Uterine signaling networks in the pathogenesis of pulmonary lymphangioleiomyomatosis (LAM)

肺淋巴管平滑肌瘤病 (LAM) 发病机制中的子宫信号网络

• 批准号: 10447086
• 负责人: YAN XU
• 金额: $ 76.13万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-21 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10447086
• 关键词: Affect; Age; Apoptosis; Attenuated; B-Cell Acute Lymphoblastic Leukemia; Binding; Breast Cancer Cell; Candidate Disease Gene; Cell Death; Cell physiology; Cells; Chromatin; Data; Development; Diagnostic tests; Disease; Disease remission; Ectopic Expression; Estrogens; FRAP1 gene; Female; Gene Expression Alteration; Genes; Genomics; Homeobox; Hormones; Immunofluorescence Microscopy; Immunohistochemistry; In Vitro; Interdisciplinary Study; Knock-out; Lead; Lesion; Life; Lung; Lung Lymphangioleiomyomatosis; Lung Neoplasms; Lung diseases; Lymphangioleiomyomatosis; Malignant Neoplasms; Mediating; Metastatic Neoplasm to the Lung; Modeling; Molecular; Morphology; Mus; Neoplasm Metastasis; Neoplasms; Pathogenesis; Patients; Population; Pre-Clinical Model; Prevalence; Progesterone; Rattus; Research; Resolution; Respiratory Failure; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Pathway; Signal Transduction; Sirolimus; Steroids; TSC2 gene; Testing; Therapeutic; Time; Tissues; Toxic effect; Transactivation; Transfection; Tuberous Sclerosis; Uterine Fibroids; Uterus; Woman; Work; antagonist; clinical diagnostics; cofactor; comparative; curative treatments; design; diagnosis design; diagnostic biomarker; diagnostic tool; epigenomics; experience; follow-up; gene network; gene regulatory network; genetic signature; in vivo; insight; loss of function mutation; lung lesion; migration; mouse model; new therapeutic target; non-invasive optical imaging; novel; novel diagnostics; novel strategies; novel therapeutics; pre-clinical; preservation; public health relevance; pulmonary function; reproductive; response; single-cell RNA sequencing; small hairpin RNA; symptomatic improvement; synthetic peptide; transcription factor; tumorigenesis

PROJECT SUMMARY Lymphangioleiomyomatosis (LAM), a lung neoplasm affecting reproductive age women, is caused by loss-of- function mutations in tuberous sclerosis 1 (TSC1) or TSC2 genes. Hyperactivation of the mechanistic target of rapamycin 1 leads to cystic lung remodeling and progressive respiratory failure. Sirolimus therapy stabilizes lung function and improves symptoms in LAM patients, but the long-term benefit and toxicity are unknown, and some patients do not respond to therapy. LAM cells are metastatic but the primary tissue of origin is unknown. Unknown mechanisms underlying lung-specific metastasis, primary pathogenesis and female prevalence of LAM are obstacles to the development of new curative therapies. Our preliminary data from LAM lung single cell RNA sequencing (scRNAseq) analysis identified a unique population of cells (LAMCORE) expressing uterine- specific homeobox (HOX) transcription factors (TFs) that are not detected in normal lung; TF profiling of LAM patient-derived cells identified activation of Pre-B-cell leukemia homeobox 1 (PBX1), a cofactor of HOXs; comparative analysis of LAM lung scRNAseq and breast cancer cell PBX1-ChIPseq revealed overly represented LAMCORE signature genes with positive PBX1 binding peaks. Collectively, this strongly suggests a uterine origin of LAM cells and a central role of HOX/PBX1 signaling in LAM pathogenesis. The objective of this proposal is to identify mechanisms by which HOXs-associated gene networks regulate LAM pathogenesis and progression. Our central hypothesis is that HOX/PBX1 orchestrates a cell-specific gene network downstream of female hormones that regulates the survival and lung metastasis of LAMCORE cells. Three specific aims are proposed to delineate the molecular mechanisms through which HOX/PBX1 gene network contributes to LAM lesion metastasis, formation and progression. Aim 1: Determine pulmonary LAM-specific genomic and epigenomic responses and mechanisms underlying HOX/PBX1-mediated LAM pathogenesis. Aim 2: Determine the functional impacts of HOX/PBX1 network genes on metastatic potentials of LAM-derived cells in vitro and in vivo. Aim 3: Determine the effect of the HOX/PBX1 antagonist, HXR9, singly or in combination with Sirolimus, on estrogen-promoted lung metastasis potentials and lung remodeling in vivo. The completion of this proposal will provide for the first time: 1) a high-resolution integrative genomic/epigenomic blueprint of the LAMCORE cells, 2) a LAMCORE cell-specific HOX/PBX1 gene network with predicted key regulatory factors and targets within the network for follow-up perturbation and clinical diagnostic tests, 3) novel mechanistic insights into HOX/PBX1 regulatory circuits and their functional impact on LAM pathogenesis, and 4) preclinical proof-of-principle evidence for targeting HOX/PBX1-regulated female hormone-mediated LAM progression as a novel remission-inducing therapeutic strategy for LAM patients. Our work has major biomedical relevance for understanding LAM pathogenesis, developing new approaches for LAM diagnosis and designing alternative remission-inducing therapeutic strategies to maximize the benefit/toxicity ratio LAM treatment.

项目摘要 淋巴血管肌瘤病（LAM）是一种影响生殖年龄女性的肺肿瘤，是由丧失 - 结节硬化症1（TS​​C1）或TSC2基因中的功能突变。机械目标的过度激活 雷帕霉素1导致囊性肺重塑和进行性呼吸衰竭。西洛里木斯疗法稳定肺 功能并改善LAM患者的症状，但长期益处和毒性尚不清楚，有些 患者对治疗没有反应。 LAM细胞是转移性的，但原始组织的主要组织尚不清楚。 肺特异性转移，主要发病机理和LAM女性患病率的未知机制 是发展新治疗疗法的障碍。我们的初步数据来自Lam肺单细胞 RNA测序（SCRNASEQ）分析确定了表达子宫 - 在正常肺中未检测到的特定同源词（HOX）转录因子（TFS）；林的TF分析 患者衍生的细胞鉴定出HOX的辅助因子的B-细胞白血病同型1（PBX1）的激活； LAM肺Scrnaseq和乳腺癌细胞PBX1-CHIPSEQ的比较分析表明过于代表 具有阳性PBX1结合峰的板岩签名基因。总体而言，这强烈暗示了子宫的起源 LAM细胞和HOX/PBX1信号传导在LAM发病机理中的核心作用。该提议的目的是 确定HOX相关基因网络调节LAM发病机理和进展的机制。 我们的核心假设是HOX/PBX1在女性下游精心策划了细胞特异性基因网络 调节叶核细胞存活和肺转移的激素。提出了三个具体目标 描述HOX/PBX1基因网络有助于LAM病变的分子机制 转移，形成和进展。 AIM 1：确定肺LAM特异性基因组和表观基因组学 HOX/PBX1介导的LAM发病机理的反应和机制。目标2：确定 HOX/PBX1网络基因的功能影响对LAM衍生细胞在体外和体内的转移电位。 目标3：确定HOX/PBX1拮抗剂HXR9，单独或与Sirolimus的效果，对 雌激素促进的肺转移电位和体内肺重塑。该提案的完成 将首次提供：1）LAMCORE的高分辨率整合基因组/表观基因组蓝图 细胞，2）具有预测的关键调节因子和靶标的lamcore细胞特异性HOX/PBX1基因网络 进行后续扰动和临床诊断测试的网络，3）对HOX/PBX1的新机械见解 调节回路及其对LAM发病机理的功能影响，4）临床前原则证据证据 用于靶向HOX/PBX1调节的雌激素介导的LAM进展，作为一种新型的缓解诱导 LAM患者的治疗策略。我们的工作具有理解LAM的主要生物医学相关性 发病机理，开发用于LAM诊断和设计替代缓解诱导的新方法 最大化益处/毒性比LAM治疗的治疗策略。