Role of the Lysosome in the Pathogenesis and Therapy of LAM

溶酶体在 LAM 发病机制和治疗中的作用

基本信息

批准号：
10633178
负责人：
Elizabeth P Henske
金额：
$ 43.95万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-15 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10633178
关键词：
Address Adult Affect Alleles Biogenesis Biological Assay Calcium Channel Cause of Death Cell Nucleus Cells Cessation of life Clinical Data Dependence Diffuse Enzymes Estrogen Receptor alpha Estrogens Exocytosis Extracellular Matrix Extracellular Space FDA approved FRAP1 gene Female Gene Expression Gene Mutation Genetic Transcription Human Immunoprecipitation In Vitro Intravenous Invaded Knowledge Lead Lung Lung diseases Lymphangioleiomyomatosis Lysosomes Mediating Membrane Metabolism NPC1 gene Nodule Organelles Oxygen Pathogenesis Pathologic Pathway interactions Patients Prevention Proliferating Proteins Role Signal Transduction Smooth Muscle Structure of parenchyma of lung TSC1 gene TSC1/2 gene TSC2 gene Testing Therapeutic Translations Tuberous Sclerosis Woman alveolar destruction cathepsin K cell growth cell type collagenase high throughput screening in vivo inhibitor lung colonization lysosomal proteins mouse model pre-clinical prevent protein complex protein expression small molecule therapeutic target transcription factor

项目摘要

Abstract Lymphangioleiomyomatosis (LAM) is a progressive, destructive lung disease of women that can lead to oxygen dependency and death. LAM cells contain bi-allelic inactivating TSC2 gene mutations. The reasons for the female predominance of LAM and the mechanisms underlying cystic lung destruction are not well understood, representing key knowledge gaps that will be addressed in this proposal. The TSC1/TSC2 protein complex inhibits mTORC1. Multiple components of the TSC signaling network can localize to the lysosomal membrane, including mTOR, Rheb, TSC1, and TSC2. Lysosomes are highly dynamic organelles with both degradation and signaling functions, thus participating in many cellular pathways. TFEB is a “master regulator” of lysosomal biogenesis and lysosomal exocytosis. We have found that TFEB is markedly elevated in the nucleus of TSC2-deficient cells and in human LAM cells. Lysosomal number is also increased in TSC2-deficient cells, and LAM cells have a striking increase in lysosomal proteins including NPC1. Lysosomal enzymes are released from the lysosomes through lysosomal exocytosis, which degrade extracellular matrix, and could be a cause of lung destruction in LAM. We have also discovered that estrogen strongly increases lysosomal gene expression in LAM patient-derived cells. Our central hypothesis is that elevated TFEB in LAM cells leads to increased lysosomal content and the release of lysosomal enzymes into the extracellular space, leading to lung destruction. We further hypothesize that these effects are enhanced by estrogen. A key translational corollary is that TFEB and/or lysosomal proteins are potential therapeutic targets for LAM. Our hypotheses will be tested in four Aims: Aim 1. To determine how TFEB impacts lysosomal exocytosis and the invasive potential of TSC2-deficient cells. Aim 2. To determine the mechanism through which estrogen affects lysosomal content and lysosomal exocytosis in TSC and LAM. Aim 3. To identify small molecules that inhibit the activity of TFEB in TSC and LAM. Aim 4. To determine how inhibition of TFEB and/or inhibition of lysosomal exocytosis impact lung destruction in a mouse model of LAM. We expect this project to have scientific and preclinical impact by elucidating the mechanisms through which LAM cells induce lung destruction and the reasons for the striking female predominance of LAM.

抽象的淋巴管平滑肌瘤病 (LAM) 是一种进行性、破坏性的女性肺部疾病，可导致缺氧 LAM 细胞含有双等位基因失活 TSC2 基因突变。 LAM 的女性优势以及囊性肺破坏的机制尚不清楚，代表本提案将解决的关键知识差距。 TSC1/TSC2 蛋白复合物可抑制 TSC 信号网络的多个成分。定位于溶酶体膜，包括 mTOR、Rheb、TSC1 和 TSC2 具有高度动态性。具有降解和信号传导功能的细胞器，从而参与许多细胞途径。 TFEB 是溶酶体生物合成和溶酶体胞吐作用的“主调节器”。 TSC2 缺陷细胞的细胞核和人类 LAM 细胞中的数量也显着升高。 TSC2 缺陷细胞中的溶酶体蛋白显着增加，而 LAM 细胞中包括 NPC1 在内的溶酶体蛋白显着增加。溶酶体酶通过溶酶体胞吐作用从溶酶体释放，降解细胞外物质基质，并且可能是 LAM 中肺部破坏的原因。我们还发现雌激素强烈。增加 LAM 患者来源细胞中溶酶体基因的表达。我们的中心假设是 LAM 细胞中 TFEB 升高导致溶酶体含量增加和释放溶酶体酶进入细胞外空间，导致肺部破坏。雌激素增强作用的一个关键翻译推论是 TFEB 和/或溶酶体蛋白是 LAM 的潜在治疗靶点将在四个目标中得到检验：目标 1. 确定 TFEB 如何影响溶酶体胞吐作用和 TSC2 缺陷细胞的侵袭潜力。目标 2. 确定雌激素影响溶酶体含量和溶酶体胞吐作用的机制在 TSC 和 LAM 中。目标 3. 鉴定抑制 TSC 和 LAM 中 TFEB 活性的小分子。目标 4. 确定抑制 TFEB 和/或抑制溶酶体胞吐作用如何影响肺损伤 LAM 小鼠模型。我们期望该项目通过阐明其机制来产生科学和临床前影响 LAM 细胞引起肺损伤以及 LAM 女性占主导地位的原因。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

TFEB drives mTORC1 hyperactivation and kidney disease in Tuberous Sclerosis Complex.

TFEB 会导致结节性硬化症中 mTORC1 过度激活和肾脏疾病。

DOI：
发表时间：
2024-01-09
期刊：
影响因子：
16.6
作者：
Alesi, Nicola;Khabibullin, Damir;Rosenthal, Dean M;Akl, Elie W;Cory, Pieter M;Alchoueiry, Michel;Salem, Samer;Daou, Melissa;Gibbons, William F;Chen, Jennifer A;Zhang, Long;Filippakis, Harilaos;Graciotti, Laura;Miceli, Caterina;Monfregola, Jl
通讯作者：
Monfregola, Jl