Citations

Abstract Pancreatic ductal adenocarcinoma (PDAC), the most common subtype of pancreatic cancer, is a deadly disease with a complex tumor microenvironment (TME). Cytotoxic combination chemotherapy treatments are the mainstay of PDAC therapy, but patients rapidly develop chemoresistance, highlighting the need to elucidate how chemotherapy alters the TME and whether these changes drive chemoresistance. Here, we examined matched pre- and post-treatment tissue specimens using single-cell RNA-sequencing and found near-universal enrichment after treatment of axonal guidance genes in cancer-associated fibroblasts (CAFs). These neural 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 recurrent lesions had the highest prevalence of neural CAFs versus primary tumors and untreated metastases. Neural 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. Additionally, in vitro studies revealed enrichment of axonal guidance genes in patient-derived CAF lines upon exposure to chemotherapy or tumor cells. Altogether, these findings implicate fibroblast-derived axonal-guidance genes in promoting PDAC invasion and point to a promising therapeutic target for this disease.

Cancer immunotherapy is a transformative technology allowing for the elimination of malignant cells through immune system stimulation; however, the large-scale clinical application of adoptive cellular therapy technologies is limited due to difficulties in ex vivo immune cell engineering, high costs, and complicated manufacturing process. Recently, in vivo immune cell engineering was discovered as an emerging approach that allows for the genetic reprogramming of immune cells in patients, thus circumventing many problems inherent to traditional cellular therapy technologies. Breakthroughs in targeted drug delivery systems such as cell-penetrating peptides, antibody- and receptor-mediated delivery approaches, lipid nanoparticles (LNPs), polymeric drug carriers, and biomimetic delivery systems allow for the more effective selective delivery of messenger RNA (mRNA), clustered regularly interspaced short palindromic repeats (CRISPR)/Cas gene editing systems, proteins, and other types of cargo to the target immune cells. In vivo immune cell engineering allows for the efficient generation of chimeric antigen receptor T-cells (CAR-T), CAR- natural killer (NK) cells, engineered macrophages, dendritic cells, and other immune cell types; moreover, transient and permanent immune reprogramming is achievable via vivo technologies. The present review discusses the biological barriers limiting immune cell-targeted delivery, including biodistribution, cellular uptake, intracellular transport, and endosomal escape of delivered drugs, and reviews the latest advancements in targeted delivery technologies addressing these obstacles. We also address the latest applications of in vivo immune cell engineering in hematological malignancies and solid tumors, including mRNA therapeutics, CRISPR-based genome editing, and advanced programmable immunotherapy technologies. Furthermore, we discuss unresolved issues related to the specificity of targeting, efficiency of drug delivery, safety of treatment, manufacturing processes, and regulation of in vivo immune engineering technologies and outline future opportunities arising from the integration of nanomedicine, synthetic biology, genome engineering, and artificial intelligence-assisted materials science.

Purpose: The purpose of this study is to characterize the clinical features, management strategies, and outcomes of endophthalmitis cases following pars plana vitrectomy using data from the American Academy of Ophthalmology (Academy) IRIS ® Registry (Intelligent Research in Sight). Methods: This is a retrospective cohort study of eyes that developed endophthalmitis following pars plana vitrectomy between 2013 and 2022. Data was collected from the Academy IRIS Registry. This study analyzed differences in visual acuity, intraocular pressure, rates of evisceration and enucleation, and rates of rhegmatogenous retinal detachment following endophthalmitis diagnosis. Results: A total of 1,223,777 unique eyes underwent a PPV during the study period, with 1,275 eyes meeting the inclusion criteria. The median time from pars plana vitrectomy to endophthalmitis diagnosis was 10 days. The majority of eyes (80.9%) were managed with intravitreal antibiotics alone, without a repeat pars plana vitrectomy. Long-term visual acuity, intraocular pressure, cup-to-disc ratio, and rate of evisceration or enucleation did not differ significantly between treatment groups. However, eyes that underwent the initial pars plana vitrectomy for retinal detachment repair had poorer long-term vision and higher rates of retinal detachment following endophthalmitis. Overall, 16.1% of eyes required evisceration or enucleation in this cohort. Conclusions: Despite its rarity, post-pars plana vitrectomy endophthalmitis can cause significant morbidity and even globe loss. There were no significant differences observed between eyes that were treated with intravitreal tap and inject versus repeat pars plana vitrectomy.

Abstract The paradigm in the field of oncolytic virotherapy proposes that tumor cell killing by an oncolytic virus (OV) culminates in the priming of antitumor CD8+ T cells. However, this ignores the impact a highly immunodominant antiviral response against the OV has on the subdominant antitumor response, particularly with regard to the naturally low affinity of tumor antigens to their cognate T-cell receptors. To investigate this experimentally, we first studied the interaction between the two responses in B16 and MC38 tumor models, both expressing ovalbumin (OVA). Contrary to the accepted paradigm, the inflammatory vesicular stomatitis virus (VSV) failed to expand OVA-specific T cells in vivo unless armed with tumor-associated antigen (TAA). In fact, treatment with the control VSV led to the ablation of antitumor T cells and a shift toward dysfunction as determined by the upregulation of markers, including LAG3 and CD39, correlating with lower rates of survival. Combination with anti–PD-1 immune checkpoint blockade therapy significantly improved survival only when VSV expressed TAA. VSV-expressing altered peptide ligand variants of tumor antigens, which evade mechanisms of central tolerance, expanded near self-antigen–specific T cells with the ability to cross-react with tolerized tumor antigens, thus providing an antitumor effector population capable of proliferating in the presence of a potent antiviral response. Taken together, these findings offer a rational approach to OV design which preserves acute antitumor therapy provided by antiviral innate and T-cell responses while promoting the induction of tumor-specific T cells. Significance: Oncolytic viruses can induce an antiviral T-cell response that occludes an antitumor response, challenging the paradigm in the field and elucidating strategies to optimize the efficacy of oncolytic viroimmunotherapy.

Abstract Background: Clostridium ramosum ( C. ramosum ) is an anaerobic gram-positive bacillus of the human intestine and a rare cause of bacteremia. This retrospective cohort study evaluated clinical characteristics and outcomes of a multicenter cohort of adults with C. ramosum bacteremia. Methods: This multicenter retrospective cohort included adults (≥ 18 years) with true C. ramosum bacteremia at Mayo Clinic hospitals in Arizona, Florida, Minnesota, and Wisconsin from September 1, 2012, through December 31, 2025. Data were collected through search for C. ramosum in microbiological culture records. Clinical characteristics assessed included immune status, source of bacteremia, presence of sepsis, neutropenic fever, monomicrobial vs. polymicrobial bacteremia, antimicrobial resistance patterns, antimicrobial therapy, length of stay and 30-day all cause mortality. Exploratory odds ratios (ORs) with 95% confidence intervals (CIs) were reported. Results: Among 90 patients with a positive C. ramosum blood culture, 17 were classified as contaminants and 73 comprised the study cohort. Median age was 66.0 years (IQR, 56.0–75.0); 44 (60.3%) were male and 33 (45.2%) were immunocompromised (including 26 with active malignancy, while the remaining 7 were received biologic immunosuppression). The most common infection sources were gastrointestinal (32/73, 43.8%) and central line-related (8/73, 11%). At presentation, 50.7% of patients met criteria for sepsis and 15/73 (20.5%), were in septic shock. Of 73 isolates tested, 31/73 (44.3%) were clindamycin resistant and 9/73 (12.9%) tested intermediate. Anaerobic-active antimicrobial therapy was received by 53/73 (72.6%). Thirty-day mortality was 11/73 (15.1%). Mortality was 30.0% (6/20) with therapy without reliable anaerobic activity and 9.4% (5/53) with anaerobic-active antimicrobial therapy (OR, 4.02; 95% CI, 0.88–19.46; P = 0.060). The median hospital LOS was 8.0 days. Among 30-day survivors, anaerobic active antimicrobial therapy was associated with a shorter median LOS compared to therapy without reliable anaerobic activity (8.5 vs. 12.0 days). Immunocompromised patients had higher odds of monomicrobial C. ramosum bacteremia than immunocompetent patients (OR 3.16, 95% CI 1.19–8.39; p = 0.03). Patients with polymicrobial bacteremia had higher odds of sepsis (OR, 3.89; 95% CI, 1.43–10.55; P = 0.008) and septic shock (OR, 5.66; 95% CI, 1.17–27.36; P = 0.020). Patients with malignancy or neutropenia demonstrated significantly lower odds of presenting in septic shock (p = 0.018; p = 0.029 retrospectively). Microbiological clearance was achieved in 91.3% (42/46) of evaluable cases. Conclusions: This largest reported cohort defines the clinical and microbiological profile of C. ramosum bacteremia. Frequent clindamycin nonsusceptibility is clinically actionable. Several clinical associations found on univariable analysis should be further investigated in larger studies.

ABSTRACT Introduction/Aims Phrenic nerve conduction studies (NCS) are regularly used in the evaluation of neuromuscular respiratory dysfunction; however, the impact of body mass index (BMI) on phrenic compound muscle action potential (CMAP) amplitudes remains poorly characterized. We evaluated the relationship between BMI and phrenic CMAP amplitudes in patients without evidence of neuromuscular diaphragmatic dysfunction. Methods We performed a retrospective secondary analysis of patients derived from two previously published cohorts from Mayo Clinic (Rochester, Minnesota) and the University of Alberta (Edmonton, Alberta) evaluating diaphragm ultrasound in suspected neuromuscular respiratory insufficiency. Patients without evidence of neuromuscular disease affecting the diaphragm who underwent bilateral phrenic NCS and diaphragm ultrasound were included. Results Seventy‐nine patients met inclusion criteria, including 36 with BMI ≥ 30 kg/m 2 . Median phrenic CMAP amplitudes were significantly lower in patients with BMI ≥ 30 kg/m 2 compared to BMI < 30 kg/m 2 (0.25 mV [ IQR 0.10–0.38] vs. 0.50 mV [0.30–0.65], p < 0.001). Low phrenic CMAP amplitudes (< 0.3 mV ) were more common in patients with obesity (58% vs. 21%, p < 0.001). BMI demonstrated a moderate negative correlation with phrenic CMAP amplitude ( ρ = −0.50, p < 0.001). Diaphragm thickness at functional residual capacity did not differ significantly between BMI groups, while diaphragm thickening ratio was modestly lower in obesity. Discussion Elevated BMI is associated with lower phrenic CMAP amplitudes in patients without evidence of neuromuscular diaphragmatic dysfunction. Reduced phrenic CMAP amplitudes in obesity may reflect technical recording limitations rather than clinically meaningful diaphragmatic dysfunction. Low‐amplitude phrenic responses in patients with obesity should be interpreted cautiously and alongside adjunctive assessments such as diaphragm ultrasound.

ABSTRACT Aims Religiosity and spirituality (R/S) are generally associated with a more favourable cardiovascular risk profile, but longitudinal evidence is limited. We examined whether multidimensional R/S predicts its future behavioural and biological components. Methods This prospective cohort study analysed 6,844 adults aged ≥50 (56% women; mean R/S score 2.1; 20% weekly attenders; 81% Christian) from waves 4–7 of the English Longitudinal Study of Ageing (ELSA). Behavioural outcomes included smoking, exercise, alcohol intake, and fruit and vegetable consumption; biological outcomes included blood pressure (BP), haemoglobin A1c (HbA1c), and the inflammatory risk markers C-reactive protein (CRP) and fibrinogen. R/S was assessed with four items from the Santa Clara Strength of Religious Faith Questionnaire. Hierarchical linear and logistic regressions adjusted for age, sex, wealth, education and ethnicity; biomarker models also adjusted for health behaviours and body mass index. Results Greater spirituality ( B =−0.019; 95%CI[−0.036,−0.003]), daily prayer/meditation ( B =−0.017; 95%CI[−0.032,−0.002]), participation in organized religion ( B =−0.017; 95%CI[−0.032,−0.002]), and importance of religious faith ( B =−0.017; 95%CI[−0.033,−0.002]) were independently associated with lower fibrinogen levels. Daily prayer/meditation also independently predicted higher fruit and vegetable intake ( B =0.004; 95%CI[0.000,0.008]). However, frequent attendance (OR=0.846; 95%CI[0.730,0.982]), the significance of faith (OR=0.935; 95%CI[0.879,0.994]), and religious purpose (OR=0.939; 95%CI[0.884,0.997]) independently reduced the odds of meeting exercise recommendations. Similarly, frequent attendance was associated with higher HbA1c ( B =0.002; 95%CI[0.000,0.005]). There were no independent associations with CRP or blood pressure. Conclusion R/S showed mixed prospective associations with the cardiovascular risk profile: favourable for fibrinogen and fruit and vegetable intake, unfavourable for physical activity and HbA1c. LAY SUMMARY We examined whether religious and spiritual involvement was related to later heart-related health behaviours and blood measurements in more than 6,800 English adults aged 50 and over. The associations ran in both directions, and the study cannot show that religion or spirituality causes these changes. Stronger religious and spiritual involvement was associated with lower fibrinogen, a blood protein linked to clotting. Daily prayer or meditation was associated with eating more fruit and vegetables. Regular attendance at religious services was associated with less physical activity and higher blood sugar. GRAPHICAL ABSTRACT