Highly effective drugs modulating the defective protein encoded by the CFTR gene have revolutionized cystic fibrosis (CF) therapy. Preclinical drug-testing on human nasal epithelial (HNE) cell cultures and 3-dimensional human intestinal organoids (3D HIO) are used to address patient-specific variation in drug response and to optimize individual treatment for people with CF. This study is the first to report comparable CFTR functional responses to CFTR modulator treatment among patients with different classes of CFTR gene variants using the three methods of 2D HIO, 3D HIO, and HNE. Furthermore, 2D HIO showed good correlation to clinical outcome markers. A larger measurable CFTR functional range and access to the apical membrane were identified as advantages of 2D HIO over HNE and 3D HIO, respectively. Our study thus expands the utility of 2D intestinal monolayers as a preclinical drug testing tool for CF.

Objective: To examine the changes over time of pediatric acute pancreatitis (AP) severity, management, and disease outcomes at our academic tertiary center. Methods: We reviewed 223 pediatric AP admissions (2002–2018) and used a time-to-event regression model to study changes over time. Disease outcomes were analyzed using a subgroup of 89 patients in whom only the AP event determined length of hospital stay and duration of opioid use. Results: There was an increase in mild, but not severe, AP episodes over the examined period. June 2014 was identified as a single cutoff point for change in AP management and disease outcomes independent of each other and of disease severity. Timing of initiating enteral nutrition decreased from 5 to 1.6 days (P < 0.0001) in the entire cohort and from 4.1 to 1.8 days in the subgroup (P = 0.0001) after June 2014. Length of hospitalization decreased from 6 to 3.3 days (P = 0.0008) and days of opioid use from 4.1 to 1.3 (P = 0.002) after June 2014. Conclusions: Timing of initiating enteral nutrition has significantly reduced at our center after June 2014. In parallel, we observed a significant improvement in disease outcomes.

Inflammatory bowel disease (IBD) is a chronic, relapsing inflammation disorder of the gastrointestinal tract characterized by disrupted intestinal epithelial barrier function. Despite advances in treatment, including biological agents, achieving sustained remission remains challenging for many patients with IBD. This highlights the urgent need for novel therapeutic strategies. Milk-derived extracellular vesicles (MDEs) have emerged as a promising therapeutic option. In this study, we isolated and characterized MDEs and evaluated their effects on the function of intestinal epithelial cells (IECs). Using a murine model of Dextran Sulfate Sodium (DSS)-induced colitis, we observed that MDEs significantly ameliorated disease symptoms. The upregulation of β-catenin, a crucial mediator of Wnt signaling, in colonic tissues suggests that MDEs may facilitate epithelial regeneration and restore barrier function. In patient-derived colon organoids (PDCOs), MDEs were internalized and modulated the expression of key signaling molecules, such as the upregulation of β-catenin, cyclin D1, and the proliferation marker Ki67, indicating their potential to promote IEC proliferation and intestinal barrier repair. Importantly, MDEs demonstrated selective activity by downregulating β-catenin and cyclin D1 in colon cancer cells, leading to reduced proliferation. This selectivity indicates a dual therapeutic potential of MDEs for promoting healthy IEC proliferation while potentially mitigating malignancy risks.

Genotype-based drug development has yielded highly effective therapies, notably the triple combinations elexacaftor/tezacaftor/ivacaftor (ETI) and vanzacaftor/tezacaftor/deutivacaftor (VTD), now approved in Europe for people with CF having at least one non-class I variant. However not all these people with CF will respond to ETI or VTD, and a few not under the label may respond. Facilitating opportunities to access for patients with rare variants has required a shift in paradigm toward functional testing-based access. This approach assesses the potential benefit of modulator therapy using in vitro functional assays, either in engineered systems expressing defined CFTR variants (theratyping) or in patient-derived tissues (theranostics). We review theranostics as a critical tool for personalized medicine in CF, highlighting its validation in in vitro models derived from patients' own cells such as human intestinal organoids and human nasal epithelial cells. We discuss the current regulatory landscape regarding modulator approval and propose strategies for improving equitable access to effective treatments for all people with CF. Importantly, we advocate for functional assays to be accepted as standalone evidence of drug efficacy for patients with rare variants. Theranostic approaches remain critical when theratyping has not been achieved, and genetic data is not available or clearly interpretable. Indeed, theranostics has emerged as an essential pillar of CF drug access, complementing genotype-driven strategies. As the field advances, continued validation, standardization, and regulatory integration of in vitro functional assays will be key to ensuring that every person with CF-regardless of their genotype-has the opportunity to benefit from precision therapies.

Background The combination of the CFTR modulators Elexacaftor (VX-445)/Tezacaftor (VX-661)/Ivacaftor (VX-770) enables the effective rescue of CFTR function in people with the F508del mutation and 177 other FDA-approved alleles. However, the effect of modulator combination treatment on many rare CFTR mutations is mostly unknown. Furthermore, rare CFTR mutations may not require all ETI components to reach maximal correction. This can be studied in intestinal organoids derived from patients carrying rare mutations. Methods Intestinal organoids were generated from six patients carrying the Q1100P and/or K163E alleles, not receiving ETI. Measurements of the response to ETI or combination of its components were performed in 3D-organoids by forskolin-induced swelling (FIS) and in 2D-monolayers by short-circuit currents (Isc). Based on these results, patients initiated off-label ETI treatment. Clinical data before and after treatment were collected. Results Functional measurements showed that both mutations are responsive to ETI. The results further showed that VX-445 had a dramatic effect on K163E function. Both Q1100P and K163E mutations achieved clinically significant CFTR activity levels with VX-661+VX-445, without benefit from VX-770. Following these results patients initiated off label ETI, resulting in significant and sustained clinical improvements, in all patients, in lung function (FEV₁, LCI), BMI and sweat chloride. Conclusion Our results support that CFTR function measurements in patient-derived intestinal organoids carrying rare CFTR alleles can detect potential responders to modulator treatment and serve as the basis for drug approval by health providers. Furthermore, this approach allows for patient-specific optimization of modulator combinations, minimizing unnecessary exposure to ineffective treatments.

Objectives: Pediatric acute pancreatitis (AP) is a growing clinical concern with a wide spectrum of severity, from mild episodes to life‐threatening conditions. Traditional diagnostic methods primarily rely on serum amylase and lipase measurements, which are invasive and can be challenging in children. This study is the first to evaluate the diagnostic accuracy of the urine trypsinogen‐2 dipstick test (UTDT) as a noninvasive test for diagnosing pediatric AP. Methods: This prospective study included 28 pediatric patients (31 episodes) presenting with acute abdominal pain at a tertiary medical center from November 2022 to October 2024. AP was diagnosed based on the International Study Group of Pediatric Pancreatitis: In Search for a Cure (INSPPIRE) criteria. Urine samples were collected either within the first 24 hours (h) or later during hospitalization. UTDT sensitivity and specificity were calculated and compared to serum amylase and lipase levels. Results: Of the 31 episodes, 19 (61%) were confirmed as AP, and 12 (39%) were attributed to other causes. The UTDT had an overall sensitivity of 68% and specificity of 100%. Sensitivity increased to 87% when urine samples were collected within 24 h of admission. In non‐AP cases, UTDT consistently produced negative results, with the high specificity supporting its reliability in distinguishing AP from other conditions. Conclusions: The UTDT demonstrates promise as a rapid, noninvasive diagnostic tool for pediatric AP, particularly when used early in the disease course. Its high specificity and ease of use suggest that it may serve as an alternative to invasive blood tests once validated through larger‐scale studies. Further research is needed to confirm these findings and establish the role of UTDT in clinical practice.