Clinical Use Research Articles

Associations of Clinical Characteristics With Sudden Cardiac Arrest in People With Type 2 Diabetes With and Without Cardiovascular Disease: A Longitudinal Case-Control Study Using Routine Primary Care Data

OBJECTIVE To assess longitudinal associations with sudden cardiac arrest (SCA) of clinical characteristics recorded in primary care in people with type 2 diabetes (T2D), both with and without cardiovascular disease (CVD). RESEARCH DESIGN AND METHODS We performed a case-control study, with SCA case subjects with T2D from the Amsterdam Resuscitation Studies (ARREST) registry of out-of-hospital resuscitation attempts in the Dutch Noord-Holland region (2010–2020) and up to five matched (age, sex, T2D, general practitioner [GP] practice) non-SCA control subjects. We collected relevant clinical measurements, medication use, and medical history from GPs’ electronic health care records. We analyzed the associations of clinical characteristics and medication use with SCA in the total sample and in subgroups with or without CVD using multivariable time-dependent Cox regression (hazard ratios, 95% confidence intervals). RESULTS We included 689 SCA case subjects and 3,230 non-SCA control subjects. In multivariable models, low fasting glucose (<4.5 mmol/mol: 1.91 [1.00–3.64]), antihypertensive (1.80 [1.39–2.33]), glucose lowering (oral only: 1.32 [1.06–1.63]; insulin only: 2.31 [1.71–3.12]; oral and insulin: 1.64 [1.21–2.22]), heart failure (1.91 [1.55–2.35]), and QTc-prolonging prokinetic (1.78 [1.27–2.50]), antibiotic (1.35 [1.05–1.73]), and antipsychotic (2.10 [1.42–3.09]) medication were associated with SCA in the total sample. In subgroup effect modification analyses, QTc-prolonging antibiotic (1.82 [1.26–2.63]) and antipsychotic (3.10 [2.09–4.59]) medication use were associated with SCA only in those without CVD. CONCLUSIONS In people with T2D, low fasting glucose and QTc-prolonging prokinetic, antibiotic, or antipsychotic medication use and a history of heart failure are associated with SCA risk. Subgroup analyses indicate antibiotic and antipsychotic medication use increases SCA risk specifically in those without CVD.

The establishment of reference intervals for the ClotPro thromboelastometry device in healthy dogs

AbstractObjectiveTo establish reference intervals using a new point‐of‐care thromboelastometry device in dogs for the extrinsically activated test (EX‐test), intrinsically activated test (IN‐test), fibrin polymerization test (FIB‐test), ecarin test (ECA‐test), and tissue plasminogen activator test (TPA‐test) and to investigate the effects of storage time on the results.DesignProspective clinical study in 2022.SettingUniversity teaching hospital.AnimalsForty‐eight healthy privately or university‐owned dogs were prospectively enrolled and included on the basis of normal physical examination and normal baseline laboratory results (CBC, biochemistry profile, prothrombin time, and activated partial thromboplastin time [aPTT]).InterventionsAfter a 30‐minute storage time, the EX‐test, IN‐test, FIB‐test, ECA‐test, and TPA‐test were performed on citrated blood samples. To determine the effect of storage time, 11 samples had the EX‐test, FIB‐test, and IN‐test repeated 90 and 150 minutes after sample collection.Measurements and Main ResultsTen thromboelastometry parameters were evaluated for each test. Reference intervals were calculated using the robust method for parametric data, and the robust Box–Cox transformed or nonparametric methods were used for nonparametric data. Increasing storage time resulted in more hypocoagulable tracings. A correlation was found between the IN‐test and aPTT (r = 0.62, P < 0.0001). Other weak to moderate correlations were seen between thromboelastometry parameters and platelet count and hematocrit.Conclusions and Clinical ImportanceThe development of reference intervals for the thromboelastometry device allows for the clinical use of this technology. Analyzing samples after a prolonged storage time of more than 30 minutes may result in erroneous results. Results may also be affected by an abnormal hematocrit or platelet count.

Parkinson’s Disease: Biomarkers for Diagnosis and Disease Progression

Parkinson’s disease (PD) is a progressive neurological disease that causes both motor and nonmotor symptoms. While our understanding of putative mechanisms has advanced significantly, it remains challenging to verify biomarkers with sufficient evidence for regular clinical use. Clinical symptoms are the primary basis for diagnosing the disease, which can be mild in the early stages and overlap with other neurological disorders. As a result, clinical testing and medical records are mostly relied upon for diagnosis, posing substantial challenges during both the initial diagnosis and the continuous disease monitoring. Recent biochemical, neuroimaging, and genetic biomarkers have helped us understand the pathophysiology of Parkinson’s disease. This comprehensive study focuses on these biomarkers, which were chosen based on their relevance, methodological excellence, and contribution to the field. Biochemical biomarkers, including α-synuclein and glial fibrillary acidic protein (GFAP), can predict disease severity and progression. The dopaminergic system is widely used as a neuroimaging biomarker to diagnose PD. Numerous genes and genome wide association study (GWAS) sites have been related to the development of PD. Recent research on the SNCA gene and leucine-rich repeat protein kinase 2 (LRRK2) has shown promising results. By evaluating current studies, this review intends to uncover gaps in biomarker validation and use, while also highlighting promising improvements. It emphasizes the need for dependable and reproducible indicators in improving PD diagnosis and prognosis. These biomarkers may open up new avenues for early diagnosis, disease progression tracking, and the development of personalized treatment programs.

Physical, biochemical, and biological characterization of olive-derived lipid nanovesicles for drug delivery applications

AbstractExtracellular vesicles (EVs) have shown great promise as drug delivery system (DDS). However, their complex and costly production limit their development for clinical use. Interestingly, the plant kingdom can also produce EV-like nanovesicles that can easily be isolated and purified from a large quantity of raw material at a high yield. In this study, olive-derived nanovesicles (ODNVs) were isolated from raw fruits using serial centrifugations and their physical and biological features characterized to demonstrate their promising potential to be used as a DDS. Nanotracking particle analysis indicated an average size of 109.5 ± 3.0 nm and yield of 1012 ODNVs/mL for the purest fraction. Microscopy imaging, membrane fluidity assay and lipidomics analysis showed the presence of a rich lipid bilayer that significantly varied between different sources of ODNVs but showed a distinct signature compared to human EVs. Moreover, ODNVs were enriched in PEN1 and TET8 compared to raw fruits, suggesting an extracellular origin. Interestingly, ODNVs size and yield stayed unchanged after exposure to high temperature (70 °C for 1 h), wide pH range (5–10), and 50–100 nm extrusion, demonstrating high resistance to physical and chemical stresses. This high resistance allowed ODNVs to stay stable in water at 4 °C for a month, or with the addition of 25 mM trehalose for long-term freezing storage. Finally, ODNVs were internalized by both 2D and 3D cell culture without triggering significant cytotoxicity and immunogenicity. Importantly, the anticancer drug doxorubicin (dox) could be loaded by passive incubation within ODNVs and dox-loaded ODNVs decreased cell viability by 90% compared to only 70% for free dox at the same concentration, indicating a higher efficiency of drug delivery by ODNVs. In addition, this high cytotoxicity effect of dox-loaded ODNVs was shown to be stable after a 2-week storage at 4 °C. Together, these findings suggested that ODNVs represent a promising candidate as drug nanocarrier for various DDS clinical applications, as demonstrated by their biocompatibility, high resistance to stress, good stability in harsh environment, and improvement of anticancer drug efficacy.

Increased cell killing effect in neutron capture enhanced proton beam therapy

AbstractThermal neutrons generated in the body during proton beam therapy (PBT) can be used to cause boron neutron capture reactions and have recently been proposed as neutron capture enhanced PBT (NCEPBT). However, the cell killing effect of NCEPBT remains underexplored. Here, we show an increase in the cell killing effect of NCEPBT. Using Monte Carlo simulations, we showed that neutrons generated by proton beam irradiation are uniformly spread on tissue culture plates. Human salivary gland tumor cell line (HSG), human osteosarcoma cell line (MG63), human tongue squamous cell carcinoma cell line (SAS), and human malignant melanoma cell line (G-361) were irradiated with X-rays, proton beams, and proton beams with 10B-enriched boronophenylalanine (boron concentration of 20 and 80 ppm). The relative biological effectiveness (RBE) values of proton beams alone, proton beams with 20 ppm boron, and proton beams with 80 ppm boron for HSG, MG63, SAS, and G-361 were 1.02, 1.07, and 1.23; 1.01, 1.08, and 1.44; 1.05, 1.09, and 1.46; and 1.04, 1.13, and 1.63, respectively. NCEPBT with high boron concentration showed high RBE and a high sensitizing effect. Our results confirm an increase in the cell killing effect of NCEPBT, should aid in its clinical use, and warrant its further investigation.

Demonstration of the Clinical Utility of a Psoriasis Precision Medicine Tool: Findings from the MATCH Study Show Altered Physician Behavior Leads to Improved Patient Outcomes

Demonstration of the Clinical Utility of a Psoriasis Precision Medicine Tool: Findings from the MATCH Study Show Altered Physician Behavior Leads to Improved Patient Outcomes

The Utility of Pre-Procedural Blood Tests in Neuraxial Blocks: A Retrospective Study in High-Risk Patients

Background/Objectives: The necessity and clinical utility of routine pre-procedural blood tests (PBTs) before neuraxial blockade remain controversial. This study evaluates the effectiveness of PBTs in identifying clinically significant conditions in an outpatient setting. Methods: This single-center retrospective study involved patients who received neuraxial blockades from January 2020 to August 2023. We extracted medical information and laboratory data from the electronic medical records during the pre-procedural period. Through a multivariate regression analysis, we identified patient factors associated with abnormal laboratory results. Results: Advanced age (OR, 1.021; p = 0.026) and a history of cancer (OR, 2.359; p = 0.016) were significantly associated with elevated CRP levels (>0.30 mg/dL). Severe hyperglycemia (≥200 mg/dL) was found in 24 patients (3.88%), with a history of cancer being a strong predictor (OR, 6.764; p < 0.001). No significant abnormalities were observed in PT or PTT. Reduced eGFR (<60 mL/min/1.73 m2) was detected in 8.62% of patients, despite no prior history of renal dysfunction. A multivariate analysis revealed that advanced age, hypertension, cancer, and coronary artery disease were significant predictors of abnormal PBT results, highlighting the importance of selective testing in high-risk patients. Conclusions: Routine PBTs are not universally required for all patients undergoing neuraxial blockade but can provide crucial information in high-risk populations. A selective testing approach based on individual risk factors is recommended to optimize patient safety and resource utilization. Future studies should aim to establish clear guidelines for targeted PBT use.

Choroidal vascularity index in health and systemic diseases: a systematic review

Abstract Background The choroid, a highly vascular structure within the eye, is significantly influenced by various systemic conditions. The advent of enhanced depth optical coherence tomography has improved our ability to evaluate choroidal pathophysiology. The choroidal vascularity index (CVI), a noninvasive and reliable tool, serves as an effective means of assessing the choroidal vascular structure. Recent studies have increasingly focused on exploring CVI alterations under different systemic conditions. This study aims to provide a comprehensive summary of the latest research findings in this area. Methods A systematic literature review was conducted on October 1, 2023, using two databases, MEDLINE (via PubMed) and Scopus. Search terms were tailored specifically for each database to ensure a thorough exploration of relevant literature. The studies identified were qualitatively assessed, with particular emphasis on outcomes related to CVI and choroidal thickness. Results A total of 48 studies were included in the review, encompassing a diverse range of systemic conditions such as diabetes, central nervous system disorders, cardiovascular diseases, autoimmune disorders, and infectious diseases. Notable reductions in CVI were observed in diabetic retinopathy, autoimmune diseases, and neurodegenerative disorders. Additionally, the review highlighted variations in CVI values related to the severity of systemic diseases, indicating its potential use as a biomarker for disease progression. Conclusion This review highlights the significant correlation between variations in the choroidal vascularity index and diverse systemic conditions affecting hemodynamics. An enhanced understanding of CVI provides deeper insights into the pathophysiological mechanisms underlying these disorders and positions CVI as a promising biomarker for early detection and monitoring. Nevertheless, its clinical utility warrants careful assessment. Future research should address the potential limitations of CVI to fully capitalize on its diagnostic and prognostic potential.

Targeted Multiplex Proteomics for the Development and Validation of Biomarkers in Primary Aldosteronism Subtyping

Abstract Objective Primary aldosteronism (PA), a significant cause of secondary hypertension affecting approximately 10% of patients with severe hypertension, exacerbates cardiovascular and cerebrovascular complications even after blood pressure control. PA is categorized into two main subtypes: unilateral aldosterone-producing adenomas (APA) and bilateral hyperaldosteronism (BHA), each requiring distinct treatment approaches. Accurate subtype classification is crucial for selecting the most effective treatment. The goal of this study was to develop novel blood-based proteomic biomarkers to differentiate between APA and BHA subtypes in patients with PA. Design and Methods Five subtyping differential protein biomarker candidates (APOC3, CD56, CHGA, KRT5, and AZGP1) were identified through targeted proteomic profiling of plasma. The subtyping efficiency of these biomarkers was assessed at both the tissue gene expression and blood protein expression levels. To explore the underlying biology of APA and BHA, significant differential pathways were investigated. Results The five-protein panel proved highly effective in distinguishing APA from BHA in both tissue and blood samples. By integrating these five protein biomarkers with aldosterone and renin, our blood-based predictive methods achieved remarkable ROC AUCs of 0.986 (95% CI: 0.963-1.000) for differentiating essential hypertension (EH) from PA, and 0.922 (95% CI: 0.846-0.998) for subtyping APA versus BHA. These outcomes surpass the performance of the existing Kobayashi score subtyping system. Furthermore, the study validated differential pathways associated with the pathophysiology of primary aldosteronism, aligning with current scientific knowledge and opening new avenues for advancing PA care. Conclusions The new blood-based biomarkers for PA subtyping hold the potential to significantly enhance clinical utility and advance the practice of PA care.

Are Nomograms Useful for Predicting Sentinel Lymph Node Status in Melanoma Patients?

ABSTRACTBackground and ObjectivesClinical nomograms have been developed to predict sentinel lymph node (SLN) status in early‐stage melanoma patients, but the clinical utility of these tools remains debatable. We created and validated a nomogram using data from a randomized clinical trial and assessed its accuracy against the well‐validated Melanoma Institute Australia (MIA) nomogram.MethodsWe developed our model to predict SLN status using logistic regression on clinicopathological patient data from the Multicenter Selective Lymphadenectomy Trial‐I. The model was externally validated using the National Cancer Database (NCDB) data set, and its performance was compared to that of the MIA nomogram.ResultsOur model had good discrimination between positive and negative SLNs, with a training set area under the curve (AUC) of 0.706 (0.661–0.751). Our model achieved an AUC of 0.715 (0.706–0.724) compared to 0.723 (0.715–0.731) with the MIA model, using the NCDB set.ConclusionOur model performed similarly to the MIA model, confirming that despite using different clinical features and data sets, no clinical nomogram is currently accurate enough for clinical use.

Abstract C049: Implementing human bone marrow organoids to interrogate microenvironmental influences on the efficacy of blood cancer immunotherapies

Abstract Cancers perpetuate immunosuppressive niches that fuel progression and hinder the clinical utility of immunotherapies. Pre-clinical validation of immunotherapies often relies on 2D models that lack spatial complexity, or mouse models that do not fully recapitulate the human tumor microenvironment (TME). We recently developed a human induced pluripotent stem cell (iPSC)-derived bone marrow organoid platform, creating a robust system to interrogate cancer-stroma cross talk. Here, we further develop the model to create the first in vitro system for evaluation of blood cancer immunotherapies in a human tissue environment. We focused on targeting myeloproliferative neoplasms (MPNs) – chronic blood cancers affecting ∼400,000 individuals in the US that are currently largely incurable. 1/3 of MPNs are driven by a mutation in calreticulin (mutCALR), and the mutCALR oncoprotein is displayed on the cell surface of disease-initiating stem cells and fibrosis-driving megakaryocytes. MPNs are characterized by pronounced inflammation and fibrosis (“myelofibrosis”). Therapeutic antibodies have entered first-in-human trials, and we recently presented pre-clinical data for the first mutCALR-directed CAR-T cell therapy. However, it is unclear how the TME may limit the efficiency of these agents. To address this, we first tested the utility of the organoid platform to evaluate target killing of leukemic cell lines and cells from patients using the anti-CD33 antibody-drug conjugate (ADC) gemtuzumab. The ADC showed good penetration and potent target cell killing in organoids, with only minimal reduction in efficacy in TGFβ-rich/fibrotic microenvironments. We then developed more sophisticated ‘chimeroids’ comprising the iPSC organoid scaffold engrafted with malignant stem cells plus CAR-Ts. Addition of mutCALR-directed CAR-Ts induced highly selective killing of mutCALR+ stem/progenitor cells with minimal toxicity to non-malignant cells including niche cells (n=5 donors), replicating the highly specific and robust killing seen in 2D cultures. To evaluate the impact of TME perturbations on CAR-T phenotype and function, we generated a scRNAseq dataset of 161,970 cells from ‘healthy’ and ‘myelofibrotic’ organoids engrafted with patient-derived mutCALR+ cells plus CAR-Ts, pre-stimulating organoids with two immunoregulatory cytokines highly abundant in myelofibrosis -TGFβ and Galectin-1. CAR- Ts exposed to mutCALR+ cells showed a 2x expansion of CD8+ effector memory cells, with significant enrichment of IFNγ, TNFα and IL-2 signaling gene-sets. The proportion of immunosuppressive Tregs increased in a high TGFβ TME, but not with Galectin-1. Ongoing analyses are further interrogating crosstalk between CAR-T, target cells, and niche components, and exploring the pro-inflammatory impact of CAR-T killing on the marrow niche – a potential mediator of delayed hematopoietic recovery. These studies demonstrate the utility of human organoids to evaluate blood cancer-targeting immunotherapies within a relevant human TME, with broad relevance for mechanistic and translational studies. Citation Format: Zoë C. Wong, Yuqi Shen, Alex Rampotas, Isaac Gannon, Charlotte Brierley, Camelia Benlabiod, Nawshad Hayder, Eleanor Murphy, Aude-Anaïs Olijnik, Claire Roddie, Martin Pulé, Adam J. Mead, Abdullah O. Khan, Bethan Psaila. Implementing human bone marrow organoids to interrogate microenvironmental influences on the efficacy of blood cancer immunotherapies [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 C049.

Abstract C042: TMEM doorway score as a predictive biomarker for therapy selection in ER+/HER2- breast cancer

Abstract Background: Endocrine therapy is the standard of care for ER+/HER- breast cancer; but some patients need additional adjuvant chemotherapy to counter the risk of metastasis. While chemotherapy can reduce metastasis, some patients still relapse. Tumor cells metastasize through TMEM (Tumor Microenvironment of Metastasis) Doorways, consisting of a Mena-expressing tumor cell, a macrophage, and a vascular endothelial cell, all in direct contact. High TMEM doorway scores correlate with increased dissemination in mice and metastatic outcomes in ER+/HER2- breast cancer patients. Recent studies show that adjuvant chemotherapy may activate growth in disseminated tumor cells (DTCs). Thus, we hypothesized that patients with high TMEM doorway scores who receive endocrine plus chemotherapy will have worse distant recurrence-free survival (DRFS) than those with lower TMEM Doorway scores, or those who receive endocrine therapy alone. Methods: We conducted a retrospective study on 119 patients with stage I-III, ER+/HER2- unilateral invasive ductal breast carcinoma (age≥18) from 2005- 2018. All underwent surgery and received adjuvant endocrine treatment alone or with chemotherapy. We excluded patients who had metastatic disease at diagnosis, received neoadjuvant chemotherapy, or lacked treatment data. Formalin-fixed paraffin-embedded primary tumor tissues were stained for TMEM doorways. A pathologist categorized the TMEM doorway scores as low, intermediate, or high. Kaplan-Meier curves and Cox-proportional hazard models compared DRFS based on treatment type and TMEM doorway scores. Multivariate logistic regression analyzed the association between macrophage density or TMEM Doorway Score and metastatic outcomes, adjusting for tumor size, grade, and node positivity. Results: Of all patients, 72 received endocrine plus chemotherapy and 47 endocrine therapy alone. In the chemotherapy group, 34 (47.2%) developed metastases, while 38 (52.8%) remained disease-free. In the endocrine alone group, 37 (78.7%) remained disease-free and 9 (19.1%) developed metastases. There was no significant difference in TMEM Doorway Scores between groups (p=0.07). In patients who received endocrine plus chemotherapy, a high TMEM Doorway Score was associated with greater odds of metastasis (OR 6.8; 95% CI 1.35–33.76; p=0.02), controlling for tumor size, grade, and node positivity but not when endocrine therapy was used alone (p=0.817). Patients with high TMEM Doorway Scores had worse DRFS in the endocrine plus chemotherapy group (HR 2.61; 95% CI 1.08–6.27; p=0.03), but not in the endocrine therapy alone group (p=0.77) adjusted similarly. Conclusions: This study highlights potential clinical utility of TMEM Doorway Scores as a predictive biomarker for chemotherapy selection in ER+/HER2- breast cancer patients. Larger trials are needed to confirm these findings and with confirmation, physicians considering giving chemotherapy to ER+/HER2- breast cancer patients might consider pre-treatment stratification using TMEM Doorway Scores to mitigate the risk of chemotherapy-induced metastasis. Citation Format: Priyanka Parmar, Andrew Miller, Burcu Karadal-Ferrena, Suryansh Shukla, Cien Huang, Mindy Ginsberg, Chedva Rosenbaum, Malka Felder, Chenxin Zhang, John S. Condeelis, Xiaonan Xue, Thomas E. Rohan, Joseph A. Sparano, Jesus D. Anampa, David Entenberg, Maja H. Oktay. TMEM doorway score as a predictive biomarker for therapy selection in ER+/HER2- breast cancer [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 C042.

Abstract B007: Identification of Asporin as a HER3 ligand exposes a therapeutic vulnerability in metastatic prostate cancer

Abstract Background: Rates of prostate cancer incidence and advanced-stage diagnoses are increasing. Despite curative-intent therapies, nearly all patients with advanced disease progress to metastatic castration-resistant prostate cancer (mCRPC), a lethal form of the disease. There is an urgent unmet need to understand the mechanisms driving prostate cancer progression and identify alternative therapies for patients with mCRPC. It is well-established that cancer-associated fibroblasts (CAF) in the tumor microenvironment secrete factors that enable cancer cells to establish metastases. Asporin (ASPN) is an extracellular protein secreted by a subset of CAF and is associated with worse oncologic outcomes, yet the mechanisms remain poorly understood. In this study, we sought to identify the molecular interactions between the CAF-secreted ASPN and metastatic prostate cancer cells. Methods: ASPN-induced HER2/HER3 activation and downstream signaling was identified by RNA-sequencing and then confirmed by immunoblotting in multiple human metastatic prostate cancer cell lines. Binding interactions were assessed computationally by AlphaFOld2 with Rosetta refinement and substantiated by co-immunoprecipitation assays. CRISPR-cas9 targeting of HER2 and HER3 and therapeutic inhibition of HER2 with Tucatinib were used to determine the role of HER2/HER3 in ASPN-induced signaling and migration. The efficacy of Tucatinib and antibody-drug conjugates (ADC) targeting HER2 (Trastuzumab-deruxtecan) and HER3 (Patritumab-deruxtecan) were assessed in multiple metastatic prostate cancer cell lines in vitro. Trastuzumab-deruxtecan was assessed in PC3 xenografts in vivo. Prostate cancer metastases from 33 patients were assessed by IHC for HER2 and HER3 and by RNAscope for ASPN. Results: We report that ASPN is a novel ligand of HER3 and induces HER3 heterodimerization with its preferred dimerization partner, HER2. ASPN activates established ErbB-associated pathways including Phosphoinositide 3-kinase (PI3K), Mitogen-activated protein kinase (MAPK), and calcium signaling, in multiple metastatic prostate cancer cell lines to promote cell migration. Genetic and molecular inhibition of HER2/HER3 mitigates ASPN-induced signaling and cell migration, suggesting these receptors are required for paracrine activation by ASPN. Importantly, small molecule and ADC therapies targeting HER2/HER3 significantly diminished prostate cancer cell growth in vitro, with enzalutamide-resistant cells showing increased sensitivity. Trastuzumab-deruxtecan nearly resolved tumors in an in vivo xenograft model. Lastly, ASPN+ CAF in the TME of HER2/HER3-expressing metastatic prostate cancer is frequently observed in patient samples, supporting the clinical relevance of these findings. Conclusions: Collectively, these findings indicate ASPN functions as a HER3 ligand to induce cellular migration, and inhibition with anti-HER2/HER3 therapies highlights potential clinical utility for patients with HER2-expressing metastatic prostate cancer. Citation Format: Amanda B. Hesterberg, Hong Yuen Wong, Jorgen Jackson, Monika Antunovic, Brenda L. Rios, Evan Watkins, Riley E. Bergman, Brad A. Davidson, Sarah E. Ginther, Diana Graves, Elliott F. Nahmias, Jared A. Googel, Lillian B. Martin, Violeta Sanchez, Paula Gonzalez Ericsson, Quanhu Sheng, Benjamin P. Brown, Jens Meiler, Ben H. Park, Kerry R. Schaffer, Jennifer B. Gordetsky, Paula J. Hurley. Identification of Asporin as a HER3 ligand exposes a therapeutic vulnerability in metastatic prostate cancer [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 B007.

A delphi-based model for prognosis of femoral head collapse in osteonecrosis: a multi-factorial approach.

Osteonecrosis of the femoral head (ONFH) is a progressive and debilitating condition characterized by the death of bone tissue due to inadequate blood supply. Despite advances in diagnostic imaging and treatment strategies, predicting the risk of femoral head collapse remains a significant clinical challenge. This study seeks to address this gap by developing a robust prognostic model that integrates clinical, imaging, and laboratory data to improve early diagnosis and guide therapeutic decision-making. We conducted a qualitative systematic review and employed the Delphi method to select key prognostic factors from clinical data, imaging findings, and laboratory indicators. The study included ONFH patients treated from January 2014 to December 2021. We used univariate and multivariate Cox regression analyses to develop a nomogram for predicting the risk of femoral head collapse. The model's performance was evaluated using the concordance index (C-index), calibration plots, and decision curve analysis (DCA). The study included 297 patients (454 hips) with ONFH. Key prognostic factors identified included pain presence (p < 0.001, RR = 0.185, 95% CI: 0.11-0.31), JIC classification (C1: p < 0.001, RR = 0.096, 95% CI: 0.054-0.171; C2: p < 0.001, RR = 0.323, 95% CI: 0.215-0.487), necrotic area (3 < MNAI < 6: p < 0.001, RR = 0.107, 95% CI: 0.061-0.190; MNAI ≥ 6: p < 0.001, RR = 0.466, 95% CI: 0.314-0.692), weight-bearing reduction (p < 0.001, RR = 0.466, 95% CI: 0.323-0.672), preservation of the anterolateral pillar (p < 0.001, RR = 0.223, 95% CI: 0.223-0.473), and subchondral bone fracture on CT (p < 0.001, RR = 0.32, 95% CI: 0.217-0.472). The nomogram demonstrated a high C-index of 0.88, indicating excellent predictive accuracy. Calibration plots showed good agreement between predicted and observed outcomes, and DCA confirmed the model's clinical utility. The prognostic model developed in this study provides a reliable tool for predicting femoral head collapse in ONFH patients. It allows for early identification of high-risk patients, guiding personalized treatment strategies to improve patient outcomes and reduce the need for invasive surgical procedures.

Prediction of Ki-67 expression and malignant potential in gastrointestinal stromal tumors: novel models based on CE-CT and serological indicators.

To identify more reliable imaging and serological indicators for predicting Ki-67 expression and malignant potential in gastrointestinal stromal tumors, as well as to develop a preoperative prediction model with clinical utility. This study retrospectively analyzed patients with gastrointestinal stromal tumors (GIST) diagnosed at the First Affiliated Hospital of Jinzhou Medical University between May 2018 and May 2024. Univariate logistic analyses, two-way stepwise regression, P-value stepwise regression, and LASSO regression were employed to screen for Ki-67 high expression and high malignant potential risk factors associated with GIST. Models were established using various regression methods; Nomograms, calibration curves, and clinical decision curves were generated for the two best prediction models. Two-way stepwise regression analysis revealed that diameter (P=0.037; OR=1.22; 95% CI: 1.01 - 1.46), growth pattern (extraluminal type: P=0.028; OR=3.54; 95% CI: 1.14 - 10.94), enhancement model (P=0.099; OR=0.39; 95% CI: 0.12 - 1.20), EVFDM (P=0.069; OR=0.43; 95% CI: 0.17 - 1.07), PLR (P=0.099; OR=3.06; 95% CI: 0.81 - 11.59), and OPNI (P=0.058; OR=2.38; 95% CI: 0.97 - 5.84) are identified as independent risk factors for Ki-67 expression. Utilizing the two-way stepwise regression model to predict Ki-67 expression, the area under the curve (AUC) for the training group was 0.865 (95% CI: 0.807-0.922), while for the validation group it was 0.784 (95% CI: 0.631-0.937). The Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC) for the training group were 153.360 and 174.619, respectively. Two-way stepwise regression analysis revealed that volume (P < .001, OR = 1.06; 95% CI: 1.03 - 1.09), contour (P = 0.066; OR = 0.17; 95% CI: 0.05 - 0.62), ulcer (P = 0.094; OR = 0.16; 95% CI: 0.03 - 0.98), IBSC (P = 0.008; OR = 5.27; 95% CI: 1.57 - 17.69), and OPNI (P = 0.045; OR = 0.22; 95% CI: 0.05 - 0.96) are independent risk factors for malignant potential. Utilizing the two-way stepwise regression model to predict malignant potential, the AUC for the training group was 0.950 (95% CI: 0.920 - 0.980), while for the validation group it was 0.936 (95% CI: 0.867 - 1.000). The AIC and BIC values for the training group were 96.330 and 114.552, respectively. Diameter, growth pattern, enhancement pattern, EVFDM, PLR, and OPNI are independent risk factors for GIST with high Ki-67 expression. Additionally, volume, contour, ulceration, IBSC, and OPNI serve as independent risk factors for GIST with high malignant potential. The preoperative models developed using CT images can predict the malignant potential and Ki-67 expression status of GIST to a certain extent. When combined with serological indicators, these models' predictive performance can be further enhanced.

Sumac liposomes/mesenchymal stem cells fight methotrexate-induced nephrotoxicity in rats via regulating Nrf-2/Keap-1/HO-1 and apoptotic signaling pathways.

Methotrexate (MTX) is commonly employed in cancer treatment, but its clinical use is restricted due to the MTX-associated renal injury. This study investigates the combined potential of Rhus coriaria (sumac) and bone marrow mesenchymal stem cells (BMMSCs) against MTX-induced nephrotoxicity in rats. The high-resolution-liquid chromatography-mass spectrometry (HR-LC-MS) of sumac extract tentatively identified 22 phytochemicals, mostly flavonoids, anthocyanins, and steroids. Preparation of sumac liposomes attained a suitable particle size of 3041.33 ± 339.42 nm, a polydispersity index of 0.208 ± 0.086, and an encapsulation efficiency of 84.92 ± 3.47%. Rat BMMSCs were injected into the tail vein of the experimental rats (0.5 × 106 cells, intravenous [iv]) of seven treated groups. The experimental design relies on either pre- or posttreatment of rats with intraperitoneal (IP) sumac liposomes (SL) (200 mg/kg, daily with a dose of MTX (300 µg/kg/14 days). The histopathological examination and serum analysis of creatinine and urea revealed good results, besides regulating levels of oxidative stress and inflammatory markers. Additionally, a significant decrease in the gene expression levels of B-Cell Lymphoma 2 (Bcl-2) and caspases-3 and -9, a remarkable increase in the Bcl-2 Associated X-Protein (Bax), nuclear factor erythroid 2-related factor 2 (Nrf2), and heme-oxygenase 1 expression, and a downregulation of Kelch-like ECH-associated protein 1 (Keap1). Collectively, the coadministration of SL with BMMSCs might be a potent therapeutic strategy for attenuation of MTX-induced renal damage. The network pharmacology analysis identified the involved key hub genes as KEAP1, Nrf2, HMOX1, mitogen-activated protein kinase (MAPK1), nuclear factor-kappa B (NF-KB), interleukin-1 beta (IL-1B), and caspase-3. The docking results revealed strong binding affinities of 7-O-methyl-cyanidin-3-O-(2″-galloyl)-galactoside with Keap1 and amentoflavone with MAPK. These insights pave the way for future experimental validation and therapeutic development of sumac-based phytoconstituents against MTX-induced nephrotoxicity.

Pathology of doxorubicin-induced organopathies under different intravenous nicotinamide riboside administration modes

Doxorubicin (DOX) is a potent chemotherapeutic drug, but its clinical use is hindered by significant side effects on vital organs like the heart, kidneys, lungs, liver, and intestines. Currently, there is a lack of effective drugs that can provide simultaneous cardioprotection and organ protection during chemotherapy. Nicotinamide riboside (NR) holds promise as a pharmacological agent capable of offering comprehensive protection against the systemic toxicity caused by DOX. This study aimed to comprehensively evaluate the morphological characteristics of vital organs (heart, lungs, liver, kidneys) in Wistar rats with chronic doxorubicin-induced cardiomyopathy using various intravenous administration modes of NR as a protective agent. Sixty male SPF Wistar rats weighing 283 ± 22 g were divided into four groups: intact, control (DOX administered intraperitoneally), combined mode (the simultaneous use of DOX and NR) and preventive mode (the preliminary use of NR to realize the cumulative effect in cells, and its further joint use together with DOX) intravenous NR administration. Animal observation spanned two months after drug administration, followed by the collection of hearts, lungs, liver, and kidneys for morphological analysis. Echocardiographic assessment confirmed DOX cardiotoxicity. The study revealed that the hearts, kidneys and lungs exhibited more pronounced toxic effects of DOX compared to the liver. Both NR administration modes demonstrated protective effects, with the preventive regime showing the greatest efficacy in safeguarding vital organs.

Efficacy and safety of Piclidenoson in the treatment of plaque psoriasis: a systematic review and meta-analysis of randomized controlled trials.

Psoriasis, an autoimmune inflammatory disease characterized by hyperproliferation of keratinocytes, affects 0.2-4.8% of the global population. The disease manifests primarily as plaque psoriasis, causing chronic physical and psychological burdens. Although numerous treatments exist, there is ongoing exploration for novel therapies due to concerns about the toxicity, efficacy, and costs of current options. An A3 adenosine receptor (A3AR) agonist called Piclidenoson has been shown to reduce inflammation and could be used to treat moderate to severe psoriasis. This meta-analysis evaluates the safety and efficacy of Piclidenoson in psoriasis treatment. This meta-analysis was conducted accordance with the PRISMA guidelines and has been registered on PROSPERO (CRD42024566459). A comprehensive search on Cochrane CENTRAL, PubMed/MEDLINE, and Google Scholar was conducted up to July 2024. Included studies were randomized controlled trials (RCTs) involving Piclidenoson. Efficacy outcomes included PASI 75 and PGA 0 or 1, while safety outcomes included adverse events. Pooled outcomes were presented as odds ratio (OR) with 95% confidence intervals (CI). Statistical analysis employed the Mantel-Haenszel random-effects model, and heterogeneity was assessed using the I2 and X2index. Three RCTs with 574 patients (313 Piclidenoson, 261 placebo) met the inclusion criteria. The pooled analysis showed no significant difference in achieving PASI 75 between Piclidenoson and placebo (OR: 1.62, 95% CI 0.70-3.75, P = 0.26, I2 = 12%). However, Piclidenoson significantly improved PGA scores (OR: 2.74, 95% CI 1.22-6.16, P = 0.01, I²=0%). Safety assessment revealed no significant differences in adverse events, including nervous system, gastrointestinal, musculoskeletal, renal, and infections, compared to placebo. Piclidenoson demonstrates a significant improvement in PGA scores and a favorable safety profile, suggesting it could be a valuable addition to plaque-like psoriasis treatment. Further research with larger, longer-term RCTs is needed to confirm its efficacy and optimize clinical use.

Submyocardial Pacing Threshold Distribution During Cold Saline Application; Exploring Reversible Arrhythmia Inhibition.

Cold saline application through an irrigation catheter may induce reversible inhibition of submyocardial excitation, which may be used to identify in advance an ideal site for radiofrequency (RF) energy delivery around delicate areas. An open irrigation catheter was positioned vertically or parallel with 10-g contact on coronary perfusing porcine hearts and the contacted myocardium was cooled by cold saline at 4°C (20mL/min). A temperature electrode was inserted 2mm below the myocardial surface (intra-myocardial temperature at approximately 2 mm below the surface [Temp-BS]). Pacing threshold inside the ventricular wall was measured using aneight-pole electrode plunge needle inserted 5-8mm close to the ablation catheter, and percent increase of the pacing threshold by the cold saline application (%increase-PT) was calculated. During cooling, pacing at 10V from the myocardial surface interrupted constant capture in 7/10 experiments in vertical and in 9/10 experiments in parallel ablation catheter contact. Minimum Temp-BS was not different in both catheter contact positions (25.9±4.0°C in vertical vs. 25.4±2.6°C in parallel). Large % increase-PT on the surface myocardium decreased as the myocardial depth became deeper, and > 150% increase was at a depth approximately 2-3mm from the surface and > 120% increase around 6-7mm from the surface. After cessation of cold saline application, the increased %increase-PT recovered to the pre-cooling values. Cold saline application through an irrigation catheter reversibly inhibited submyocardial excitation. This simple method may have the potential to pre-determine the ideal ablation site in sensitive areas of the heart, although further studies and technological adjustments are required before clinical use.

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.