will help in the design of novel treatment options. Citation Format: Marina Bury, Volker Blank. Role of the transcription factor NFE2L3 in TNFα signaling pathway 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 2086. doi:10.1158/1538-7445.AM2015-2086
Prognostic Significance of Bone Marrow Involvement At Diagnosis and At the Time of Autologous Stem Cell Transplantation for Lymphoma Relapse,
Authors: Aleksandr Lazaryan, John Sweetenham, Lisa Rybicki, Robert Dean, Brad Pohlman, Brian T. Hill, Hien Duong, Ronald Sobecks, Steven Andresen, Shawnda Tench, Matt Kalaycio, Brian J. Bolwell, Edward A. Copelan
Journal: Blood (2011)
222. AST-VAC2: An Embryonic Stem Cell-Derived Dendritic Cell Cancer Immunotherapy
Authors: Casey C. Case, Kevin Nishimoto, Erik Whiteley, Rashi Srivastava, Jane Lebkowski
Journal: Molecular Therapy (2015)

AST-VAC2 is a cancer immunotherapy product comprising embryonic stem cell-derived dendritic cells electroporated with an mRNA encoding a telomerase/lysosome-associated membrane protein 1 (LAMP hTERT) chimeric tumor antigen. Previous studies have shown that AST-VAC2 can elicit a telomerase-specific T-cell response from partially HLA-matched donor peripheral blood mononuclear cells. A related product (AST-VAC1), an autologous dendritic cell/LAMP hTERT cancer immunotherapy, has been tested in clinical trials on patients with prostate cancer or acute myeloid leukemia. The AST-VAC1 treatment was well tolerated and generated anti-telomerase immune responses. AST-VAC2 is being developed as an off-the-shelf allogeneic cancer vaccine for clinical testing in partially HLA matched patients with non-small cell lung carcinoma. The AST-VAC2 product is derived from the H1 human embryonic stem cell line. The stem cells are differentiated into mature dendritic cells in a multi-step process via embryoid bodies, prepared growth surfaces and specific growth factors. The differentiation process requires 34 days in culture. The cells are then electroporated with the LAMP hTERT mRNA and irradiated prior to cryopreservation. In preparation for clinical testing the manufacturing process has been scaled up from T-flasks to large surface area cell stacks. This has required a number of novel process modifications including large scale embryoid body filtration and flow-through electroporation. The final product will be rendered incapable of replication by gamma irradiation. The radiation dose required for halting replication while still preserving antigen presentation was determined using a combination colony formation/potency assay. It is anticipated that the proposed Phase 1/2a clinical trial will enroll approximately 30 patients and will include both a dose escalation and a broadening of inclusion criteria to permit an assessment of safety, toxicity and immunogenicity in patients with advanced disease. Asterias has recently partnered with Cancer Research United Kingdom (CRUK) and the University of Southampton for the GMP manufacturing and initial clinical testing of AST-VAC2 in the UK.