Citations

Abstract B cell maturation antigen (BCMA)-targeted chimeric antigen receptor T (CART) cell therapies have emerged as a potentially curative treatment for multiple myeloma (MM). However, 50-70% of patients relapse within 3 years. The MM tumor microenvironment (TME) is highly immunosuppressive with an abundance of bone marrow (BM) cancer-associated fibroblasts (CAFs), which contribute to BCMA-CART cell failure. Therapeutic targeting of BM-CAFs has been beset by 1) an incomplete understanding of CAF interactions with CART cells and other BM cells and 2) toxicity risks due to prolonged depletion of healthy fibroblasts and inefficient drug delivery to the BM. To elucidate how BM-CAFs drive BCMA-CART resistance, we performed single cell RNA sequencing (scRNAseq) of baseline BM aspirates from responders (R, n = 6) and non-responders (NR, n = 6) to BCMA-CART cell therapy. NR BMs were enriched for inhibitory CAFs (iCAFs) that highly express fibroblast activation protein (FAP). Spatial analyses of BM cells revealed greater iCAF-iCAF and iCAF-inhibitory macrophage interactions in NR. These findings suggest that FAP+ iCAFs in NR are abundant in the BM and exhibit significant interactions with immunosuppressive macrophages. Next, we sought to validate our scRNAseq data in preclinical models. Xenograft mice were intravenously (IV) engrafted with luciferase+ (luc+) BCMA+ OPM2 cells and/or patient-derived (pd) BM-CAFs (1x106 each). Following engraftment, mice received 1x106 BCMA-CART cells (IV). BCMA-CART cells eradicated tumors in mice engrafted with OPM2 cells, but not in mice engrafted with pdBM-CAFs and OPM2 cells (p<0.01). In NSG and humanized mice (NSG mice engrafted with human CD34+ cells in the first 48 hours of life), tumor growth was significantly accelerated in the presence of pdBM-CAFs (NSG: p<0.01; humanized: p<0.001). BCMA-CART, OPM2, and pdBM-CAF co-cultures also showed significant increases in immunosuppressive cytokines including IL-1Rα, IL-4 and IL-10 and inhibited CART cell proliferation compared to co-cultures without pdBM-CAFs (IL-1Rα, IL-10: p<0.01; IL-4: p<0.001; proliferation: p<0.05). To overcome CAF-mediated CART dysfunction, we engineered BCMA-CART cells to secrete molecules to target BM FAP+ iCAFs and remodel the MM-TME (STriKEs). We designed two BCMA-CART-STriKE strategies: 1) a tri-specific composed of IL-15, anti-FAP, and anti-CD16 molecules to activate innate immune cells to kill CAFs and 2) bi-specific anti-FAP, anti-SIRPα molecules linked to an antibody Fc region (Fc fusion) targeting the CD47/SIRPα axis to induce phagocytosis of FAP+ iCAFs. We hypothesized that BCMA-CART-STriKEs would surmount CAF-mediated immunosuppression without toxicity. We generated BCMA-CART-STriKEs from healthy human donor T cells. CAR expression, STriKE secretion, and binding were confirmed via flow cytometry. Co-cultures of BCMA-CART-STriKEs, donor-matched macrophages, OPM2 cells or BCMA+ MM1S cells, and pdBM-CAFs showed significantly higher OPM2 or MM1S killing (p<0.001), immune cell activation (p<0.01), and phagocytosis (p<0.05), compared to control BCMA-CART cells secreting IL-15 or Fc fusions targeting only FAP or only SIRPα. Next, we assessed BCMA-CART-STriKE efficacy in vivo. NOD-SCID-γ-/- (NSG) mice received 1x106 pdBM-CAFs and 1x106 luc+ OPM2 or luc+ MM1S cells via IV or subcutaneous (SQ) injection. Mice were randomized after confirming tumor engraftment (via bioluminescence imaging for IV or tumor caliper for SQ) and received IV treatment with 1x106 BCMA-CART-STriKEs (or controls) and 5x105 donor-matched macrophages or CD16+ immune cells. BCMA-CART-STriKE-treated mice achieved total tumor clearance. Treatment with BCMA-CART cells secreting IL-15 led to lethal weight loss despite tumor clearance while BCMA-CART cells secreting Fc fusions targeting only FAP or SIRPα failed to eradicate tumors. Lastly, we assessed the potential toxicities of BCMA-CART-STriKEs in humanized mice engrafted with OPM2 cells and pdBM-CAFs (1x106 each). CART cells expressing BCMA and FAP CARs (BCMA-FAP CART) served as a control. BCMA-FAP CART treatment resulted in significant toxicity and weight loss while BCMA-CART-STriKE treatment did not (p<0.01). In summary, we have identified iCAF interactions with immunosuppressive immune cells as key contributors to BCMA-CART cell failure and designed novel BCMA-CART-STriKEs that overcome this failure in MM preclinical models. These findings support the translation of BCMA-CART-STriKEs to a phase I clinical trial.

Abstract Background Due to the high mortality of immune thrombotic thrombocytopenic purpura (iTTP), rapid clinical evaluation and early intervention are crucial. The PLASMIC score remains a valuable and widely used tool for predicting severe ADAMTS13 deficiency. However, recent studies have shown variable performance across different populations. In addition, its variables may often be missing during initial presentation. Refining the accuracy of early iTTP evaluation could improve timely recognition and treatment, while reducing unnecessary interventions in those eventually ruled out for iTTP. Machine learning advancements offer several advantages for enhancing clinical prediction models: the ability to assess continuous variables, capture complex interactions between them, and handle missing data. We previously utilized XGBoost (extreme gradient boosting) to develop a novel machine learning tool, TTP-14, which improved upon the PLASMIC score's predictive performance in repeat 10-fold cross-validation achieving an AUC of 0.91 (Blood. 2024;144[Suppl 1]:2617). TTP-14 generates a predicted probability for iTTP, expressed as a continuous percentage estimate of iTTP diagnostic likelihood, and is also able to do so with missing variables. However, it has not previously been externally validated. Objectives To externally validate the ability of TTP-14 to predict iTTP diagnosis and improve upon the PLASMIC score. Methods We performed an external validation of TTP-14 using an independent cohort of patients with suspected iTTP at the Mayo Clinic. These patients were not part of the original multi-center development cohort of 106 iTTP patients from the University of Utah, Case Western Reserve University, Rochester Regional Health, and University of Illinois. For the validation cohort, we identified patients who underwent ADAMTS13 activity testing between 2010 and 2025 for evaluation of thrombotic microangiopathy at the Mayo Clinic. iTTP was defined as ADAMTS13 activity <10%, and controls as ADAMTS13 activity >20%. TTP-14 variables were collected at initial presentation: platelet count, lactate dehydrogenase, creatinine, international normalized ratio, reticulocyte percentage, active cancer, indirect bilirubin, mean corpuscular volume, total bilirubin, haptoglobin, neurologic deficit, hemoglobin, age, and history of transplant. Receiver operating characteristic curve (AUC) was reported with 95% confidence intervals (CI). While the model provides a continuous estimate and is not categorical, we evaluated clinically meaningful probability thresholds to illustrate performance in decision-relevant zones. Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) were reported at these select thresholds and analyzed for patients with and without missing variables. Result We included 50 iTTP patients and 56 controls in the external validation cohort. 13 patients had missing PLASMIC variables. TTP-14 achieved an AUC of 0.96 (95% CI: 0.93–0.99), outperforming the PLASMIC score's AUC of 0.90 (95% CI: 0.83–0.96) in the same dataset, and improving upon TTP-14's previously reported cross-validated AUC of 0.91 (95% CI: 0.90–0.92) in the development cohort. Among patients with complete data (n = 93), applying select thresholds for clinical relevance demonstrated: Low risk cut-off (<5% predicted iTTP risk): Sensitivity = 98% Specificity = 71% PPV = 76% NPV = 97% Accuracy = 84% High risk cut-off (≥50% predicted iTTP risk): Sensitivity = 71% Specificity = 98% PPV = 97% NPV = 78% Accuracy = 85% In the full cohort (n = 106) that included those with missing PLASMIC variables, TTP-14 maintained excellent predictive performance with AUC = 0.95 (95% CI: 0.92–0.99). Performance metrics analyzed using the above thresholds also yielded similar results. Conclusion External validation of TTP-14 demonstrated high predictive accuracy for iTTP diagnosis (AUC = 0.96). TTP-14 outperformed the PLASMIC score in the validation cohort (AUC = 0.96 vs 0.90, respectively). TTP-14 offers more flexibility than the PLASMIC score by generating predictions even with incomplete data, and maintained excellent performance in the cohort that included those with missing variables (AUC = 0.95) Its refined accuracy and ability to predict iTTP despite missing data provide a key advantage, offering an alternative tool to improve rapid iTTP diagnosis. TTP-14 may help enhance early recognition and treatment of iTTP while reducing unnecessary interventions in non-TTP patients.

Background The Woven EndoBridge (WEB) device is an effective treatment for wide‐neck aneurysms (WNA). This study aims to identify and compare anatomical and procedural factors that predicts the aneurysm occlusion, both adequate and inadequate after WEB device treatment. Methods A systematic review and meta‐analysis were conducted using PubMed, Embase, Scopus, and Web of Science, including studies up to October 25, 2024. Data on angiographic outcomes of WEB occlusion were extracted and analyzed using R version 4.4.2 Results Nine studies (790 patients) were included. Female patients were 1.64 times more likely to achieve adequate occlusion than males (OR: 1.64, 95% CI: [1.06, 2.52], p = 0.026). Regular‐shaped aneurysms had a 2.44‐fold higher likelihood of adequate occlusion compared to irregular ones (OR: 0.41, 95% CI: [0.23, 0.74]). Aneurysm neck size was significantly smaller in the adequate occlusion group (3.9 mm vs. 4.4 mm, MD: ‐0.48 mm, 95% CI: [‐0.78, ‐0.18], p = 0.002). Maximum aneurysm diameter was also smaller in the adequate occlusion group (5.7 mm vs. 6.8 mm, MD: ‐1.13 mm, 95% CI: [‐1.69, ‐0.57], p < 0.001). Aneurysm height was also significantly lower than the inadequate occlusion group (5.4 mm vs. 6.3 mm, MD: ‐0.92 mm, 95% CI: [‐1.55, ‐0.29], p = 0.004). Conclusion Smaller aneurysm dimensions, regular shape, and female gender were associated with higher adequate occlusion rates after WEB treatment. These findings may guide patient selection strategies for optimizing aneurysm occlusion. Further high‐quality studies are needed to confirm these associations. image image

Objective To evaluate the safety and efficacy of Grayson's presurgical nasoalveolar molding (PNAM) technique on nasal and alveolar morphology in infants with unilateral cleft lip and palate (UCLP). Design Systematic review and meta-analysis. Setting : Multicenter international literature across clinical and academic institutions. Patients, Participants Infants with UCLP reported in 21 eligible studies (n = 534), of whom 368 received PNAM. Interventions Presurgical nasoalveolar molding (PNAM) according to the Grayson protocol. Main Outcome Measure(s) : Changes in nasal symmetry (nostril width ratio, columellar angle) and alveolar morphology (alveolar gap dimensions, intertuberosity distance). Results PNAM was performed for an average duration of 96 days with 11 adjustments per patient. Pooled analyses demonstrated significant improvements in nasal symmetry: nostril width ratio decreased from 2.4 (95% CI, 1.7–3.1; I² = 98%) to 1.8 (95% CI, 0.9–2.6; I² = 97%), and the columellar angle increased from 52° (95% CI, 42.0–62.8; I² = 100%) to 70° (95% CI, 59.1–80.6; I² = 98%). Alveolar gap narrowed from 10.7 mm (95% CI, 9.5–11.9; I² = 92%) to 3.9 mm (95% CI, 3.1–4.8; I² = 93%). Intertuberosity distance increased slightly, from 31.3 mm (95% CI, 29.3–33.3; I² = 95%) to 31.8 mm (95% CI, 28.9–34.8; I² = 98%). Conclusions Grayson's PNAM appears to improve nasal and alveolar morphology in infants with UCLP prior to definitive surgery. Standardized outcome measures and long-term prospective studies are needed to confirm these findings and refine clinical protocols.

Abstract Medulloblastoma (MB) is one of the most prevalent malignant brain tumors in children, with tremendous cognitive and neuroendocrine disability among survivors. Group 3 (G3) MBs have poor overall survival at &lt;50%, higher frequency of metastasis, and no targeted therapies. Amplification of MYC and activation of TGFβ signaling are frequent in G3MB. Remarkably, some tumors have no reported mutations, suggesting roles for chromatin-driven mechanisms in this disease. We have generated novel humanized models for G3MB by transducing neuroepithelial stem cells (NESC) derived from human induced pluripotent stem cells with TGFβ effectors alone and/or in combination with MYC, prioritizing combinations observed in patients. We determined that both MYC and TGFβ effectors drove tumor formation in vivo with the combination driving more aggressive tumors and higher frequency of metastasis. Importantly, NESC-derived tumors clustered with human G3MB, indicating our models recapitulated human disease. To decipher mechanisms of resistance, we integrated CUT&RUN for MYC genomic localization and histone PTMs with RNA-sequencing and discovered MYC-bound neural developmental genes repressed in MYC and TGFβR1-driven lines. These gene signatures were associated with the Polycomb Repressive Complex (PRC) and demarcated with H3K27me3; the histone mark directly regulated by PRC. Further interrogation identified JARID1B, a MYC binding partner and H3K4me3 demethylase, as a regulator of these repressed neural genes. This is further reinforced by the observation of higher H3K27me3 levels concurrent with increased JARID1B expression in human G3MB patient samples, compared to other subtypes. We postulate that deregulation of MYC and context-specific targeting to the chromatin template culminates in transcriptional alterations that promote aggressive G3MB. These studies support a critical role of MYC in repressing target genes to drive tumor formation and JARID1B as a new target for therapy. Collectively, we present a critical framework for understanding mechanisms of resistance and the chromatin landscape of MYC- and TGFβ-driven G3MB.

Background For large vessel occlusion (LVO) ischemic strokes, three major techniques of mechanical thrombectomy (MT) have demonstrated efficacy. Here we compare the safety and efficacy of stent retrievers (SR), contact aspiration (Asp), and a combined mechanical thrombectomy (cMT) for LVO ischemic stroke. Methods Using Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, we queried the PubMed, EMBASE, Scopus, and Web of Science databases for studies published prior to December 2024 describing patients with ischemic stroke who underwent MT using SR, Asp, or cMT. Results Twnety-four studies comprising a total of 12 845 patients were included in the analysis. Compared with cMT, Asp was associated with a lower rate of first pass effect (FPE) (risk ratio (RR) 0.67; 95% CI 0.46 to 0.96), shorter procedure duration (mean difference (MD) −13.35 min; 95% CI −20.83 to –5.87), and lower risk of arterial dissection (RR 0.50; 95% CI 0.29 to 0.87). Additionally, SR use was linked to a higher procedure duration compared with Asp (MD 13.39 min; 95% CI 5.55 to 21.22) and a lower symptomatic intracranial hemorrhage rate (RR 0.75; 95% CI 0.59 to 0.96). No significant differences were observed between groups in terms of number of passes, successful recanalization, 90-day functional outcomes, mortality, arterial perforation, or embolization to new territory. Conclusions All three techniques demonstrated similar recanalization rates and functional outcomes. However, Asp was associated with shorter procedure times and fewer arterial dissection complications, while cMT showed a higher FPE. Given the comparable overall efficacy, the choice of technique should be guided by the physician’s preference, expertise, and confidence, tailored to the individual patient’s profile.

Abstract Disclosure: M. Kakakhel: None. S. Cheema: None. R. Haider: None. M.H. Alsaadi: None. M. Ikram: None. W. Ftih: None. H. Jawad: None. M.U. Rehman: None. R. Akhtar: None. A. Qadeer: None. M. Khawar: None. A. Ullah: None. H. Ahmad: None. F. Ahmed: None. R. Alcaraz: None. Introduction: Conditions that disrupt the adrenal glands' typical function are known as adrenal gland ailments or diseases. This study examines the distribution of adrenal disorders across various demographic and geographic groups in the United States, including gender, census region, race, age groups, and urbanization levels. Our objective was to identify disparities and trends to inform targeted interventions and resource allocation. Methods: From 1999 to 2020, mortality statistics for adrenal diseases were extracted from the CDC-WONDER database. Age, sex, race, location, urbanization status, and place of death were used to stratify the age-adjusted mortality rates (AAMRs) and crude mortality rates (CMRs) per 100,000. We calculated the average annual percentage change (APC) using Joinpoint regression software. Results: A total of 31,028 fatalities were reported due to adrenal disorders from 1999-2020 (AAPC: 1.277; 95% CI, -0.032 to 2.6032, p=0.055916). An initial rising trend was observed from 1999-2003 (APC: 2.4799), followed by a declining trend from 2003-2013 (APC: -3.0094), then a modest rise from 2013-2018 (APC: -0.9188), and finally a sharp increase from 2018-2020 (APC: 2.9445). Males had a higher AAMR (6.8) than females (6.4). The West census region had the highest AAMR (7.2), followed by the Midwest (7.1), South (6.3), and Northeast (5.6). The highest crude mortality rate was observed in individuals aged 85+ years (52.2). Several disparities were noted when stratified by urbanization and race, including the highest AAMR in Non Core (Nonmetro) regions (7.9) and among American Indian or Alaska Native populations (8.2). The highest percentage of deaths occurred in medical facilities. Conclusions: The findings underscore the need for tailored public health strategies to address disparities in adrenal disorders among vulnerable populations. Specific attention should be directed towards the male gender, West census region, American Indian or Alaska Native populations, and the geriatric population. Future research should explore underlying causes and interventions to mitigate these disparities. Presentation: Monday, July 14, 2025

Background: Acute heart failure affects ~9% of patients with hematological malignancies. This study aimed to evaluate the impact of acute heart failure on in-hospital mortality among patients with hematological malignancies and to identify independent predictors of mortality in this population. Methods: The National Inpatient Sample (2016–2021) was queried for adults (⩾18 years of age) with hematological malignancies and those who experienced in-hospital mortality. Among these, we identified patients who were hospitalized with a diagnosis of acute heart failure. Univariate analysis and multivariate regression were employed to analyze the mortality risk. A p ⩽ 0.05 was considered significant. Results: Among 40,246 patients with hematological malignancies who died in the hospital, the univariate analysis showed a higher in-hospital mortality in the acute heart failure cohort compared to the non-acute heart failure group (8.08% versus 4.52%, p < 0.001). However, after adjusting for sociodemographic factors and comorbidities, acute heart failure was associated with a lower risk of in-hospital mortality (OR 0.87, 95% CI 0.84–0.9, p < 0.001). Predictors of mortality in the acute heart failure cohort were age >60 years, hematopoietic stem cell transplant, severe sepsis, arrhythmias, infective endocarditis, bleeding from any source, acute respiratory failure, hepatic dysfunction, metabolic encephalopathy, and invasive ventilation (all p < 0.01). On subgroup analysis, acute myeloid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, and multiple myeloma had a higher in-hospital mortality in the acute heart failure cohort compared to the non-acute heart failure group ( p < 0.001). Conclusion: Acute heart failure in patients with hematological malignancies was associated with poor outcomes in the presence of severe sepsis, organ dysfunction, and the need for mechanical ventilation. Close monitoring and a multidisciplinary team approach are recommended to optimize the treatment plan for this population.

Background: Doxorubicin (DOX) is an effective chemotherapy but limited by severe cardiotoxicity. Chemokine receptors CXCR4 and CXCR7 are involved in regulating cardiac survival, inflammation, and fibrosis, yet their specific roles in DOX-induced cardiotoxicity (DICT) remain unclear. Objective: We investigated the cardioprotective effects of TC14012, a dual-target CXCR4 antagonist and CXCR7 agonist, against DICT. Methods: Male C57BL/6J mice received chronic or acute DOX administration with twice-weekly TC14012 treatment. Cardiac function was evaluated by echocardiography. Markers of cardiac fibrosis and inflammation were assessed. RNA-seq analysis was performed to explore associated molecular pathways. Key pathways were validated in cardiac cells and bone marrow-derived mast cells. Results: TC14012 significantly improved cardiac function, indicated by preservation of ejection fraction and fractional shortening, and increased survival rates optimally at 5 mg/kg, which were accompanied with reductions in cardiac collagen deposition and infiltration of macrophages and neutrophils. RNA-seq revealed that TC14012 protecting against DICT was associated with a remarkable stimulation of signals involving in cardiac contraction and energy reserve metabolism processes, particularly glycogen and glucan metabolism that are crucial for meeting the cardiac energy demand in response to DOX induction. These signals could be further enriched into AMPK signalling pathway and validated by enhancement of cardiac AMPK phosphorylation both in vivo and in vitro . Meanwhile, TC14012’s cardioprotective effects were accompanied by a reduction in signals involving in cardiac mast cell activation and degranulation, which could be enriched into Rap1 signalling pathway and validated by TC14012 inhibiting mast cell activation and Rap1-GTP activity in vitro . Importantly, TC14012 did not compromise DOX’s antitumor efficacy in EL-4 lymphoma-bearing mice. Conclusions: TC14012 may prevent DICT by activating CXCR7-AMPK signalling to reprogram cardiomyocyte metabolism and antagonizing CXCR4-Rap1 signalling to blunt mast cell-mediated sentinel effects without compromising the anticancer efficacy DOX.