@misc{hill_murphy_polkoff_edwards_walker_moatti_greenbaum_piedrahita_2024, title={A gene edited pig model for studying LGR5<SUP>+</SUP> stem cells: implications for future applications in tissue regeneration and biomedical research}, volume={6}, ISSN={["2673-3439"]}, DOI={10.3389/fgeed.2024.1401163}, abstractNote={Recent advancements in genome editing techniques, notably CRISPR-Cas9 and TALENs, have marked a transformative era in biomedical research, significantly enhancing our understanding of disease mechanisms and helping develop novel therapies. These technologies have been instrumental in creating precise animal models for use in stem cell research and regenerative medicine. For instance, we have developed a transgenic pig model to enable the investigation of LGR5-expressing cells. The model was designed to induce the expression of H2B-GFP under the regulatory control of the LGR5 promoter via CRISPR/Cas9-mediated gene knock-in. Notably, advancements in stem cell research have identified distinct subpopulations of LGR5-expressing cells within adult human, mouse, and pig tissues. LGR5, a leucine-rich repeat-containing G protein-coupled receptor, enhances WNT signaling and these LGR5 + subpopulations demonstrate varied roles and anatomical distributions, underscoring the necessity for suitable translational models. This transgenic pig model facilitates the tracking of LGR5-expressing cells and has provided valuable insights into the roles of these cells across different tissues and species. For instance, in pulmonary tissue, Lgr5 + cells in mice are predominantly located in alveolar compartments, driving alveolar differentiation of epithelial progenitors via Wnt pathway activation. In contrast, in pigs and humans, these cells are situated in a unique sub-basal position adjacent to the airway epithelium. In fetal stages a pattern of LGR5 expression during lung bud tip formation is evident in humans and pigs but is lacking in mice. Species differences with respect to LGR5 expression have also been observed in the skin, intestines, and cochlea further reinforcing the need for careful selection of appropriate translational animal models. This paper discusses the potential utility of the LGR5 + pig model in exploring the role of LGR5 + cells in tissue development and regeneration with the goal of translating these findings into human and animal clinical applications.}, journal={FRONTIERS IN GENOME EDITING}, author={Hill, Amanda B. T. and Murphy, Yanet M. and Polkoff, Kathryn M. and Edwards, Laura and Walker, Derek M. and Moatti, Adele and Greenbaum, Alon and Piedrahita, Jorge A.}, year={2024}, month={Jun} }
 @article{moatti_cai_li_sattler_edwards_piedrahita_ligler_greenbaum_2020, title={Three-dimensional imaging of intact porcine cochlea using tissue clearing and custom-built light-sheet microscopy}, volume={11}, ISSN={["2156-7085"]}, url={http://dx.doi.org/10.1364/boe.402991}, DOI={10.1364/BOE.402991}, abstractNote={Hearing loss is a prevalent disorder that affects people of all ages. On top of the existing hearing aids and cochlear implants, there is a growing effort to regenerate functional tissues and restore hearing. However, studying and evaluating these regenerative medicine approaches in a big animal model (e.g. pigs) whose anatomy, physiology, and organ size are similar to a human is challenging. In big animal models, the cochlea is bulky, intricate, and veiled in a dense and craggy otic capsule. These facts complicate 3D microscopic analysis that is vital in the cochlea, where structure-function relation is time and again manifested. To allow 3D imaging of an intact cochlea of newborn and juvenile pigs with a volume up to ∼ 250 mm3, we adapted the BoneClear tissue clearing technique, which renders the bone transparent. The transparent cochleae were then imaged with cellular resolution and in a timely fashion, which prevented bubble formation and tissue degradation, using an adaptive custom-built light-sheet fluorescence microscope. The adaptive light-sheet microscope compensated for deflections of the illumination beam by changing the angles of the beam and translating the detection objective while acquiring images. Using this combination of techniques, macroscopic and microscopic properties of the cochlea were extracted, including the density of hair cells, frequency maps, and lower frequency limits. Consequently, the proposed platform could support the growing effort to regenerate cochlear tissues and assist with basic research to advance cures for hearing impairments.}, number={11}, journal={BIOMEDICAL OPTICS EXPRESS}, publisher={The Optical Society}, author={Moatti, Adele and Cai, Yuheng and Li, Chen and Sattler, Tyler and Edwards, Laura and Piedrahita, Jorge and Ligler, Frances S. and Greenbaum, Alon}, year={2020}, month={Nov}, pages={6181–6196} }
 @article{lennon_borst_edwards_moeser_2018, title={Mast cells exert anti-inflammatory effects in an IL10(-/-) model of spontaneous colitis}, journal={Mediators of Inflammation}, author={Lennon, E. M. and Borst, L. B. and Edwards, L. L. and Moeser, A. J.}, year={2018} }
 @article{pohl_medland_mackey_edwards_bagley_dewilde_williams_moeser_2017, title={Early weaning stress induces chronic functional diarrhea, intestinal barrier defects, and increased mast cell activity in a porcine model of early life adversity}, volume={29}, ISSN={["1365-2982"]}, DOI={10.1111/nmo.13118}, abstractNote={AbstractBackgroundEarly life adversity (ELA) is a risk factor for development of gastrointestinal disorders later in life. The underlying mechanisms through which ELA and sex interact to influence disease susceptibility remains poorly understood.MethodsUtilizing a porcine early weaning stress (EWS) model to mimic ELA, we investigated the long‐term effects of EWS on functional diarrhea, ileal permeability, mast cell activity and mast cell relationship with enteric ganglia.Key ResultsJuvenile and adult EWS pigs exhibited chronic, functional diarrhea (EWS 43.6% vs late wean control(LWC) 4.8%, P<.0001), increased intestinal permeability (2 fold increase EWS vs LWC, P<.0001), and mast cell numbers (at 7 weeks and 20 weeks ~1.6 fold increase EWS vs LWC, P<.05). Compared with EWS male castrates (Male‐C), females EWS pigs exhibited more frequent diarrhea (58.8% vs 29.9%, P=.0016), and increased intestinal permeability (1‐2 fold higher in EWS females, P<.001). Increased mast cell numbers and their enhanced co‐localization with neuronal ganglia were observed in both Male‐C and female EWS pigs; however, female pigs exhibited greater release of mast cell tryptase upon activation with c48/80 (~1.5 fold increase, P<.05), compared with Male‐C pigs.Conclusions and InferencesThese data demonstrate that pigs exposed to ELA exhibit increased vulnerability to functional diarrhea, intestinal permeability and mast cell activity. Further, these studies also showed that EWS female and Male‐C pigs exhibited dimorphic responses to EWS with female piglets exhibited greater susceptibility and severity of diarrhea, intestinal permeability and mast cell tryptase release. Together, these findings mimic some of the key pathophysiologic findings in human functional GI disorders functional gastrointestinal disorders (FGIDs) suggesting that the EWS porcine model could be a valuable preclinical translational model for FGID research associated with ELA.}, number={11}, journal={NEUROGASTROENTEROLOGY AND MOTILITY}, author={Pohl, C. S. and Medland, J. E. and Mackey, E. and Edwards, L. L. and Bagley, K. D. and DeWilde, M. P. and Williams, K. J. and Moeser, A. J.}, year={2017}, month={Nov} }
 @article{medland_pohl_edwards_frandsen_bagley_li_moeser_2016, title={Early life adversity in piglets induces long-term upregulation of the enteric cholinergic nervous system and heightened, sex-specific secretomotor neuron responses}, volume={28}, ISSN={["1365-2982"]}, DOI={10.1111/nmo.12828}, abstractNote={AbstractBackgroundEarly life adversity (ELA) is a risk factor for the later‐life onset of gastrointestinal (GI) diseases such as irritable bowel syndrome (IBS); however, the mechanisms are poorly understood. Here, we utilized a porcine model of ELA, early weaning stress (EWS), to investigate the influence of ELA on the development and function of the enteric nervous system (ENS).MethodsFemale and castrated male (Male‐C) piglets were weaned from their sow either at 15 days of age (EWS) or 28 days of age (late weaning control, LWC). At 60 and 170 days of age, ileal mucosa‐submucosa preparations were mounted in Ussing chambers and veratridine‐ and corticotropin releasing factor (CRF)‐releasing factor‐evoked short circuit current (Isc) responses were recorded as indices of secretomotor neuron function. Enteric neuron numbers and the expression of select neurotransmitters and their receptors were also measured.Key ResultsCompared with LWC pigs, female, but not Male‐C EWS, pigs exhibited heightened veratridine‐induced Isc responses at 60 and 170 days of age that were inhibited with tetrodotoxin and atropine. Ileum from EWS pigs had higher numbers of enteric neurons that were choline acetyltransferase positive. Markers of increased cholinergic signaling (increased acetylcholinesterase) and downregulated mucosal muscarinic receptor 3 gene expression were also observed in EWS pigs.Conclusions & InferencesThis study demonstrated that EWS in pigs induces lasting and sex‐specific hypersensitivity of secretomotor neuron function and upregulation of the cholinergic ENS. These findings may represent a mechanistic link between ELA and lifelong susceptibility to GI diseases such as IBS.}, number={9}, journal={NEUROGASTROENTEROLOGY AND MOTILITY}, author={Medland, J. E. and Pohl, C. S. and Edwards, L. L. and Frandsen, S. and Bagley, K. and Li, Y. and Moeser, A. J.}, year={2016}, month={Sep}, pages={1317–1329} }
 @article{boyer_s. d'costa_edwards_milloway_susick_borst_thakur_campbell_crenshaw_polo_et al._2015, title={Early-life dietary spray-dried plasma influences immunological and intestinal injury responses to later-life Salmonella typhimurium challenge}, volume={113}, ISSN={["1475-2662"]}, DOI={10.1017/s000711451400422x}, abstractNote={Increasing evidence supports the concept that early-life environmental influences, including nutrition and stress, have an impact on long-term health outcomes and disease susceptibility. The objective of the present study was to determine whether dietary spray-dried plasma (SDP), fed during the first 2 weeks post-weaning (PW), influences subsequent immunological and intestinal injury responses toSalmonellatyphimuriumchallenge. A total of thirty-two piglets (age 16–17 d) were weaned onto nursery diets containing 0, 2·5 % SDP (fed for 7 d PW) or 5 % SDP (fed for 14 d PW), and were then fed control diets (without SDP), for the remainder of the experiment. At 34 d PW (age 50 d), pigs were challenged with 3 × 109colony-forming units ofS. typhimurium. A control group (non-challenged) that was fed 0 % SDP in the nursery was included. At 2 d post-challenge, the distal ileum was harvested for the measurement of inflammatory, histological and intestinal physiological parameters.S.typhimuriumchallenge induced elevated ileal histological scores, myeloperoxidase (MPO), IL-8 and TNF, and increased intestinal permeability (indicated by reduced transepithelial voltage (potential difference) and elevated 4 kDa fluorescein isothiocyanate dextran (FD4) flux rates). Compared withS.typhimurium-challenged controls (0 % SDP), pigs fed the 5 % SDP-14 d diet exhibited reduced ileal histological scores, MPO levels, IL-8 levels and FD4 flux rates. Pigs fed the 5 % SDP-14 d nursery diet exhibited increased levels of plasma and ileal TNF-α in response to the challenge, compared with the other treatments. These results indicate that inclusion of SDP in PW diets can have an influence on subsequent immunological and intestinal injury responses induced by later-lifeS.typhimuriumchallenge.}, number={5}, journal={BRITISH JOURNAL OF NUTRITION}, author={Boyer, P. E. and S. D'Costa and Edwards, L. L. and Milloway, M. and Susick, E. and Borst, L. B. and Thakur, S. and Campbell, J. M. and Crenshaw, J. D. and Polo, J. and et al.}, year={2015}, month={Mar}, pages={783–793} }