Rehman, T. | bioRxiv Science

bioRxiv2026

In Utero CFTR Modulation Alleviates Disease in G551D Cystic Fibrosis Pigs

Previous studies indicate that pigs with CFTR-null and CFTR-{Delta}F508 mutations develop multiorgan disease similar to that in people with cystic fibrosis (CF). At birth, their airways exhibit host defense defects that predispose to airway infection, inflammation, and mucus accumulation. The CFTR-G551D mutation causes CF by producing CFTR channels that localize correctly but have reduced channel activity. Ivacaftor (VX-770) is a small molecule drug developed to potentiate CFTR activity. To test the phenotype of the CFTR-G551D mutation in pigs and determine whether ivacaftor can rescue CF abnormalities, we developed CFTRG551D/G551D (CF-G551D) pigs through homologous recombination in fetal fibroblasts and somatic cell nuclear transfer. Newborn CF-G551D piglets exhibited phenotypes similar to CF-null piglets, including meconium ileus, exocrine pancreatic destruction, micro-gallbladder, vas deferens destruction, and airway structural abnormalities. Compared to wild-type pigs, CF-G551D pigs had reduced forskolin-stimulated short-circuit current in airway and intestinal tissues. Ivacaftor increased the single-channel open state probability of CFTR-G551D and increased short-circuit current to near wild-type levels. Similar to our other CF pig models, we found that 100% of CF-G551D pigs were born with meconium ileus. To test whether in utero ivacaftor treatment could prevent or alleviate meconium ileus, pregnant sows were treated with ivacaftor beginning at day 35 of gestation and continuing until delivery. This treatment rescued the pancreas, gallbladder, and vas deferens phenotype in the majority of CF-G551D pigs. Animals that were spared from meconium ileus were able to survive without ivacaftor treatment. Airway disease developed similar to other CF pig models. These findings indicate that this model may be useful for studies in which CFTR function can be reversed, for investigating in utero CFTR correction strategies, and for longitudinal studies in CF pigs.

physiology↗

bioRxiv2025

Phenotypic and Molecular characterization of thermophlic bacterial isolates from the rizosphere of Panicum antidotale in Cholistan desert, Pakistan

The major aim of this study was to isolate, characterize, and determine the distribution of culturable bacteria in the rhizosphere of Panicum antidotale (PAMA) from the Cholistan desert, Pakistan. A total of 109 thermophilic bacterial isolates were obtained from PAMA and subjected to morphological, biochemical, and molecular characterization at The Islamia University of Bahawalpur. Standard protocols were used for physical and biochemical analysis, while molecular identification involved 16S rRNA profiling, restriction fragment length polymorphism (RFLP), and DNA sequence analysis. PCR amplicons of selected isolates with distinct RFLP patterns were sequenced, and the DNA sequences were analyzed using BLAST to determine species identity. Results revealed the presence of Bacillus subtilis, Brevibacillus borstelensis, and Paenibacillus dendritiformis in the majority of analyzed rhizosphere samples. Bacillus licheniformis and Bacillus subtilis were the most dominant species in PAMA, accounting for 45% and 30% of the isolates, respectively. In addition, lower frequencies of other species, such as Paenibacillus dendritiformis (10-15%), were also observed. These findings demonstrate that Panicum antidotale supports a unique rhizospheric bacterial community dominated by thermophilic Bacillus species. This study represents the first comprehensive investigation of PAMA microbiota in the Cholistan desert. The results provide novel insights into bacterial diversity in extreme environments and highlight the potential role of these communities in plant growth promotion and stress adaptation. Further studies are warranted to explore the ecological significance and possible biotechnological applications of PAMA-associated thermophilic bacteria for sustainable agriculture and environmental management. ImportanceThis study presents the first molecular-level investigation of thermophilic bacterial communities inhabiting the rhizosphere of Panicum antidotale in the Cholistan Desert, Pakistan. Through combined phenotypic, biochemical, and molecular analyses, diverse thermophilic Bacillus, Brevibacillus, and Paenibacillus species were identified as dominant members of this extreme arid ecosystem. These bacteria possess plant growth-promoting and stress-alleviating traits, underscoring their ecological significance in supporting desert plant resilience. Importantly, this first molecular characterization of P. antidotale-associated microbiota reveals that these isolates may also serve as promising sources of valuable enzymes and bioactive metabolites. The findings broaden our understanding of microbial diversity in arid rhizospheres and highlight the potential applications of these thermophilic bacteria in sustainable agriculture, biotechnology, and environmental management.

microbiology↗