Corrigendum to “The development of a sensitive fluorescent protein-based transcript reporter for high throughput screening of negative modulators of lncRNAs” [Genes & Diseases 5 (2018) 62–74]

The authors regret having an image assembly error in Figure 5Ca,in which the image for the "Oh dBiFP-AdRFp"group was erroneously duplicated with an overlapping image from the"36h BiFP dBIFP-AdR-simH19"group.We confirm the error is restricted to the image assembly,and the underlying data and conclusions are correct and unchanged.The authors would like to apologize for any inconvenience caused.

Adenovirus-mediated gene delivery:Potential applications for gene and cell-based therapies in the new era of personalized medicine

With rapid advances in understanding molecular pathogenesis of human diseases in the era of genome sciences and systems biology,it is anticipated that increasing numbers of therapeutic genes or targets will become available for targeted therapies.Despite numerous setbacks,efficacious gene and/or cell-based therapies still hold the great promise to revolutionize the clinical management of human diseases.It is wildly recognized that poor gene delivery is the limiting factor for most in vivo gene therapies.There has been a long-lasting interest in using viral vectors,especially adenoviral vectors,to deliver therapeutic genes for the past two decades.Among all currently available viral vectors,adenovirus is the most efficient gene delivery system in a broad range of cell and tissue types.The applications of adenoviral vectors in gene delivery have greatly increased in number and efficiency since their initial development.In fact,among over 2000 gene therapy clinical trials approved worldwide since 1989,a significant portion of the trials have utilized adenoviral vectors.This review aims to provide a comprehensive overview on the characteristics of adenoviral vectors,including adenoviral biology,approaches to engineering adenoviral vectors,and their applications in clinical and preclinical studies with an emphasis in the areas of cancer treatment,vaccination and regenerative medicine.Current challenges and future directions regarding the use of adenoviral vectors are also discussed.It is expected that the continued improvements in adenoviral vectors should provide great opportunities for cell and gene therapies to live up to its enormous potential in personalized medicine.

The development of a sensitive fluorescent protein-based transcript reporter for high throughput screening of negative modulators of lncRNAs

While the human genome is pervasively transcribed,<2%of the human genome is transcribed into protein-coding mRNAs,leaving most of the transcripts as noncoding RNAs,such as microRNAs and long-noncoding RNAs(lncRNAs),which are critical components of epigenetic regulation.lncRNAs are emerging as critical regulators of gene expression and genomic stability.However,it remains largely unknown about how lncRNAs are regulated.Here,we develop a highly sensitive and dynamic reporter that allows us to identify and/or monitor negative modulators of lncRNA transcript levels in a high throughput fashion.Specifically,we engineer a fluorescent fusion protein by fusing three copies of the PEST destruction domain of mouse ornithine decarboxylase(MODC)to the C-terminal end of the codon-optimized bilirubin-inducible fluorescent protein,designated as dBiFP,and show that the dBiFP protein is highly destabilized,compared with the commonly-used eGFP protein.We further demonstrate that the dBiFP signal is effectively down-regulated when the dBiFP and mouse lncRNA H19 chimeric transcript is silenced by mouse H19-specific siRNAs.Therefore,our results strongly suggest that the dBiFP fusion protein may serve as a sensitive and dynamic transcript reporter to monitor the inhibition of lncRNAs by microRNAs,synthetic regulatory RNA molecules,RNA binding proteins,and/or small molecule inhibitors so that novel and efficacious inhibitors targeting the epigenetic circuit can be discovered to treat human diseases such as cancer and other chronic disorders.

Clinical and economic benefits of a pharmacist in a multidisciplinary team for Chinese outpatients with gastrointestinal stromal tumors

The multidisciplinary approach is beneficial to the treatment of gastrointestinal stromal tumors(GISTs).However,pharmacists are seldom incorporated into a multidisciplinary team(MDT)of GIST.In the present study,we evaluated the clinical and economic benefits of a pharmacist in an MDT of GIST.This was a retrospective study that included 240 GIST patients receiving imatinib therapy.The GIST MDT pharmacist developed and validated an HPLC method to monitor the trough concentrations of imatinib in GIST patients.Besides,the pharmacist also provided patient education and pharmaceutical care services and collected the data for analysis.The 240 GIST patients received the services provided by the pharmacist.The trough concentrations in 25.42%of the 240 patients were less than 1100 ng/m L.The main genotypes of 121 in the 240 patients were KIT exon 11 mutations,wild type,and KIT exon 9 mutations.Moreover,13 GIST patients with trough concentrations less than 1100 ng/m L were confirmed to have low compliance.The adverse reactions were primarily mild and tolerable,except that 13 GIST patients were adjusted to lower doses because of the intolerable adverse reactions.The daily cost could be lowered by monitoring the trough concentrations and dose reductions.Collectively,a pharmacist included in a GIST MDT could increase the compliance of Chinese GIST patients to imatinib therapy and improve the efficacy,safety,and economy of imatinib therapy.

GAPDH suppresses adenovirus-induced oxidative stress and enables a superfast production of recombinant adenovirus

Recombinant adenovirus(rAdV)is a commonly used vector system for gene transfer.Efficient initial packaging and subsequent production of rAdV remains time-consuming and labor-intensive,possibly attributable to rAdv infection-associated oxidative stress and reactive oxygen species(ROS)production.Here,we show that exogenous GAPDH expression mitigates adenovirus-induced ROS-associated apoptosis in HEK293 cells,and expedites adenovirus production.By stably overexpressing GAPDH in HEK293(293G)and 293pTP(293GP)cells,respectively,we demonstrated that rAdV-induced RoS production and cell apoptosis were significantly suppressed in 293G and 293GP cells.Transfection of 293G cells with adenoviral plasmid pAd-G2Luc yielded much higher titers of Ad-G2Luc at day 7 than that in HEK293 cells.Similarly,Ad-G2Luc was amplified more efficiently in 293G than in HEK293 cells.We further showed that transfection of 293GP cells with pAd-G2Luc produced much higher titers of Ad-G2Luc at day 5 than that of 293pTP cells.293GP cells amplified the Ad-G2Luc much more efficiently than 293pTP cells,indicating that exogenous GAPDH can further augment pTP-enhanced adenovirus production.These results demonstrate that exogenous GAPDH can effectively suppress adenovirus-induced ROS and thus accelerate adenovirus production.Therefore,the engineered 293GP cells represent a superfast rAdV production system for adenovirus-based gene transfer and gene therapy.