Molecular and Immunologic Techniques in a Genetically Engineered Mouse Model of Gastrointestinal Stromal Tumor.

Autor: Tieniber AD; Department of Surgery, Hospital of the University of Pennsylvania., Hanna AN; Department of Surgery, Hospital of the University of Pennsylvania., Do K; Department of Surgery, Hospital of the University of Pennsylvania., Wang L; Department of Surgery, Hospital of the University of Pennsylvania., Rossi F; Department of Surgery, Hospital of the University of Pennsylvania., DeMatteo RP; Department of Surgery, Hospital of the University of Pennsylvania; Ronald.DeMatteo@pennmedicine.upenn.edu.
Jazyk: angličtina
Zdroj: Journal of visualized experiments : JoVE [J Vis Exp] 2022 May 02 (183). Date of Electronic Publication: 2022 May 02.
DOI: 10.3791/63853
Abstrakt: Gastrointestinal stromal tumor (GIST) is the most common human sarcoma and is typically driven by a single mutation in the KIT receptor. Across tumor types, numerous mouse models have been developed in order to investigate the next generation of cancer therapies. However, in GIST, most in vivo studies use xenograft mouse models which have inherent limitations. Here, we describe an immunocompetent, genetically engineered mouse model of gastrointestinal stromal tumor harboring a Kit V558Δ/+ mutation. In this model, mutant KIT, the oncogene responsible for most GISTs, is driven by its endogenous promoter leading to a GIST which mimics the histological appearance and immune infiltrate seen in human GISTs. Furthermore, this model has been used successfully to investigate both targeted molecular and immune therapies. Here, we describe the breeding and maintenance of a Kit V558Δ/+ mouse colony. Additionally, this paper details the treatment and procurement of GIST, draining mesenteric lymph node, and adjacent cecum in Kit V558Δ/+ mice, as well as sample preparation for molecular and immunologic analyses.
Databáze: MEDLINE