背景:Glanzmann血栓性(GT)是一种先天性血小板疾病,影响全球约1:1000000人,其特征是血小板聚集和凝块收缩受损。常染色体隐性遗传,失去功能,αIIbβ3受体的ITGA2B或ITGB3变体在人类中引起该疾病。一只受格兰兹曼和大型血小板减少症影响的猫被送到加州大学戴维斯分校的VMTH。
目的:这只猫的严重血栓病具有潜在的遗传病因。
方法:一名受影响的患者,2个年龄匹配的临床健康对照,和正常猫的老年群体(n=20)。
方法:对患者进行体格检查和临床病理检查。对患者表型进行流式细胞术和血小板聚集分析。提取患者和验证组群gDNA样品用于先前鉴定的ITGA2B(c.1986delC)变体的Sanger测序。对患者和健康对照PRP样品进行逆转录酶PCR以验证ITGA2B变体结果。
结果:在ITGA2B中发现了一个新的c.1986_1987insCC常染色体隐性变异。在44个不同品种的194只无关猫的种群中不存在这种变体。通过RT-PCR和流式细胞术验证ITGA2B转录本和蛋白质表达的完全丧失,解释GT的潜在病因,和可能的大型血小板减少症,在这只猫。
结论:这项研究强调了精准医学在猫心血管疾病中的作用,并确定了另一种可能用于猫人群筛查的变体。这项研究提供了一个小体积,标准化,成功的方案进行适当的血小板RNA分离和随后的基因表达的分子评估在猫。
BACKGROUND: Glanzmann\'s thrombasthenia (GT) is a congenital platelet disorder affecting approximately 1:1 000 000 people globally and characterized by impaired platelet aggregation and clot retraction. Autosomal recessive, loss-of-function, variants in ITGA2B or ITGB3 of the αIIbβ3 receptor cause the disease in humans. A cat affected by Glanzmann\'s and macrothrombocytopenia was presented to the UC Davis VMTH.
OBJECTIVE: Severe thrombopathia in this cat has an underlying genetic etiology.
METHODS: A single affected patient, 2 age-matched clinically healthy controls, and a geriatric population (n = 20) of normal cats.
METHODS: Physical examination and clinical pathology tests were performed on the patient. Flow cytometry and platelet aggregometry analyses for patient phenotyping were performed. Patient and validation cohort gDNA samples were extracted for Sanger sequencing of a previously identified ITGA2B (c.1986delC) variant. Reverse transcriptase PCR was performed on patient and healthy control PRP samples to verify ITGA2B variant consequence.
RESULTS: A novel c.1986_1987insCC autosomal recessive variant in ITGA2B was identified. This variant was absent in a population of 194 unrelated cats spanning 44 different breeds. Complete loss of ITGA2B transcript and protein expression was verified by RT-PCR and flow cytometry, explaining the underlying etiology of GT, and likely macrothrombocytopenia, in this cat.
CONCLUSIONS: This study emphasizes the role of precision medicine in cardiovascular disease of cats and identified yet another variant that may be of utility for screening in the feline population. This study provides a small-volume, standardized, successful protocol for adequate platelet RNA isolation and subsequent molecular assessment of gene expression in cats.