Citations

Abstract The success of Chimeric Antigen Receptor (CAR) T cell therapy is heavily dependent on the quality of the final cellular product. Current expansion protocols often rely on reagents that require removal from cell culture media, posing logistical challenges in manufacturing, and can also lead to terminal differentiation. Here, we evaluate the use of a soluble, bead-free T cell activator, T cell expansion protein (T-CEP), as a streamlined alternative for generating potent CAR-T cells. Human T cells were activated with T-CEP or known T cell activators (Dynabeads and TransAct) and transduced with either CD19 or interleukin-13 (IL-13) mutein (tetravariant-13; TV-13)-based CAR lentiviral vectors. Our results demonstrate that T-CEP supports robust CAR-T cell expansion and achieves transduction efficiencies comparable to commercial reagents for both types of CAR-T cells. Notably, T-CEP significantly favored the expansion of CD8 + T cells, yielding an enhanced CD27 + phenotype and a lower CD4:CD8 ratio compared to TransAct. Cytotoxicity assays confirmed that T-CEP-expanded CAR-T cells possess cytolytic function equivalent to commercial reagents for both CARs, while exhibiting lower levels of inflammatory cytokine secretion. In summary, T-CEP represents a competitive alternative to existing expansion agents, as it does not require its removal during CAR-T manufacturing and generates a CD8 + dominant, less-differentiated phenotype without compromising efficacy. Key Points T-CEP is a soluble, bead-free activator that streamlines CAR-T manufacturing by eliminating the need for reagent removal from cultures. T-CEP consistently yields a desirable CD27 + CD8 + phenotype with a moderate inflammatory profile compared to industry-standard expanders.

Background: Hepatic stellate cells (HSCs) play a pivotal role in driving fibrosis during chronic liver injury. HSCs produce vast amounts of fibrotic proteins, causing endoplasmic reticulum (ER) stress and initiating the unfolded protein response (UPR). While the UPR is important for fibrogenesis, how signaling through UPR transducer Protein Kinase R-like ER Kinase (PERK) and its effectors impact HSC activation and fibrogenesis is unclear. Here, we sought to uncover the role of PERK and its effector GADD45A in liver fibrosis. Methods: PERK–GADD45A signaling was assessed in primary and immortalized HSCs treated with TGFβ, and mouse models of fibrosis. Genetic and pharmacological disruption of PERK or GADD45A was used to assess the role of PERK or GADD45A in HSC activation, proliferation, and fibrogenesis. HSC-specific Gadd45a -null mice were utilized to investigate the role of GADD45A on CCl 4 -induced fibrosis. Results: We found that TGFβ-induction of collagen I drives activation of PERK signaling in HSCs. Furthermore, loss or inhibition of PERK limits long-term HSC activation as illustrated by reduced levels of collagen I and fibronectin, impaired collagen I deposition, and reduced cell proliferation in vitro. Next, we show that PERK signaling induces expression of GADD45A during HSC activation, and loss of GADD45A disrupts HSC activation and expression of proliferation and cell-cycle-associated genes in immortalized and primary HSCs. Finally, HSC-specific GADD4Aa loss limits CCl 4 -driven fibrogenesis in vivo. Conclusion: PERK signaling is critical for HSC activation, and loss of the PERK downstream effector GADD45A limits fibrosis progression. Disruption of HSC activation and proliferation, coupled with dysregulation of cell-cycle-associated genes upon PERK or GADD45A loss, suggests that PERK–GADD45A signaling impacts multiple facets of HSC pathophysiology.

Abstract Endoscopic retrograde cholangiopancreatography (ERCP)-based tissue acquisition is the cornerstone for assessment of suspected malignant biliary strictures. Conventional techniques, including biliary brush cytology and transpapillary forceps biopsy, are highly specific but are limited by modest sensitivity for detection of malignancy, often leaving a substantial proportion of individuals without a definitive diagnosis. To improve diagnostic sensitivity for cancer, several adjunctive approaches have been integrated into ERCP-based sampling. Peroral cholangioscopy (POC)-guided biopsies have become a reliable ERCP-based adjunctive modality, allowing direct visualization and targeted sampling, particularly in individuals with prior nondiagnostic procedures. Fluorescence in situ hybridization (FISH) is a technique used to improve diagnostic yield with biliary brushings by detecting chromosomal abnormalities, but is limited to specialized centers due to the need for specialized processing and training. Next-generation sequencing provides high sensitivity and can enable identification of actionable genomic alterations, while methylated DNA markers represent a promising, scalable molecular approach with strong diagnostic performance. Additional techniques include artificial intelligence–augmented direct visualization by POC and bile-based liquid biopsy. Current guidelines recommend multimodal ERCP-based sampling, combining at least two modalities where feasible, to optimize diagnostic accuracy. This review highlights the clinical performance, strengths, and limitations of established and emerging ERCP-guided techniques, emphasizing their evolving role in improving the diagnosis and management of malignant biliary strictures.

Abstract Background Adipose tissue influences blood pressure regulation through depot-specific endocrine, inflammatory, and exosome-dependent mechanisms. While obesity is a well-established risk factor for hypertension, emerging evidence suggests that distinct fat depots may contribute to vascular dysfunction independent of total adiposity. Methods: This review synthesizes post-2020 literature on 6 anatomically defined depots: visceral, perivascular, epicardial, bone marrow, brown, and beige adipose tissue, with an emphasis on their mechanistic roles, imaging biomarkers, and translational relevance. Results Alterations in adipose depot function can impair nitric oxide signaling, promote oxidative stress, and modulate sympathetic tone, even in the absence of obesity. Exosomal microRNAs from dysfunctional adipose tissue further disrupt vascular homeostasis. Although clinical tools for assessing depot dysfunction are limited, advances in imaging and molecular profiling may enable targeted therapeutic strategies. Conclusions Reframing adipose tissue as a modular regulator of blood pressure, rather than a passive consequence of obesity, may unlock new venues for precision management of hypertension.

Background: Pediatric tibial fractures are common long-bone injuries, yet uncertainty remains in the optimal immobilization strategy. The comparative effectiveness and safety of short leg versus long leg casts in achieving stable healing and functional recovery have not been clearly established. Methods: A literature search was conducted across Medline, CINAHL, Embase, and Cochrane Library, from inception to August 2025, following PRISMA guidelines and registered with the International Prospective Register of Systematic Reviews (PROSPERO). Statistical analyses were performed using a random-effects model. Results: The initial search yielded 1236 studies; 6 studies with 413 pediatric patients met the inclusion criteria. A total of 163 underwent short leg casting (SLC) and 250 long leg casting (LLC). No differences were observed between SLC and LLC for coronal angulation (MD=0.12 degrees, 95% CI=−0.89 to 1.12, I 2 =58%) or sagittal angulation (MD=0.18 degrees, 95% CI=−0.71 to 1.07, I 2 =56%). Subgroup analyses showed no differences for tibial shaft fractures or Salter-Harris distal tibial fractures. Cast duration was similar between groups (MD=−0.74 wk, 95% CI=−1.49 to 0.01, I 2 =92%). Overall complication rates did not differ (OR=0.76, 95% CI=0.25 to 2.34, I 2 =0%), with no differences in Salter-Harris subgroup analysis (OR=0.67, 95% CI=0.05 to 9.91; I 2 =54%). SLC was associated with shorter time to weight-bearing (MD=−2.29, 95% CI=−4.49 to −0.08, I 2 =68%) and faster time to union (MD=−1.21, 95% CI=−2.16 to −0.25, I 2 =30%). Conclusion: SLCs may be considered as an alternative to LLCs in pediatric tibial shaft and distal tibial physeal fractures based on currently available retrospective comparative evidence. However, the evidence is limited by the retrospective design, clinical heterogeneity, and risk of confounding. Further prospective comparative studies are needed to define optimal indications for SLC use in this population. Level of Evidence: Level III, systematic review and meta-analysis of retrospective comparative studies.