Pulmonary Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda.
Flow Cytometry Core Facility, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda.
Chest. 2019 Aug;156(2):298-307. doi: 10.1016/j.chest.2019.03.040. Epub 2019 Apr 26.
Lymphangioleiomyomatosis (LAM) is a destructive metastasizing neoplasm of the lung characterized by proliferation of LAM cells in specialized lung nodules. LAM cells are characterized by expression of the prometastatic and cancer-initiating hyaluronan receptor CD44v6, and loss of heterozygosity (LOH) of TSC1 and TSC2. The circulating neoplastic LAM cells are thought to be involved in metastasis. Because LAM cells display properties of neoplastic, metastatic, and stem cell-like cancer cells, we hypothesized that elevated aldehyde dehydrogenase (ALDH) activity, characteristic of cancer and stem cells, is a property of LAM cells.
We performed an in silico search of ALDH genes in microdissected LAM lung nodules. To identify circulating LAM cells, we osmotically removed red blood cells from whole blood to obtain peripheral blood mononuclear cells, which were then sorted by fluorescence-activated cell sorting based on their level of ALDH activity.
Microdissected LAM lung nodules possess a distinctive ALDH gene profile. The cell subpopulation with high ALDH activity, isolated from circulating cells, possessed TSC2 LOH in 8 of 14 patients with LAM. Approximately 60% of the circulating cells with high ALDH activity expressed CD44v6. Cells with TSC2 LOH from patients with LAM and LAM/TSC exhibited different properties in different body locations, but all cell types showed high ALDH activity.
This new procedure allows for isolation of circulating LAM cells from cultured cells, blood, and chylous effusions and shows that circulating LAM cells are heterogeneous with neoplastic, metastatic, and cancer-stem cell-like properties.
淋巴管平滑肌瘤病(LAM)是一种破坏性转移的肺部肿瘤,其特征是在专门的肺结节中增殖 LAM 细胞。LAM 细胞的特征是表达促转移和起始癌症的透明质酸受体 CD44v6,并失去 TSC1 和 TSC2 的杂合性(LOH)。循环肿瘤性 LAM 细胞被认为参与转移。由于 LAM 细胞表现出肿瘤性、转移性和干细胞样癌细胞的特性,我们假设升高的醛脱氢酶(ALDH)活性,这是癌症和干细胞的特征,是 LAM 细胞的一个特性。
我们对微切割的 LAM 肺结节中的 ALDH 基因进行了计算机搜索。为了鉴定循环的 LAM 细胞,我们通过渗透压去除全血中的红细胞,以获得外周血单核细胞,然后根据其 ALDH 活性水平通过荧光激活细胞分选进行分选。
微切割的 LAM 肺结节具有独特的 ALDH 基因谱。从循环细胞中分离出的高 ALDH 活性细胞亚群在 14 名 LAM 患者中有 8 名存在 TSC2 LOH。大约 60%的具有高 ALDH 活性的循环细胞表达 CD44v6。来自 LAM 患者和 LAM/TSC 的具有 TSC2 LOH 的细胞在不同的身体部位表现出不同的特性,但所有细胞类型均显示出高 ALDH 活性。
这项新程序允许从培养细胞、血液和乳糜性渗出液中分离出循环的 LAM 细胞,并表明循环的 LAM 细胞具有肿瘤性、转移性和干细胞样特性的异质性。