Genetics of LAM

LAM 遗传学

• 批准号: 10318188
• 负责人: DAVID J. KWIATKOWSKI
• 金额: $ 44.97万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318188
• 关键词: AKT1 gene; AKT2 gene; AKT3 gene; Adult; Affect; Alleles; Allelic Imbalance; Angiofibroma; Angiomyolipoma; Bilateral; Binding; Biological Assay; Biological Markers; Biopsy Specimen; Blood; Cell Line; Cells; ChIP-seq; Chromosome 15; Clinical; Cyst; DNA; Data; Detection; Development; Diagnosis; Enhancers; Epigenetic Process; Event; FRAP1 gene; Face; Family member; Freezing; Frequencies; Gene Family; Gene Frequency; Genes; Genetic; Genetic Screening; Genetic Transcription; Growth; Human; Individual; Lesion; Lung; Lung Lymphangioleiomyomatosis; Lymphangioleiomyomatosis; Massive Parallel Sequencing; Mosaicism; Mutation; Mutation Analysis; Mutation Detection; Neoplasms; Pathogenesis; Pathologic; Pathway interactions; Patients; Plasma Cells; Population; Prognosis; Renal Angiomyolipoma; Reporting; Research Personnel; Respiratory Failure; Role; Sampling; Series; Skin Manifestations; Somatic Mutation; Specimen; Subgroup; Suggestion; TFE3 gene; TSC1 gene; TSC1/2 gene; TSC2 gene; Testing; Time; Tretinoin; United States National Institutes of Health; analog; apoAI regulatory protein-1; cell free DNA; cell type; clinical center; clinical phenotype; diagnostic criteria; genome wide association study; genome-wide; high standard; inhibitor; lung lesion; new technology; new therapeutic target; novel; novel marker; overexpression; transcription factor; transcriptome sequencing; treatment response; variant detection

Abstract Lymphangioleiomyomatosis is a low grade neoplasm that causes progressive lung destruction, lung cyst formation, and respiratory failure. Bi-allelic mutations in TSC2 (or much less commonly TSC1) have been known as the main genetic driver of LAM in both individuals with TSC as well as sporadic LAM. It has also been thought that the distinction between TSC-LAM and sporadic LAM was well-defined. However, recent studies by PI Kwiatkowski and co-investigator Darling have shown that mosaicism for TSC1/TSC2 is common in adults with TSC-LAM, and associated with a milder clinical phenotype that may be missed in some apparent sporadic LAM patients. In addition, detailed analyses of LAM lung lesions have been able to identify TSC1 or TSC2 mutations in only a fraction of sporadic LAM patients, suggesting the involvement of other genes. Furthermore, a LAM GWAS led by the PI has identified SNPs on chromosome 15 near the transcription factor NR2F2 as having alleles that show association with sporadic LAM. In this proposal, we examine all three of these issues in greater detail. Two of the Aims will use massively parallel sequencing (MPS) and a novel technology we have developed, Multiplex High-sensitivity PCR Assay (MHPA), that is capable of highly sensitive variant detection in TSC2, down to an allele frequency of 0.05%, 10-fold lower than our previous targeted capture assay. In Aim 1, we will determine whether mutations in other mTOR pathway genes and/or MITF family member translocation or amplification cause sporadic LAM in a set of 100 LAM patients. In Aim 2, we will determine whether the presence of TSC2 mutations in cell free (cf) DNA is a biomarker of LAM; and examine the frequency of genetic mosaicism in selected subsets of apparent sporadic LAM patients; simultaneously, also in 100 LAM patients. We will enrich the patients studied for those with singleton TSC lesions, such as hypomelanotic macule (HMM) or facial angiofibroma, or bilateral angiomyolipoma. We will use our new MHPA assay for this analysis. In Aim 3, we will examine the role of NR2F2 in LAM development, by examining allelic imbalance in the H3K27ac ChIP-Seq data, performing NR2F2 ChIP-Seq, using Binding and expression target analysis to infer the genes most likely to have their expression driven by NR2F2, determine if NR2F2 is part of the Core transcription Regulatory Circuitry (CRC) in angiomyolipoma, and assess effects of NR2F2 expression modulation, and treatment with activators and inhibitors in the human angiomyolipoma cell line 621-101.

抽象的 淋巴血管瘤瘤病是一种低级肿瘤，会导致进行性肺部破坏，肺囊肿 形成和呼吸衰竭。 TSC2中的双性突变（或较少常见的TSC1）已是 在TSC和零星的LAM中，被称为LAM的主要遗传驱动器。它也有 人们认为TSC-LAM和零星LAM之间的区别是明确的。但是，最近 Pi Kwiatkowski和共同研究器的研究表明，TSC1/TSC2的镶嵌性很常见 在患有TSC-LAM的成年人中，并且与温和的临床表型相关 零星的LAM患者。此外，对Lam肺部病变的详细分析已经能够识别TSC1或 只有一小部分零星LAM患者的TSC2突变，表明其他基因的参与。 此外，由PI领导的LAM GWAS已在转录因子附近鉴定在15号染色体上的SNP NR2F2的等位基因显示与零星的LAM相关。在此提案中，我们检查了所有三个 这些问题更详细。其中两个目标将使用大规模平行测序（MP）和一个新颖 我们开发的技术，多重高敏性PCR分析（MHPA），能够高度 TSC2中的敏感变体检测到0.05％的等位基因频率，比以前低10倍 有针对性的捕获测定法。在AIM 1中，我们将确定其他MTOR途径基因中的突变和/或 MITF家庭成员的易位或扩增会导致一组100名LAM患者偶发的LAM。在AIM 2中， 我们将确定无细胞（CF）DNA中TSC2突变是否是LAM的生物标志物。和 检查明显零星LAM患者选定子集中遗传镶嵌的频率； 同时，也有100名LAM患者。我们将丰富针对Singleton TSC研究的患者 病变，例如低黄内毛毛细血管（HMM）或面部血管纤维瘤或双侧血管肌莫洛膜瘤。我们将使用 我们的新MHPA测定法进行了此分析。在AIM 3中，我们将研究NR2F2在LAM开发中的作用， 使用绑定和 表达靶向分析以推断最有可能由NR2F2驱动其表达的基因，确定是否是否 NR2F2是血管肌瘤中核心转录调节电路（CRC）的一部分，并评估了影响的影响 NR2F2表达调制，并在人血管肌瘤细胞中用激活剂和抑制剂进行处理 第621-101号线。