Development and Validation of a Reporter Cell Line for Rapid AAV Quality Control Assessment

Publication date: Available online 23 January 2020Source: MethodsXAuthor(s): Huira C. Kopera, Roland S. Hilgarth, Tonya L. Kopas, Thomas M. LaniganAbstractAdenovirus-associated virus is a powerful vector system for transducing cells in vivo. It is widely used in animal systems due to high transduction efficiency of non-dividing cells with more than a dozen serotypes that have preferential tissue tropism. The viral genome remains episomal in the nucleus but maintains sustained expression in terminally differentiated cells for several weeks to months. Despite the popularity of recombinant AAV (rAAV) vectors, quality control testing of the virus after production is largely limited to physical characteristic such as viral genomes/ml determinations and silver staining acrylamide gels to determine purity. Functional testing, in vivo, is not practical due to high cost and restricted access of animal care and long duration of the assay (2-3 weeks). Some functional testing can be accomplished in cultured cells such as HEK293 cells, but HEK293 cells limit the types of rAAV constructs that can be tested. Many rAAV constructs are designed to study neurons in the brain with neural-specific promoters and many are floxed with loxp sites to be “activated” only in Cre expressing neurons in transgenic animals. To develop a reporter cell line for rapid rAAV quality control assessment of these neural-specific, floxed rAAV constructs, we used the lentiviral system to stably express Cr...
Source: MethodsX - Category: Science Source Type: research

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Childhood cancer survivors (CCSs) from high-grade malignancies, such as high-risk neuroblastoma, have been increased, and second malignant neoplasm, becomes a serious problem for CCSs. However, detailed reports about rare types of second cancer such as gastric cancer remain limited. We herein reported a female patient who developed diffuse type gastric carcinoma after 21 years from completion of treatment to high-risk neuroblastoma. We reviewed the previous cohort studies for second gastrointestinal cancer in CCSs and the case reports with second gastric carcinoma for CCSs. We presumed second gastric cancer was refractory ...
Source: Journal of Pediatric Hematology Oncology - Category: Hematology Tags: Clinical and Laboratory Observations Source Type: research
Publication date: Available online 19 February 2020Source: International Journal of PharmaceuticsAuthor(s): Subrat K. Bhattamisra, Angeline S. Tzeyung, Lim Wen Xi, Nurul Hasyanazifa Safian, Hira Choudhury, Lim Wei Meng, Naiyer Shahzad, Nabil A. Alhakamy, Md khalid Anwer, Ammu K. Radhakrishnan, Shadab MdAbstractRotigotine, a non-ergoline dopamine agonist, has been shown to be highly effective for the treatment of Parkinson's disease (PD). However, despite its therapeutic potential, its’ clinical applications were hindered due to low aqueous solubility, first-pass metabolism and low bioavailability. Therefore, we devel...
Source: International Journal of Pharmaceutics - Category: Drugs & Pharmacology Source Type: research
We present here a murine model that overcomes many of these limitations. We found that carrier cells outcompeted intravenous administration of naked particles in delivering the oncolytic virus into the tumor masses. The protection that MSCs could provide to the oncolytic adenovirus did not preclude the development of an antiadenoviral immune response. However, the presence of circulating antiadenoviral antibodies did not prevent changes detected at the tumor masses: increased infiltration and changes in the quality of immune cells per unit of tumor volume, and a less protumoral and more inflammatory profile of the tumor mi...
Source: Oncotarget - Category: Cancer & Oncology Tags: Oncotarget Source Type: research
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Source: Pediatric Hematology and Oncology - Category: Pediatrics Authors: Source Type: research
Publication date: 18 February 2020Source: Cell Reports, Volume 30, Issue 7Author(s): Christin Suenkel, Daniel Cavalli, Simone Massalini, Federico Calegari, Nikolaus RajewskySummarycircSLC45A4 is the main RNA splice isoform produced from its genetic locus and one of the highest expressed circRNAs in the developing human frontal cortex. Knockdown of this highly conserved circRNA in a human neuroblastoma cell line is sufficient to induce spontaneous neuronal differentiation, measurable by increased expression of neuronal marker genes. Depletion of circSlc45a4 in the developing mouse cortex causes a significant reduction ...
Source: Cell Reports - Category: Cytology Source Type: research
MYCN-amplified neuroblastoma (NB) is characterized by poor prognosis, and directly targeting MYCN has proven challenging. Here, we showed that aldehyde dehydrogenase family 18 member A1 (ALDH18A1) exerts profound impacts on the proliferation, self-renewal, and tumorigenicity of NB cells and is a potential risk factor in patients with NB, especially those with MYCN amplification. Mechanistic studies revealed that ALDH18A1 could both transcriptionally and posttranscriptionally regulate MYCN expression, with MYCN reciprocally transactivating ALDH18A1 and thus forming a positive feedback loop. Using molecular docking and scree...
Source: Science Translational Medicine - Category: Biomedical Science Authors: Tags: Research Articles Source Type: research
In this study, two novel benzimidazolium salts consisting propane sulfonate order were integrated in high yields as N-heterocyclic carbene (NHC) precursors.The silver-NHC buildings were set up by responding 1-(2,2-dimethoxyethyl)- 3-(propyl-3-sulfonate)benzimidazolium and 1-(2,2-diethoxyethyl)- 3-(propyl-3-sulfonate)benzimidazolium with silver oxide on average state. Gold-NHC edifices were acquired by means of transmetallation of their silver precursors with [AuCl{S(CH3)2}]. The structures of all mixes were completely portrayed by spectroscopic techniques (1H-NMR, 13C{1H} NMR , IR and elemental and further analyzed by DFT/...
Source: Inorganica Chimica Acta - Category: Chemistry Source Type: research
(Rice University) Rice University neurodevelopmental biologist Rosa Uribe has won a prestigious CAREER Award from the National Science Foundation to study neural crest cells, specialist stem cells that form the enteric nervous system and also cause neuroblastoma, the third most prevalent form of pediatric cancer in the U.S.
Source: EurekAlert! - Biology - Category: Biology Source Type: news
Publication date: Available online 17 February 2020Source: Materials Today: ProceedingsAuthor(s): Michelina Catauro, Federico Barrino, Ignazio Blanco, Giovanni Dal Poggetto, Simona Piccolella, Giuseppina Crescente, Severina PacificoAbstractSol-gel chemistry is an attractive approach in the design of new antioxidant biomaterials. Its versatility was exploited to obtain organic-inorganic hybrid materials in which a natural antioxidant compound, namely chlorogenic acid (CGA), was entrapped, in different percentage, in matrices variously constituted in silica and PEG400.The synthetized hybrids were chemically characterized by ...
Source: Materials Today: Proceedings - Category: Materials Science Source Type: research
We report our clinical experience of motion mitigation with volumetric rescanning (vRSC) and outcomes of children with neuroblastoma.
Source: Clinical Oncology - Category: Radiology Authors: Tags: Original Article Source Type: research
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