In vivo AAV-SB-CRISPR screens of tumor-infiltrating primary NK cells identify genetic checkpoints of CAR-NK therapy.

Natural killer (NK) cells have clinical potential against cancer; however, multiple limitations hinder the success of NK cell therapy. Here, we performed unbiased functional mapping of tumor-infiltrating NK (TINK) cells using in vivo adeno-associated virus (AAV)-SB (Sleeping Beauty)-CRISPR (clustered regularly interspaced short palindromic repeats) screens in four solid tumor mouse models. In parallel, we characterized single-cell transcriptomic landscapes of TINK cells, which identified previously unexplored subpopulations of NK cells and differentially expressed TINK genes. As a convergent hit, CALHM2-knockout (KO) NK cells showed enhanced cytotoxicity and tumor infiltration in mouse primary NK cells and human chimeric antigen receptor (CAR)-NK cells. CALHM2 mRNA reversed the CALHM2-KO phenotype. CALHM2 KO in human primary NK cells enhanced their cytotoxicity, degranulation and cytokine production. Transcriptomics profiling revealed CALHM2-KO-altered genes and pathways in both baseline and stimulated conditions. In a solid tumor model resistant to unmodified CAR-NK cells, CALHM2-KO CAR-NK cells showed potent in vivo antitumor efficacy. These data identify endogenous genetic checkpoints that naturally limit NK cell function and demonstrate the use of CALHM2 KO for engineering enhanced NK cell-based immunotherapies.

Can dissociative symptoms be explained by coping and emotion regulation? A longitudinal investigation.

OBJECTIVE: Dissociative symptoms are prevalent and are associated with considerable impairments. There are ongoing debates regarding how to conceptualize these phenomena. This study examined whether dissociative symptoms could be explained by coping and emotion regulation strategies. METHOD: We conducted a secondary analysis of data from a randomized controlled trial. A total of 115 Hong Kong Chinese adults completed standardized self-report assessments of trauma exposure, dissociative symptoms, coping, and emotion regulation at baseline. They provided data regarding dissociative symptoms again after 2 months. RESULTS: After controlling for baseline dissociative symptoms and trauma exposure, baseline active coping (ß = -.157, p = .035) and dysfunctional coping (ß = .227, p = .005) predicted more subsequent dissociative symptoms at follow-up. Furthermore, baseline dysfunctional coping mediated the relationship between trauma exposure and subsequent dissociative symptoms. CONCLUSIONS: This study provides insights into the ongoing debate concerning factors that could contribute to dissociation. As dysfunctional coping could particularly predict dissociative symptoms, we call for future studies to evaluate whether coping skills training could effectively prevent and treat dissociative symptoms. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Hurting myself to cope: Using network analysis to understand the association between perceived stigma and non-suicidal self-injury functions among men who have sex with men in China.

Prior studies have demonstrated that perceived stigma is robustly associated with risky and life-threatening behaviors, including non-suicidal self-injury (NSSI), among men who have sex with men (MSM). However, studies regarding the relationship between perceived stigma and NSSI are limited. The present study aimed to investigate the network structure of perceived stigma and NSSI functions, along with bridge nodes, to elucidate how they co-exist. A sample of 2610 Chinese MSM (mean age = 23.99, age range: 18-68 years) was recruited from an online survey platform. All participants completed a web-based survey with measures of perceived stigma and NSSI functions. Results indicated that 'negative attitudes towards homosexuality' and 'disappointment in gay son' were identified as the most central nodes in the perceived stigma network, whereas 'seeking attention' and 'influencing others to change' ranked highest on centrality in the NSSI network. Two bridge connections were exhibited within the combined perceived stigma and NSSI network model: 'unwelcoming in public' and 'avoiding unpleasant tasks' from perceived stigma and NSSI communities, respectively. This is among the first studies investigating the co-occurrence between perceived stigma and NSSI from the network approach. Our findings provide an empirically-based perspective on the importance of family- and community-based interventions, with potential clinical implications for reducing NSSI among sexual and gender minority groups.

Trauma, Emotional Regulation, and Coping Styles in Individuals with and without Probable Dissociative Disorders in Hong Kong.

Previous studies showed that dissociation and dissociative disorders (DDs) are prevalent and are associated with considerable individual and social consequences. There are ongoing debates regarding whether dissociation is a response to betrayal trauma across cultures and whether dissociation can be explained by maladaptive coping. Additionally, little is known about the clinical features of individuals with DDs in the Chinese context. This study aimed to investigate the relationship between trauma, emotional regulation, coping, and dissociation. We analyzed baseline data from a randomized controlled trial (N = 101). Participants with dissociative symptoms in Hong Kong completed self-report assessments. Structured interviews were also conducted subsequently. Participants with probable DDs reported more traumatic events (p = .009 to .017) and exhibited significantly higher levels of dysfunctional coping (p < .001) compared to those who reported dissociative symptoms but did not have a DD. Dissociative symptoms were more strongly associated with betrayal trauma than with non-betrayal trauma. Among different emotion regulation and coping strategies, dysfunctional coping was the only significant factor associated with dissociative symptoms (ß = .309, p = .003). Dysfunctional coping was a statistically significant mediator that may explain the relationship between betrayal trauma and dissociative symptoms. Although other mediation paths are also possible and further longitudinal studies are required, our findings highlight the strong link between dysfunctional coping and dissociative symptoms and suggest that coping skills training should be incorporated into interventions for betrayal trauma survivors with dissociative symptoms. Additionally, this study provides evidence for the cross-cultural validity of the betrayal trauma theory. Further studies, however, are required.

A prospective study of nightmare disorder among Chinese adults in Hong Kong: Persistence and mental health outcomes.

OBJECTIVES: Previous studies showed that nightmares are prevalent and are associated with negative health outcomes. However, no empirical data is available demonstrating the extent to which nightmare disorder persists over time. Current literature provides a limited understanding of the trajectory and wider mental health outcomes of nightmare disorder. This longitudinal study examined the persistence and mental health outcomes of nightmare disorder. METHODS: A total of 230 Hong Kong Chinese adults completed standardized assessments twice with an interval of about 6 months. RESULTS: Over half (66.7%) of the participants with probable nightmare disorder at baseline remained to meet the DSM-5 criteria for the disorder at follow-up. Participants with probable nightmare disorder at baseline were significantly more likely to screen positive for PTSD (82.1% vs 18.3%) (p < .001) (p < .001), and they reported higher rates of mental health service usage at both timepoints (p = .001 to .003). Baseline nightmare disorder severity was negatively associated with subsequent self-rated mental health (ß = -.151, p = .010) and self-esteem (ß = -.141, p = .009) and it also predicted subsequent PTSD symptoms (ß = .122, p = .012). CONCLUSIONS: This study provides first empirical data showing that nightmare disorder could be persistent over time. Nightmare disorder symptoms are associated not only with PTSD symptoms but also with a broader range of mental health issues. This study points to the public health importance of identifying and managing nightmare disorder symptoms in the community. Additionally, the presence of nightmare disorder symptoms may be a helpful indicator for identifying post-traumatic stress.

A Culturally Specific Community Supported Agriculture (CSA) Program to Improve Diet in Immigrant Communities in Brooklyn, New York.

Anti-Asian and anti-immigrant sentiment has surged in the country in the last 3 years. Food insecurity is also on the rise; in our local needs assessment of n = 1,270 Asian American adults in New York City, accessing food was cited as the number 1 priority among those who needed help. Finally, racial discrimination and food access are related to fear of being attacked-driving feelings of safety and therefore willingness to travel for food. To combat these narratives and leveraging pivots by our community partners, we implemented a community-supported agriculture pilot program (n = 38) to assess whether culturally appropriate food access can improve diet and foster cross-cultural learning among immigrant families in Brooklyn, NY. Over a 20-week period from June to October 2022, participants received Chinese-specific produce and nutrition education. Participants reported eating more and a greater variety of vegetables and had higher vegetable intake measured via skin carotenoid scores. This pilot may inform the adaptation of nutrition interventions to reduce inequities in chronic diseases in immigrant communities.

A Comparison of Precision and Safety using Three Recognized Ultrasound-Guided Approaches to Cervical Medial Branch Blocks: A Cadaver Study.

BACKGROUND: Ultrasound (US) guidance is widely used for needle positioning for cervical medial branch blocks (CMBB) and radiofrequency ablation, however, limited research is available comparing different approaches. OBJECTIVE: We aimed to assess the accuracy and safety of 3 different US-guided approaches for CMBB. STUDY DESIGN: A cadaveric study divided into ultrasound-guided needle placement and fluoroscopy evaluation stages. SETTING: Department of Pathology, Forensic, and Insurance Medicine, Semmelweis University. METHODS: Sonographically guided third occipital nerve (TON), C3, C4, C5 and C6 medial branch injections and radiology evaluations were performed.The 3 approaches compared were:1. ES (published by Eichenberger-Siegenthaler): US probe in the coronal plane to visualize the cervical articular pillars, needle approach out of the plane, from anterior to posterior.2. Fi (published by Finlayson): US probe in the transverse plane to visualize a cervical articular pillar and its lamina, needle approach in the plane, from posterior to anterior.3. FiM (Modified Finlayson approach): Needles are placed as in Fi, but then adjusted with a coronal view of the cervical articular pillars.Fluoroscopy images were taken and later evaluated, for "crude", "high precision" and "dangerous" placement. RESULTS: One hundred and fifty-five needle placements were assessed (10 were excluded, as no anterior-posterior fluoroscopy images were saved). Interobserver agreement on position of needle placement between the 5 observers was very high; the Fleiss' Kappa was 0.921. For crude placement, no significant differences were identified between various approaches; (77.6%, 79.5%, and 75.6% for the ES, Fi, and FiM respectively). However, for placement in predefined high-precision zones, ES resulted in significantly more success (ES: 42.9%, Fi: 22.7%, and FiM: 24.4%, P = 0.032). Fi and FiM resulted in no dangerous placements, while ES led to the potential compromise of the exiting nerve root and vertebral artery on three occasions. In 10% of the placements, the levels were identified wrongly, with no difference between the various approaches. LIMITATIONS: Feedback from a live patient, may prevent some existing nerve root injections, unlike in a cadaver. Though a higher number of needles were placed in this study than in most available publications, the number is still low at each individual medial branch level. CONCLUSION: Fi proved safer than ES. Fi was equally successful in targeting the articular pillar, however, ES proved the most successful in placing the needle in the center of the articular pillar. Adding another, (coronal) US view to check needle position in FiM did not improve safety or precision. Identifying CMB levels with the US is challenging with all approaches, therefore we still recommend using fluoroscopy for level identification. While there were pros and cons with either procedure, the efficacy findings of previous papers were not replicated on elderly cadavers with arthritic necks.

Reliability and Mental Health Correlates of a Single-Item Measure of Self-Rated Mental Health (SRMH) in the Chinese Context.

The use of single-item measures of self-rated mental health (SRMH) has been increasingly valued in epidemiologic research. However, little is known about the reliability and mental health correlates of SRMH in Chinese populations. This study examined the reliability and mental health correlates of SRMH in three Chinese samples. We analyzed data collected from two convenience samples of Chinese adults from Hong Kong and/or Taiwan (Sample 1: N = 205; Sample 2: N = 377), and a random sample of Taiwan psychiatric inpatients (Sample 3: N = 100). Our results showed that the single-item measure of SRMH had moderate to good test-retest reliability (intraclass correlation [ICC] = 0.75) in Sample 1 and acceptable reliability between the self-report and interviewer-administered versions (ICC = 0.58) in Sample 3. It had a high positive correlation with self-esteem and a moderately high negative correlation with depression. It also had a consistently negative correlation with borderline personality disorder symptoms and post-traumatic stress disorder symptoms. The SRMH score was also associated with psychiatric service usage. These findings contribute to the body of knowledge regarding the use of a single-item measure of SRMH to assess overall self-perceived mental health in Chinese communities.

COULD FAMILY WELL-BEING MODERATE THE RELATIONSHIP BETWEEN ADVERSE CHILDHOOD EXPERIENCES AND SOMATOFORM DISSOCIATION? A PRELIMINARY INVESTIGATION.

The impacts of adverse childhood experiences (ACEs) have been well documented. One possible consequence of ACEs is dissociation, which is a major feature of post-traumatic psychopathology and is also associated with considerable impairment and health care costs. Although ACEs are known to be associated with both psychoform and somatoform dissociation, much less is known about the mechanisms behind this relationship. Little is known about whether social and interpersonal factors such as family environments would moderate the relationship between ACEs and somatoform dissociation. This paper discusses the importance of having a positive and healthy family environment in trauma recovery. We then report the findings of a preliminary study in which we examined whether the association between ACEs and somatoform dissociation would be moderated by family well-being in a convenience sample of Hong Kong adults (N = 359). The number of ACEs was positively associated with somatoform dissociative symptoms, but this association was moderated by the level of family well-being. The number of ACEs was associated with somatoform dissociation only when the family well-being scores were low. These moderating effects were medium. The findings point to the potential importance of using family education and intervention programs to prevent and treat trauma-related dissociative symptoms, but further investigation is needed.

A Preliminary Longitudinal Analysis of Symptom Management, Post-traumatic Stress, and Depressive Symptoms in Chinese Adults with Dissociative Symptoms.

People with dissociative symptoms are generally poly-symptomatic and require high levels of healthcare resources. Post-traumatic stress disorder (PTSD) and depressive symptoms are two major disabling comorbid symptoms in people with dissociative symptoms. While the sense of control over symptoms may be associated with PTSD and dissociative symptoms, the interplay among these factors over time remains unexplored. This study examined the predictors of PTSD and depressive symptoms in people with dissociative symptoms. Longitudinal data from 61 participants with dissociative symptoms were analyzed. Participants completed self-report measures of dissociative, depressive, and PTSD symptoms and the sense of control over symptoms two times (T1 & T2) with an interval of over one month. PTSD and depressive symptoms were not transient or time-specific, but they persisted over time in our sample. Hierarchical multiple regression analyses revealed that, after controlling for age, treatment usage and baseline symptom severity, T1 symptom management scores (ß = -.264, p = .006) negatively predicted T2 PTSD symptoms, while T1 PTSD symptoms (ß = .268, p = .017) positively predicted T2 depressive symptoms. T1 depressive symptoms (ß = -.087, p = .339) did not predict T2 PTSD symptoms. The findings highlight the importance of improving symptom management skills and treating comorbid PTSD symptoms when working with people with dissociative symptoms.

AAV-mediated delivery of a Sleeping Beauty transposon and an mRNA-encoded transposase for the engineering of therapeutic immune cells.

Engineering cells for adoptive therapy requires overcoming limitations in cell viability and, in the efficiency of transgene delivery, the duration of transgene expression and the stability of genomic integration. Here we report a gene-delivery system consisting of a Sleeping Beauty (SB) transposase encoded into a messenger RNA delivered by an adeno-associated virus (AAV) encoding an SB transposon that includes the desired transgene, for mediating the permanent integration of the transgene. Compared with lentiviral vectors and with the electroporation of plasmids of transposon DNA or minicircle DNA, the gene-delivery system, which we named MAJESTIC (for 'mRNA AAV-SB joint engineering of stable therapeutic immune cells'), offers prolonged transgene expression, as well as higher transgene expression, therapeutic-cell yield and cell viability. MAJESTIC can deliver chimeric antigen receptors (CARs) into T cells (which we show lead to strong anti-tumour activity in vivo) and also transduce natural killer cells, myeloid cells and induced pluripotent stem cells with bi-specific CARs, kill-switch CARs and synthetic T-cell receptors.

Association Between Psychotic and Dissociative Symptoms: Further Investigation Using Network Analysis.

The association and overlap between psychotic and dissociative phenomena have been increasingly recognized. Previous studies found that psychotic symptoms are closely associated with post-traumatic and dissociative symptoms and that these trauma-related phenomena may mediate the relationship between trauma and psychotic symptoms. It remained less explored which specific post-traumatic and dissociative symptom clusters are particularly associated with psychotic symptoms. This cross-sectional study used a data-driven approach (network analysis) to explore the associations among different psychotic and post-traumatic/dissociative symptom clusters in an online convenience predominantly female sample (N = 468)(59.2% had ever seen a psychiatrist). Participants completed well-established multidimensional measures that assessed different symptom clusters of psychosis, dissociation, and PTSD. In addition, multiple mediation analysis was conducted to examine which post-traumatic/dissociative symptoms could mediate the relationship between childhood and adulthood trauma and different psychotic symptoms. Our results confirmed previous findings that PTSD and dissociative symptoms are closely associated with psychotic symptoms. More importantly, both data-driven and multiple mediation analysis results indicated that identity dissociation was particularly associated with perceptual anomalies and bizarre experiences, while emotional constriction was particularly associated with negative symptoms. It is important to screen for trauma and dissociation and provide trauma-and dissociation-informed care when working with people at risk of or experiencing psychosis. Further longitudinal studies using more representative samples are needed.

Substance abuse among mothers in Taiwan: Investigating its prevalence and testing the trauma model.

Substance abuse is a serious problem in our society. In particular, household or parental substance abuse could lead to adverse childhood experiences for the affected children. However, little is known about the prevalence of substance abuse among parents, especially in Asian societies. Moreover, although the trauma model of addiction has significant implications for the prevention and treatment of substance abuse, most evidence comes from Western samples. To address these knowledge gaps, we examined the prevalence of substance abuse in a sample of mothers in Taiwan and tested the trauma model of substance abuse. A total of 867 Taiwanese mothers completed standardized self-report assessments in 2023. The screening results indicated that the 12-month prevalence of drug abuse and alcohol abuse was 12.3% and 7.3%, respectively; 15.5% had either or both drug and alcohol abuse. Most types of trauma exposure were associated with drug abuse severity, while interpersonal stress and dissociative symptoms were associated with alcohol abuse severity, even after controlling for a variety of demographic and health variables as well as general psychopathology (i.e., depressive symptoms). The relatively high prevalence of substance abuse among Taiwanese mothers in our sample is concerning, particularly when compared to the rates reported in other Asian samples. We also provide preliminary cross-cultural evidence supporting the self-medication theory and the trauma model of substance abuse in the Asian context. We discuss the potential importance of addressing trauma and stress to prevent substance abuse and call for follow-up studies.

Dissociative symptoms among community health service users in Hong Kong: a longitudinal study of clinical course and consequences.

Background: Previous studies have demonstrated the high prevalence of dissociative symptoms and their association with considerable healthcare costs. However, there is a lack of studies that describe whether dissociative symptoms persist and lead to other clinical outcomes over time in the community.Objectives: This study investigated the persistence, predictors, and consequences of dissociative symptoms in the community.Methods: We analyzed longitudinal data in a sample of community health service users in Hong Kong (N = 173).Results: A relatively high proportion (63.6%) of participants with baseline dissociative symptoms continued to exhibit dissociative symptoms after approximately 9 months. Baseline non-betrayal trauma predicted subsequent dissociative symptoms (ß = .141, p = .024). Participants with baseline dissociative symptoms were more likely to have received subsequent emergency mental health services (9.1% vs 0.7%, p = .005). Baseline dissociative symptoms significantly predicted subsequent post-traumatic symptoms (ß = .165 to .191, p < .05) and difficulty in social and occupational participation (ß = -.152 to -.182, p < .05) even after controlling for baseline scores, trauma exposure, and use of professional support. The predictive role of dissociative symptoms on subsequent disturbances in self-organization symptoms and social participation difficulty remained significant after applying the Bonferroni correction.Conclusions: This is one of the very few studies showing that dissociative symptoms are persistent to a certain degree and could predict other symptoms and subsequent impairments even in community settings. Factors that affect the trajectory of dissociative symptoms should be further investigated. Regular screening for dissociative symptoms is recommended. Considering its prevalence, persistence, and clinical and social consequences, dissociation should be given greater public health attention.

Dissociative symptoms have been linked to considerable healthcare costs.The persistence and consequences of dissociation in the community had not been previously reported.This study showed that dissociative symptoms persisted to a certain degree and predicted subsequent impairments after approximately 9 months.Dissociation should be given greater public health attention.

Persistence and clinical consequences of post-traumatic and dissociative symptoms in people with depressive symptoms: a one-year follow-up study.

BACKGROUND: Recent studies found that post-traumatic and dissociative symptoms are common in people with depressive symptoms. Although a trauma-related subtype of depression has been proposed, little is known about the persistence and clinical consequences of these symptoms. OBJECTIVE: This one-year follow-up study investigated the persistence and clinical consequences of post-traumatic and dissociative symptoms in people with depressive symptoms. METHODS: We analyzed longitudinal data from an international sample of people self-reporting depressive emotions (N = 152) (mean Patient Health Questionnaire-9 score = 17.27; SD = 6.31). RESULTS: More than half (58.4%) of participants with baseline post-traumatic stress disorder (PTSD) still met the criteria for PTSD after one year. Participants with dissociative symptoms at baseline were significantly more likely to report lifetime psychiatric hospitalization (31.2% vs 14.7%), past-year use of psychiatric hospitalization (10.4% vs 0%) and emergency services (16.9% vs 4%) than those without dissociative symptoms. All post-traumatic and dissociative symptom clusters were cross-sectionally (r = .286 to .528, p < .001) and longitudinally (r = .181 to .462, p < .001) correlated with depressive symptoms. A sense of current threat (ß = .146, p < .05) and negative self-concept (ß = .173, p < .05) at baseline significantly predicted depressive symptoms after one year. CONCLUSIONS: These findings contribute to the increasing body of knowledge regarding the PTSD/dissociation-depression comorbidity. Given their persistence and clinical consequences, we recommend that post-traumatic and dissociative symptoms be regularly screened for in clinical settings. The existence of a possible trauma-related subtype of depression should receive more attention in both research and clinical practice.

Post-traumatic and dissociative symptoms are common in people with depressive symptoms.These symptoms generally persist over one year and predict more depressive symptoms at follow-up.Trauma-related symptoms should be regularly screened for in clinical settings.

Borderline personality disorder features and their relationship with trauma and dissociation in a sample of community health service users.

BACKGROUND: Previous studies have indicated that borderline personality disorder (BPD) is closely associated with trauma and dissociation. Nevertheless, BPD is a heterogeneous condition, and not all people with BPD have severe dissociation. This study examined whether the relationship of BPD features with trauma and dissociation would remain significant after controlling for some general non-specific mental health distress. We also made the first attempt to explore which specific BPD features would be particularly associated with dissociation. METHODS: We analyzed survey data from a sample of community health service users in Hong Kong (N = 376). Hierarchical multiple regression and data-driven network analysis were used. RESULTS: The lifetime prevalence of DSM-5 BPD was 16.0% in our sample. Of participants who met criteria for BPD, 43.3% scored above cutoff on the dissociation measures, thus possibly having clinically significant dissociative symptoms. BPD features were associated with adulthood trauma and psychoform dissociation even after controlling for age, depression and self-esteem. Network analysis showed that some BPD features - including impulsivity, identity disturbance and suicidal/self-mutilation behaviors - were particularly associated with dissociation; other BPD features such as interpersonal-related problems had relatively weak to no connection with dissociation. CONCLUSIONS: Our results suggested that some particular BPD features might be dissociative in nature, although further longitudinal research is required. We argue that a trauma-informed perspective should be employed when working with clients presenting with BPD features despite these features being commonly stigmatized. Further research on the intervention needs of the people with BPD who suffer from high levels of dissociation is required.

The effects of a family-focused dyadic psychoeducational intervention for stroke survivors and their family caregivers: A randomised controlled trial.

BACKGROUND: Stroke can cause a variety of physical and psychosocial disturbances for both survivors and their family caregivers (i.e., stroke dyads). Dyadic psychoeducation appears to be a promising approach for providing knowledge of stroke and self-care or caregiving skills to improve stroke dyads' health outcomes. Therefore, a family-focused psychoeducation intervention was designed and tested to improve the health outcomes of stroke dyads. OBJECTIVE: To examine the effects of a family-focused dyadic psychoeducational intervention on the functional and psychosocial outcomes of stroke survivors and family caregivers. DESIGN: A single-blinded, parallel-group randomised controlled trial with repeated-measures design. SETTINGS: Two general hospitals and one rehabilitation facility in Jinan, China. PARTICIPANTS: Stroke survivors and family caregivers (N = 162 dyads). METHODS: The dyads were randomly allocated to either psychoeducation or control group with usual care only (N = 81 dyads per group). The intervention included three structured face-to-face education sessions (one hour per session) in hospital pre-discharge and four weekly telephone counselling calls post-discharge. Study outcomes included survivor functioning and caregiver burden (primary outcomes), and other secondary outcomes (i.e., caregiving competence, dyads' coping, depressive and anxiety symptoms, family functioning, and dyadic relationship, as well as survivor healthcare utilisation and caregiving-related injury). Data were collected at baseline (T0) and immediately (T1) and 3 months post-intervention (T2). The intervention effects were estimated using generalised estimating equation models. RESULTS: Participants in the psychoeducation group revealed significantly greater reductions on caregiver burden than the control group at T1 (ß = -6.01, p = 0.026) and T2 (ß = -6.73, p = 0.039), but non-significant effects on survivor functioning, except in emotion domain at T1 (ß = 7.22, p = 0.015). In addition, the intervention demonstrated significantly greater improvements on caregiving competence (ß = 0.98, p = 0.013; ß = 1.58, p < 0.001), survivors' depressive symptoms (ß = -1.56, p = 0.007; ß = -2.06, p = 0.005), and dyadic relationship (ß = 0.26, p = 0.012; ß = 0.27, p = 0.022) at T1 and T2, as well as on survivor coping at T2 (ß = 6.73, p = 0.008). CONCLUSIONS: Our study added values on the benefits of family-focused dyadic psychoeducation to routine stroke rehabilitation and family care. Future research can evaluate its long-term effects for families of stroke survivors with diverse socio-demographic and stroke-related characteristics. REGISTRATION: Chinese Clinical Trial Registry (ChiCTR2100042684). Recruitment: March to June 2021.

Perturbomics of tumor-infiltrating NK cells.

Natural killer (NK) cells are an innate immune cell type that serves at the first level of defense against pathogens and cancer. NK cells have clinical potential, however, multiple current limitations exist that naturally hinder the successful implementation of NK cell therapy against cancer, including their effector function, persistence, and tumor infiltration. To unbiasedly reveal the functional genetic landscape underlying critical NK cell characteristics against cancer, we perform perturbomics mapping of tumor infiltrating NK cells by joint in vivo AAV-CRISPR screens and single cell sequencing. We establish a strategy with AAV-SleepingBeauty(SB)- CRISPR screening leveraging a custom high-density sgRNA library targeting cell surface genes, and perform four independent in vivo tumor infiltration screens in mouse models of melanoma, breast cancer, pancreatic cancer, and glioblastoma. In parallel, we characterize single-cell transcriptomic landscapes of tumor-infiltrating NK cells, which identifies previously unexplored sub-populations of NK cells with distinct expression profiles, a shift from immature to mature NK (mNK) cells in the tumor microenvironment (TME), and decreased expression of mature marker genes in mNK cells. CALHM2, a calcium homeostasis modulator that emerges from both screen and single cell analyses, shows both in vitro and in vivo efficacy enhancement when perturbed in chimeric antigen receptor (CAR)-NK cells. Differential gene expression analysis reveals that CALHM2 knockout reshapes cytokine production, cell adhesion, and signaling pathways in CAR- NKs. These data directly and systematically map out endogenous factors that naturally limit NK cell function in the TME to offer a broad range of cellular genetic checkpoints as candidates for future engineering to enhance NK cell-based immunotherapies.