Phenotypic Characterization of Disseminated Cells with TSC2 Loss of Heterozygosity in Patients with Lymphangioleiomyomatosis
Phenotypic Characterization of Disseminated Cells with TSC2 Loss of Heterozygosity in Patients with Lymphangioleiomyomatosis
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DOI：
10.1164/rccm.201003-0489oc
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发表时间：
2010-12-01
影响因子：
24.7
通讯作者：
Moss, Joel
中科院分区：
文献类型：
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作者：
Cai, Xiong;Pacheco-Rodriguez, Gustavo;Moss, Joel
基本原理：偶发性 (S-LAM) 或结节性硬化症 (TSC) 患者中发生的淋巴管平滑肌瘤病 (LAM)，是由于 TSC 基因 TSC1 或 TSC2 发生突变或杂合性缺失 (LOH) 的 LAM 细胞异常增殖所致。 目的：鉴定可用于从体液中分离 LAM 细胞的分子标记，并确定其发生频率 TSC1 或TSC2 LOH。方法：使用人类TSC2(-/-) 细胞的基因微阵列分析来鉴定候选细胞表面标记。根据与针对这些蛋白质（例如 CD9、CD44v6）的抗体的反应性对来自支气管肺泡灌洗液 (BALF)、尿液、乳糜积液和血液的细胞进行分类，并使用 TSC1 和 TSC2 相关微卫星标记以及 TSC2 基因中的单核苷酸多态性分析 LOH。 测量和主要结果： 来自 BALF、尿液和乳糜的 CD44v6(+)CD9(+) 细胞分别在 80%、69% 和 50% 的患者样本中显示 TSC2 LOH。在超过 90% 的血液样本中检测到带有 TSC2 LOH 的 LAM 细胞。在大多数情况下，来自同一患者不同体液的 LAM 细胞表现出相同的 LOH 模式，即相同微卫星位点的等位基因丢失。在少数患有 S-LAM 的患者中，来自不同体液的 LAM 细胞的 LOH 模式有所不同。没有发现患有 TSC1 LOH 的 S-LAM 患者，这表明 TSC2 异常是绝大多数 S-LAM 病例的原因，并且 TSC1 疾病可能是亚临床的。结论：我们的数据支持大多数 S-LAM 患者的 LAM 细胞具有共同的遗传起源，与转移模型一致。然而，在某些情况下，有证据表明不同位点或同一位点内的 LAM 细胞之间存在遗传异质性。
Rationale: Lymphangioleiomyomatosis (LAM), occurring sporadically (S-LAM) or in patients with tuberous sclerosis complex (TSC), results from abnormal proliferation of LAM cells exhibiting mutations or loss of heterozygosity (LOH) of the TSC genes, TSC1 or TSC2.Objectives: To identify molecular markers useful for isolating LAM cells from body fluids and determine the frequency of TSC1 or TSC2 LOH.Methods: Candidate cell surface markers were identified using gene microarray analysis of human TSC2(-/-) cells. Cells from bronchoalveolar lavage fluid (BALF), urine, chylous effusions, and blood were sorted based on reactivity with antibodies against these proteins (e.g., CD9, CD44v6) and analyzed for LOH using TSC1- and TSC2-related microsatellite markers and single nucleotide polymorphisms in the TSC2 gene.Measurements and Main Results: CD44v6(+)CD9(+) cells from BALF, urine, and chyle showed TSC2 LOH in 80%, 69%, and 50% of patient samples, respectively. LAM cells with TSC2 LOH were detected in more than 90% of blood samples. LAM cells from different body fluids of the same patients showed, in most cases, identical LOH patterns, that is, loss of alleles at the same microsatellite loci. In a few patients with S-LAM, LAM cells from different body fluids differed in LOH patterns. No patients with S-LAM with TSC1 LOH were identified, suggesting that TSC2 abnormalities are responsible for the vast majority of S-LAM cases and that TSC1-disease may be subclinical.Conclusions: Our data support a common genetic origin of LAM cells in most patients with S-LAM, consistent with a metastatic model. In some cases, however, there was evidence for genetic heterogeneity between LAM cells in different sites or within a site.