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.

项目概要 淋巴管平滑肌瘤病 (LAM) 是一种影响育龄妇女的肺部肿瘤，是由丧失 结节性硬化症 1 (TSC1) 或 TSC2 基因的功能突变。机械目标的过度激活 雷帕霉素 1 会导致囊性肺重塑和进行性呼吸衰竭。西罗莫司治疗可稳定肺部 功能并改善 LAM 患者的症状，但长期获益和毒性尚不清楚，一些 患者对治疗没有反应。 LAM 细胞具有转移性，但原发组织来源未知。 LAM 肺部特异性转移、主要发病机制和女性患病率的未知机制 是开发新疗法的障碍。我们来自 LAM 肺单细胞的初步数据 RNA 测序 (scRNAseq) 分析确定了表达子宫的独特细胞群 (LAMCORE) 在正常肺中未检测到的特定同源框 (HOX) 转录因子 (TF)； LAM 的 TF 分析 患者来源的细胞鉴定出前 B 细胞白血病同源盒 1 (PBX1)（HOX 的辅助因子）的激活； LAM 肺 scRNAseq 和乳腺癌细胞 PBX1-ChIPseq 的比较分析显示过度代表性 LAMCORE 特征基因具有阳性 PBX1 结合峰。总的来说，这强烈表明子宫起源 LAM 细胞的作用以及 HOX/PBX1 信号在 LAM 发病机制中的核心作用。该提案的目标是 确定 HOX 相关基因网络调节 LAM 发病机制和进展的机制。 我们的中心假设是 HOX/PBX1 协调女性下游的细胞特异性基因网络 调节 LAMCORE 细胞存活和肺转移的激素。提出了三个具体目标 描述 HOX/PBX1 基因网络导致 LAM 损伤的分子机制 转移、形成和进展。目标 1：确定肺部 LAM 特异性基因组和表观基因组 HOX/PBX1 介导的 LAM 发病机制的反应和机制。目标 2：确定 HOX/PBX1网络基因对LAM衍生细胞体外和体内转移潜能的功能影响。 目标 3：确定 HOX/PBX1 拮抗剂 HXR9 单独或与西罗莫司联合使用对 雌激素促进肺转移潜力和体内肺重塑。本提案的完成情况 将首次提供：1) LAMCORE 的高分辨率综合基因组/表观基因组蓝图 细胞，2) LAMCORE 细胞特异性 HOX/PBX1 基因网络，其中包含预测的关键调控因子和靶标 用于后续扰动和临床诊断测试的网络，3）对 HOX/PBX1 的新颖机制见解 调节回路及其对 LAM 发病机制的功能影响，以及 4) 临床前原理验证证据 用于靶向 HOX/PBX1 调节的女性激素介导的 LAM 进展作为一种新的缓解诱导药物 LAM 患者的治疗策略。我们的工作对于理解 LAM 具有重要的生物医学意义 发病机制，开发 LAM 诊断的新方法并设计替代的缓解诱导方法 最大化 LAM 治疗效益/毒性比的治疗策略。