AAV-NRIP gene therapy ameliorates motor neuron degeneration and muscle atrophy in ALS model mice.

BACKGROUND: Amyotrophic lateral sclerosis (ALS) is characterized by progressive motor neuron (MN) degeneration, leading to neuromuscular junction (NMJ) dismantling and severe muscle atrophy. The nuclear receptor interaction protein (NRIP) functions as a multifunctional protein. It directly interacts with calmodulin or α-actinin 2, serving as a calcium sensor for muscle contraction and maintaining sarcomere integrity. Additionally, NRIP binds with the acetylcholine receptor (AChR) for NMJ stabilization. Loss of NRIP in muscles results in progressive motor neuron degeneration with abnormal NMJ architecture, resembling ALS phenotypes. Therefore, we hypothesize that NRIP could be a therapeutic factor for ALS. METHODS: We used SOD1 G93A mice, expressing human SOD1 with the ALS-linked G93A mutation, as an ALS model. An adeno-associated virus vector encoding the human NRIP gene (AAV-NRIP) was generated and injected into the muscles of SOD1 G93A mice at 60 days of age, before disease onset. Pathological and behavioral changes were measured to evaluate the therapeutic effects of AAV-NRIP on the disease progression of SOD1 G93A mice. RESULTS: SOD1 G93A mice exhibited lower NRIP expression than wild-type mice in both the spinal cord and skeletal muscle tissues. Forced NRIP expression through AAV-NRIP intramuscular injection was observed in skeletal muscles and retrogradely transduced into the spinal cord. AAV-NRIP gene therapy enhanced movement distance and rearing frequencies in SOD1 G93A mice. Moreover, AAV-NRIP increased myofiber size and slow myosin expression, ameliorated NMJ degeneration and axon terminal denervation at NMJ, and increased the number of α-motor neurons (α-MNs) and compound muscle action potential (CMAP) in SOD1 G93A mice. CONCLUSIONS: AAV-NRIP gene therapy ameliorates muscle atrophy, motor neuron degeneration, and axon terminal denervation at NMJ, leading to increased NMJ transmission and improved motor functions in SOD1 G93A mice. Collectively, AAV-NRIP could be a potential therapeutic drug for ALS.

Interoceptive sensibility differentiates the predictive pattern of emotional reactivity on depression.

The role of emotional reactivity in the psychopathology of depression has been studied widely but not comprehensively. Inconsistencies in existing literature indicate the presence of other factors may affect this dynamic. An individual's method of processing their physiological sensations is a third variable because emotions are psychophysiological. This study identified the predictiveness of ease of activation, intensity, and duration of negative and positive emotions on depressive symptoms differentiated by interoceptive sensibility (IS). A total of 270 community participants filled-in questionnaires assessing their IS, habitual emotional reactivity, depressive severity, and response bias. A two-step clustering analysis identified the IS characteristics. Negative and positive reactivity models among each IS cluster were tested using bootstrapping regression, controlling for gender and response bias. IS can be clustered into "high IS," "low IS," and "worriers." Both positive and negative reactivity's predictiveness patterns on depression were different between IS clusters. Lower positive reactivity predicted depression among individuals with low IS (harder to activate positive emotions) and worriers (shorter duration of positive emotions) but not among individuals with high IS. Those with high IS also exhibited the highest positive reactivity. Ease of activating negative emotions predicted depression among high IS individuals, and a longer duration of negative emotions predicted depression among worriers. IS may affect the psychopathology of depression through subjective emotional reactivity. Thus, IS characteristics can be incorporated into intervention plans.

Do alexithymia and negative affect predict poor sleep quality? The moderating role of interoceptive sensibility.

OBJECTIVES: Emotion-related hyperarousal is an important core pathology of poor sleep. Studies investigating the interplay of alexithymia and affective experiences in determining sleep quality have yielded mixed results. To disentangle the inconsistency, this study examined the concurrent predictive power of alexithymia, and negative and positive affect, while incorporating interoceptive sensibility (IS) as a possible moderator. METHODS: A sample of 224 (70.10% were female) participants completed the Toronto Alexithymia Scale, Positive and Negative Affect Schedule, Pittsburgh Sleep Quality Index, Multidimensional Assessment of Interoceptive Awareness (MAIA), and Marlowe-Crowne Social Desirability Scale (for controlling response bias) using paper and pencil. A two-stage cluster analysis of the MAIA was used to capture IS characteristics. Stepwise regression was conducted separately for each IS cluster. RESULTS: A three-group structure for IS characteristics was found. Higher alexithymia was predictive of poor sleep quality in the low IS group, while higher negative affect predicted poor sleep quality in the moderate and high IS groups. Additionally, alexithymia and positive affect were significantly different in the three IS groups, while negative affect and sleep quality were not. CONCLUSIONS: Emotion and cognitive arousal may impact sleep quality differently in individuals with different levels of internal focusing ability, depending on physiological versus emotional self-conceptualization. The implications on pathological research, clinical intervention, study limitations and future directions are discussed.

Predicting Hypnotic Use among Insomnia Patients with the Theory of Planned Behavior and Craving.

While long-term hypnotic use is very common in clinical practice, the associated factors have been understudied. This study aims to explore the cognitive factors that might influence the long-term use of hypnotics based on the theory of planned behavior (TPB), and examines the moderating effect of craving between cognitive intention and actual hypnotic-use behavior at follow-up. A total of 139 insomnia patients completed a self-constructed TPB questionnaire to measure their attitude, subjective norm, perceived behavioral control, and behavioral intention of hypnotic use, as well as the Hypnotic-Use Urge Scale (HUS) to measure their craving for hypnotics. They were then contacted through phone approximately three months later to assess their hypnotic use. Hierarchical regression showed that perceived behavioral control was the most significant determinant for behavioral intention of hypnotic use. Behavioral intention, in turn, can predict the frequency of hypnotic use after three months. However, this association was moderated by hypnotic craving. The association was lower among the participants with higher cravings for hypnotic use. The findings suggest that the patients' beliefs about their control over sleep and daily life situations, and their craving for hypnotics should be taken into consideration in the management of hypnotic use.

Role of anterior midcingulate cortex in self-reward representation and reward allocation judgments within social context.

Evaluating rewards for the self and others is essential for social interactions. Previous research has probed the neural substrates signaling rewards in social decision-making tasks as well as the differentiation between self- and other-reward representations. However, studies with different designs have yielded mixed results. After analyzing and comparing previous designs, we differentiated three components in this study: task (reward representation vs. social judgment of reward allocation), agency (self vs. other), and social context (without vs. within). Participants were asked to imagine various share sizes as a proposer in a dictator game during fMRI, and then rated their willingness and preference for these offers in a post-scan behavioral task. To differentiate the regions involved in processing rewards without and within context, we presented the reward to each agent in two sequential frames. Parametric analyses showed that, in the second frame (i.e., within social context), the anterior midcingulate cortex (aMCC) signaled self-reward and preferences for the offer, whereas the right insula tracked the likelihood of proposing the offer. Belief in a just world is positively associated with aMCC responses to self-reward. These results shed light on the role of the aMCC in coding self-reward within the social context to guide social behaviors.

Autoantibody of NRIP, a novel AChR-interacting protein, plays a detrimental role in myasthenia gravis.

BACKGROUND: Nuclear receptor interaction protein (NRIP) co-localizes with acetylcholine receptor (AChR) at the neuromuscular junction (NMJ), and NRIP deficiency causes aberrant NMJ architecture. However, the normal physiological and pathophysiological roles of NRIP in NMJ are still unclear. METHODS: We investigated the co-localization and interaction of NRIP with AChR-associated proteins using immunofluorescence and immunoprecipitation assay, respectively. The binding affinity of AChR-associated proteins was analysed in muscle-restricted NRIP knockout mice and NRIP knockout muscle cells (C2C12). We further collected the sera from 43 patients with myasthenia gravis (MG), an NMJ disorder. The existence and features of anti-NRIP autoantibody in sera were studied using Western blot and epitope mapping. RESULTS: NRIP co-localized with AChR, rapsyn and α-actinin 2 (ACTN2) in gastrocnemius muscles of mice; and α-bungarotoxin (BTX) pull-down assay revealed NRIP with rapsyn and ACTN2 in complexes from muscle tissues and cells. NRIP directly binds with α subunit of AChR (AChRα) in vitro and in vivo to affect the binding affinity of AChR with rapsyn and rapsyn with ACTN2. In 43 patients with MG (age, 58.4 ± 14.5 years; female, 55.8%), we detected six of them (14.0%) having anti-NRIP autoantibody. The presence of anti-NRIP autoantibody correlated with a more severe type of MG when AChR autoantibody existed (P = 0.011). The higher the titre of anti-NRIP autoantibody, the more severe MG severity (P = 0.032). The main immunogenic region is likely on the IQ motif of NRIP. We also showed the IgG subclass of anti-NRIP autoantibody mainly to be IgG1. CONCLUSIONS: NRIP is a novel AChRα binding protein and involves structural NMJ formation, which acts as a scaffold to stabilize AChR-rapsyn-ACTN2 complexes. Anti-NRIP autoantibody is a novel autoantibody in MG and plays a detrimental role in MG with the coexistence of anti-AChR autoantibody.

Muscle-restricted nuclear receptor interaction protein knockout causes motor neuron degeneration through down-regulation of myogenin at the neuromuscular junction.

BACKGROUND: Nuclear receptor interaction protein (NRIP) is a calcium/calmodulin (CaM) binding protein. Nuclear receptor interaction protein interacts with CaM to activate calcineurin and CaMKII signalling. The conventional NRIP knockout mice (global knockout) showed muscular abnormality with reduction of muscle oxidative functions and motor function defects. METHODS: To investigate the role of NRIP on neuromuscular system, we generated muscle-restricted NRIP knockout mice [conditional knockout (cKO)]. The muscle functions (including oxidative muscle markers and muscle strength) and lumbar motor neuron functions [motor neuron number, axon denervation, neuromuscular junction (NMJ)] were tested. The laser-captured microdissection at NMJ of skeletal muscles and adenovirus gene therapy for rescued effects were performed. RESULTS: The cKO mice showed muscular abnormality with reduction of muscle oxidative functions and impaired motor performances as global knockout mice. To our surprise, cKO mice also displayed motor neuron degeneration with abnormal architecture of NMJ. Specifically, the cKO mice revealed reduced motor neuron number with small neuronal size in lumbar spinal cord as well as denervating change, small motor endplates, and decreased myonuclei number at NMJ in skeletal muscles. To explore the mechanisms, we screened various muscle-derived factors and found that myogenin is a potential candidate that myogenin expression was lower in skeletal muscles of cKO mice than wild-type mice. Because NRIP and myogenin were colocalized around acetylcholine receptors at NMJ, we extracted RNA from synaptic and extrasynaptic regions of muscles using laser capture microdissection and showed that myogenin expression was especially lower at synaptic region in cKO than wild-type mice. Notably, overexpression of myogenin using intramuscular adenovirus encoding myogenin treatment rescued abnormal NMJ architecture and preserved motor neuron death in cKO mice. CONCLUSIONS: In summary, we demonstrated that deprivation of NRIP decreases myogenin expression at NMJ, possibly leading to abnormal NMJ formation, denervation of acetylcholine receptor, and subsequent loss of spinal motor neuron. Overexpression of myogenin in cKO mice can partially rescue abnormal NMJ architecture and motor neuron death. Therefore, muscular NRIP is a novel trophic factor supporting spinal motor neuron via stabilization of NMJ by myogenin expression.

Quick screening tool for patients with severe negative emotional reactions to chronic illness: psychometric study of the negative emotions due to chronic illness screening test (NECIS).

Background: Severe negative emotional reactions to chronic illness are maladaptive to patients and they need to be addressed in a primary care setting. Objective: The psychometric properties of a quick screening tool-the Negative Emotions due to Chronic Illness Screening Test (NECIS)-for general emotional problems among patients with chronic illness being treated in a primary care setting was investigated. Method: Three studies including 375 of patients with chronic illness were used to assess and analyze internal consistency, test-retest reliability, criterion-related validity, a cut-off point for distinguishing maladaptive emotions and clinical application validity of NECIS. Self-report questionnaires were used. Results: Internal consistency (Cronbach's α) ranged from 0.78 to 0.82, and the test-retest reliability was 0.71 (P < 0.001). Criterion-related validity was 0.51 (P < 0.001). Based on the 'severe maladaptation' and 'moderate maladaptation' groups defined by using the 'Worsening due to Chronic Illness' index as the analysis reference, the receiver-operating characteristic curve analysis revealed an area under the curve of 0.81 and 0.82 (ps < 0.001), and a cut-off point of 19/20 was the most satisfactory for distinguishing those with overly negative emotions, with a sensitivity and specificity of 83.3 and 69.0%, and 68.5 and 83.0%, respectively. The clinical application validity analysis revealed that low NECIS group showed significantly better adaptation to chronic illness on the scales of subjective health, general satisfaction with life, self-efficacy of self-care for disease, illness perception and stressors in everyday life. Conclusion: The NECIS has satisfactory psychometric properties for use in the primary care setting.