Metabolomic analysis of Mdx mouse tumors

肌营养不良蛋白（Duchenne型肌营养不良症的责任基因）也被视为肿瘤抑制因子，因为一些DMD患者以及mdx小鼠自发地发展各种肿瘤，如横纹肌肉瘤和神经胶质瘤。在2022财年，为了了解肌营养不良蛋白缺陷与肿瘤发展的关系，申请人成功建立了6只mdx小鼠，并观察它们分别自发地发展各种肿瘤。通过免疫组织化学染色和FISH分析对六个肿瘤进行表征，并将其分为两组：三个横纹肌肉瘤和三个梭形细胞肉瘤。申请人接下来用从肿瘤建立的培养细胞通过气相色谱/质谱法对亲水代谢物组学谱进行综合分析，并将结果与来自骨骼肌的代谢物进行比较，并在两种肿瘤类型之间进行比较。鉴定了76种代谢物，其中20种代谢物显示出显著更高的倍数变化。在鉴定的代谢物中，有许多氨基酸在糖酵解中作为替代碳源，如丙氨酸、丝氨酸、天冬氨酸和GABA，以及糖。途径分析确定了丙氨酸、天冬氨酸和谷氨酸代谢途径;牛磺酸和亚牛磺酸途径; TCA途径;丙酮酸代谢途径;戊糖和葡萄糖醛酸相互转化途径等。

Dystrophin the responsible gene for Duchenne muscular dystrophy is also regarded as a tumor suppressor because some DMD patients as well as mdx mouse spontaneously develop various tumors such as rhabdomyosarcoma and glioma.In FY2022, to understand the relationship of dystrophin defect and tumor development, the applicants have successfully established six mdx mice and observed them to spontaneously develop various tumors, respectively. The six tumors were characterized by immunohistochemical staining, and FISH analysis and divided into two groups: three rhabdomyosarcoma and three spindle cell sarcomas.The applicants next performed a comprehensive analysis of the hydrophilic metabolomic profile by Gas chromatography/Mass Spectrometry with cultured cells established from the tumors and compared the result to metabolites from skeletal muscle and among the two tumor types. 76 metabolites were identified with 20 metabolites that demonstrated a significantly higher fold change. Amongst the identified metabolites were many amino acids that function as alternative carbon source in glycolysis such as alanine, serine, aspartate and GABA, as well as sugars. Pathway analysis identified alanine, aspartate and glutamate metabolic pathway; taurine and hypotaurine pathway; TCA pathway; pyruvate metabolic pathway, pentose and glucuronic interconversion pathways etc.