Michelini S, Vettori A, Maltese P E, Cardone M, Bruson A, Fiorentino A, Cappellino F, Sainato V, Guerri G, Marceddu G, Tezzele S, Bertelli M
Lymphology. 2016 Jun;49(2):57-72.
Primary lymphedema is a rare inherited condition characterized by swelling of body tissues caused by accumulation of fluid, especially in the lower limbs. In many patients, primary lymphedema has been associated with variations in a number of genes involved in the development and maintenance of the lymphatic system. In this study, we performed a genetic screening in patients affected by primary lymphedema using a next generation sequencing (NGS) approach. With this technology, based on a custom-made oligonucleotide probe library, we were able to analyze simultaneously in each patient all the coding exons of 10 genes (FLT4, FOXC2, CCBE1, GJC2, MET, HGF, GATA2, SOX18, VEGFC, KIF11) associated with primary lymphedema. In the study population, composed of 45 familial and 71 sporadic cases, we identified the presence of rare variants with a potential pathogenic effect in 33% of subjects. Overall, we found a total of 36 different rare nucleotidic alterations, 30 of which had not been previously described. Among these, we identified 23 mutations that we considered most likely to be disease causing. Patients with an FLT4 or FOXC2 alteration accounted for the largest percentage of the sample, followed by MET, HGF, KIK11, GJC2 and GATA2. No alterations were identified in SOX18, VEGFC, and CCBE1 genes. In conclusion, we showed that NGS technology can be successfully applied to perform molecular screening of lymphedema-associated genes in large cohort of patients with a reasonable effort in terms of cost, work, and time.
原发性淋巴水肿是一种罕见的遗传性疾病，其特征是身体组织因液体蓄积而肿胀，尤其是下肢。在许多患者中，原发性淋巴水肿与一些参与淋巴系统发育和维持的基因变异有关。在本研究中，我们采用下一代测序（NGS）方法对原发性淋巴水肿患者进行了基因筛查。利用这项技术，基于定制的寡核苷酸探针文库，我们能够在每位患者中同时分析与原发性淋巴水肿相关的10个基因（FLT4、FOXC2、CCBE1、GJC2、MET、HGF、GATA2、SOX18、VEGFC、KIF11）的所有编码外显子。在由45例家族性病例和71例散发性病例组成的研究人群中，我们在33%的受试者中发现了具有潜在致病作用的罕见变异。总体而言，我们共发现了36种不同的罕见核苷酸改变，其中30种此前未被描述。在这些改变中，我们鉴定出23种最有可能导致疾病的突变。FLT4或FOXC2改变的患者在样本中占比最大，其次是MET、HGF、KIK11、GJC2和GATA2。在SOX18、VEGFC和CCBE1基因中未发现改变。总之，我们表明NGS技术可以成功应用于对大量淋巴水肿患者进行淋巴水肿相关基因的分子筛查，在成本、工作量和时间方面付出合理的努力即可。
