A Huntingtin Knockin Pig Model Recapitulates Features of Selective Neurodegeneration in Huntington's Disease.

Huntington's disease (HD) is characterized by preferential loss of the medium spiny neurons in the striatum. Using CRISPR/Cas9 and somatic nuclear transfer technology, we established a knockin (KI) pig model of HD that endogenously expresses full-length mutant huntingtin (HTT). By breeding this HD pig model, we have successfully obtained F1 and F2 generation KI pigs. Characterization of founder and F1 KI pigs shows consistent movement, behavioral abnormalities, and early death, which are germline transmittable. More importantly, brains of HD KI pig display striking and selective degeneration of striatal medium spiny neurons. Thus, using a large animal model of HD, we demonstrate for the first time that overt and selective neurodegeneration seen in HD patients can be recapitulated by endogenously expressed mutant proteins in large mammals, a finding that also underscores the importance of using large mammals to investigate the pathogenesis of neurodegenerative diseases and their therapeutics.

Loss of TDP-43 mediates severe neurotoxicity by suppressing PJA1 gene transcription in the monkey brain.

The nuclear loss and cytoplasmic accumulation of TDP-43 (TAR DNA/RNA binding protein 43) are pathological hallmarks of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Previously, we reported that the primate-specific cleavage of TDP-43 accounts for its cytoplasmic mislocalization in patients' brains. This prompted us to investigate further whether and how the loss of nuclear TDP-43 mediates neuropathology in primate brain. In this study, we report that TDP-43 knockdown at the similar effectiveness, induces more damage to neuronal cells in the monkey brain than rodent mouse. Importantly, the loss of TDP-43 suppresses the E3 ubiquitin ligase PJA1 expression in the monkey brain at transcriptional level, but yields an opposite upregulation of PJA1 in the mouse brain. This distinct effect is due to the species-dependent binding of nuclear TDP-43 to the unique promoter sequences of the PJA1 genes. Further analyses reveal that the reduction of PJA1 accelerates neurotoxicity, whereas overexpressing PJA1 diminishes neuronal cell death by the TDP-43 knockdown in vivo. Our findings not only uncover a novel primate-specific neurotoxic contribution to the loss of function theory of TDP-43 proteinopathy, but also underscore a potential therapeutic approach of PJA1 to the loss of nuclear TDP-43.

Distinctive Patterns of 5-Methylcytosine and 5-Hydroxymethylcytosine in Schizophrenia.

Schizophrenia is a highly heritable neuropsychiatric disorder characterized by cognitive and social dysfunction. Genetic, epigenetic, and environmental factors are together implicated in the pathogenesis and development of schizophrenia. DNA methylation, 5-methycytosine (5mC) and 5-hydroxylcytosine (5hmC) have been recognized as key epigenetic elements in neurodevelopment, ageing, and neurodegenerative diseases. Recently, distinctive 5mC and 5hmC pattern and expression changes of related genes have been discovered in schizophrenia. Antipsychotic drugs that affect 5mC status can alleviate symptoms in patients with schizophrenia, suggesting a critical role for DNA methylation in the pathogenesis of schizophrenia. Further exploring the signatures of 5mC and 5hmC in schizophrenia and developing precision-targeted epigenetic drugs based on this will provide new insights into the diagnosis and treatment of schizophrenia.

Huntington's Disease: Complex Pathogenesis and Therapeutic Strategies.

Huntington's disease (HD) arises from the abnormal expansion of CAG repeats in the huntingtin gene (HTT), resulting in the production of the mutant huntingtin protein (mHTT) with a polyglutamine stretch in its N-terminus. The pathogenic mechanisms underlying HD are complex and not yet fully elucidated. However, mHTT forms aggregates and accumulates abnormally in neuronal nuclei and processes, leading to disruptions in multiple cellular functions. Although there is currently no effective curative treatment for HD, significant progress has been made in developing various therapeutic strategies to treat HD. In addition to drugs targeting the neuronal toxicity of mHTT, gene therapy approaches that aim to reduce the expression of the mutant HTT gene hold great promise for effective HD therapy. This review provides an overview of current HD treatments, discusses different therapeutic strategies, and aims to facilitate future therapeutic advancements in the field.

Aberrant splicing of mutant huntingtin in Huntington's disease knock-in pigs.

Huntington's disease (HD) is an autosomal-dominant inherited neurodegenerative disease caused by a CAG repeat expansion in exon1 of the huntingtin gene (HTT). This expansion leads to the production of N-terminal mutant huntingtin protein (mHtt) that contains an expanded polyglutamine tract, which is toxic to neurons and causes neurodegeneration. While the production of N-terminal mHtt can be mediated by proteolytic cleavage of full-length mHtt, abnormal splicing of exon1-intron1 of mHtt has also been identified in the brains of HD mice and patients. However, the proportion of aberrantly spliced exon1 mHTT in relation to normal mHTT exon remains to be defined. In this study, HTT exon1 production was examined in the HD knock-in (KI) pig model, which more closely recapitulates neuropathology seen in HD patient brains than HD mouse models. The study revealed that aberrant spliced HTT exon1 is also present in the brains of HD pigs, but it is expressed at a much lower level than the normally spliced HTT exon products. These findings suggest that careful consideration is needed when assessing the contribution of aberrantly spliced mHTT exon1 to HD pathogenesis, and further rigorous investigation is required.

Lack of association of somatic CAG repeat expansion with striatal neurodegeneration in HD knock-in animal models.

Our previous work has established a huntingtin knock-in (KI) pig model that displays striatal neuronal loss, allowing us to examine if somatic CAG expansion in striatum accounts for the preferential neurodegeneration in Huntington disease (HD). We found that HD KI pigs do not display somatic CAG expansion in striatum as HD KI mice and that the majority of polyQ repeats in exon 1 HTT in the striatum of HD KI mice are fairly stable. We also found that striatal MSH2 and MLH3, which are involved in DNA repair, are more abundant in mouse brains than pig brains. Consistently inhibiting MSH2 and MLH3 reduced the somatic CAG expansion in HD KI mouse striatum with no influence on neuropathology. Our findings suggest that somatic CAG expansion is species-dependent, occurs in a small fraction of the HD gene in mice, and does not critically contribute to HD neuropathology.

Participation of protein metabolism in cancer progression and its potential targeting for the management of cancer.

Cancer malignancies may broadly be described as heterogeneous disorders manifested by uncontrolled cellular growth/division and proliferation. Tumor cells utilize metabolic reprogramming to accomplish the upregulated nutritional requirements for sustaining their uncontrolled growth, proliferation, and survival. Metabolic reprogramming also called altered or dysregulated metabolism undergoes modification in normal metabolic pathways for anabolic precursor's generation that serves to continue biomass formation that sustains the growth, proliferation, and survival of carcinogenic cells under a nutrition-deprived microenvironment. A wide range of dysregulated/altered metabolic pathways encompassing different metabolic regulators have been described; however, the current review is focused to explain deeply the metabolic pathways modifications inducing upregulation of proteins/amino acids metabolism. The essential modification of various metabolic cycles with their consequent outcomes meanwhile explored promising therapeutic targets playing a pivotal role in metabolic regulation and is successfully employed for effective target-specific cancer treatment. The current review is aimed to understand the metabolic reprogramming of different proteins/amino acids involved in tumor progression along with potential therapeutic perspective elucidating targeted cancer therapy via these targets.

CO2-Repurification Microemulsion Detergent for Oil-Based Slurry Cleaning.

In order to solve problems such as environmental pollution and pipeline blockage caused by oily wastewater after washing, N,N-dimethylcyclohexylamine (DMCHA) with CO2 response was selected as the oil phase, and an O/W microemulsion wellbore cleaning fluid with CO2 switching characteristics was successfully prepared with erucamide propyl betaine (EAB-40), sodium dodecyl benzene sulfonate (SDBS), n-butanol, silicone defoamer, and water. The water content of the microemulsion was 89.99%, and it had good stability at 40 and -5 °C. The emulsion was rapidly demulsified after being injected with CO2 in the CO2-repurification microemulsion detergent, and CO2 was removed with a N2 detergent. The emulsion was restored to its original state, which demonstrated the CO2/N2 switching properties of the emulsion. It is proven that the switching microemulsion has a good wetting transformation ability by cleaning the steel sheet and quartz sheet contaminated by oil-based slurry. The switching microemulsion system can clean the simulated wellbore contaminated by oil-based slurry, and the cleaning efficiency is above 99%. CO2 can be used at room temperature to separate oil and water from oily wastewater after cleaning.

CO2-Responsive Hydrophobic Deep Eutectic Solvent Based on Surfactant-Free Microemulsion-Mediated Synthesis of BaF2 Nanoparticles.

A new form of surfactant-free microemulsion (SFME) including hydrophobic deep eutectic solvent (HDES)/ethanol/water was constructed based on its CO2 response, and three regions, that is, HDES-in-water (HDES/W), bicontinuous (B.C.), and water-in-HDES (W/HDES) regions, were recognized. It is anticipated that SFMEs with tunable microstructures have outstanding applications as nanoreactors in reaction processes. The feasibility of preparing nanoparticles from HDES/ethanol/water SFME using barium fluoride (BaF2) as a model nanoparticle was investigated. HDES-based microemulsions benefit from HDES's excellent properties (novel, low toxicity, CO2-responsive, easy availability) and have potential in universal reactions, drug delivery, advanced material fabrication, etc. In this research, HDES-based microemulsions were prepared using HDES as the oil phase. Phase equilibria and microstructure were investigated using a ternary phase diagram, UV spectrophotometry, and the conductivity method. The CO2 switchable characteristics of the HDES-based microemulsions were investigated. HDES-based microemulsions were proposed as nanoreactors for the synthesis of barium fluoride nanomaterials. The microemulsion structure can modulate the size, morphology, and physicochemical properties of the nanoparticles through the CO2 switchable properties. It is argued that nanoreactors constructed with versatile HDES will offer a new direction for creation of cutting-edge scientific applications.

Multiple-Stimuli-Responsive Surfactant-Free Microemulsions Based on Hydrophobic Deep Eutectic Solvents.

Hydrophobic deep eutectic solvents (HDESs) have been applied to colloidal systems such as microemulsions, despite the development of stimulus-responsive HDESs still being in a preliminary stage. Here, menthol and indole were hydrogen bonded to form CO2-responsiveness HDES. A surfactant-free microemulsion constituted of HDES (menthol-indole) as the hydrophobic phase, water as the hydrophilic phase, and ethanol as the double solvent was demonstrated to be CO2- and temperature-responsive. Dynamic light scattering (DLS) proved the single-phase region of the phase diagram, while conductivity and polarity probing techniques confirmed the kind of microemulsion. The ternary phase diagram and DLS methods were used to investigate the responsiveness of CO2 and effect temperature on the microemulsion drop size and behavior of the phase of the HDES/water/ethanol microemulsion. The findings revealed that when temperature increased, the homogeneous phase region increased. The droplet size in the homogeneous phase region of the associated microemulsion may be reversibly and accurately adjusted by adjusting the temperature. Surprisingly, a slight temperature change can cause a significant phase inversion. Furthermore, in the system, there was no demulsification in time for the CO2/N2 responsiveness process but rather the production of a homogeneous and pellucid aqueous solution.

Differential expression and roles of Huntingtin and Huntingtin-associated protein 1 in the mouse and primate brains.

Huntingtin-associated protein 1 (HAP1) is the first identified protein whose function is affected by its abnormal interaction with mutant huntingtin (mHTT), which causes Huntington disease. However, the expression patterns of Hap1 and Htt in the rodent brain are not correlated. Here we found that the primate HAP1, unlike the rodent Hap1, is correlatively expressed with HTT in the primate brains. CRISPR/Cas9 targeting revealed that HAP1 deficiency in the developing human neurons did not affect neuronal differentiation and gene expression as seen in the mouse neurons. However, deletion of HAP1 exacerbated neurotoxicity of mutant HTT in the organotypic brain slices of adult monkeys. These findings demonstrate differential HAP1 expression and function in the mouse and primate brains, and suggest that interaction of HAP1 with mutant HTT may be involved in mutant HTT-mediated neurotoxicity in adult primate neurons.

Lack of RAN-mediated toxicity in Huntington's disease knock-in mice.

Identification of repeat-associated non-AUG (RAN) translation in trinucleotide (CAG) repeat diseases has led to the emerging concept that CAG repeat diseases are caused by nonpolyglutamine products. Nonetheless, the in vivo contribution of RAN translation to the pathogenesis of CAG repeat diseases remains elusive. Via CRISPR/Cas9-mediated genome editing, we established knock-in mouse models that harbor expanded CAG repeats in the mouse huntingtin gene to express RAN-translated products with or without polyglutamine peptides. We found that RAN translation is not detected in the knock-in mouse models when expanded CAG repeats are expressed at the endogenous level. Consistently, the expanded CAG repeats that cannot be translated into polyglutamine repeats do not yield the neuropathological and behavioral phenotypes that were found in knock-in mice expressing expanded polyglutamine repeats. Our findings suggest that RAN-translated products do not play a major role in the pathogenesis of CAG repeat diseases and underscore the importance in targeting polyglutamine repeats for therapeutics.

Loss of Hap1 selectively promotes striatal degeneration in Huntington disease mice.

Huntington disease (HD) is an ideal model for investigating selective neurodegeneration, as expanded polyQ repeats in the ubiquitously expressed huntingtin (HTT) cause the preferential neurodegeneration in the striatum of the HD patient brains. Here we report that adeno-associated virus (AAV) transduction-mediated depletion of Hap1, the first identified huntingtin-associated protein, in adult HD knock-in (KI) mouse brains leads to selective neuronal loss in the striatum. Further, Hap1 depletion-mediated neuronal loss via AAV transduction requires the presence of mutant HTT. Rhes, a GTPase that is enriched in the striatum and sumoylates mutant HTT to mediate neurotoxicity, binds more N-terminal HTT when Hap1 is deficient. Consistently, more soluble and sumoylated N-terminal HTT is presented in HD KI mouse striatum when HAP1 is absent. Our findings suggest that both Rhes and Hap1 as well as cellular stress contribute to the preferential neurodegeneration in HD, highlighting the involvement of multiple factors in selective neurodegeneration.

Emerging Roles for DNA 6mA and RNA m6A Methylation in Mammalian Genome.

Epigenetic methylation has been shown to play an important role in transcriptional regulation and disease pathogenesis. Recent advancements in detection techniques have identified DNA N6-methyldeoxyadenosine (6mA) and RNA N6-methyladenosine (m6A) as methylation modifications at the sixth position of adenine in DNA and RNA, respectively. While the distributions and functions of 6mA and m6A have been extensively studied in prokaryotes, their roles in the mammalian brain, where they are enriched, are still not fully understood. In this review, we provide a comprehensive summary of the current research progress on 6mA and m6A, as well as their associated writers, erasers, and readers at both DNA and RNA levels. Specifically, we focus on the potential roles of 6mA and m6A in the fundamental biological pathways of the mammalian genome and highlight the significant regulatory functions of 6mA in neurodegenerative diseases.

Huntingtin Interacting Proteins and Pathological Implications.

Huntington's disease (HD) is caused by an expansion of a CAG repeat in the gene that encodes the huntingtin protein (HTT). The exact function of HTT is still not fully understood, and previous studies have mainly focused on identifying proteins that interact with HTT to gain insights into its function. Numerous HTT-interacting proteins have been discovered, shedding light on the functions and structure of HTT. Most of these proteins interact with the N-terminal region of HTT. Among the various HTT-interacting proteins, huntingtin-associated protein 1 (HAP1) and HTT-interacting protein 1 (HIP1) have been extensively studied. Recent research has uncovered differences in the distribution of HAP1 in monkey and human brains compared with mice. This finding suggests that there may be species-specific variations in the regulation and function of HTT-interacting proteins. Understanding these differences could provide crucial insights into the development of HD. In this review, we will focus on the recent advancements in the study of HTT-interacting proteins, with particular attention to the differential distributions of HTT and HAP1 in larger animal models.

Carboxylate Group-Based Phase-Selective Organogelators with a pH-Triggered Recyclable Property.

Phase-selective organogelators (PSOGs) have recently attracted more attention because of their advantages in handling oil spills and leaked organic solvents. However, it is difficult to separate and recover the organic phase and PSOGs from organic gels due to the strong interaction between them. Aiming to enhance the separation and recovery performance of the organic phase and PSOGs, we synthesized a series of pH-responsive PSOGs by using itaconic anhydride and fatty amines with carbon chain lengths of C12-C18. Here, PSOGs have an excellent gelation ability in that amounts of organic solvents and fuel oil can be solidified at a low concentration (<3 wt %). It is worth noting that these gels are stronger, which is more convenient for removal by a salvage operation. More importantly, compared with traditional organogelators, pH-responsive PSOGs can easily recover the organic phase and fuel oil with an adjustment of the pH without extraction or distillation. Because of the transformation between the hydrophilicity and hydrophobicity of PSOGs by pH stimulation, 83.15% PSOGs are recovered in three-cycle experiments. In addition, the recycled PSOGs can be used to realize the removal of the organic phase again. Herein, we find that pH-responsive PSOGs could be used as promising and sustainable materials for separating and recovering organic solvents/oils and PSOGs.

pH-Responsive Regulation of a Surfactant-Free Microemulsion Based on Hydrophobic Deep Eutectic Solvents.

Microemulsions containing a responsive hydrophobic deep eutectic solvent (HDES) as the oil phase that can replace conventional organic solvents are considered to be a green strategy. It is anticipated that a pH-responsive HDES is synthesized to prepare rapid responsive surfactant-free microemulsions (SFMEs), which enable the transition from SFMEs to nanoemulsions. Menthol and n-octanoic acid (OA) were assembled into HDES by hydrogen bonding at a molar ratio of 1:2. The pH-responsive HDES as the oil phase and isopropyl alcohol (IPA) as the double solvent could form HDES/IPA/water SFMEs, which have unique responsiveness. Specifically, from the nuclear magnetic resonance hydrogen spectrum, pH, thermogravimetry, and Fourier transform infrared spectroscopy investigations, the excellent switchability and stability of menthol-OA were demonstrated. On the basis of these complexes, microemulsions were successfully prepared. Electrical conductivity and pH measurements were used to determine the structures of microemulsions and the phase inversion process. The effects of the contents of water and HDES, NaCl concentration, and pH of the system were investigated. Nanoemulsions were successfully prepared on the basis of the pH response of the microemulsions. In addition, the prepared nanoemulsion has a unique pH-responsive behavior that can be controllably regulated among nanoemulsions, microemulsions, and phase separation systems.

Phosphorylation of myelin regulatory factor by PRKG2 mediates demyelination in Huntington's disease.

Demyelination is a common pathological feature of a large number of neurodegenerative diseases including multiple sclerosis and Huntington's disease (HD). Laquinimod (LAQ) has been found to have therapeutic effects on multiple sclerosis and HD. However, the mechanism underlying LAQ's therapeutic effects remains unknown. Using HD mice that selectively express mutant huntingtin in oligodendrocytes and show demyelination, we found that LAQ reduces the Ser259 phosphorylation on myelin regulatory factor (MYRF), an oligodendrocyte-specific transcription factor promoting the expression of myelin-associated genes. The reduced MYRF phosphorylation inhibits MYRF's binding to mutant huntingtin and increases the expression of myelin-associated genes. We also found that PRKG2, a cGMP-activated protein kinase subunit II, promotes the Ser259-MYRF phosphorylation and that knocking down PRKG2 increased myelin-associated protein's expression in HD mice. Our findings suggest that PRKG2-regulated phosphorylation of MYRF is involved in demyelination and can serve as a potential therapeutic target for reducing demyelination.

pH-Induced reversible conversion between non-Pickering and Pickering high internal phase emulsion.

Solid-stabilized high internal phase emulsions have received extensive attention. Many previous studies have confirmed that solid emulsifiers in high internal phase Pickering emulsions (HIPPEs) provide a great interface mechanical barrier. With the development of research, novel solid-stabilized emulsions have emerged. These emulsions are stabilized by the electrostatic repulsion between the surfactants and hydrophilic solid particles. They are distinct from Pickering emulsions in that the solid particles do not exist at the oil-water interface, but are dispersed in the continuous phase, so it is called a non-Pickering emulsion. However, high internal phase non-Pickering emulsions (HIPNPEs) are rarely reported. Herein, HIPNPEs that are synergistically stabilized by anionic surfactants with dynamic covalent bonds and negatively charged nano-SiO2 particles were prepared. In the presence of dodecylamine, the acidity causes the dynamic covalent bonds to break and the surfactant to be inactivated. Additionally, the long-chain amine is protonated and adsorbed on nano-SiO2 particles to form a new surfactant for stabilizing HIPPEs. However, alkalinity causes the HIPNPEs to form again. In addition, rheological tests confirmed that the HIPNPEs and HIPPEs had similar rheological behaviors, which were typical gel-like fluids. The emulsion can quickly respond to realize the conversion between the different types of high internal phase emulsion by simple stimulation, which provides a new direction for stimulus-responsive high internal phase emulsions.

Exposure to urban green space may both promote and harm mental health in socially vulnerable neighborhoods: A neighborhood-scale analysis in New York City.

BACKGROUND: There is growing evidence that exposure to green space can impact mental health, but these effects may be context dependent. We hypothesized that associations between residential green space and mental health can be modified by social vulnerability. METHOD: We conducted an ecological cross-sectional analysis to evaluate the effects of green space exposure on mental disorder related emergency room (ER) visits in New York City at the level of census tract. To objectively represent green space exposure at the neighborhood scale, we calculated three green space exposure metrics, namely proximity to the nearest park, percentage of green space, and visibility of greenness. Using Bayesian hierarchical spatial Poisson regression models, we evaluated neighborhood social vulnerability as a potential modifier of greenness-mental disorder associations, while accounting for the spatially correlated structures. RESULTS: We found significant associations between green space exposure (involving both proximity and visibility) and total ER visits for mental disorders in neighborhoods with high social vulnerability, but no significant associations in neighborhoods with low social vulnerability. We also identified specific neighborhoods with particularly high ER utilization for mental disorders. CONCLUSIONS: Our findings suggest that exposure to green space is associated with ER visits for mental disorders, but that neighborhood social vulnerability can modify this association. Future research is needed to confirm our finding with longitudinal designs at the level of individuals.