Citations

Abstract Background: Hemorrhagic necrotizing pancreatitis (HNP) is a highly lethal form of pancreatitis that lacks mechanism-based therapy. Intra-acinar enzyme activation is fundamental to pancreatitis initiation, yet it does not explain why only a subset of episodes progress to catastrophic hemorrhagic necrosis. Objective: To identify determinants of progression to lethal HNP and test a mechanism-based preventive intervention. Design: Using humanized trypsinogen mouse models and secretagogue regimens, we compared high-dose cerulein with low/moderate-dose cerulein and bombesin. We quantified pancreatic protease activity, performed blinded histopathologic scoring, assessed hemorrhage/vascular disruption, and evaluated enzyme localization and ductal injury by PRSS1 immunostaining. De-identified human pancreatic histology from HNP were examined. We tested a repurposed strategy using clinically approved agents for other indications: secretin to stimulate ductal fluid secretion and isosorbide mononitrate (ISMN) to promote ductal outflow via sphincter of Oddi relaxation. Results: High-dose cerulein induced robust intrapancreatic protease activation but predominantly caused edematous pancreatitis. Paradoxically, bombesin and low/moderate-dose cerulein elicited lower total pancreatic protease activity yet produced severe, frequently lethal HNP with vascular disruption and lobular ischemic necrosis. HNP was associated with intraductal enzyme accumulation and ductal injury. Secretin or ISMN alone showed limited protection, whereas combined secretin+ISMN prevented hemorrhagic necrotizing injury. Conclusion: Pancreatitis severity is determined not only by the magnitude of enzyme activation but also by where enzyme activity is exerted, implicating intraductal enzyme accumulation as a mechanistic determinant and therapeutic target in HNP. Coordinated enhancement of ductal secretion and outflow with clinically approved agents prevents HNP in vivo, supporting a translatable prevention strategy.

ABSTRACTTissue‐specific gene manipulation using Cre/loxP or Flp/frt recombination systems is a cornerstone of genetically engineered mouse models. In this study, we aim to develop a novel hepatocyte‐specific, tamoxifen‐inducible Flp mouse line. BAC (bacterial artificial chromosome)‐Alb(albumin)‐FlpER2(estrogen receptor ligan binding domain) was developed by inserting IRES‐FlpER2 cDNA between the translation stop codon and 3′‐UTR of the mouse albumin gene in a bacterial artificial chromosome. Upon tamoxifen induction in mice crossed with reporter lines, western blotting, immunohistochemistry, immunofluorescence staining, and X‐gal staining (5‐bromo‐4‐chloro‐3‐indolyl‐β‐D‐galactopyranoside Staining) were used to verify the recombination efficiency and specificity of this mouse model. Recombination was highly efficient and specific in hepatocytes, with no recombination detected in intrahepatic cholangiocytes or other organs in this mouse model. We generated a new tamoxifen‐induced hepatocyte‐specific mouse model with highly efficient recombination specifically in hepatocytes, and this model can be used to generate tumor model lines.

AbstractA deeper understanding of pathophysiology, from the initiation of tumors of epithelial cells to the outcomes associated with it, is imperative to test potential interventions for these diseases. However, few models can accomplish this because of the complexity of origin cells and their transformation. We established an epithelial cell original tumor mouse model by conditional KrasG12D activation and tumor suppressor gene p53 deletion specifically in epithelial cells of the whole body using Cytokeratin 19 (CK19) promoter via Cre‐ERT mediation. Multiple tumors derived from epithelial cells developed in this novel model. This model can recapitulate the progress of intestinal, pancreatic, liver, lung, and subcutaneous tumors which derive from epithelial cells. This model is suitable for studying the pathophysiology of tumors derived from epithelial cells and may have the potential to test interventions.

This study presents a streamlined, cost-effective genotyping workflow for clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) edited mouse models, focusing on trypsinogen genes. It simplifies initial F0 mouse screening using PCR and Sanger sequencing without needing exact sequences. For F1 mice, precise editing is identified through Sanger sequencing and algorithm decoding. The workflow includes a novel PCR strategy for distinguishing homozygous and heterozygous statuses in subsequent generations, effective for small deletions, multiple-gene knockouts, and knockins.