Synaptic dysregulation in a human iPS cell model of mental disorders.

Dysregulated neurodevelopment with altered structural and functional connectivity is believed to underlie many neuropsychiatric disorders, and 'a disease of synapses' is the major hypothesis for the biological basis of schizophrenia. Although this hypothesis has gained indirect support from human post-mortem brain analyses and genetic studies, little is known about the pathophysiology of synapses in patient neurons and how susceptibility genes for mental disorders could lead to synaptic deficits in humans. Genetics of most psychiatric disorders are extremely complex due to multiple susceptibility variants with low penetrance and variable phenotypes. Rare, multiply affected, large families in which a single genetic locus is probably responsible for conferring susceptibility have proven invaluable for the study of complex disorders. Here we generated induced pluripotent stem (iPS) cells from four members of a family in which a frameshift mutation of disrupted in schizophrenia 1 (DISC1) co-segregated with major psychiatric disorders and we further produced different isogenic iPS cell lines via gene editing. We showed that mutant DISC1 causes synaptic vesicle release deficits in iPS-cell-derived forebrain neurons. Mutant DISC1 depletes wild-type DISC1 protein and, furthermore, dysregulates expression of many genes related to synapses and psychiatric disorders in human forebrain neurons. Our study reveals that a psychiatric disorder relevant mutation causes synapse deficits and transcriptional dysregulation in human neurons and our findings provide new insight into the molecular and synaptic etiopathology of psychiatric disorders.

G9a and Jhdm2a regulate embryonic stem cell fusion-induced reprogramming of adult neural stem cells.

Somatic nuclei can be reprogrammed to pluripotency through fusion with embryonic stem cells (ESCs). The underlying mechanism is largely unknown, primarily because of a lack of effective approaches to monitor and quantitatively analyze transient, early reprogramming events. The transcription factor Oct4 is expressed specifically in pluripotent stem cells, and its reactivation from somatic cell genome constitutes a hallmark for effective reprogramming. Here we developed a double fluorescent reporter system using engineered ESCs and adult neural stem cells/progenitors (NSCs) to simultaneously and independently monitor cell fusion and reprogramming-induced reactivation of transgenic Oct4-enhanced green fluorescent protein (EGFP) expression. We demonstrate that knockdown of a histone methyltransferase, G9a, or overexpression of a histone demethylase, Jhdm2a, promotes ESC fusion-induced Oct4-EGFP reactivation from adult NSCs. In addition, coexpression of Nanog and Jhdm2a further enhances the ESC-induced Oct4-EGFP reactivation. Interestingly, knockdown of G9a alone in adult NSCs leads to demethylation of the Oct4 promoter and partial reactivation of the endogenous Oct4 expression from adult NSCs. Our results suggest that ESC-induced reprogramming of somatic cells occurs with coordinated actions between erasure of somatic epigenome and transcriptional resetting to restore pluripotency. These mechanistic findings may guide more efficient reprogramming for future therapeutic applications of stem cells. Disclosure of potential conflicts of interest is found at the end of this article.

Monodisperse ordered indium-palladium nanoparticles: synthesis and role of indium for boosting superior electrocatalytic activity for ethanol oxidation reaction.

The slow kinetics of ethanol oxidation reaction (EOR) has limited its widespread use for fuel cells. Bimetallic catalysts with optimized surface compositions can considerably govern rate-determining steps through selectivity for CH3COOH formation or by facilitating the adsorption of OHadsvia the bifunctional effect of an alloy to increase the EOR's kinetic rates. Here, we reported monodisperse ordered In-Pd nanoparticles as new bimetallic high-performance catalysts for EOR. In-Pd nanoparticles, i.e., In3Pd2 and In3Pd5 were prepared using arrested precipitation in solution, and their composition, structures, phase and crystallinity were confirmed using a variety of analyses including TEM, XPS, EDS and XRD. In-Pd nanoparticles were loaded on carbon black (Vulcan XC-72) as electrocatalysts for EOR in alkaline media. In3Pd2 and In3Pd5 nanoparticles exhibited 5.8 times and 4.0 times higher mass activities than commercial Pd/C, which showed that the presence of indium greatly boosts electrocatalytic reactivity for EOR of Pd catalysts. This performance is the best among those of bimetallic nanoparticles reported to date. Such high performance of In-Pd nanoparticles may be attributed to the following two reasons. First, In-Pd nanoparticles exhibited excellent CO anti-poison ability, as confirmed by CO striping experiments. Second, as revealed by DFT calculations of metals with OHads adsorption, In atoms on In3Pd2 surface exhibited the lowest energy (-1.659 eV) for OHads adsorption as compared to other common oxophilic metals including Sn, SnPt, Ag, Ge, Co, Pb, and Cu. We propose that the presence of indium sites promoted efficient free OH radical adsorption on indium sites and resulted in a faster reaction rate of acetate formation from acetaldehyde (the rate determining step for EOR on Pd sites). Finally, a single direct ethanol fuel cell (DEFC) with Pd/C anode was prepared. Compared to the results for a commercial Pd/C anode, the open circuit voltage (OCV) of In3Pd2/C improved by 0.25 V (from 0.64 to 0.89 V) and the power density improved by ∼80% (from 3.7 to 6.7 mW cm-2), demonstrating its practical uses as Pt or Pd catalyst alternatives for DEFC.

Human Parechovirus 3 Meningitis and Fatal Leukoencephalopathy.

Human parechovirus 3 (HPeV3) is a picornavirus associated with neurologic disease in neonates. Human parechovirus 3 infection of preterm and term infants is associated with seizures and destructive periventricular white matter lesions. Despite unremarkable cerebrospinal fluid (CSF), HPeV3 RNA can be amplified from CSF and nasopharyngeal and rectal swabs. We report pathologic findings in 2 autopsy cases of infants with active HPeV3 infection. Both children were born approximately 1 month premature and were neurologically intact but, after a few weeks, developed seizures and radiologic evidence of white matter lesions. Neuropathologic examination demonstrated classic severe periventricular leukomalacia in the absence of an immune response. Human parechovirus 3 sequences were identified in RNA extracted from CSF, sera, and tissues. Human parechovirus 3 in situ hybridization detection of infected cells was limited to meninges and associated blood vessels in addition to smooth muscle of pulmonary vessels. Ultrastructural evaluation of meninges demonstrated dense core structures compatible with picornavirus virions. These findings suggest that encephalopathic changes are secondary to infection of meninges and potential compromise of vascular perfusion. Thus, parechovirus infection of vascular smooth muscle may be a more general pathogenic process.

Astrocytes generated from patient induced pluripotent stem cells recapitulate features of Huntington's disease patient cells.

BACKGROUND: Huntington's Disease (HD) is a devastating neurodegenerative disorder that clinically manifests as motor dysfunction, cognitive impairment and psychiatric symptoms. There is currently no cure for this progressive and fatal disorder. The causative mutation of this hereditary disease is a trinucleotide repeat expansion (CAG) in the Huntingtin gene that results in an expanded polyglutamine tract. Multiple mechanisms have been proposed to explain the preferential striatal and cortical degeneration that occurs with HD, including non-cell-autonomous contribution from astrocytes. Although numerous cell culture and animal models exist, there is a great need for experimental systems that can more accurately replicate the human disease. Human induced pluripotent stem cells (iPSCs) are a remarkable new tool to study neurological disorders because this cell type can be derived from patients as a renewable, genetically tractable source for unlimited cells that are difficult to acquire, such as neurons and astrocytes. The development of experimental systems based on iPSC technology could aid in the identification of molecular lesions and therapeutic treatments. RESULTS: We derived iPSCs from a father with adult onset HD and 50 CAG repeats (F-HD-iPSC) and his daughter with juvenile HD and 109 CAG repeats (D-HD-iPSC). These disease-specific iPSC lines were characterized by standard assays to assess the quality of iPSC lines and to demonstrate their pluripotency. HD-iPSCs were capable of producing phenotypically normal, functional neurons in vitro and were able to survive and differentiate into neurons in the adult mouse brain in vivo after transplantation. Surprisingly, when HD-iPSCs were directed to differentiate into an astrocytic lineage, we observed the presence of cytoplasmic, electron clear vacuoles in astrocytes from both F-HD-iPSCs and D-HD-iPSCs, which were significantly more pronounced in D-HD-astrocytes. Remarkably, the vacuolation in diseased astrocytes was observed under basal culture conditions without additional stressors and increased over time. Importantly, similar vacuolation phenotype has also been observed in peripheral blood lymphocytes from individuals with HD. Together, these data suggest that vacuolation may be a phenotype associated with HD. CONCLUSIONS: We have generated a unique in vitro system to study HD pathogenesis using patient-specific iPSCs. The astrocytes derived from patient-specific iPSCs exhibit a vacuolation phenotype, a phenomenon previously documented in primary lymphocytes from HD patients. Our studies pave the way for future mechanistic investigations using human iPSCs to model HD and for high-throughput therapeutic screens.