Progesterone/Progesterone Receptor Axis in Lymphangioleiomyomatosis

淋巴管平滑肌瘤病中的孕酮/孕酮受体轴

• 批准号: 9231019
• 负责人: Lucia Schuger
• 金额: $ 39.69万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-15 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9231019
• 关键词: Adjuvant Therapy; Affect; Alkaline Phosphatase; Antibodies; Behavior; Biological Assay; Biopsy; Blood; Cell Death; Cell Line; Cell Proliferation; Cell model; Cells; Cessation of life; Chemicals; Chylothorax; Clinical Trials; Complement; Cyst; Data; Development; Disease; Dose; Enzyme-Linked Immunosorbent Assay; Epithelioid Cells; Estrogen Antagonists; Estrogen Receptors; Estrogens; Etiology; Extracellular Matrix; FRAP1 gene; Female; Female of child bearing age; Genetic Transcription; Growth; Histones; Hormones; Human; In Vitro; Infection; Lesion; Link; Lung; Lung Lymphangioleiomyomatosis; Lymphangioleiomyomatosis; Malignant neoplasm of lung; Mediating; Mifepristone; Modeling; Modification; Mus; Mutation; Neoplasms; Nodule; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Pilot Projects; Postmenopause; Pregnancy; Production; Progesterone; Progesterone Receptors; Proteolysis; Protocols documentation; Receptor Cell; Reporter; Respiratory Insufficiency; Role; Series; Serum; Sirolimus; TSC1 gene; TSC2 gene; Testing; Transcriptional Activation; Treatment Protocols; Uncertainty; Ursidae Family; base; cancer cell; chromatin immunoprecipitation; gene repression; in vivo; mouse model; neoplastic; prevent; progesterone receptor negative; progesterone receptor positive; promoter; receptor expression; young woman

ABSTRACT Lymphangioleiomyomatosis (LAM) is a rare, low grade neoplasia affecting primarily women of childbearing age. LAM is characterized by the nodular proliferation of SMA and HMB-45 positive LAM cells throughout the lung, producing cystic destruction and respiratory insufficiency. Mutations in the tsc2 or tsc1 genes leading to activation of the mTOR pathway underlie the disease. LAM cells express mainly progesterone receptors (PRs) rather than estrogen receptors; however the role of PRs in the disease has not been investigated. Preliminary studies using a new, well characterized, sporadic-LAM-derived LAM cell line (LAM1) which expresses PRs suggested that the progesterone (P4)/PR axis stimulates LAM cell proliferation, extracellular matrix (ECM) proteinases expression, and pericellular proteolysis, all independently of mTOR activation, whereas mifepristone, a P4 antagonist, abolishes these effects. In a separate pilot study we found that TSC2 transfer suppresses PR expression in LAM1 cells. Additionally, we found that a LAM cell subpopulation expresses PLAP (placental alkaline phosphatase) and preliminary studies suggested a direct correlation between PR positivity and level of secreted PLAP. Finally, intratracheal delivery of LAM1 cells to ovariectomized immunodeficient female mice receiving P4 resulted in LAM-like features undistinguishable from human pulmonary LAM. Based on these observations we hypothesize that: A. the P4/PRs axis contributes to the pathogenesis of LAM by stimulating LAM cell malignant behavior and lung cystic destruction; B. PR expression in LAM results from lack of TSC2-mediated PR transcription repression. C. PLAP levels in serum are a marker for PR positive (PR+) LAM cell burden; and D. concomitant blockage of mTOR and the P4/PR axis is more effective in preventing/ameliorating LAM manifestations in an orthotopic mouse model that blockage of mTOR alone. To test these hypotheses we specifically propose: 1. A To determine the involvement of the P4/PR axis in the promotion of LAM cellular features associated with a low grade neoplastic behavior, including LAM1 cell proliferation, death, production of ECM proteolytic enzymes, etc. 1. B To determine the involvement of the P4/PR axis in the development of pulmonary LAM-like disease in an orthotopic mouse model by quantifying LAM cell burden, cystic destruction, and metastatic dissemination. 1. C To determine the effect of the P4 antagonists mifepristone and lonaprisan on LAM cell low grade neoplastic behavior in vitro and in vivo using the same read outs presented in sub-aims A and B. 2. To determine the effect of TSC2 transfer on pr transcription by LAM1 cells using qPCR, promoter reporter assays and ChIP analysis. 3. To assess whether PLAP level is a marker of PR+ LAM cell burden by determining the potential in vitro and in vivo correlation between PR+ LAM cell burden and PLAP levels in medium and serum. 4. To compare the effectivity of rapamycin alone versus rapamycin plus a P4 antagonist in the treatment of LAM-like disease in an orthotopic mouse model. The experimental protocol will be modeled on that presented in aim 1 B, with same read outs.

抽象的 淋巴血管瘤瘤病（L​​AM）是一种罕见的低级肿瘤，主要影响育儿的妇女 年龄。 LAM的特征是SMA和HMB-45阳性LAM细胞的结节增殖。 肺部，产生囊性破坏和呼吸功能不全。 TSC2或TSC1基因的突变导致 MTOR途径的激活是该疾病的基础。 LAM细胞主要表达孕酮受体（PRS） 而不是雌激素受体；但是，尚未研究PRS在该疾病中的作用。初步的 使用新的，富有特征的，零星衍生的LAM细胞系（LAM1）的研究，该细胞系（LAM1）表达PRS 建议孕酮（P4）/PR轴刺激LAM细胞增殖，细胞外基质（ECM） 蛋白酶表达和细胞周围蛋白水解，全部独立于mTOR激活 P4拮抗剂Mifepristone废除了这些影响。在另一项试点研究中，我们发现TSC2转移 抑制LAM1细胞中的PR表达。此外，我们发现LAM细胞亚群表示 PLAP（胎盘碱性磷酸酶）和初步研究表明PR是直接相关的 阳性和分泌PLAP的水平。最后，气管内递送LAM1细胞向卵巢切除 接收P4的免疫缺陷雌性小鼠产生了类似于人类的LAM样特征 肺部。基于这些观察 我们假设：A。P4/PRS轴有助于 通过刺激LAM细胞恶性行为和肺囊性破坏来使LAM发病； B. PR表达 在LAM中，由于缺乏TSC2介导的PR转录抑制作用。 C.血清中的PLAP水平是标记 用于PR阳性（PR+）LAM细胞负担；和D. MTOR和P4/PR轴的伴随阻塞更多 有效预防/改善LAM表现在原位小鼠模型中，MTOR的阻塞 独自的。 为了测试这些假设，我们特别提出：1。a确定P4/PR轴的参与 在促进与低级肿瘤行为相关的LAM细胞特征中，包括LAM1细胞 增殖，死亡，ECM蛋白水解酶的产生等。1。b确定参与 P4/PR轴通过量化原位小鼠模型中肺Lam样疾病的发展 LAM细胞负担，囊性破坏和转移性传播。 1。C确定P4的效果 在体外和体内使用拮抗剂的米非司证和lonaprisan对LAM细胞低级肿瘤行为 Sub-aims a和B中介绍的相同读取结果。2。确定TSC2转移对PR的影响 使用QPCR，启动子报告测定和芯片分析通过LAM1细胞转录。 3。评估是否 PLAP水平是通过确定体外和体内相关性的电势来标记Pr+ LAM细胞负担的标志 在培养基和血清中的Pr+ LAM细胞负担和PLAP水平之间。 4。比较 雷帕霉素与雷帕霉素以及P4拮抗剂在原位的治疗中的LAM样疾病中 鼠标模型。实验协议将以相同的读取为AIM 1 B中呈现的实验协议。