Screening for harmful substance use in emergency departments: a systematic review.

BACKGROUND: Substance use-related emergency department (ED) visits have increased substantially in North America. Screening for substance use in EDs is recommended; best approaches are unclear. This systematic review synthesizes evidence on diagnostic accuracy of ED screening tools to detect harmful substance use. METHODS: We included derivation or validation studies, with or without comparator, that included adult (≥ 18 years) ED patients and evaluated screening tools to identify general or specific substance use disorders or harmful use. Our search strategy combined concepts Emergency Department AND Screening AND Substance Use. Trained reviewers assessed title/abstracts and full-text articles for inclusion, extracted data, and assessed risk of bias (QUADAS-2) independently and in duplicate. Reviewers resolved disagreements by discussion. Primary investigators adjudicated if necessary. Heterogeneity precluded meta-analysis. We descriptively summarized results. RESULTS: Our search strategy yielded 2696 studies; we included 33. Twenty-one (64%) evaluated a North American population. Fourteen (42%) applied screening among general ED patients. Screening tools were administered by research staff (n = 21), self-administered by patients (n = 10), or non-research healthcare providers (n = 1). Most studies evaluated alcohol use screens (n = 26), most commonly the Alcohol Use Disorders Identification Test (AUDIT; n = 14), Cut down/Annoyed/Guilty/Eye-opener (CAGE; n = 13), and Rapid Alcohol Problems Screen (RAPS/RAPS4/RAPS4-QF; n = 12). Four studies assessing six tools and screening thresholds for alcohol abuse/dependence in North American patients (AUDIT ≥ 8; CAGE ≥ 2; Diagnostic and Statistical Manual of Mental Disorders, 4th Edition [DSM-IV-2] ≥ 1; RAPS ≥ 1; National Institute on Alcohol Abuse and Alcoholism [NIAAA]; Tolerance/Worry/Eye-opener/Amnesia/K-Cut down [TWEAK] ≥ 3) reported both sensitivities and specificities ≥ 83%. Two studies evaluating a single alcohol screening question (SASQ) (When was the last time you had more than X drinks in 1 day?, X = 4 for women; X = 5 for men) reported sensitivities 82-85% and specificities 70-77%. Five evaluated screening tools for general substance abuse/dependence (Relax/Alone/Friends/Family/Trouble [RAFFT] ≥ 3, Drug Abuse Screening Test [DAST] ≥ 4, single drug screening question, Alcohol, Smoking and Substance Involvement Screening Test [ASSIST] ≥ 42/18), reporting sensitivities 64%-90% and specificities 61%-100%. Studies' risk of bias were mostly high or uncertain. CONCLUSIONS: Six screening tools demonstrated both sensitivities and specificities ≥ 83% for detecting alcohol abuse/dependence in EDs. Tools with the highest sensitivities (AUDIT ≥ 8; RAPS ≥ 1) and that prioritize simplicity and efficiency (SASQ) should be prioritized.

Artificial-intelligence-driven measurements of brain metastases' response to SRS compare favorably with current manual standards of assessment.

Background: Evaluation of treatment response for brain metastases (BMs) following stereotactic radiosurgery (SRS) becomes complex as the number of treated BMs increases. This study uses artificial intelligence (AI) to track BMs after SRS and validates its output compared with manual measurements. Methods: Patients with BMs who received at least one course of SRS and followed up with MRI scans were retrospectively identified. A tool for automated detection, segmentation, and tracking of intracranial metastases on longitudinal imaging, MEtastasis Tracking with Repeated Observations (METRO), was applied to the dataset. The longest three-dimensional (3D) diameter identified with METRO was compared with manual measurements of maximum axial BM diameter, and their correlation was analyzed. Change in size of the measured BM identified with METRO after SRS treatment was used to classify BMs as responding, or not responding, to treatment, and its accuracy was determined relative to manual measurements. Results: From 71 patients, 176 BMs were identified and measured with METRO and manual methods. Based on a one-to-one correlation analysis, the correlation coefficient was R2 = 0.76 (P = .0001). Using modified BM response classifications of BM change in size, the longest 3D diameter data identified with METRO had a sensitivity of 0.72 and a specificity of 0.95 in identifying lesions that responded to SRS, when using manual axial diameter measurements as the ground truth. Conclusions: Using AI to automatically measure and track BM volumes following SRS treatment, this study showed a strong correlation between AI-driven measurements and the current clinically used method: manual axial diameter measurements.

Radiation and Immune Checkpoint Inhibitors: Combination Therapy for Treatment of Hepatocellular Carcinoma.

The liver tumor immune microenvironment has been thought to possess a critical role in the development and progression of hepatocellular carcinoma (HCC). Despite the approval of immune checkpoint inhibitors (ICIs), such as programmed cell death receptor 1 (PD-1)/programmed cell death ligand 1 (PD-L1) and cytotoxic T lymphocyte associated protein 4 (CTLA-4) inhibitors, for several types of cancers, including HCC, liver metastases have shown evidence of resistance or poor response to immunotherapies. Radiation therapy (RT) has displayed evidence of immunosuppressive effects through the upregulation of immune checkpoint molecules post-treatment. However, it was revealed that the limitations of ICIs can be overcome through the use of RT, as it can reshape the liver immune microenvironment. Moreover, ICIs are able to overcome the RT-induced inhibitory signals, effectively restoring anti-tumor activity. Owing to the synergetic effect believed to arise from the combination of ICIs with RT, several clinical trials are currently ongoing to assess the efficacy and safety of this treatment for patients with HCC.

Quality of Life Before and After Radiation Therapy for Early-Stage Keratinocyte Carcinoma: A Longitudinal Analysis.

PURPOSE: Little is known about patient-reported quality of life (QOL) before and after radiotherapy (RT) for early keratinocyte carcinoma (KC). The present study tested the hypothesis that QOL is hypothesized to deteriorate soon after RT, with subsequent improvement of a magnitude similar to other KC treatments. METHODS AND MATERIALS: This observational study assessed prospectively collected longitudinal patient-reported QOL data at a single center. Patients undergoing external beam RT for early-stage (T0-2, <4 cm) KC that completed the Skindex-16 (S16) or Skin Cancer Index (SCI) before and at least once after RT were included. These indices include subscales of symptoms, emotions, and functioning for S16 and emotional, social, and appearance for SCI. RESULTS: 81 patients (median age 79 years) were treated for early KC, most being basal cell carcinomas (87%) on the face (80%) an average of 1.3 cm in diameter. Composite S16 scores and total SCI scores showed significant, clinically meaningful improvements at 3 months and 6 weeks after RT, respectively. QOL declined at 6 weeks after treatment on the S16 symptoms subscale. Concurrently, S16 and SCI emotional subscales significantly improved, and this trend persisted for 2+ years. Improvements in S16 functioning and SCI social and appearance QOL occurred after RT. The magnitude of improvement in QOL 3-12 months after RT was comparable to what has been previously reported after other types of KC treatments, like surgery. CONCLUSIONS: QOL related to symptoms deteriorates during the first 6 weeks after RT for early KC, but it improves by 3 months. QOL related to emotions, functioning, social, and appearance all improve after RT. These changes in QOL are similar to those observed after other treatments of early KC.

Racialization, colonialism, and imperialism: a critical autoethnography on the intersection of forced displacement and race in a settler colonial context.

Migration has been identified as a priority area for policy responses by both the federal and provincial/territorial governments yet, much of our knowledge about migration is not premised on addressing current xenophobic and racist narratives about migrants. The purpose of this research is an interrogation of Canada's colonialism, imperialism, and racialization, which produce specific oppressive policies and practices that have impacted my family. This research is premised on the understanding that in the space between what is known about migration in Canada and what is not, a great deal of narrative and interpretive work is done that makes assumptions about migrants, specifically forcibly displaced people from the Global South. Through a critical autoethnography focused on my lived experiences as a descendant of forcibly displaced Chinese-Vietnamese people living in a settler colonial nation state, this study critiques to what extent these assumptions are founded, and to what extent they represent a socio-political climate in which migration is set out as particular problems requiring a legal and policing solution. In particular, my analysis centers anti-colonialism and anti-racism, shifting to resistance to systemic violence and liberation, while considering the discursive and on-the-ground effects of racist, colonial, and imperial policies and practice. Set against the backdrop of the rise of white nationalism, xenophobia, and racism across all levels of government and academia, and the general public, the results of this study produce a counter-narrative focused on the intersection of forced displacement and race in a settler colonial context, which is both timely and urgent.

Ethnic/racial differences in risk factors and clinical outcomes among patients with amyloidosis.

BACKGROUND: Cardiac amyloidosis is caused by abnormal extracellular deposition of insoluble fibrils in cardiac tissue. It can be fatal when untreated and is often underdiagnosed. Understanding the ethnic/racial differences in risk factors is critical for early diagnosis and treatment to improve clinical outcomes. METHODS: We performed a retrospective cross-sectional study utilizing the National Inpatient Sample database from 2015 to 2018 using ICD-10-CM codes. The primary variables of interest were race/ethnicity and amyloidosis subtypes, while the primary outcomes were in-hospital mortality, gastrointestinal bleeding, renal failure, and hospital length-of-stay. RESULTS: Amyloidosis was reported in 0.17% of all hospitalizations (N  =  19,678,415). Of these, 0.09% were non-Hispanic whites, 0.04% were non-Hispanic blacks, and 0.02% were Hispanic. Hospitalizations with ATTR amyloidosis subtype were frequently observed in older individuals and males with coronary artery disease, whereas AL amyloidosis subtype was associated with non-Hispanic whites, congestive heart failure, and longer hospital length of stay. Renal failure was associated with non-Hispanic blacks (adjusted relative risk [RR]  =  1.31, p < 0.001), Hispanics (RR  =  1.08, p  =  0.028) and had an increased risk of mortality. Similarly, the hospital length of stay was longer with non-Hispanic blacks (RR  =  1.19, p < 0.001) and Hispanics (RR  =  1.05, p  =  0.03) compared to non-Hispanic whites. Hispanics had a reduced risk of mortality (RR  =  0.77, p  =  0.028) compared to non-Hispanic whites and non-Hispanic blacks, and no significant difference in mortality was seen between non-Hispanic whites and non-Hispanic blacks (RR  =  1.00, p  =  0.963). CONCLUSIONS: Our findings highlight significant ethnic/racial differences in risk factors and outcomes among amyloidosis-related US hospitalizations that can possibly be used for early detection, treatment, and better clinical outcomes.

The effects of immediate recall and subsequent retrieval strategy on eyewitness memory.

The current study investigated the effects of immediate recall and subsequent interview on mock witnesses' memory of a crime. After watching a video of a crime, 111 participants were randomly allocated to an immediate recall condition (the iWitnessed mobile app, free recall). One week later, participants were randomly allocated to a retrieval condition (timeline aid, category clustering recall, free recall). Results showed that iWitnessed elicited more correct details than free recall at immediate recall (Time 1), without compromising accuracy. However, the immediate recall tool did not affect memory performance at a one-week delay (Time 2). At Time 2, participants with retrieval support (timeline aid, category clustering recall) reported more details than those with no retrieval support (free recall), without compromising accuracy. The findings highlight that high-quality immediate recall and retrieval support during a subsequent interview are necessary to elicit more complete and accurate accounts.

Genomic epidemiology of Mycobacterium abscessus in a Canadian cystic fibrosis centre.

The Mycobacterium abscessus complex causes significant morbidity and mortality among patients with Cystic Fibrosis (CF). It has been hypothesized that these organisms are transmitted from patient to patient based on genomics. However, few studies incorporate epidemiologic data to confirm this hypothesis. We longitudinally sampled 27 CF and 7 non-CF patients attending a metropolitan hospital in Ontario, Canada from 2013 to 2018. Whole genome sequencing along with epidemiological data was used to evaluate the likelihood of transmission. Overall, the genetic diversity of M. abscessus was large, with a median pairwise distance (IQR) of 1,279 (143-134) SNVs between all Ontario M. abscessus isolates and 2,908 (21-3,204) single nucleotide variants (SNVs) between M. massiliense isolates. This reflects the global diversity of this pathogen, with Ontario isolates widely dispersed throughout global phylogenetic trees of each subspecies. Using a maximum distance of 25 SNVs as a threshold to identify possible transmission, we identified 23 (of 276 total) pairs of closely-related isolates. However, transmission was probable for only one pair based on both genomic and epidemiological data. This suggests that person-to-person transmission of M. abscessus among CF patients is indeed rare and reinforces the critical importance of epidemiological data for inferences of transmission.

Consumer Perspectives on the Use of Artificial Intelligence Technology and Automation in Crisis Support Services: Mixed Methods Study.

BACKGROUND: Emerging technologies, such as artificial intelligence (AI), have the potential to enhance service responsiveness and quality, improve reach to underserved groups, and help address the lack of workforce capacity in health and mental health care. However, little research has been conducted on the acceptability of AI, particularly in mental health and crisis support, and how this may inform the development of responsible and responsive innovation in the area. OBJECTIVE: This study aims to explore the level of support for the use of technology and automation, such as AI, in Lifeline's crisis support services in Australia; the likelihood of service use if technology and automation were implemented; the impact of demographic characteristics on the level of support and likelihood of service use; and reasons for not using Lifeline's crisis support services if technology and automation were implemented in the future. METHODS: A mixed methods study involving a computer-assisted telephone interview and a web-based survey was undertaken from 2019 to 2020 to explore expectations and anticipated outcomes of Lifeline's crisis support services in a nationally representative community sample (n=1300) and a Lifeline help-seeker sample (n=553). Participants were aged between 18 and 93 years. Quantitative descriptive analysis, binary logistic regression models, and qualitative thematic analysis were conducted to address the research objectives. RESULTS: One-third of the community and help-seeker participants did not support the collection of information about service users through technology and automation (ie, via AI), and approximately half of the participants reported that they would be less likely to use the service if automation was introduced. Significant demographic differences were observed between the community and help-seeker samples. Of the demographics, only older age predicted being less likely to endorse technology and automation to tailor Lifeline's crisis support service and use such services (odds ratio 1.48-1.66, 99% CI 1.03-2.38; P<.001 to P=.005). The most common reason for reluctance, reported by both samples, was that respondents wanted to speak to a real person, assuming that human counselors would be replaced by automated robots or machine services. CONCLUSIONS: Although Lifeline plans to always have a real person providing crisis support, help-seekers automatically fear this will not be the case if new technology and automation such as AI are introduced. Consequently, incorporating innovative use of technology to improve help-seeker outcomes in such services will require careful messaging and assurance that the human connection will continue.

Help-seeker expectations and outcomes of a crisis support service: Comparison of suicide-related and non-suicide-related contacts to lifeline Australia.

Lifeline Australia aims to prevent suicide and support community members in personal crisis via the provision of free anonymous telephone, online chat and text message services. This study aimed to identify the expectations and outcomes of Lifeline help-seekers, including whether there are differences between suicide-related and non-suicide-related contacts. Help-seekers (N = 553) who had previously contacted Lifeline via telephone, online chat, or text message crisis services were recruited via social media and a link provided after Lifeline service use, who completed an online survey about their awareness, expectations and outcomes of Lifeline's services. The responses from help-seekers who self-reported suicide-related and non-suicide-related reasons for contact were compared. Participants were highly aware of Lifeline's services, particularly the phone service. The main expectations of all help-seekers were to feel heard and listened to, feel less upset and feel understood. There were 59.5% of the sample that reported suicidality as a reason for contact. Suicide-related contacts endorsed more reasons for contact than non-suicide-related contacts. Expectations of suicide-related help-seekers were greater, but they were less likely to report that their expectations were met. The high expectations and complexity of suicide-related contacts reveal the challenges in meeting the needs of this high-priority group, particularly within the context of the multiple demands on crisis support services.

Emergency department return visits and hospital admissions in trauma team assessed patients initially discharged from the emergency department: A population-based cohort study.

BACKGROUND: Many injured patients are transported directly to trauma centers, found to be minimally injured, and discharged directly home from the emergency department (ED). Our objectives were to characterize the short-term outcomes in this discharged patient population and to identify patient factors predictive of ED return visits. METHODS: We conducted a retrospective population-based cohort study using linked administrative data sets involving patients assessed at trauma centers in Ontario, Canada between April 1, 2009, and March 31, 2020. Patients who were assessed by a trauma team and discharged directly home from ED were included. The primary outcome was the percentage of patients with an ED return visit within 14 days. We used multivariate logistic regression analyses to identify patient characteristics predictive of at least one ED return visit. RESULTS: There were 5,550 patients included in the study. A total of 1,004 (18.1%) of patients had at least one ED return visit, but only 100 patients (1.8%) were admitted to hospital following initial discharge. Common reasons for ED return visits included wound care concerns (17.2%), head injury complaints (15.6%), and substance misuse (6.8%). Rural residence (odds ratio [OR], 1.83; 95% CI, 1.45-2.29), history of anxiety disorder (OR, 2.05; 95% CI, 1.54-2.73), high baseline ED usage (OR, 2.58; 95% CI, 2.03-3.28), penetrating injury (OR, 1.42; 95% CI, 1.20-1.68), and extremity fracture (OR, 1.52; 95% CI, 1.24-1.88) predicted return visits. CONCLUSION: Patients discharged directly have high rates of ED return visits but low rates of hospital admission or delayed surgical intervention. Trauma services should expand quality assurance initiatives to capture return visits, understand any gaps in clinical service provision, and aim to minimize unnecessary ED return visits. LEVEL OF EVIDENCE: Prognostic/Epidemiological; Level IV.

PD-1 and TIGIT downregulation distinctly affect the effector and early memory phenotypes of CD19-targeting CAR T cells.

CD19-targeting chimeric antigen receptor (CAR) T cells have become an important therapeutic option for patients with relapsed and refractory B cell malignancies. However, a significant portion of patients still do not benefit from the therapy owing to various resistance mechanisms, including high expression of multiple inhibitory immune checkpoint receptors. Here, we report a lentiviral two-in-one CAR T approach in which two checkpoint receptors are downregulated simultaneously by a dual short hairpin RNA cassette integrated into a CAR vector. Using this system, we evaluated CD19-targeting CAR T cells in the context of four different checkpoint combinations-PD-1/TIM-3, PD-1/LAG-3, PD-1/CTLA-4, and PD-1/TIGIT-and found that CAR T cells with PD-1/TIGIT downregulation uniquely exerted synergistic antitumor effects. Importantly, functional and phenotypic analyses suggested that downregulation of PD-1 enhances short-term effector function, whereas downregulation of TIGIT is primarily responsible for maintaining a less differentiated/exhausted state, providing a potential mechanism for the observed synergy. The PD-1/TIGIT-downregulated CAR T cells generated from diffuse large B cell lymphoma patient-derived T cells also showed robust antitumor activity and significantly improved persistence in vivo. The efficacy and safety of PD-1/TIGIT-downregulated CD19-targeting CAR T cells are currently being evaluated in adult patients with relapsed or refractory large B cell lymphoma (ClinicalTrials.gov: NCT04836507).

Community expectations and anticipated outcomes for crisis support services-Lifeline Australia.

Crisis lines provide a critical first line of mental wellbeing support for community members in distress. Given the increasing referral to such services, there is a need to understand what the expectations of the community are around the role of such services in our public health responses. A computer assisted telephone interview was undertaken between 28th October and 30th November 2019. The aim was to explore expectations and anticipated outcomes of Lifeline Australia's crisis support services from a nationally representative community sample (N = 1,300). Analysis was undertaken to determine if demographic variables (age, gender, indigenous status, country of birth, culturally and linguistically diverse (CALD) status, sexual orientation, household composition, region and State/territory) and past service use affected community expectations. Results showed that a majority of respondents expected Lifeline to listen and provide support, recommend other services, and provide information. Help-seekers were expected to feel heard and listened to, receive safety advice or support to stay safe, and feel more hopeful. Lifeline was expected to prioritise people feeling suicidal, in immediate personal crisis, and experiencing domestic violence. Findings reveal that community members hold expectations for Lifeline Australia to serve as a suicide prevention and general crisis support service, which are congruent with the service's aims. There was little variation in community expectations of crisis support services based on demographic factors and past service use. The results show that the community has extensive and diverse expectations for this national crisis service to meet both short and longer-term needs for all vulnerable members of the community-entailing a very substantial public health service responsibility.

Instructor facilitation mediates students' negative perceptions of active learning instruction.

Studies have demonstrated students' resistance to active learning, despite evidence illustrating that their learning is improved relative to students in lectures. Specifically, while active learning and group work are effective at engaging students in their learning process, studies report that students' perceptions of active learning approaches are not always positive. What remains underexplored is whether students' perceptions of active learning improve with effective instructor facilitation and whether there exists differential perceptions between racially minoritized students and represented students. Here, we estimate students' perceptions of effective instructor facilitation as the mediator in the relationship between active learning and perceptions of learning and perceived utility for class activities (task value). Then, we examine differences by racial identification. We collected classroom observation data to empirically categorize courses as active learning or lecture-based and surveyed 4,257 college students across 25 STEM classrooms at a research-intensive university. We first examined the relationship between active learning on student perceptions and found a negative relationship between active learning and perceptions of learning and task value for both racially minoritized students and represented students. Next, we assessed whether students' perceptions of instructor effectiveness in facilitating group activities mediate these negative relationships. We found that, on average, students of all races were more likely to positively perceive instructor facilitation in active learning classes relative to lectures. In turn, the positive perceptions of instructor facilitation partially suppressed the negative relationship between active learning and perceptions of learning and task value. These results demonstrate that effective instructor facilitation can influence both students' self-assessment of learning and perceived utility of the learning activities, and underscores the importance of developing pedagogical competence among college instructors.

Associations between Combined Influenza and Pneumococcal Pneumonia Vaccination and Cardiovascular Outcomes.

BACKGROUND: In 2017, the CDC listed heart disease as the leading cause of death, with pneumonia and influenza being the eighth cause of death. Several studies have suggested the protective effects of influenza vaccination on myocardial infarction (MI). Available evidence supports the use of influenza vaccination in decreasing cardiovascular events, and the Joint Commission considers influenza vaccination a metric of quality care for hospitalized patients. Our specific aim was to evaluate the combined use of pneumococcal pneumonia vaccine (PPV) and influenza vaccine on cardiovascular outcomes and mortality. METHODS: A retrospective observational study was conducted using the 2012-2015 US National Inpatient Sample (NIS) database, to compare cardiovascular events in adult patients who did and did not receive vaccination during their hospitalization. ICD-9 codes were used to extract data for specific variables. The outcomes included MI, transient ischemic attacks, cardiac arrest, stroke, heart failure, and death. Adjusted relative risks (RR) were calculated using survey-weighted generalized linear models after adjusting for gender, race, socioeconomic status, diabetes, hypertension, hyperlipidemia, smoking status, prior coronary artery disease, and cerebrovascular disease. The effect of vaccination on in-hospital mortality was assessed in each subgroup of cardiovascular events using RR regressions. RESULTS: This study included 22,634,643 hospitalizations, of which 21,929,592 did not receive immunization. Vaccination solely against influenza was associated with lower MI (RR = 0.84, 95% CI: 0.82-0.87, p < 0.001), TIA (RR = 0.93, 95% CI: 0.9-0.96, p < 0.001), cardiac arrest (RR = 0.36, 95% CI: 0.33-0.39, p < 0.001), stroke (RR = 0.94, 95% CI: 0.91-0.97, p < 0.001), and mortality (RR = 0.38, 95% CI: 0.36-0.4, p < 0.001). Vaccination with PPV alone was associated with MI (RR = 1.13, 95% CI: 1.11-1.16, p < 0.001), TIA (RR = 1.28, 95% CI: 1.26-1.31, p < 0.001), stroke (RR = 1.21, 95% CI: 1.18-1.24, p < 0.001), and lower mortality (RR = 0.47, 95% CI: 0.45-0.49, p < 0.001). Combined PPV and influenza vaccine was associated with lower mortality (2.21% vs. 1.03%, p < 0.001) and lower cardiac arrest (0.61% vs. 0.51%, p < 0.001). In the adjusted analysis, the RR was 0.46 (95% CI: 0.43, 0.49) for mortality in the combined vaccinated cohort. The combined vaccination group also had a significantly reduced risk of mortality among those admitted with MI (RR = 0.46), transient ischemic attacks (RR = 0.58), and stroke (RR = 0.42) compared to the nonvaccinated group. CONCLUSIONS: Our study shows a significantly reduced risk of mortality with influenza vaccine and PPV and with combined pneumococcal and influenza vaccination. These data suggest that in-hospital administration of pneumonia and influenza vaccines appears safe and supports the use of combined vaccination during hospitalization due to their cardiovascular benefits.

A PSMA-targeted bispecific antibody for prostate cancer driven by a small-molecule targeting ligand.

Despite the development of next-generation antiandrogens, metastatic castration-resistant prostate cancer (mCRPC) remains incurable. Here, we describe a unique semisynthetic bispecific antibody that uses site-specific unnatural amino acid conjugation to combine the potency of a T cell-recruiting anti-CD3 antibody with the specificity of an imaging ligand (DUPA) for prostate-specific membrane antigen. This format enabled optimization of structure and function to produce a candidate (CCW702) with specific, potent in vitro cytotoxicity and improved stability compared with a bispecific single-chain variable fragment format. In vivo, CCW702 eliminated C4-2 xenografts with as few as three weekly subcutaneous doses and prevented growth of PCSD1 patient-derived xenograft tumors in mice. In cynomolgus monkeys, CCW702 was well tolerated up to 34.1 mg/kg per dose, with near-complete subcutaneous bioavailability and a PK profile supporting testing of a weekly dosing regimen in patients. CCW702 is being evaluated in a first in-human clinical trial for men with mCRPC who had progressed on prior therapies (NCT04077021).

Microdroplet-based one-step RT-PCR for ultrahigh throughput single-cell multiplex gene expression analysis and rare cell detection.

Gene expression analysis of individual cells enables characterization of heterogeneous and rare cell populations, yet widespread implementation of existing single-cell gene analysis techniques has been hindered due to limitations in scale, ease, and cost. Here, we present a novel microdroplet-based, one-step reverse-transcriptase polymerase chain reaction (RT-PCR) platform and demonstrate the detection of three targets simultaneously in over 100,000 single cells in a single experiment with a rapid read-out. Our customized reagent cocktail incorporates the bacteriophage T7 gene 2.5 protein to overcome cell lysate-mediated inhibition and allows for one-step RT-PCR of single cells encapsulated in nanoliter droplets. Fluorescent signals indicative of gene expressions are analyzed using a probabilistic deconvolution method to account for ambient RNA and cell doublets and produce single-cell gene signature profiles, as well as predict cell frequencies within heterogeneous samples. We also developed a simulation model to guide experimental design and optimize the accuracy and precision of the assay. Using mixtures of in vitro transcripts and murine cell lines, we demonstrated the detection of single RNA molecules and rare cell populations at a frequency of 0.1%. This low cost, sensitive, and adaptable technique will provide an accessible platform for high throughput single-cell analysis and enable a wide range of research and clinical applications.

Pathogenic ATM Mutations in Cancer and a Genetic Basis for Radiotherapeutic Efficacy.

BACKGROUND: Radiation therapy is one of the most commonly used cancer therapeutics but genetic determinants of clinical benefit are poorly characterized. Pathogenic germline variants in ATM are known to cause ataxia-telangiectasia, a rare hereditary syndrome notable for marked radiosensitivity. In contrast, somatic inactivation of ATM is a common event in a wide variety of cancers, but its clinical actionability remains obscure. METHODS: We analyzed 20 107 consecutively treated advanced cancer patients who underwent targeted genomic sequencing as part of an institutional genomic profiling initiative and identified 1085 harboring a somatic or germline ATM mutation, including 357 who received radiotherapy (RT). Outcomes of irradiated tumors harboring ATM loss-of-function (LoF) mutations were compared with those harboring variants of unknown significance. All statistical tests were 2-sided. RESULTS: Among 357 pan-cancer patients who received 727 courses of RT, genetic inactivation of ATM was associated with improved radiotherapeutic efficacy. The 2-year cumulative incidence of irradiated tumor progression was 13.2% vs 27.5% for tumors harboring an ATM LoF vs variant of unknown significance allele, respectively (hazard ratio [HR] = 0.51, 95% confidence interval [CI] = 0.34 to 0.77, P = .001). The greatest clinical benefit was seen in tumors harboring biallelic ATM inactivation (HR = 0.19, 95% CI = 0.06 to 0.60, P = .005), with statistically significant benefit also observed in tumors with monoallelic ATM inactivation (HR = 0.57, 95% CI = 0.35 to 0.92, P = .02). Notably, ATM LoF was highly predictive of outcome in TP53 wild-type tumors but not among TP53-mutant tumors. CONCLUSIONS: We demonstrate that somatic ATM inactivation is associated with markedly improved tumor control following RT. The identification of a radio-sensitive tumor phenotype across multiple cancer types offers potential clinical opportunities for genomically guided RT.