AML/T cell interactomics uncover correlates of patient outcomes and the key role of ICAM1 in T cell killing of AML.

T cells are important for the control of acute myeloid leukemia (AML), a common and often deadly malignancy. We observed that some AML patient samples are resistant to killing by human-engineered cytotoxic CD4+ T cells. Single-cell RNA-seq of primary AML samples and CD4+ T cells before and after their interaction uncovered transcriptional programs that correlate with AML sensitivity or resistance to CD4+ T cell killing. Resistance-associated AML programs were enriched in AML patients with poor survival, and killing-resistant AML cells did not engage T cells in vitro. Killing-sensitive AML potently activated T cells before being killed, and upregulated ICAM1, a key component of the immune synapse with T cells. Without ICAM1, killing-sensitive AML became resistant to killing by primary ex vivo-isolated CD8+ T cells in vitro, and engineered CD4+ T cells in vitro and in vivo. While AML heterogeneity implies that multiple factors may determine their sensitivity to T cell killing, these data show that ICAM1 acts as an immune trigger, allowing T cell killing, and could play a role in AML patient survival in vivo.

Epigenetic signature and key transcriptional regulators of human antigen-specific type 1 regulatory T cells.

Human adaptive immunity is orchestrated by effector and regulatory T (Treg) cells. Natural Tregs arise in the thymus where they are shaped to recognize self-antigens, while type 1 Tregs or Tr1 cells are induced from conventional peripheral CD4 + T cells in response to peripheral antigens, such as alloantigens and allergens. Tr1 cells have been developed as a potential therapy for inducing antigen-specific tolerance, because they can be rapidly differentiated in vitro in response to a target antigen. However, the epigenetic landscape and the identity of transcription factors (TFs) that regulate differentiation, phenotype, and functions of human antigen-specific Tr1 cells is largely unknown, hindering Tr1 research and broader clinical development. Here, we reveal the unique epigenetic signature of antigen-specific Tr1 cells, and TFs that regulate their differentiation, phenotype and function. We showed that in vitro induced antigen-specific Tr1 cells are distinct both clonally and transcriptionally from natural Tregs and other conventional CD4 + T cells on a single-cell level. An integrative analysis of Tr1 cell epigenome and transcriptome identified a TF signature unique to antigen-specific Tr1 cells, and predicted that IRF4, BATF, and MAF act as their transcriptional regulators. Using functional genomics, we showed that each of these TFs play a non-redundant role in regulating Tr1 cell differentiation, suppressive function, and expression of co-inhibitory and cytotoxic proteins. By using the Tr1-specific TF signature as a molecular fingerprint, we tracked Tr1 cells in peripheral blood of recipients of allogeneic hematopoietic stem cell transplantation treated with adoptive Tr1 cell therapy. Furthermore, the same signature identified Tr1 cells in resident CD4 + T cells in solid tumors. Altogether, these results reveal the epigenetic signature and the key transcriptional regulators of human Tr1 cells. These data will guide mechanistic studies of human Tr1 cell biology and the development and optimization of adoptive Tr1 cell therapies.

Evidence-based approach to mitigate cumulative stress in pediatric nurses through the development of respite rooms.

BACKGROUND: The cumulative stress toll on nurses increased during the COVID-19 pandemic. An evidence-based practice (EBP) project was conducted to understand what is known about the impacts of cumulative stress within nursing and if there are ways to mitigate stress during a nurse's shift. AIM/IMPLEMENTATION: A project team from three clinical units completed an extensive literature review and identified the need to promote detachment while supporting parasympathetic recovery. Based on this review, leaders from three pediatric clinical units (neonatal intensive care unit, cardiovascular intensive care unit, and acute pulmonary floor) implemented respite rooms. OUTCOMES: Follow-up outcomes showed a statistically significant stress reduction. For all shifts combined, the Wilcoxon Signed-Rank Test revealed that perceived stress scores from an 11-point Likert scale (0 = no stress and 10 = maximum perceived stress) were significantly lower in the post-respite room (Md = 3, n = 68) compared to in the pre-respite room (Md = 6, n = 68), Z = -7.059, p < .001, with a large effect size, r = .605. Nurses and other staff frequently utilized respite rooms during shifts. IMPLICATIONS FOR PRACTICE: Clinical inquiry and evidence-based practice processes can mitigate cumulative stress and support staff wellbeing. Respite rooms within the hospital can promote a healthy work environment among nurses and promote a self-care culture change. Evidence-based strategies to mitigate cumulative stress using respite rooms are a best practice to promote nurse wellbeing and mitigate cumulative stress.

AML/T cell interactomics uncover correlates of patient outcomes and the key role of ICAM1 in T cell killing of AML.

T cells are important for the control of acute myeloid leukemia (AML), a common and often deadly malignancy. We observed that some AML patient samples are resistant to killing by human engineered cytotoxic CD4 + T cells. Single-cell RNA-seq of primary AML samples and CD4 + T cells before and after their interaction uncovered transcriptional programs that correlate with AML sensitivity or resistance to CD4 + T cell killing. Resistance-associated AML programs were enriched in AML patients with poor survival, and killing-resistant AML cells did not engage T cells in vitro . Killing-sensitive AML potently activated T cells before being killed, and upregulated ICAM1 , a key component of the immune synapse with T cells. Without ICAM1, killing-sensitive AML became resistant to killing to primary ex vivo -isolated CD8 + T cells in vitro , and engineered CD4 + T cells in vitro and in vivo . Thus, ICAM1 on AML acts as an immune trigger, allowing T cell killing, and could affect AML patient survival in vivo . SIGNIFICANCE: AML is a common leukemia with sub-optimal outcomes. We show that AML transcriptional programs correlate with susceptibility to T cell killing. Killing resistance-associated AML programs are enriched in patients with poor survival. Killing-sensitive, but not resistant AML activate T cells and upregulate ICAM1 that binds to LFA-1 on T cells, allowing immune synapse formation which is critical for AML elimination.

Dynamic phenotypic heterogeneity and the evolution of multiple RNA subtypes in hepatocellular carcinoma: the PLANET study.

Intra-tumor heterogeneity (ITH) is a key challenge in cancer treatment, but previous studies have focused mainly on the genomic alterations without exploring phenotypic (transcriptomic and immune) heterogeneity. Using one of the largest prospective surgical cohorts for hepatocellular carcinoma (HCC) with multi-region sampling, we sequenced whole genomes and paired transcriptomes from 67 HCC patients (331 samples). We found that while genomic ITH was rather constant across stages, phenotypic ITH had a very different trajectory and quickly diversified in stage II patients. Most strikingly, 30% of patients were found to contain more than one transcriptomic subtype within a single tumor. Such phenotypic ITH was found to be much more informative in predicting patient survival than genomic ITH and explains the poor efficacy of single-target systemic therapies in HCC. Taken together, we not only revealed an unprecedentedly dynamic landscape of phenotypic heterogeneity in HCC, but also highlighted the importance of studying phenotypic evolution across cancer types.

Alloantigen-specific type 1 regulatory T cells suppress through CTLA-4 and PD-1 pathways and persist long-term in patients.

Type 1 regulatory T (Tr1) cells are inducible, interleukin (IL)-10+FOXP3− regulatory T cells that can suppress graft-versus-host disease (GvHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). We have optimized an in vitro protocol to generate a Tr1-enriched cell product called T-allo10, which is undergoing clinical evaluation in patients with hematological malignancies receiving a human leukocyte antigen (HLA)­mismatched allo-HSCT. Donor-derived T-allo10 cells are specific for host alloantigens, are anergic, and mediate alloantigen-specific suppression. In this study, we determined the mechanism of action of T-allo10 cells and evaluated survival of adoptively transferred Tr1 cells in patients. We showed that Tr1 cells, in contrast to the non-Tr1 population, displayed a restricted T cell receptor (TCR) repertoire, indicating alloantigen-induced clonal expansion. Tr1 cells also had a distinct transcriptome, including high expression of cytotoxic T lymphocyte­associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1). Blockade of CTLA-4 or PD-1/PD-L1 abrogated T-allo10­mediated suppression, confirming that these proteins, in addition to IL-10, play key roles in Tr1-suppressive function and that Tr1 cells represent the active component of the T-allo10 product. Furthermore, T-allo10­derived Tr1 cells were detectable in the peripheral blood of HSCT patients up to 1 year after T-allo10 transfer. Collectively, we revealed a distinct molecular phenotype, mechanisms of action, and in vivo persistence of alloantigen-specific Tr1 cells. These results further characterize Tr1 cell biology and provide essential knowledge for the design and tracking of Tr1-based cell therapies.

Electroconvulsive Therapy for Trauma-Related Nightmares: A Case Report and Commentary.

Trauma-related nightmares (TRN), one of the most reported symptoms of posttraumatic stress disorder (PTSD), may not always respond to current pharmacologic and therapeutic treatments. Validity of electroconvulsive therapy (ECT), which is used worldwide in clinical treatment for a broad range of neuropsychiatric conditions, is investigated as a potential therapeutic option for TRN in this report. A case of a 39-year-old male with a history of severe combat-related PTSD, major depressive disorder, history of traumatic brain injury, suicidal ideations, and persistent TRN is discussed here. Successful treatment outcome of this case with six sessions of right unilateral ECT is presented. On initial presentation, the patient had a Patient Health Questionnaire-9 (PHQ-9) score of 27 and a Posttraumatic Stress Disorder Checklist for DSM-5 (PCL-5) score of 77. After six sessions of ECT, the patient had a PHQ-9 score of 3 and a PCL-5 score of 45. Furthermore, the rationale and potential mechanisms of action underlying the ECT treatment for treatment-resistant PTSD and TRN are also reviewed in this report.

Diagnosing Urothelial Carcinoma from Delirium: A Near Miss.

Urinary tract infection (UTI) is a common cause of delirium in the elderly. Although diagnosis of delirium secondary to UTI is relatively straightforward, a lack of thorough investigation could result in missing underlying factors and medical conditions that may require immediate clinical/surgical intervention. Case of a 77-year-old male with delirium diagnosis and multiple psychiatric hospital admissions is reported here. This patient with multiple medical disorders and anxiety was admitted to psychiatric facilities on three different occasions with multiple psychiatric diagnoses including delirium. After a month of hospital stay and thorough medical and radiological examinations, the cause of refractory delirium was identified as multifactorial including urothelial carcinoma. Although UTI and urinary retention are common in the elderly, this case shows the importance of multifactorial diagnoses in cases of prolonged or refractory delirium to avoid delays in appropriate treatment.

Mania Induced by Epidural Steroid Injection in an Elderly Female With No Psychiatric History.

The psychiatric risks of epidural steroid injections for chronic pain in a geriatric patient with no prior psychiatric history are presented here. A 76-year old Caucasian female presented to the emergency department with her family for an inability to sleep, confusion, and behavioral outbursts. The mood instability and psychosis were reported as having started a week after her third epidural steroid injection for low-back pain associated with a prior fall. After 12 days of mixed treatment outcomes and increasing paranoia without any localized neurological findings, the patient was transferred to the geriatric psychiatry unit. Upon admission to the inpatient unit, she was loud, grandiose, verbally aggressive, unable to sleep, hyper-religious, paranoid, and identified her husband and daughter as demons. The patient was started on risperidone and valproic acid for the management of psychosis and manic symptoms. Hyper-religiosity and paranoia greatly improved within a week, though the patient remained very talkative and tangential, with a disorganized thought process. Valproic acid was titrated to 500 mg twice a day, yielding a level of 56.2 ug/ml, accompanied by improvement to mild talkativeness and circumstantiality. She was able to interact appropriately, with minimal lorazepam requirement, and discharged with a linear thought process and absence of psychosis. On outpatient follow up, there were minimal residual mania and no recurrence of psychosis, allowing her to be weaned off valproic acid and to discontinue risperidone. Two months later, symptoms resolved completely. The persistence of this patient's psychosis for nearly one month, and mania for about three months, underscores the importance of careful risk-benefit analysis before initiating epidural steroids. This is particularly important in elderly patients who may be more susceptible to psychiatric adverse effects that can outlast any analgesic benefits.

Attempted Suicides in the Elderly With Existing DNRs: An Emerging Geriatric Ethical Dilemma.

Several critical clinical and ethical issues, including immediate treatment decisions, emerged in a case of a double suicide attempt by an elderly couple with a suicide pact and existing do-not-resuscitate (DNR) documentation. This case was complicated by the advanced age of both patients and their family's expectations and perception of mental illness in the geriatric population. In addition to the myriad of legal and ethical challenges that frequent the end-of-life care, the emerging trend of suicide pacts among the elderly, particularly with existing DNR documentation, warrants further exploration.

Genomic landscape of lung adenocarcinoma in East Asians.

Lung cancer is the world's leading cause of cancer death and shows strong ancestry disparities. By sequencing and assembling a large genomic and transcriptomic dataset of lung adenocarcinoma (LUAD) in individuals of East Asian ancestry (EAS; n = 305), we found that East Asian LUADs had more stable genomes characterized by fewer mutations and fewer copy number alterations than LUADs from individuals of European ancestry. This difference is much stronger in smokers as compared to nonsmokers. Transcriptomic clustering identified a new EAS-specific LUAD subgroup with a less complex genomic profile and upregulated immune-related genes, allowing the possibility of immunotherapy-based approaches. Integrative analysis across clinical and molecular features showed the importance of molecular phenotypes in patient prognostic stratification. EAS LUADs had better prediction accuracy than those of European ancestry, potentially due to their less complex genomic architecture. This study elucidated a comprehensive genomic landscape of EAS LUADs and highlighted important ancestry differences between the two cohorts.

Complications and adequacy of transplant kidney biopsies: A comparison of techniques.

INTRODUCTION: Kidney biopsies are an essential tool in the diagnosis and management of kidney diseases, particularly in kidney transplant recipients. Biopsies carry a risk for serious complications and not all biopsies achieve adequate tissue. We examined the impact of kidney biopsy technique on complications and biopsy adequacy. METHODS: The cohort consisted of consecutive kidney transplant patients undergoing biopsy by one of three techniques: ultrasound localization, real-time ultrasound guidance, and ultrasound-guided trocar placement. Variables of interest included patient characteristics and procedural characteristics. The primary outcome was serious complication attributable to kidney biopsy, and the secondary outcome was biopsy adequacy as defined by Banff criteria. RESULTS: Among 263 patients undergoing biopsy, 27 (10.3%) had a complication (14 with gross hematuria, 10 requiring blood transfusion, 3 requiring an unplanned interventional radiology procedure, 1 kidney loss; no deaths). Complications were more common among patients biopsied using ultrasound-guided trocar compared to real-time ultrasound and ultrasound localization (21.4% vs 7.9% vs 7.1%, respectively, p = 0.008). After adjusting for patient and procedure characteristics, technique was no longer significantly associated with complication. Biopsy adequacy was significantly higher when using ultrasound localization and real-time ultrasound compared to ultrasound-guided trocar (84.6% vs 86.8% vs 69.6%, p = 0.029), and this finding persisted in adjusted analysis. CONCLUSION: Kidney biopsy complications appear to be similar when using any of the three techniques examined in our study. However, ultrasound-guided trocar technique may yield lower biopsy adequacy when compared to non-trocar techniques.

Gender Trends in Academic Radiology Publication in the United States Revisited.

RATIONALE AND OBJECTIVES: Although substantial increases in publications by female academic radiologists have appeared over the last several decades, it is possible that the rate of increase is decreasing. We examined temporal trends in gender composition for full-time radiology faculty, radiology residents, and medical students over a 46-year period. METHODS: We examined authorship gender trends to determine if the increases in female authorship seen since 1970 have been sustained in recent years and whether female radiologists continue to publish in proportion to their numbers in academic departments. Original articles for selected years in Radiology and in the American Journal of Roentgenology between 1970 and 2016 were examined to determine the gender of first, corresponding, and last authors. Generalized linear models evaluated (1) changes in proportions of female authorship over time and (2) associations between proportions of female authorship and female radiology faculty representation. RESULTS: While linear increases in first, corresponding, and senior authorships were observed for female radiologists from 1970 to 2000, the rate of increase in female first and corresponding authorships then changed, with the slope of the first author relationship decreasing from 0.81 to 0.34, corresponding to 47% fewer female first authors added per year. In contrast, the proportion of female last authorship continued to increase at the same rate. The proportion of female first authorship was linearly related to the proportion of female radiology faculty from 1970 to 2016. CONCLUSIONS: Annual increases in first author academic productivity of female radiologists have lessened in the past 16 years, possibly related to reductions in the growth of female radiology faculty and trainees. As mixed, compared to homogeneous gender, authorship teams are associated with more citations, efforts to encourage more women to pursue careers in academic radiology could benefit the radiology research community.

Eye Pain in Children.

Primary care physicians are often the first to see a child with the complaint of eye pain. The eye examination in children is not easy, and the entities that can cause pain in children range from a foreign body in the cornea to the most serious amblyogenic (inducing decreased vision) and life-threatening conditions. Eye pain is a red flag for ocular and systemic conditions, either in a "quiet eye" or "red eye." A detailed history of present illness; pertinent review of systems; close attention to the past medical, family, and social histories; and meticulous physical examination can help to determine the cause, establish the correct treatment, and serve as the basis for referral to a pediatric ophthalmologist if necessary.

Detection of chromosomal breakpoints in patients with developmental delay and speech disorders.

Delineating candidate genes at the chromosomal breakpoint regions in the apparently balanced chromosome rearrangements (ABCR) has been shown to be more effective with the emergence of next-generation sequencing (NGS) technologies. We employed a large-insert (7-11 kb) paired-end tag sequencing technology (DNA-PET) to systematically analyze genome of four patients harbouring cytogenetically defined ABCR with neurodevelopmental symptoms, including developmental delay (DD) and speech disorders. We characterized structural variants (SVs) specific to each individual, including those matching the chromosomal breakpoints. Refinement of these regions by Sanger sequencing resulted in the identification of five disrupted genes in three individuals: guanine nucleotide binding protein, q polypeptide (GNAQ), RNA-binding protein, fox-1 homolog (RBFOX3), unc-5 homolog D (C.elegans) (UNC5D), transmembrane protein 47 (TMEM47), and X-linked inhibitor of apoptosis (XIAP). Among them, XIAP is the causative gene for the immunodeficiency phenotype seen in the patient. The remaining genes displayed specific expression in the fetal brain and have known biologically relevant functions in brain development, suggesting putative candidate genes for neurodevelopmental phenotypes. This study demonstrates the application of NGS technologies in mapping individual gene disruptions in ABCR as a resource for deciphering candidate genes in human neurodevelopmental disorders (NDDs).

Genome-wide analysis reveals methyl-CpG-binding protein 2-dependent regulation of microRNAs in a mouse model of Rett syndrome.

MicroRNAs (miRNAs) are a class of small, noncoding RNAs that function as posttranscriptional regulators of gene expression. Many miRNAs are expressed in the developing brain and regulate multiple aspects of neural development, including neurogenesis, dendritogenesis, and synapse formation. Rett syndrome (RTT) is a progressive neurodevelopmental disorder caused by mutations in the gene encoding methyl-CpG-binding protein 2 (MECP2). Although Mecp2 is known to act as a global transcriptional regulator, miRNAs that are directly regulated by Mecp2 in the brain are not known. Using massively parallel sequencing methods, we have identified miRNAs whose expression is altered in cerebella of Mecp2-null mice before and after the onset of severe neurological symptoms. In vivo genome-wide analyses indicate that promoter regions of a significant fraction of dysregulated miRNA transcripts, including a large polycistronic cluster of brain-specific miRNAs, are DNA-methylated and are bound directly by Mecp2. Functional analysis demonstrates that the 3' UTR of messenger RNA encoding Brain-derived neurotrophic factor (Bdnf) can be targeted by multiple miRNAs aberrantly up-regulated in the absence of Mecp2. Taken together, these results suggest that dysregulation of miRNAs may contribute to RTT pathoetiology and also may provide a valuable resource for further investigations of the role of miRNAs in RTT.