Citations

Hypoplastic left heart syndrome (HLHS) is the most lethal congenital heart disease (CHD) whose genetic basis remains elusive, likely due to oligogenic complexity. To identify regulators of cardiomyocyte (CM) proliferation relevant to HLHS, we performed a genome-wide siRNA screen in human iPSC-derived CMs, revealing ribosomal protein (RP) genes as the most prominent effectors of CM proliferation. Whole-genome sequencing of 25 HLHS proband–parent trios similarly showed enrichment of rare RP gene variants, including a damaging RPS15A promoter variant shared in a familial CHD case. Cross-species functional analyses demonstrated that perturbation of RP genes impairs cardiac growth: knockdown of RPS15A, RPS17, RPL26L1, RPL39, or RPS15 reduced CM proliferation, caused cardiac malformations in Drosophila , and produced hypoplastic or dysfunctional hearts in zebrafish. Genetic interactions between RP genes and key cardiac transcription factors (TBX5 and NKX2–7) further support their developmental role. Importantly, p53 suppression or Hippo activation partially rescued RP deficiency phenotypes. Together, these findings implicate RP genes as critical regulators of cardiogenesis and candidate contributors to HLHS.

Objectives: In the United States, training for physicians who manage critically ill adult patients (intensivists) evolved through parallel subspecialty critical care medicine (CCM) pathways with significant commonality. The Society of Critical Care Medicine Adult Critical Care Physician Core Knowledge and Skills Task Force aimed to delineate the common core knowledge and skills required of all intensivists. Design: A master list of content areas and procedural skills was compiled from all CCM subspecialty program requirements and blueprints of the certification examinations. Using a modified Delphi approach, participants were asked to categorize the knowledge items as “advanced knowledge is essential,” “general, but not advanced, knowledge is essential,” or “knowledge is not essential.” Procedures were categorized as “intensivist performs routinely,” “intensivist only performs in an emergency,” or “intensivist knows” about the procedure. Setting: Representatives from CCM stakeholder organizations, including accreditation and certification organizations, critical care societies, and program directors’ societies, were invited to participate. SUBJECTS: Members of the Adult Critical Care Physician Core Knowledge and Skills Task Force of the Society of Critical Care Medicine. Interventions: For the first two rounds of the modified Delphi process, Research Electronic Data Capture was used. For the third and fourth rounds, the process was completed through online meetings with Zoom (Zoom Video Corporations, San Jose, CA) utilizing Zoom’s polling feature. Measurements and Main Results: A total of 541 items were determined to be essential, with 145 requiring advanced knowledge and 323 requiring general knowledge. For 73 items, consensus regarding advanced vs. general could not be achieved, but they remained essential. Only eight items were felt to be nonessential. Of the 16 procedures, most were categorized as “intensivist performs.” Conclusions: The large number of items included in the list of essential knowledge and skills demonstrates the complexity of modern CCM. Utilization of a common framework across the subspecialties of CCM could lead to greater harmonization among the fellowship program requirements and certification examinations.

Abstract Pancreatic ductal adenocarcinoma (PDAC), the most common subtype of pancreatic cancer, is a deadly disease with a complex tumor microenvironment (TME). How chemotherapy alters the TME, and whether these changes drive chemoresistance, is poorly understood. We examined matched pre- and post-treatment tissue specimens and found near-universal enrichment of axonal guidance genes in cancer-associated fibroblasts (CAFs) after treatment. These CAFs were enriched near sites of perineural invasion, coinciding with regions of increased tumor cell proliferation, and were enriched in tumor areas distant from nerves after chemotherapy. Metastatic recurrence lesions had the highest prevalence of these CAFs versus primary tumors and untreated metastasis. These CAFs showed elevated non-canonical Wnt mediators and axonal guidance genes, which complemented matching cognate binding partners in tumor epithelial cells, suggesting a role in tumor-stroma crosstalk. Our findings implicate fibroblast-derived axonal-guidance genes in promoting PDAC invasion and point to a promising target for this disease. Statement of Significance Therapeutic resistance remains a major challenge in pancreatic cancer. Our findings shed light on the changes in the complex tumor microenvironment in response to chemotherapy and identify fibroblasts high in axonal-guidance genes that may drive tumor progression and chemoresistance, thus uncovering a potential avenue for targeting pancreatic cancer.

Abstract Valvular heart disease (VHD) remains significantly underdiagnosed and undertreated. This review examines an artificial intelligence (AI)-enhanced ‘spoke–hub–node’ care model designed to improve the early detection, risk stratification, and treatment of VHD. In this model, AI tools—such as automated ECG interpretation, digital stethoscopes, and point-of-care ultrasound—facilitate decentralized screening and referral for cardiac imaging at the community level. During the transition from outpatient settings to tertiary care centres, AI-integrated echocardiography, cardiac tomography, and magnetic resonance imaging facilitate advanced diagnostic evaluation and inform procedural planning. We review emerging innovations that can enhance this model of care delivery—including unsupervised machine learning to uncover novel VHD phenotypes, generative AI for automated reporting, the use of digital twins to simulate interventions, and the integration of multiple AI agents to support heart team meetings. These advances are followed by the emerging use of AI in robotic transoesophageal and intracardiac echocardiography, as well as in fusion fluoroscopy imaging, to guide valve interventions. While outlining the challenges inherent in this rapidly evolving field, the review’s central contribution is its vision to connect the continuum—from AI-enabled community screening to personalized, image-guided therapies at tertiary care centres—offering a scalable and equitable model for VHD care.

Abstract Chimeric Antigen Receptor (CAR) T cell therapy is a transformative treatment for hematologic malignancies. However, ex vivo autologous CAR T cell manufacturing presents substantial logistical and clinical challenges. Direct in vivo CAR-T cell engineering offers a new paradigm for CAR T delivery. We recently developed the CD3-targeted lentiviral vector LV-169 which can deliver a CAR transgene to resting niche-resident T cells in lymphoid tissues. LV169 is pseudotyped with a mix of chimeric and nonchimeric VSV-G proteins detargeted (blinded) from the natural LDL receptor and re-targeted to T cells through the display of a CD3-specific scFv. Here we demonstrate that LV169 can specifically deliver a functional aBCMA-CAR transgene to resting human T cells in PBMC cultures from the healthy volunteers and from multiple myeloma patients. For clinical translation, we selected an αBCMA-CAR construct comprising a fully human single heavy chain nanobody linked to 41BB and CD3z cytoplasmic signaling domains. Human T cells expressing this CAR construct have low tonic signaling and efficiently kill BCMA-positive myeloma target cells with similar or lower associated cytokine release compared to a clinically relevant benchmark BCMA-CAR. To further improve LV169 and to minimize CAR protein incorporation in the LV envelope which may lead undesirably to myeloma cell transduction, we placed the CAR transgene under a T-cell specific Lck promoter in reverse orientation relative to the 5' LTR. The transcriptional activity of the Lck promoter was comparable to that of a constitutive EF1a promoter in primary T cells and, in contrast to EF1a had minimal activity in a variety of other tested cell types including the HEK293T producer cell line and the THP-1 monocytic cell line. Western blotting confirmed the absence of CAR protein in LV particles (LVP) generated using the Lck promoter-driven reverse orientation construct. In contrast, high levels of CAR protein were detected in LVP generated using the parental EF1a promoter-driven construct. The LV-incorporated CAR protein mediated off-target gene delivery to BCMA-positive myeloma cells. The therapeutic efficacy of intravenously administered LV169 LV particles carrying BCMA-CAR constructs was tested in the OPM-2 human multiple myeloma xenograft model. NSG mice with knockout of MHC-I and MHC-II, and with advanced disseminated OPM-2-Luc tumors were engrafted with human PBMC one week prior to intravenous LV169-BCMA-CAR administration. Mice were treated with single intravenous injection of LV169 at different doses 3 weeks after tumor engraftment. OPM-2-Luc tumor burden was monitored by regular luciferase imaging. All treated mice cleared their tumors within 28-days following LV administration and the rate of clearance was correlated to the LV particle dose. Even at the 4e8 dose level (by p24) tumors were completely cleared. Eighty four days post-treatment the treated mice remained tumor-free and resisted re-challenge with OPM-2 tumor cells. Circulating CAR-T cells, monitored by flow cytometry, became detectable by 14 dpi and peaked between 21-35 dpi. Following tumor clearance, the number of circulating CAR-T cells declined dramatically. The treatment was well tolerated and analysis of cytokines associated with CAR-T cell cytotoxicity confirmed the absence of a severe cytokine storm. IFNγ was the only cytokine which was elevated during the expansion period of CAR-T cells, while IL-2, IL-6, TNFα and GM-CSF were barely detectable. In summary the characteristics of our new developed LV169-based platform make it suitable for further clinical translation which is underway.

Abstract Direct in vivo T cell engineering offers a new paradigm for CAR T delivery. We have evaluated several approaches to engineer lentiviral (LV) vectors capable of delivering CAR transgenes directly and selectively to T cells in vivo, within the lymphoid niche. Our goal was to develop an LV delivery platform that following intravenous administration can selectively bind and activate T cells, deliver and integrate the (CAR) transgene into genomic DNA, and drive CAR expression on the T cell surface. Further requirements were that the retargeted LV should be associated with minimal off-target transduction of tumor cells and normal host tissues and should be manufacturable at titers sufficient for human application. Our strategy for retargeting has been to modify the trimeric VSV-G protein, which is well established for efficient LV pseudotyping. Of the approaches evaluated, three retargeting platforms have proven to be effective: direct covalent display, indirect covalent display, and G-capping. Direct covalent display. This (LV-169) approach uses chimeric VSV-G protein that contains a CD3-targeting ligand displayed as a N-terminal extension of VSV-G protein that has been mutated to ablate (blind) its interaction with the LDL receptor. Chimeric and nonchimeric blinded G proteins are co- incorporated into LV particles within mixed envelopes which bind to CD3. Upon binding to cell surface CD3, the engineered particles activate T-cells and enter acidifying endosomes, whereupon they trigger membrane fusion and deliver their payloads. LV-169 displaying an anti-human CD3 scFv selectively transduced resting human T cells in PBMC cultures, in fresh human whole blood and following intravenous administration to humanized mice. In immunocompetent Cre reporter mice, surrogate vectors displaying anti-murine CD3 scFv selectively transduced lymph node resident T cells, with off-target transduction being limited to tissue-resident macrophages. To eliminate the incorporation of CAR protein into the LV-169 envelope, which can lead to highly undesirable transduction of tumor cells expressing the CAR target, we placed the CAR transgene in reverse to 5'LTR orientation, downstream of a T-cell specific promoter/enhancer. In vivo efficacy of LV-169-BCMA-CAR was demonstrated in humanized mice bearing disseminated OPM-2 myeloma tumors. Clinical translation of this construct is now underway. Indirect covalent display. In this G-protein engineering approach, a short peptide (SPY-tag) is displayed as an N-terminal extension of the LDLR-blinded VSV-G protein. The SPY-tagged LV vectors are then covalently modified to display a CD3-targeting ligand by simply mixing them at room temperature with a bifunctional fusion protein consisting of the SPY-catcher and the CD3 ligand. Under these conditions, the SPY-catcher and the SPY-tag spontaneously form a covalent isopeptide bond. The CD3 ligand-modified LV particles were shown to specifically activate and transduce T cells ex vivo within human PBMC cultures. G-capping. In the final approach, the VSV-G protein is not genetically modified. Instead, a “capping” protein crosslinker that binds directly to the unmodified G protein on preformed LV particles is used to mask their LDLR binding sites and redirect them to interact with CD3. Mixing of the CD3-targeted capping protein with traditional G-pseudotyped LV particles leads to efficient binding of the cap to the trimeric G proteins. The cap-modified LV are unable to interact with LDLR but interact efficiently with CD3. CD3-capped LV particles were highly specific and efficient in human T cell activation and transduction in PBMC cultures. The association between the G protein and the bound cap is highly stable during freeze-thaw cycles and at room temperature, a critical attribute of this platform to facilitate the manufacture of a clinically and commercially viable LV product. In summary, we have developed three translationally relevant platform technologies for the generation of LV particles capable of delivering CAR transgenes specifically and efficiently to resting niche-resident T cells after intravenous administration. Manufacturing studies are underway and, in the case of the LV-169 platform, clinical translation is already in progress.

Abstract Introduction: Older adults (OAs) with classic Hodgkin lymphoma (cHL) have inferior survival compared with younger patients (pts). NAVD has been widely adopted in clinical practice for frontline (1L) treatment of OAs with advanced stage cHL based on the subset analysis of S1826 study (Rutherford et al, JCO 2025) showing a 1-year progression-free survival (PFS) of 93% and 1-year overall survival (OS) of 95% after NAVD (n = 50) which remained durable on longer-term follow up. Another phase 2 study (Torka et al, JCO 2025) showed that NAVD is a highly effective and well-tolerated 1L regimen in OAs with cHL (n=40) across a wide range of geriatric impairments. Adverse events (AEs) and efficacy have not been reported in OAs treated with NAVD outside of a clinical trial. Hence, we sought to evaluate safety profile and efficacy outcomes of OAs with cHL treated with 1L NAVD in a real-world setting (RWS). Methods This is a multicenter retrospective cohort study of adults (≥18 years) with cHL treated with 1L NAVD between 2023 and 2025 at 20 US academic sites. Baseline characteristics and outcomes of pts ≥60 years were evaluated as a pre-planned subset analysis. The safety evaluable cohort received at least 1 cycle of NAVD and included pts who had not yet completed treatment. The efficacy evaluable cohort had completed 6 cycles of treatment including end of treatment (EOT) PET-CT and included pts who intended to complete treatment but died or discontinued due to progressive disease or AEs. The primary endpoint was EOT overall response rate (ORR). Secondary endpoints included EOT complete response rate (CRR), PFS and OS. PFS and OS were analyzed using the Kaplan Meier method. Results Among 311 pts, 81 were OAs (26%). Median age of the OA cohort was 70 (range 60-88) yrs, 65% male, 84% Caucasian, 71% nodular sclerosis subtype, 58% stage 4 disease, 59% B symptoms, 14% bulky disease (>7.5 cm), 69% extranodal sites, 41% EBV positive, 54% hypoalbuminemia, 49% IPS ≥ 4 and10% had an autoimmune disease. G-CSF was given in 81% pts. All 81 pts were evaluable for safety; 74% experienced a grade 3+ (≥G3) AE, 54% had ≥G3 hematological AE, 48% had ≥G3 neutropenia, 17% had febrile neutropenia. Infection rate was 28% with 10% ≥G3, neuropathy rate was 23% with 4% ≥G3. One infusion reaction was noted. Treatment related hospitalizations rate was 23% with median duration of 7 (range 1-16) days. There were 24 immune related AEs (IrAE) (30%) of which 5 were ≥G3. Most common IrAE was hypothyroidism (n=5), followed by inflammatory arthritis (n=4), pneumonitis (n=2, ≥G3, n=1), hepatitis (n=2, both ≥G3), adrenal insufficiency (n=2, ≥G3, n=1), colitis (n=2), and rash (n=2). One patient each had autoimmune neutropenia, sarcoidosis of lung/skin and myocarditis (≥G3). Overall, 15% pts required steroids with resolution of IrAEs in 56% cases, 30% (n=7) pts required continued therapy (levothyroxine in 4 pts, steroids in 3 pts, medical tx for heart failure in 1 pt). Treatment was interrupted in 22%, reduced in 22%, and discontinued in 23% pts. Nivolumab was discontinued in 14% pts. Among all pts (n=81), 81% had an interim PET-CT after 2-4 cycles. The ORR and CR rate at interim PET-CT were 100% and 81%, respectively. In the efficacy evaluable pts (n=52; 24 still on therapy), the EOT ORR and CRR were 93.2% and 84% in OAs compared to 96.8% and 91.7% in pts <60yrs, respectively. At a median follow up of 10.2 mo (range 0.27-28.52), the 1-yr PFS was 90.6% (95% CI: 82.7%-99.3%) in OAs and 95.5% (95% CI: 91.6%- 99.6%) in pts <60 yrs (p=0.044). 1-yr OS was 97.5% (95% CI: 94.1-100%) and not evaluable in pts <60yrs (p=0.017). None of the baseline features were predictive of response, additional analysis to understand impact on AEs and survival is ongoing. Among the 5 pts with progression/relapse, 4 were primary refractory. 4 pts received 2nd line (2L) therapy.79 pts are alive at data cut off, 2 deaths occurred- 1 infection-related and other unknown; both after the 1st cycle. Conclusions In this largest RWS cohort of OAs treated with NAVD we showed comparable response rates, AEs and 1-year survival outcomes to those in the pivotal S1826 trial. Notably, rates of ≥G3 AEs including infections, neuropathy and IrAEs remained low and manageable. Although survival significantly improved with NAVD compared to historical data, outcomes of OAs remained inferior compared to pts <60 yrs. Our data reinforces the use of NAVD as the preferred 1L option for OAs with advanced stage cHL.

Abstract Introduction: Lutetium-177 DOTATATE peptide receptor radionuclide therapy (PRRT) is an effective treatment for gastroenteropancreatic neuroendocrine tumors (GEP-NETs). While generally well tolerated, PRRT can result in hematologic toxicity ranging from mild cytopenias to therapy-related myeloid neoplasms (t-MNs). Understanding the incidence, severity, and clinical consequences of hematologic toxicity is essential for optimizing treatment decisions and establishing effective monitoring strategies. This study aimed to characterize hematologic toxicity patterns, severity, and recovery profiles in GEP-NET patients treated with Lutetium-177 DOTATATE PRRT and to determine the frequency of cytopenia-related treatment discontinuation. Methods: We conducted a retrospective analysis of 160 consecutive GEP-NET patients who underwent PRRT. Hematologic parameters (hemoglobin, white blood cell count, absolute neutrophil count, platelet count, and lymphocyte count) were assessed at baseline, mid-treatment (post-cycle 2), and post-treatment (post-cycle 4). Cytopenias were graded using Common Terminology Criteria for Adverse Events version 4.03 and categorized as never (grade <2 throughout), transient (grade ≥2 mid-treatment, resolving post-treatment), persistent (grade ≥2 at both timepoints), or late-onset (grade <2 mid-treatment, grade ≥2 post-treatment). Treatment completion rates and discontinuation reasons were documented. Results: Overall, 73% of patients (117/160) completed all planned PRRT cycles. Among the 43 patients who discontinued treatment early, only 4 (2.5% of total cohort) discontinued specifically due to cytopenia, while the remainder discontinued for other reasons. Post-treatment grade ≥2 cytopenias were uncommon: lymphocytes (4.4%), hemoglobin (2.5%), neutrophils (1.9%), white blood cells (1.3%), and platelets (1.3%). The majority of patients (≥85%) never experienced grade ≥2 cytopenia throughout treatment. When cytopenias occurred, they were predominantly transient (8-10% of patients) with complete recovery by treatment completion. Persistent cytopenias were rare (<5% across all cell lines), while late-onset cytopenia occurred most frequently with neutrophils (14%). Conclusion: Lutetium-177 DOTATATE PRRT demonstrates an excellent hematologic safety profile in GEP-NET patients. Severe cytopenias are uncommon, predominantly transient, and rarely necessitate treatment discontinuation. These findings support the continued use of PRRT in appropriate candidates, with cytopenia-related discontinuation occurring in fewer than 3% of cases. Long-term follow-up remains essential to monitor delayed hematologic effects, including t-MNs. Future research should evaluate baseline predictors such as prior chemotherapy, renal function, and marrow reserve to identify patients at higher risk who may benefit from closer hematologic surveillance.

Abstract Background: Recent approvals and clinical trials have established covalent BTK inhibitors (BTKis), including ibrutinib, acalabrutinib, and zanubrutinib, as effective agents for the treatment of mantle cell lymphoma. Multiple BTKi-based regimens are recommended by experts and clinical guidelines in both the first-line and later-line settings. Additional approvals of the noncovalent BTKi pirtobrutinib and multiple CAR T-cell therapies have also advanced the management of patients who have progressed on covalent BTKi therapy. In light of these developments in treatment strategies for mantle cell lymphoma (MCL), this study compares trends in treatment choice among experts and healthcare professionals (HCPs) in an online treatment decision support tool. Methods: An online decision support tool, developed with 5 US-based experts in MCL, was launched in April 2023 and updated in April 2025. The tool provided treatment recommendations for various clinical scenarios involving newly diagnosed (ND) or relapsed/refractory (R/R) MCL, incorporating factors such as age, fitness, previous therapies, and response duration. HCPs entered specific patient characteristics along with their intended treatment choice. The tool then displayed expert recommendations for that scenario, and HCPs were asked whether these recommendations influenced their treatment decision. HCP-entered data and expert recommendations were analyzed to identify common trends and divergence in current BTKi therapy use. Results: The 2023 online tool was used by 119 HCPs who entered 148 patient scenarios, and the 2025 online tool was used by 69 HCPs who entered 96 patient scenarios. In 2023, for patients with ND MCL who had later-stage disease or were not eligible for radiation therapy (n = 75), 64% of expert-recommended treatments included BTKis, with the ECHO and TRIANGLE regimens representing 12% and 17% of recommendations, respectively. In 2025, 81% of expert recommendations were BTKi-based regimens, with ≥3 experts recommending a BTKi regimen in 66 of 67 unique ND case scenarios. The ECHO and TRIANGLE regimen each represented 28% to 29% of expert recommendations. In addition, 51% of recommendations for patients with 17p deletions or TP53 mutations consisted of the BOVen regimen. The percentage of HCPs who indicated that they were considering a BTKi-based regimen in the setting of ND MCL was 17% (13 of 75 ND cases) in 2023 and 66% (44 of 67 ND cases) in 2025. The use of both the ECHO and TRIANGLE regimens by HCPs increased from 5% and 4%, respectively, in 2023 to 19% for each in 2025. In 2025, 6% of HCPs considered the BOVen regimen for patients with TP53 mutation (n = 17). In 2023 and 2025, approximately 50% of the HCP-entered patient cases for previously treated MCL had not received a BTKi (n = 48 and n = 18, respectively). For these scenarios, 94% and 98% of expert recommendations consisted of covalent BTKis in 2023 and 2025, respectively. HCPs selected covalent BTKis for 68% of cases in 2023 and 89% of cases in 2025. For patients with R/R MCL previously treated with a BTKi, CAR T-cell therapy represented 36% of expert recommendations in 2023 (23 scenarios) and 60% in 2025 (18 scenarios). By contrast, 4% and 11% of HCPs considered CAR T-cell therapy for these patients in 2023 and 2025, respectively. For HCPs reporting on the tool's clinical impact (n = 96), 35% indicated expert recommendations changed their intended therapy, 41% stated their treatment plan matched the recommendations, 10% cited barriers to implementing recommendations, and 14% remained undecided on optimal treatment. Conclusions:This analysis highlights the ongoing adoption of expert-recommended treatments into real-world clinical practice. Our data suggest that expert and HCP use of BTKi therapy in the ND MCL setting has increased in recent years. In R/R MCL, CAR T-cell therapy appears to be considered infrequently by HCPs. Barriers to implementation of expert recommendations warrant further investigation. Enhanced decision support tools and targeted educational initiatives could help increase the use of expert treatment recommendations and potentially improve patient outcomes.