Immunogenicity and Vaccine Potential of InsB, an ESAT-6-Like Antigen Identified in the Highly Virulent Mycobacterium tuberculosis Beijing K Strain

Our group recently identified InsB, an ESAT-6-like antigen belonging to the Mtb9.9 subfamily within the Esx family, in the Mycobacterium tuberculosis Korean Beijing strain (Mtb K) via a comparative genomic analysis with that of the reference Mtb H37Rv and characterized its immunogenicity and its induced immune response in patients with tuberculosis (TB). However, the vaccine potential of InsB has not been fully elucidated. In the present study, InsB was evaluated as a subunit vaccine in comparison with the most well-known ESAT-6 against the hypervirulent Mtb K. Mice immunized with InsB/MPL-DDA exhibited an antigen-specific IFN-γ response along with antigen-specific effector/memory T cell expansion in the lungs and spleen upon antigen restimulation. In addition, InsB immunization markedly induced multifunctional Th1-type CD4+ T cells coexpressing TNF-α, IL-2, and IFN-γ in the lungs following Mtb K challenge. Finally, we found that InsB immunization conferred long-term protection against Mtb K comparable to that conferred by ESAT-6 immunization, as evidenced by a similar level of CFU reduction in the lung and spleen and reduced lung inflammation. These results suggest that InsB may be an excellent vaccine antigen component for developing a multiantigenic Mtb subunit vaccine by generating Th1-biased memory T cells with a multifunctional capacity and may confer durable protection against the highly virulent Mtb K

Nanovesicles derived from iron oxide nanoparticles-incorporated mesenchymal stem cells for cardiac repair

Because of poor engraftment and safety concerns regarding mesenchymal stem cell (MSC) therapy, MSC-derived exosomes have emerged as an alternative cell-free therapy for myocardial infarction (MI). However, the diffusion of exosomes out of the infarcted heart following injection and the low productivity limit the potential of clinical applications. Here, we developed exosome-mimetic extracellular nanovesicles (NVs) derived from iron oxide nanoparticles (IONPs)-incorporated MSCs (IONP-MSCs). The retention of injected IONP-MSC-derived NVs (IONP-NVs) within the infarcted heart was markedly augmented by magnetic guidance. Furthermore, IONPs significantly increased the levels of therapeutic molecules in IONP-MSCs and IONP-NVs, which can reduce the concern of low exosome productivity. The injection of IONP-NVs into the infarcted heart and magnetic guidance induced an early shift from the inflammation phase to the reparative phase, reduced apoptosis and fibrosis, and enhanced angiogenesis and cardiac function recovery. This approach can enhance the therapeutic potency of an MSC-derived NV therapy.

Preclinical assessment of a new live attenuated Mycobacterium tuberculosis Beijing-based vaccine for tuberculosis.

Tuberculosis still claims more lives than any other pathogen, and a vaccine better than BCG is urgently needed. One of the challenges for novel TB vaccines is to protect against all Mycobacterium tuberculosis lineages, including the most virulent ones, such as the Beijing lineage. Here we developed a live attenuated M. tuberculosis mutant derived from GC1237, a Beijing strain responsible for tuberculosis outbreaks in the Canary Islands. The mutant strain is inactivated both in the Rv1503c gene, responsible for surface glycolipid synthesis, and in the two-component global regulator PhoPR. This double mutant is as safe as BCG in immunodeficient SCID mice. In immune-competent mice and guinea pigs, the mutant is as protective as BCG against M. tuberculosis strains of common lineage 4 (Euro-American). By contrast, in mice the vaccine is protective against a M. tuberculosis strain of lineage 2 (East-Asian, Beijing), while BCG is not. These results highlight differences in protection efficacy of live attenuated M. tuberculosis-derived vaccine candidates depending on their genetic background, and provide insights for the development of novel live vaccines against TB, especially in East-Asian countries where M. tuberculosis strains of the Beijing family are highly dominant

Generation of homogeneous midbrain organoids with in vivo-like cellular composition facilitates neurotoxin-based Parkinson\u27s disease modeling

Recent studies have demonstrated the generation of midbrain-like organoids (MOs) from human pluripotent stem cells. However, the low efficiency of MO generation and the relatively immature and heterogeneous structures of the MOs hinder the translation of these organoids from the bench to the clinic. Here we describe the robust generation of MOs with homogeneous distribution of midbrain dopaminergic (mDA) neurons. Our MOs contain not only mDA neurons but also other neuronal subtypes as well as functional glial cells including astrocytes and oligodendrocytes. Furthermore, our MOs exhibit mDA neuron-specific cell death upon treatment with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, indicating that MOs could be a proper human model system for studying the in vivo pathology of Parkinson\u27s disease (PD). Our optimized conditions for producing homogeneous and mature MOs might provide an advanced patient-specific platform for in vitro disease modeling as well as for drug screening for PD

Pharmacogenomic profiling reveals molecular features of chemotherapy resistance in IDH wild-type primary glioblastoma

Background Although temozolomide (TMZ) has been used as a standard adjuvant chemotherapeutic agent for primary glioblastoma (GBM), treating isocitrate dehydrogenase wild-type (IDH-wt) cases remains challenging due to intrinsic and acquired drug resistance. Therefore, elucidation of the molecular mechanisms of TMZ resistance is critical for its precision application. Methods We stratified 69 primary IDH-wt GBM patients into TMZ-resistant (n = 29) and sensitive (n = 40) groups, using TMZ screening of the corresponding patient-derived glioma stem-like cells (GSCs). Genomic and transcriptomic features were then examined to identify TMZ-associated molecular alterations. Subsequently, we developed a machine learning (ML) model to predict TMZ response from combined signatures. Moreover, TMZ response in multisector samples (52 tumor sectors from 18 cases) was evaluated to validate findings and investigate the impact of intra-tumoral heterogeneity on TMZ efficacy. Results In vitro TMZ sensitivity of patient-derived GSCs classified patients into groups with different survival outcomes (P = 1.12e−4 for progression-free survival (PFS) and 3.63e−4 for overall survival (OS)). Moreover, we found that elevated gene expression of EGR4, PAPPA, LRRC3, and ANXA3 was associated to intrinsic TMZ resistance. In addition, other features such as 5-aminolevulinic acid negative, mesenchymal/proneural expression subtypes, and hypermutation phenomena were prone to promote TMZ resistance. In contrast, concurrent copy-number-alteration in PTEN, EGFR, and CDKN2A/B was more frequent in TMZ-sensitive samples (Fishers exact P = 0.0102), subsequently consolidated by multi-sector sequencing analyses. Integrating all features, we trained a ML tool to segregate TMZ-resistant and sensitive groups. Notably, our method segregated IDH-wt GBM patients from The Cancer Genome Atlas (TCGA) into two groups with divergent survival outcomes (P = 4.58e−4 for PFS and 3.66e−4 for OS). Furthermore, we showed a highly heterogeneous TMZ-response pattern within each GBM patient usingin vitro TMZ screening and genomic characterization of multisector GSCs. Lastly, the prediction model that evaluates the TMZ efficacy for primary IDH-wt GBMs was developed into a webserver for public usage (http://www.wang-lab-hkust.com:3838/TMZEP) Conclusions We identified molecular characteristics associated to TMZ sensitivity, and illustrate the potential clinical value of a ML model trained from pharmacogenomic profiling of patient-derived GSC against IDH-wt GBMs

Health Care Burden of Spinal Diseases in the Republic of Korea: Analysis of a Nationwide Database From 2012 Through 2016

Objective This study aimed to determine the incidence and analyze trends related to spinal diseases based on a national database in the Republic of Korea (ROK) and to elucidate the healthcare burden that will serve as a useful resource for researchers, clinicians, and patients. Methods This study was a retrospective analysis of data obtained from Healthcare Bigdata Hub, the Korean Statistical Information Service, and Open Data Portal from 2012 through 2016. The main disease codes for spinal diseases (M40–M54) were used for identification of these conditions. Results The overall annual incidence rates for spinal disease in the ROK was median 15,877 (men, 13,181; women, 18,588) per 100,000 population, and sex ratio was 1:1.41 (p<0.01). The incidence rate and annual costs per patient increased by 7.6% and 14.7% over 5 years continuously, respectively. The age-adjusted incidence rate increased with age; the highest rates were 42.6% in the 75–79 years group. Patients older than 65 years old accounted for median 31.0% of number of patients and 40.1% of medical expenses over 5 years. Lumbar disc herniation (M51) and spinal stenosis (M48) might accounted for both the highest incidence and medical expenses in patients under the age of 60 and over 60 years, respectively. Conclusion The incidence and medical expenditures of spinal disease increased continuously. As the population of ROK in aging, the incidence and medical expenditures due to spondylosis and stenosis (M48) for the old are also increasing. The social burden of spinal diseases in elder patients needs to be prudently considered in health policy makers

KMEDIA-UNIX: An Object-Oriented Hypermedia System Based on the Dexter Reference Model and the MHEG Standard

In this paper, we describe the design and implementation of a hypermedia system that has the following characteristics. First, being designed according to the Dexter hypertext reference model, it has a layered architecture and thus maintains commonality with other hypermedia systems based on the Dexter model. Second, being designed based on the MHEG standard, it has data structures that are inherently suitable for data interchange and synchronization. Third, adopting the MIME protocol, it provides multimedia mail services. Finally, being built on top of an object-oriented DBMS, it makes it easy to represent Dexter and MHEG models and also provides efficient storage and search capabilities. The contribution of this paper is combining these characteristics to build an integrated hypermedia system reflecting reference architectures and international standard efforts. Keywords: hypermedia systems, object-oriented model, Dexter hypertext reference model, MHEG standard, multimedia mail 1 In..

Time- dependent changes of the infarct volume in a rat stroke model: a comparison of the use of MRI and TTC- staining as monitoring tools

Objectives : Serial changes of focal ischemic lesions as seen on magnetic resonance (MR) images and triphenyltetrazolium chloride (TTC)-stained samples of transient middle cerebral artery occlusion in a rat model were evaluated to investigate the natural course of the lesions and the feasibility of the use of each method as a monitoring tool. Methods : Transient middle cerebral artery occlusion (MCAO) was induced in fifteen adult female Sprague Dawley rats using the method of intraluminal vascular occlusion. Two hours after MCAO was induced, reperfusion was performed. Serial MR images were obtained and the volume of the brain infarct was estimated. For macroscopic and microscopic evaluation of the ischemic lesions, the ten animals were sacrificed at different times after MCAO. The rat brains were then removed and six coronal sections were made. Each section was incubated at 37℃ in 2% TTC solution for 15 minutes. Results : Postischemic injury evaluations that were made periodically for eight weeks revealed that the lesion volume as determined from T2 maps had reached a peak on the second day after ischemic injury and the volume decreased afterwards for one week; by the fourth week, the lesion volume again increased to stabilize initial lesion development. There were considerable discrepancies between the infarct area of the samples determined by TTC staining and the in vivo infarct area estimated from the MR images, especially for late stages. Conclusion : T2 map MR images, with a careful consideration of the natural course of infarction development, can provide an adequate and noninvasive means to evaluate the degree of ischemic injury under diverse experimental circumstances.This work was supported by grant from Korean Ministry of Health & Welfare (Grant number: A04-0018-AY1204-06A3-00050B)