Validation Study Research Articles

Abstract A027: Acceleration of drug metabolism mediated by CYP3A4 enzyme activation provides a bona fide environmental resistance mechanism to targeted therapies

Abstract Background: Targeted therapies against oncogenic signaling addictions, including one resulting from EML4-ALK fusions, can induce strong and durable clinical responses. However, these therapies are not curative in advanced cancers, as a subset of tumor cells avoid elimination, and these persisting populations evolve resistance that drives tumor relapse. Using a drug screen, we found that the evolving ALK inhibitor (ALKi) persisters display exquisite sensitivity to the histone demethylase inhibitor, JIB04 in vitro. Paradoxically, validation studies in mice revealed that instead of enhancing tumor sensitivity, JIB04 desensitizes tumors to ALKi, alectinib and lorlatinib, suggesting that it activates an unknown environmental modifier. While JIB04 is not used in the clinics, we reasoned that deciphering the underlying mechanism could uncover environment-mediated resistance to ALKi that might be triggered by clinically relevant factors. Results: We found that the resistance-promoting effect of JIB04 in mice is mediated by reducing systemic ALKi concentrations. This reduction is attributable to accelerated pharmacokinetics, due to the hyperactivation of the P450 family member, CYP3A4. Higher CYP3A4 activity leads to faster drug clearance and reduced drug exposure to tumors. Importantly, JIB04 triggers a similar resistance to other clinically relevant CYP3A4 substrates, the frontline EGFR inhibitor, osimertinib and the recently introduced KRASG12C inhibitor, sotorasib. Finally, the resistance- promoting effects of JIB04 could be phenocopied by chemically unrelated activators of CYP3A4, indicating a wide generalizability of this macro-environmental therapy resistance mechanism. Conclusions: Our study uncovered that acceleration of drug metabolism, triggered by CYP3A4 enzyme activators is a bona fide environmental resistance mechanism. Since P450 family liver detoxifying enzymes mediate the metabolism of many drugs, this resistance mechanism is likely to be relevant to a broad range of therapeutic contexts. Significance: While the importance of pharmacokinetic considerations is widely appreciated, modulation of drug metabolism is not typically considered as a candidate mechanism when therapy resistance is encountered in research and clinical settings. Multiple dietary, pharmacological and genetic modifiers are known to modulate P450 family enzymes. Moreover, substantial variability in drug plasma concentrations was documented in patients treated with identical doses of targeted therapies, and this variability strongly correlated with variability in clinical responses. While the direct link between variability in CYP3A4 activity and clinical outcomes in human patients has yet to be established, the mechanism uncovered in our study might represent a substantial but entirely overlooked contributor to resistance. Consequently, consideration of this mechanism and monitoring of CYP3A4 activity and systemic drug levels might improve our understanding of the biology of therapy resistance and enable an actionable path to improve clinical outcomes. Citation Format: Pragya Kumar, Robert Vander Velde, Malgorzata Weh, Andriy Marusyk. Acceleration of drug metabolism mediated by CYP3A4 enzyme activation provides a bona fide environmental resistance mechanism to targeted therapies [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A027.

Abstract PR017: Anti-metastatic immunotherapy discovery using ex vivo lung tissue cultures and high-throughput single-cell chemical transcriptomics

Abstract Introduction: Tumors systemically remodel the immune system during metastasis. Developing anti-metastatic immunotherapies that target these tumor-immune interactions to make the metastatic niche hostile colonization would represent a paradigm shift in cancer treatment. However, traditional high-throughput screening (HTS) platforms that use simple read-outs and contrived in vitro systems are ill-suited to identify such therapies. In contrast, HTS platforms that accurately model the metastatic niche and use high-dimensional read-outs such as multiplexed single-cell RNA-sequencing (scRNA-seq) can accurately profile nuanced perturbation responses in individual immune cell types and therefore have the potential to discover anti-metastatic immunotherapies. In this study, we describe the development of an ex vivo lung tissue culture HTS platform that preserves immune gene expression signatures observed in the in vivo metastatic niche and is amenable to single-cell chemical transcriptomic analysis using MULTI- seq (McGinnis et al., Nature Methods, 2019). Inspired by our recent description of myeloid cell TLR-NFκB inflammation during breast cancer lung metastasis (McGinnis et al., Cancer Cell, 2024), we then performed the first anti-metastatic immunotherapy drug screen and identified TLR-NFκB inhibitors that effectively operate on all desired myeloid cell types in the lung metastatic niche. Methods and Results: Building upon existing techniques for culturing precision-cut lung slices (Wu et al., Journal of Experimental Medicine, 2024) and patient- derived tumor fragments (Voabil et al., Nature Medicine, 2021), we established and optimized an ex vivo lung tissue culture system that enables 48-hour culturing of lung slices isolated from 4T1 tumor-bearing mice. scRNA-seq profiling revealed successful capture of all relevant immune cell types and retention of metastasis-associated gene expression programs (e.g., myeloid TLR-NFκB inflammation and neutrophil degranulation) following culture. We then scaled our platform to address how myeloid cells in the lung metastatic niche respond to perturbations targeting every component of the canonical TLR-NFκB signaling cascade. Cell-type-specific perturbation modeling and quantification of TLR-NFκB signaling inhibition identified compounds that optimally perturb all intended myeloid cell types (e.g., tissue-resident macrophages, neutrophils, and monocytes) and represent prioritized candidates for future in vivo validation studies to assess impact on metastatic disease progression. Conclusions: In this study, we describe the development and application of a novel HTS platform that couples ex vivo lung tissue cultures with single-cell chemical transcriptomics to identify anti-metastatic immunotherapy candidates. We demonstrate how our platform can identify TLR-NFκB signaling inhibitors that optimally operate in the lung metastatic niche, paving the way for future interrogation of additional metastasis-associated immune cell signaling pathways in different disease and tissue backgrounds. Citation Format: Chris McGinnis, Winnie Yao, Ansuman Satpathy. Anti-metastatic immunotherapy discovery using ex vivo lung tissue cultures and high-throughput single-cell chemical transcriptomics [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr PR017.

Abstract C035: Spatially limited stroma-mediated resistance potentiates targeted therapy resistance through an integration of multiple juxtacrine and paracrine mechanisms

Abstract Despite the ability to induce strong clinical responses, therapies that target mutational drivers of oncogenic signaling are not curative in advanced cancers, as a subset of cancer cells is capable of surviving therapy and evolving resistance. While therapy resistance is commonly viewed as a cell-intrinsic phenomenon, multiple factors produced by tumors can also confer strong therapy resistance. In contrast to the detailed elucidation of molecular mediators of cell-intrinsic and stroma-mediated (SM) resistance, the relative importance of cell-intrinsic and SM resistance to in vivo therapeutic responses remains poorly defined. To address this gap of knowledge, we used a well-characterized experimental model of ALK+ lung cancer H3122 cell line that exemplifies a duality observed across common models of targetable cancers. Under standard stroma-free in vitro cultures, H3122 cells can survive under therapeutically relevant concentrations of ALK inhibitors (ALKi), eventually acquiring cell-intrinsic therapy resistance. At the same time, co-culture with stromal fibroblasts drastically reduces the sensitivity of H3122 cells to ALKi, indicating the relevance of SM resistance. To understand the impact of SM resistance on in vivo responses, we sought to identify the mechanism(s) responsible for the therapy-protective effects of stromal fibroblasts, subsequently interrogating the impact of the disruption of this mechanism on in vivo therapeutic responses. Our in vitro studies with a large panel of primary stromal fibroblast isolates identified the HGF-cMET axis as the major mechanism responsible for ALKi desensitization. Surprisingly, xenograft validation studies demonstrated a weak effect of HGF-cMET modulation. Histological analyses of tumor tissues revealed that this relative weakness is attributable to a strong, spatially limited HGF-independent component of SM resistance. Mechanistic follow-up demonstrated that the HGF-independent component of SM resistance integrates the effect of multiple well-known juxtacrine and paracrine-acting mechanisms. Whereas the multifactorial nature of stroma-mediated resistance prevented a clear assessment of its relative contribution to in vivo therapy responses, our spatial analyses indicate that SM resistance dominates the ability of tumors to avoid therapeutic elimination. Surprisingly, we found that SM resistance was also a substantial contributor to tumor relapse, indicating that in vivo therapy resistance can integrate both cell-intrinsic and cell-extrinsic effects. This duality of therapy resistance and the multifactorial underpinning of both cell-intrinsic and stroma-mediated resistance present a challenge to therapeutic strategies focused on identifying and targeting specific resistance mechanisms. Our studies suggest that this limitation can be overcome by shifting the therapeutic focus towards shared orthogonal therapeutic sensitivities of tumor cells within residual disease. Citation Format: Bina Desai, Tatiana Miti, Sandhya Prabhakaran, Daria Miroshnychenko, Menkara Henry, Viktoriya Marusyk, Chandler Gatenbee, Marilyn Bui, Jacob Scott, Philipp M. Altrock, Eric Haura, Alexander R.A. Anderson, David Basanta, Andriy Marusyk. Spatially limited stroma-mediated resistance potentiates targeted therapy resistance through an integration of multiple juxtacrine and paracrine mechanisms [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr C035.

The perioperative frailty index derived from the Chinese hospital information system: a validation study.

There are various frailty assessment tools in the world, and the application choice of frailty assessment tools for the elderly perioperative population varies. It remains unclear which frailty assessment tool is more suitable for the perioperative population in China. To validate the Perioperative Frailty Index (FI-32) derived from the Chinese Hospital Information System by investigating the impact of preoperative frailty on postoperative outcomes, and ascertain the diagnostic value of FI-32 for predicting postoperative complications through comparing with the FRAIL scale and the modified Frailty Index (mFI-11). A prospective cohort study was conducted in a tertiary hospital. Elderly patients who were 60 years or older and underwent selective operation were included. The FI-32, FRAIL scale, and mFI-11 were assessed. Demographic, surgical variables and outcome variables were extracted from medical records. The data of readmission and mortality within 30 days and 90 days of surgery were ascertained by Telephone follow-up by professionally trained researchers. Multiple logistic regression was used to examine the association between frailty and complications. Receiver operating characteristic curves(ROC) were used to compare FI-32 with mFI-11 and FRAIL, to explore the predictive ability of frailty. 335 patients qualified for the inclusion criteria and were enrolled in the study, and among them, 201 (60.0%) were females, and the Median(P25, P75)age at surgery was 69 (65,74) years. The prevalence of frailty in the study population was 16.4% (assessed by FI-32). After adjusting for concomitant variables including demographic characteristics (such as gender, BMI, smoking, drinking, average monthly income and educational level) and surgical factors (such as surgical approach, surgical site, anesthesia method, operation time, intraoperative bleeding, and intraoperative fluid intake), there was a statistically significant association between frailty and the development of postoperative complication after surgery (OR = 3.051, 95% CI:1.460-6.378, P = 0.003). There were also significant differences in mortality within 30 days of surgery, the length of hospital stay (LOS) and the hospitalization costs. FI-32, FRAIL and mFI-11 showed a moderate predictive ability for postoperative complications, the Area Under Curves (AUCs) were 0.582, 0.566 and 0.531, respectively. With adjusting concomitant variables associated with postoperative complications, the AUCs of FI-32, FRAIL and mFI-11 in the adjusted prediction models were 0.824, 0.827 and 0.820 respectively. The FI-32 has a predictive effect on postoperative adverse outcomes in elderly Chinese patients. Compared to FRAIL and mFI-11, the FI-32 had the same ability to predict postoperative complications, and FI-32 can be extracted directly from HIS, which greatly saves the time for clinical medical staff to evaluate perioperative frailty.

Digital trust in China’s digital transformation process: methodology and results of an empirical research

The article is devoted to the topical issues of China's digital transformation and the role of digital trust in it. Digital transformation is a long — term comprehensive and rapid process, accompanied by rapid development of digital technologies, institutional changes and appropriate adaptation of people to new conditions. At the same time, it is emphasized that the drastic changes caused by digital transformation bring a lot of uncertainty and risks to society, which, without proper government response, can lead to social unrest and social unrest, and reduce confidence in this process. In the process of digital transformation, high trust between people can reduce suspicions, doubts and reduce social tension in society. In this regard, digital trust comes to the fore of sociological analysis — a new complex social phenomenon that combines sufficient confidence in the reliability of individuals, processes and technologies related to the security and benefits of the digital world. The author shows that the structure of digital trust is multifaceted and sets various objects for research: they can be technologies, processes, individuals, etc., therefore, those factors that affect these three components of the trust structure can be considered as factors affecting digital trust as a whole. However, the topic of digital trust has not been studied much, both from a theoretical and practical point of view, and its valid empirical studies are clearly insufficient. Indeed, some scientists have used factor analysis in their research to study the elements that affect digital trust. However, due to the lack of a well-developed theoretical framework, the mechanism of interaction between influencing factors and characteristics of digital trust cannot be reliably described. The article presents an original methodology for the sociological analysis of digital trust, based on the methodology of factor analysis. As a result of its testing, factors affecting the level of digital trust have been identified, which are of theoretical and practical importance.

Expression of TSPAN1 and its link to thyroid nodules: one step forward on the path to thyroid tumorigenesis biomarkers.

Thyroid cancer is ranked as the most common malignancy within the endocrine system and the seventh most prevalent cancer in women globally. Thyroid malignancies require evaluating biomarkers capable of distinguishing between them for accurate diagnosis. We examined both mRNA and protein levels of TSPAN1 in plasma and tissue samples from individuals with thyroid nodules to aid this endeavour. In this case-control study, TSPAN1 was assessed at both protein and mRNA levels in 90 subjects, including papillary thyroid cancer (PTC; N = 60), benign (N = 30), and healthy subjects (N = 26) using enzyme-linked immunosorbent assay (ELISA) and SYBR-Green Real-Time PCR, respectively. TSPAN1 plasma levels were decreased in PTC and benign compared to healthy subjects (P = 0.002). TSPAN1 mRNA levels were also decremented in the tumoral compared to the paired normal tissues (P = 0.012); this drop was also observed in PTC patients compared to benign patients (P = 0.001). Further, TSPAN1 had an appropriate diagnostic value for detecting PTC patients from healthy plasma samples with a sensitivity of 76.7% and specificity of 65.4% at the cutoff value < 2.7 (ng/ml). TSPAN1 levels are significantly reduced in patients with benign and PTC, demonstrating its potential value as a diagnostic biomarker. Additionally, the significant reduction in TSPAN1 mRNA expression within PTC tumor tissues may suggest its involvement in tumor progression and development. Further studies, including larger-scale validation studies and mechanistic investigations, are imperative to clarify the molecular mechanisms behind TSPAN1 and, ultimately, its clinical utility for treating thyroid disorders.

The Development of a Food Frequency Questionnaire for the Assessment of Ultra-Processed Food Consumption in the Italian Adult Population: Protocol for a Validity and Reproducibility Study

Background: Over the last decade, while studies on the detrimental effects of ultra-processed food (UPF) consumption have increased, methodological limitations on the quality of available evidence have emerged. Starting from a critical reassessment of the NOVA classification, this project will aim to develop and validate a food frequency questionnaire (FFQ), which is based on the processing of consumed foods and specifically designed to estimate the UPF consumption and total dietary intake of macro- and micronutrients in the Italian adult population. Methods: This study will take place in selected workplaces and include healthy males and females aged ≥18 years, residing in Italy and with Italian citizenship. The FFQ will be online, voluntary, self-administered, semi-quantitative, and designed to assess food intake over the past year and distinguish between industrial, artisanal, and home-made products. This project will consist of two phases. First, a pilot study will be conducted to obtain the final version of the FFQ. The current food consumption of the target population will be investigated, through a 24 h dietary recall, and the face validity of the new tool will be tested. The second phase will involve at least 436 participants. To assess reproducibility, the FFQ will be administered twice (at an interval of 3–10 months), and the test–retest method will be used. A 7-day weighed dietary record (WDR) will also be completed after each FFQ administration. To evaluate criterion validity, data from the two WDRs will be compared against those from the first FFQ administration. Conclusions: The results will provide a new valid tool focused on food processing, potentially useful for future studies.

Cross-cultural validation of the Clinician-Administered PTSD scale for DSM-5, child and adolescent version in Japan

ABSTRACT Background: The establishment of a formal diagnosis of post-traumatic stress disorder (PTSD) for children and adolescents is the foundation for advancing pertinent clinical research and formulating proper treatment and management. However, a validated diagnostic tool for PTSD in children and adolescents is lacking in Japan. Objective: To examine the cross-cultural validity and reliability of the Japanese Clinician-Administered PTSD Scale for the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5), child and adolescent version (CAPS-CA-5-J). Method: Overall, 73 children who had experienced potentially traumatic events were recruited from three medical facilities. The linguistically validated CAPS-CA-5-J was conducted by trained psychiatrists and psychologists. Additionally, children and their caretakers completed questionnaires such as the UCLA PTSD reaction index for DSM-5 (PTSD-RI-5), Depression Self-Rating Scale for Children (DSRDC), and Strength and Difficulties Questionnaire (SDQ); 16 participants were randomly selected for the inter-rater reliability assessment. Results: Owing to missing values in the CAPS-CA-5-J and PTSD-RI-5, 68 children were included in the final analysis. Regarding reliability, the CAPS-CA-5-J showed excellent internal consistency (Cronbach’s alpha coefficient = 0.90) and interrater agreement (kappa coefficient = 0.88). Convergent validity was supported by a strong correlation between the total severity scale of the CAPS-CA-5-J and the PTSD-RI-5 scores (Pearson’s correlation coefficient = 0.82). Divergent validity was indicated by a moderate correlation between the CAPS-CA-5-J and DSRDC, and no correlation was found with the SDQ scores. Conclusions: This study is the first to validate a structured clinical interview for children and adolescents with PTSD in Japan. The psychometric properties of the CAPS-CA-5-J were good enough and comparable to those reported in previous validation studies. Therefore, the CAPS-CA-5-J can be considered reliable and valid for use in Japan.

Next step of molecular pathology: next-generation sequencing in cytology

The evolving landscape of precision oncology underscores the pivotal shift from morphological diagnosis to treatment decisions driven by molecular profiling. Recent guidelines from the European Society for Medical Oncology recomend the use of next-generation sequencing (NGS) across a broader range of cancers, reflecting its superior efficiency and clinical value. NGS not only updates oncology testing by offering quicker, sample-friendly, and sensitive analysis but also reduces the need for multiple individual tests. Cytology samples, often obtained through less invasive methods, can yield high-quality genetic material suitable for molecular analysis. This article focuses on optimizing the use of cytology samples in NGS, and outlines their potential benefits in identifying actionable molecular alterations for targeted therapies across various solid tumors. It also addresses the need for validation studies and the strategies to incorporate or combine different types of samples into routine clinical practice. Integrating cytological and liquid biopsies into routine clinical practice, alongside conventional tissue biopsies, offers a comprehensive approach to tumor genotyping, early disease detection, and monitoring of therapeutic responses across various solid tumor types. For comprehensive biomarker characterization, all patient specimens, although limited, is always valuable.

Creating a content validation process to advance a campus-wide leadership framework

PurposeA content validation process of an institutional leadership framework is described for leadership educators in higher education. We created this process to further integrate our leadership framework across campus, maintain alignment with advancements in leadership research and ensure it is broadly inclusive and culturally responsive.Design/methodology/approachOur approach included seven essential design elements and was informed by a review of leadership frameworks in practice and the literature, validation studies and a comprehensive document review.FindingsOur approach yielded a validated leadership framework with modifications to its principles, values, competencies and outcomes. Modifications addressed pre-determined criteria and were deemed relevant to leadership research and our institutional context.Originality/valueThe external content validation process of our leadership framework is novel and serves as a valuable guide for those considering opportunities to strengthen their own institutional approaches to leadership education.

Making plaque assessment easier - a validation study of simplified versions of the Marginal Plaque Index.

The assessment of plaque indices may be time-consuming and error-prone. Simplification of these indices may increase their utility without compromising their validity. The aim of this study was to evaluate the validity of two simplified versions of the Marginal Plaque Index (MPI). Two simplified versions of the MPI as well as the Plaque Control Record (PCR) were derived from full-scale MPI assessments in two studies with four age groups (N = 42 10-year-olds; N = 24 15 year-olds; N = 53 university students (18y-33y); N = 66 parents (32y-57y). Correlations with the Turesky modification of the Quigley-Hein Index (TQHI) and the Papillary Bleeding Index (PBI) were calculated. The indices derived from the MPI showed high convergence with each other (all r ≥ 0.94) and with the TQHI (r ≥ 0.80). The concurrent validity of the MPI with the PBI was equal to that of the TQHI in all age groups. The simplified versions of the MPI and the PCR show a lower convergent validity with the PBI than the MPI within parents (p < 0.05). In the other age groups, their convergent validity was equal to that of the MPI. Simplification of the MPI does not affect its convergent validity with other plaque indices but may reduce its concurrent validity with the PBI in middle-aged adults.

Hybrid 3D bioprinting for advanced tissue-engineered trachea: merging fused deposition modeling (FDM) and top-down digital light processing (DLP).

In this present study, we introduce an innovative hybrid 3D bioprinting methodology that integrates fused deposition modeling (FDM) with top-down digital light processing (DLP) for the fabrication of an artificial trachea. Initially, polycaprolactone (PCL) was incorporated using an FDM 3D printer to provide essential mechanical support, replicating the structure of tracheal cartilage. Subsequently, a chondrocyte-laden glycidyl methacrylated silk fibroin (Sil-MA) hydrogel was introduced via top-down DLP into the PCL scaffold (PCL-Sil scaffold). &#xD;The mechanical evaluation of PCL-Sil scaffolds showed that they have greater flexibility than PCL scaffolds, with a higher deformation rate (PCL-Sil scaffolds: 140.9±5.37% vs. PCL scaffolds: 124.3±6.25%) and ability to withstand more force before fracturing (3.860±0.140 N for PCL-Sil scaffolds vs. 2.502±0.126 N for PCL scaffolds, ***P < 0.001). Both types of scaffolds showed similar axial compressive strengths (PCL-Sil scaffolds: 4.276±0.127 MPa vs. PCL scaffolds: 4.291±0.135 MPa). Additionally, PCL-Sil scaffolds supported fibroblast proliferation, indicating good biocompatibility. In vivo testing of PCL-Sil scaffolds in a partial tracheal defect rabbit model demonstrated effective tissue regeneration. The scaffolds were pre-cultured in the omentum for two weeks to promote vascularization before transplantation. Eight weeks after transplantation into the animal, bronchoscopy and histological analysis confirmed that the omentum-cultured PCL-Sil scaffolds facilitated rapid tissue regeneration and maintained the luminal diameter at the anastomosis site without signs of stenosis or inflammation. Validation study to assess the feasibility of our hybrid 3D bioprinting technique showed that structures, not only the trachea but also the vertebral bone-disc and trachea-lung complex, were successfully printed. &#xD.

Enhancing detection of various pancreatic lesions on endoscopic ultrasound through artificial intelligence: a basis for computer-aided detection systems.

Endoscopic ultrasound (EUS) is the most sensitive method for evaluation of pancreatic lesions but is limited by significant operator dependency. Artificial intelligence (AI), in the form of computer-aided detection (CADe) systems, has shown potential in increasing accuracy and bridging operator dependency in several endoscopic domains. However, the complexity of integrating AI into EUS is far more challenging. This aims to develop and test the basis for a CADe system for real-time detection and segmentation of all pancreatic lesions. In this single-center study EUS studies of pancreatic findings were included. Lesions were outlined by two expert (>5years performing EUS) endoscopists, and the two leading types of models were benchmarked. The models' performance was evaluated through per-pixel intersection over union (IoU). A total of 1497 EUS images from 165 patients were evaluated. The dataset included malignancies, neuroendocrine tumors, benign cysts, chronic and acute pancreatitis, normal fatty pancreas, and benign lesions. The best model demonstrated detection and segmentation on the test set with a mean IoU of 0.73, achieving a PPV, NPV, total accuracy, and ROC of 0.82, 0.96, 0.95, and 0.95, respectively. The algorithm is adaptable for real-time processing. We developed and tested deep learning models for real-time detection and segmentation of pancreatic lesions during EUS with promising results. This constitutes the basis for a CADe system for EUS, which could be valuable in future detection and evaluation of pancreatic lesions. Further studies for validation and generalization are underway.

Gadoxetic Acid-Enhanced Magnetic Resonance Imaging Features Can Predict Immune-Excluded Phenotype of Hepatocellular Carcinoma

Introduction: Immunotherapy is the first-line treatment for intermediate-advanced stage hepatocellular carcinoma (HCC), although its outcomes vary. This study aimed to identify imaging biomarkers of immunotherapy susceptibility linked to gadoxetic acid-enhanced magnetic resonance imaging (EOB-MRI) and immune phenotypes, particularly immune-excluded phenotypes, with a tumor immune barrier. Methods: We performed immunohistochemical staining with a CD8+ antibody, and samples were classified into immune-inflamed, -intermediate, -excluded, and -ignored phenotypes. We assessed EOB-MRI findings obtained from 104 patients who underwent hepatectomy for HCC and evaluated the relationship between MRI findings and immune phenotype. Spatial transcriptome analysis of tumor tissues in each immune phenotype was performed to characterize the MRI findings. For validation, we analyzed the treatment effect on 60 nodules in another cohort of 27 patients who received combined immunotherapy using anti-programmed death-ligand 1 and anti-vascular endothelial growth factor (VEGF) antibodies. Results: HCCs with rim arterial phase hyperenhancement (APHE) (odds ratio [OR] 17.3 P=0.009), peritumoral enhancement on the arterial phase (OR 8.6, P&lt;0.004), and intermediate intensity on the hepatobiliary phase (HBP) measured with a visual 3-point scale (OR 28.2, P=0.002) were associated with immune-excluded phenotype, where tumors tended to be larger and of the single nodular type with extranodular growth and confluent multinodular rather than the simple nodular type. Spatial transcriptome analysis revealed a spatial relationship among cytotoxic T lymphocytes, VEGF signals, and cancer-associated fibroblasts at the tumor-invasive margins in this phenotype. From the validation study, nodules with any one of these three imaging findings had a significantly prolonged time to-nodular progression (P=0.007, median not reached vs. 226 days). Conclusion: HCCs with rim APHE, peritumoral enhancement on arterial phase, and intermediate intensity on HBP with visual 3-point scale could be non-invasive biomarkers to predict the immune-excluded phenotype with tumor immune barrier. These HCCs were most likely to respond to the combined immunotherapy.

Visual inspection time as an accessible measure of processing speed: A validation study in children with cerebral palsy.

This study examined the validity of a visual inspection time (IT) task as a measure of processing speed (PS) in a sample of children with and without cerebral palsy (CP). IT tasks measure visualization speed without focusing on the motor response time to indicate decision making about the properties of those stimuli. Participants were 113 children ages 8-16, including 45 with congenital CP, and 68 typically developing peers. Measures were a standard visual IT task that required dual key responding and a modified version using an assistive technology button with response option scanning. Performance on these measures was examined against traditional Wechsler PS measures (Coding, Symbol Search). IT performance shared considerable variance with traditional paper-pencil PS measures for the group with CP, but not necessarily in the typically developing group. Concurrent validity was found for both IT task versions with traditional PS measures in the group with CP. IT classification accuracy for lowered PS showed modest sensitivity and good specificity particularly for the modified IT task. As measures of PS in children with CP who are unable to validly participate in traditional PS tasks, IT tasks demonstrate adequate concurrent validity and may serve as a beneficial alternative measure of PS in this population.

An Exercise-Based Precision Medicine Tool and Smartphone App for Managing Achilles Tendinopathy (the 'PhysViz' System): User-Centered Development Study.

People with Achilles tendinopathy (AT) experience persistent pain that can limit engagement with daily occupations and negatively impact mental health. Current therapeutic exercise approaches vary in success, with many people experiencing reinjury, leading to a cycle of chronic tendinopathy often lasting years. High-magnitude precision loading may help people exit this feedback cycle, but applying these principles clinically is challenging. This user-centered design case study aims to provide an overview on how the PhysViz (a prototype for a novel remote rehabilitation intervention for AT management) was developed and evaluated following the development phase of the Framework for Accelerated and Systematic Technology-Based Intervention Development and Evaluation Research (FASTER). The development process engaged a multidisciplinary team comprising people with AT experiences, clinicians, and engineers. It followed the 5 stages within the FASTER development phase: empathize, define, ideate, prototype, and test. The PhysViz development and evaluation were informed by needs assessments, surveys, literature reviews, validation studies, case studies, roundtable discussions, and usability testing (some of which have been published previously). The FASTER systematically guided the integration of evidence-based features and behavior change theory. By using the FASTER and ensuring that the PhysViz system was underpinned by diverse stakeholder needs, this work resulted in the development of a working prototype for both the PhysViz physical exercise tool and the accompanying PhysViz software package (mobile app and web application). A variety of study designs informed user-desired features that were integrated into the PhysViz prototype, including real-time biofeedback in the form of precision load monitoring, customizable exercise programs, and pain tracking. In addition, clinicians can visualize client data longitudinally and make changes to client exercise prescriptions remotely based on objective data. The identified areas for improvement, such as upgrading the user interface and user experience and expanding clinical applications, provide valuable insights for future PhysViz iterations. Further research is warranted to assess the long-term efficacy and feasibility of the PhysViz in diverse clinical settings and its potential to improve AT symptoms. Being one of the first technology development initiatives guided by the FASTER, this study exemplifies a systematic and multidisciplinary approach to creating a remote rehabilitation intervention. By incorporating stakeholder feedback and evidence-based features, the PhysViz addresses key challenges in AT rehabilitation, offering a novel solution for precision loading and therapeutic exercise engagement. Positive feedback from users and clinicians underscores the potential impact of the PhysViz in improving AT management outcomes. The PhysViz serves as a model for technology-based intervention development, with potential implications for other tendinopathies and remote rehabilitation strategies.

Developing and Validating the Health Literacy Scale for Migrant Workers: Instrument Development and Validation Study.

Research concerning health literacy among migrant workers in South Korea has been limited, especially given the lack of validated instruments and the lack of focus on the cultural diversity of migrant workers. This study aimed to develop and validate a health literacy scale for unskilled migrant workers (HLS-MW) in South Korea. We first generated a pool of potential items based on a literature review and in-depth interviews with 23 migrant workers. Subsequently, we reviewed empirical referents from the first step to select relevant medical terminologies and passages, ultimately choosing 709 words. The study team initially generated 35 items with 709 health-related terms through empirical referent reviews. After content validity testing by an expert panel, 28 items comprising 89 terms on the 2 subscales of prose and documents were selected for psychometric testing. Overall, 402 unskilled migrant workers in South Korea completed a web-based survey between August and September 2021, with 334 responses included in the final analysis. We used multiple analytic approaches, including exploratory factor analysis, Rasch analysis (item response theory), and descriptive analysis, to examine the new scale's validity and reliability. The final sample primarily included young male workers from South Asian countries. The HLS-MW yielded 2 factors: prose and documents. The item difficulty scores ranged from -1.36 to 2.56. The scale was reduced to 13 items (10 prose and 3 document items), with the final version exhibiting good internal reliability (Kuder-Richardson index=0.88; intraclass correlation coefficient=0.94, 95% CI 0.93-0.95) and test-retest reliability (r=0.74, 95% CI 0.57-0.92). HLS-MW scores differed significantly by Korean language proficiency (F2,331=3.54, P=.004). The HLS-MW is a reliable and valid measure to assess health literacy among migrant workers in South Korea. Further studies are needed to test the psychometric properties of the HLS-MW in diverse migrant groups in South Korea while also establishing cutoffs to help identify those in need of health literacy support.

Abstract A001: Analytical validation of a novel multiomic metastatic breast cancer liquid biopsy test that combines ctDNA analysis with CTC HER2-ultralow and ER co-expression, as well as single-cell chromosomal instability

Abstract Background: Metastatic breast cancer (MBC) subtype characterization is critical for effective treatment. The DefineMBC™ multiomic blood biopsy clinical diagnostic combines ctDNA and circulating tumor cell (CTC) analyses to characterize MBC patients when a tissue biopsy is not feasible. Innovation in the DefineMBC 2.0 5-channel immunofluorescent (IF) CTC characterization assay now measures ultralow (UL) levels of HER2 protein expression, ER protein expression, and chromosomal instability (CI) as well as ERBB2 amplification via single- cell sequencing. Cell-level HER2/ER co-detection enables monitoring of receptor conversion during a patient’s MBC treatment journey. As trastuzumab-deruxtecan (T-DxD) has become standard of care for HER2-low disease, distinguishing HER2-UL from HER2[-] by DefineMBC’s liquid biopsy approach may identify additional patients who can benefit from such therapy. This addresses an unmet need given the known shortcomings of tissue biopsy IHC in the metastatic setting as recently noted by communications from NCCN, ASCO, and the College of American Pathologists. Methods: Assay development including analytical validation studies utilized biologically relevant cell lines of epithelial origin and known expression levels of HER2 and ER (MDA-MB-453: HER2[+] &amp; ER[-]; MCF-7: HER2-low &amp; ER[+]; and MCF-7/ERBB2 knockout: HER2[-]) that were spiked into healthy donor blood. Additionally, patients with various forms of MBC were characterized with the new assay. As with all samples, following red blood cell lysis, nucleated cells were deposited on glass slides at high density (∼3E6 WBC/slide). Slides underwent IF staining for cytokeratins (CK), CD31, CD45, HER2, ER, and Hoechst to localize nuclear DNA, followed by scanning and machine learning-based rare cell detection. CK[+], CD31[-]/CD45[-] cells were classified as CTC candidates and assessed for HER2/ER expression. Pathologist-selected CTCs were isolated for single-cell whole genome sequencing and analyzed using a proprietary CTC DNA copy number pipeline to detect CI and amplification at the ERBB2 locus. Results: Analytical validation studies demonstrated the limit of detection at 1 CTC per 12E6 WBC (∼1.6 mL of blood) with a linear range of 1- 300 CTCs per slide. Sensitivity, specificity, accuracy, and precision metrics of the HER2 and ER IF assays were 96%, 96%, 96%, 2% CV; 91%, 95%, 93%, 3% CV; respectively. CTC enumeration precision was validated at greater than 80% for enumeration bins of 0, 1-10, 11-50, and&amp;gt;50 CTCs per sample. During patient testing 7 patients previously called HER2[-] are now designated as consistent with HER2-low by the pathologist, suggesting improved test performance in future clinical validation studies is likely. Conclusions: Advancement of our comprehensive cancer profiling platform now includes ER/HER2-UL/CI co-detection on enumerated CTCs in MBC patients. Clinical studies currently in progress will reveal whether HER2-UL/HER2[+] detection on CTCs will identify patients better suited for T-DxD or trastuzumab, respectively. Citation Format: Raj Srikrishnan, Jordan Ritchie, Alessandra Cunsolo, Andrew Kunihiro, Brandon Guillory, Sara Tin, Zhuo Meng, Ernest T Lam, Bharat Annaldas, Evan Schwab, Nilesh Dharajiya, Timothy Pluard, Martin Blankfard, David Bourdon. Analytical validation of a novel multiomic metastatic breast cancer liquid biopsy test that combines ctDNA analysis with CTC HER2-ultralow and ER co-expression, as well as single-cell chromosomal instability [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A001.

Comprehensive Symptom Prediction in Inpatients With Acute Psychiatric Disorders Using Wearable-Based Deep Learning Models: Development and Validation Study.

Assessing the complex and multifaceted symptoms of patients with acute psychiatric disorders proves to be significantly challenging for clinicians. Moreover, the staff in acute psychiatric wards face high work intensity and risk of burnout, yet research on the introduction of digital technologies in this field remains limited. The combination of continuous and objective wearable sensor data acquired from patients with deep learning techniques holds the potential to overcome the limitations of traditional psychiatric assessments and support clinical decision-making. This study aimed to develop and validate wearable-based deep learning models to comprehensively predict patient symptoms across various acute psychiatric wards in South Korea. Participants diagnosed with schizophrenia and mood disorders were recruited from 4 wards across 3 hospitals and prospectively observed using wrist-worn wearable devices during their admission period. Trained raters conducted periodic clinical assessments using the Brief Psychiatric Rating Scale, Hamilton Anxiety Rating Scale, Montgomery-Asberg Depression Rating Scale, and Young Mania Rating Scale. Wearable devices collected patients' heart rate, accelerometer, and location data. Deep learning models were developed to predict psychiatric symptoms using 2 distinct approaches: single symptoms individually (Single) and multiple symptoms simultaneously via multitask learning (Multi). These models further addressed 2 problems: within-subject relative changes (Deterioration) and between-subject absolute severity (Score). Four configurations were consequently developed for each scale: Single-Deterioration, Single-Score, Multi-Deterioration, and Multi-Score. Data of participants recruited before May 1, 2024, underwent cross-validation, and the resulting fine-tuned models were then externally validated using data from the remaining participants. Of the 244 enrolled participants, 191 (78.3%; 3954 person-days) were included in the final analysis after applying the exclusion criteria. The demographic and clinical characteristics of participants, as well as the distribution of sensor data, showed considerable variations across wards and hospitals. Data of 139 participants were used for cross-validation, while data of 52 participants were used for external validation. The Single-Deterioration and Multi-Deterioration models achieved similar overall accuracy values of 0.75 in cross-validation and 0.73 in external validation. The Single-Score and Multi-Score models attained overall R² values of 0.78 and 0.83 in cross-validation and 0.66 and 0.74 in external validation, respectively, with the Multi-Score model demonstrating superior performance. Deep learning models based on wearable sensor data effectively classified symptom deterioration and predicted symptom severity in participants in acute psychiatric wards. Despite lower computational costs, Multi models demonstrated equivalent or superior performance than Single models, suggesting that multitask learning is a promising approach for comprehensive symptom prediction. However, significant variations were observed across wards, which presents a key challenge for developing clinical decision support systems in acute psychiatric wards. Future studies may benefit from recurring local validation or federated learning to address generalizability issues.

Validity and Reliability Study of the Turkish Version of the "Uncivil Behavior in Clinical Nursing Education Scale" for Nursing Students.

Background and Purpose: Nursing undergraduate students may encounter unkind behaviors during their clinical education. This study aimed to test the psychometric properties of the Turkish version of the "Uncivil Behavior in Clinical Nursing Education (UBCNE) Questionnaire," which measures nursing students' perceived levels of uncivil behavior in the clinical learning environment. Methods: The study was conducted in four steps: translation, face and content validity, confirmatory factor analysis, and reliability assessment. Data were collected from 216 nursing students from Turkey. Results: In the exploratory factor analysis (EFA), a unidimensional factor structure with an eigenvalue above 1 was detected, and therefore the scale was analyzed as a single dimension. In the EFA with 13 items, the Kaiser-Meyer-Olkin value was 0.943, and the Barlett test result was 1862.965 (p <05). The fit indices for the class were calculated as χ2/df = 2.218, root-mean-square error of approximation = .075, and comparative fit index = 0.961. Cronbach's coefficient α (alpha) for Turkish culture was .942. Conclusion: The study showed that the Turkish version of the UBCNE scale is a valid and reliable measurement tool for assessing perceived incivil behaviours in nursing students' clinical education.