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KLF5 Shapes Developing Respiratory Tubules by Inhibiting Actin Asymmetry in Epithelial Cells. KLF5 通过抑制上皮细胞中肌动蛋白的不对称性来塑造发育中的呼吸小管
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-18 DOI: 10.1165/rcmb.2024-0140OC
Qing Li, Yong Liao, Junwei Zeng, Silu Hu, Chunjie Li, Jeffrey A Whitsett, Yi Zheng, Fengming Luo, Chang Xu, Taozhen He, Xinhua Lin, Huajing Wan

Tubulogenesis depends on precise cell shape changes driven by asymmetric tension from the actin cytoskeleton. How actin asymmetry is dynamically controlled to coordinate epithelial cell shape changes required for respiratory tubulogenesis remains unknown. Herein, we unveiled a critical role for the transcription factor KLF5, regulating actin asymmetry, inducing epithelial cell shape changes by balancing RHOA and CDC42 GTPase activity via RICH2. Conditional Klf5 expression or deletion in pulmonary epithelial cells affected apical actin organization and the positioning of apical polarity proteins in cell membranes, disrupting branching and sacculation of respiratory tubules during mouse lung morphogenesis. Increased KLF5 levels were observed in epithelial cells lining dilated tubules in lungs from patients with congenital pulmonary airway malformation (CPAM). Together, our study demonstrates that dynamic regulation of apical actin organization by KLF5 is essential for respiratory tubulogenesis, providing a mechanistic framework for comprehending the morphogenesis of respiratory tubules.

肾小管的生成依赖于由肌动蛋白细胞骨架的不对称张力驱动的精确的细胞形状变化。肌动蛋白的不对称性是如何被动态控制以协调呼吸管生成所需的上皮细胞形状变化的,目前仍是未知数。在这里,我们揭示了转录因子 KLF5 在调节肌动蛋白不对称性方面的关键作用,它通过 RICH2 平衡 RHOA 和 CDC42 GTPase 的活性,诱导上皮细胞形状的改变。肺上皮细胞中条件性 Klf5 表达或缺失会影响顶端肌动蛋白的组织和顶端极性蛋白在细胞膜中的定位,从而在小鼠肺形态发生过程中破坏呼吸小管的分支和囊状结构。在先天性肺气道畸形(CPAM)患者肺部扩张的肺小管内衬上皮细胞中观察到 KLF5 水平升高。总之,我们的研究表明,KLF5对顶端肌动蛋白组织的动态调控对呼吸小管的发生至关重要,为理解呼吸小管的形态发生提供了一个机理框架。
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引用次数: 0
Spatiotemporal Clusters of ERK Activity Coordinate Cytokine-induced Inflammatory Responses in Human Airway Epithelial Cells. ERK活动的时空集群协调细胞因子诱导的人类气道上皮细胞炎症反应
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-18 DOI: 10.1165/rcmb.2024-0256OC
Nicholaus L DeCuzzi, Daniel Oberbauer, Kenneth J Chmiel, Michael Pargett, Justa M Ferguson, Devan Murphy, Marion Hardy, Abhineet Ram, Amir A Zeki, John G Albeck

Spatially coordinated ERK signaling events ("SPREADs") transmit radially from a central point to adjacent cells via secreted ligands for EGFR and other receptors. SPREADs maintain homeostasis in non-pulmonary epithelia, but it is unknown whether they play a role in the airway epithelium or are dysregulated in inflammatory disease. To address these questions, we measured SPREAD activity with live-cell ERK biosensors in human bronchial epithelial cell lines (HBE1 and 16HBE) and primary human bronchial epithelial (pHBE) cells, in both submerged and biphasic Air-Liquid Interface (ALI) culture conditions (i.e., differentiated cells). Airway epithelial cells were exposed to pro-inflammatory cytokines relevant to asthma and chronic obstructive pulmonary disease (COPD). Type 1 pro-inflammatory cytokines significantly increased the frequency of SPREADs, which coincided with epithelial barrier breakdown in differentiated pHBE cells. Furthermore, SPREADs correlated with IL-6 peptide secretion and the appearance of localized clusters of phospho-STAT3 immunofluorescence. To probe the mechanism of SPREADs, cells were co-treated with pharmacological treatments (gefitinib, tocilizumab, hydrocortisone) or metabolic modulators (insulin, 2-deoxyglucose). Hydrocortisone, inhibitors of receptor signaling, and suppression of metabolic function decreased SPREAD occurrence, implying that pro-inflammatory cytokines and glucose metabolism modulate SPREADs in human airway epithelial cells via secreted EGFR and IL6R ligands. We conclude that spatiotemporal ERK signaling plays a role in barrier homeostasis and dysfunction during inflammation of the airway epithelium. This novel signaling mechanism could be exploited clinically to supplement corticosteroid treatment for asthma and COPD. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/).

空间协调的 ERK 信号事件("SPREADs")通过表皮生长因子受体和其他受体的分泌配体从中心点向邻近细胞进行辐射传输。SPREADs 可维持非肺部上皮细胞的平衡,但它们是否在气道上皮细胞中发挥作用或在炎症性疾病中失调尚不清楚。为了解决这些问题,我们使用活细胞ERK生物传感器测量了人支气管上皮细胞系(HBE1和16HBE)和原代人支气管上皮细胞(pHBE)在浸没和双相气液界面(ALI)培养条件下(即分化细胞)的SPREAD活性。气道上皮细胞暴露于与哮喘和慢性阻塞性肺病(COPD)相关的促炎细胞因子。1 型促炎细胞因子显著增加了 SPREADs 的频率,这与分化 pHBE 细胞的上皮屏障破坏相吻合。此外,SPREADs与IL-6肽分泌和局部磷-STAT3免疫荧光群的出现相关。为了探究SPREADs的机制,细胞与药物治疗(吉非替尼、替西珠单抗、氢化可的松)或代谢调节剂(胰岛素、2-脱氧葡萄糖)共同处理。氢化可的松、受体信号转导抑制剂和代谢功能抑制剂可减少 SPREAD 的发生,这意味着促炎细胞因子和葡萄糖代谢可通过分泌的表皮生长因子受体和 IL6R 配体调节人气道上皮细胞中的 SPREAD。我们的结论是,时空 ERK 信号在气道上皮细胞炎症期间的屏障稳态和功能障碍中发挥作用。临床上可以利用这种新型信号机制来辅助皮质类固醇治疗哮喘和慢性阻塞性肺病。
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引用次数: 0
Increased Circulating Extracellular Superoxide Dismutase Attenuates Platelet-Neutrophil Interactions. 增加循环中的细胞外超氧化物歧化酶可减轻血小板与中性粒细胞的相互作用
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-12 DOI: 10.1165/rcmb.2024-0292OC
Christina Sul, Caitlin V Lewis, Janelle Posey, Mariah Jordan, Daniel Colon Hidalgo, Timothy Porfilio, Hanan Elajaili, Genevieve McCormack, Samuel Burciaga, Cassidy Delaney, Eva S Nozik

Acute respiratory distress syndrome (ARDS) is a serious illness accounting for 10% of ICU admissions and high mortality of 31-45% with a paucity of pharmacologic treatment options. Dysregulated inflammation and oxidative stress are hallmark features of ARDS. We previously showed that transgenic mice expressing a naturally occurring polymorphism of the antioxidant enzyme extracellular superoxide dismutase (EC-SOD), are protected against Staphylococcus aureus (S. aureus) pneumonia, acute lung injury, and pulmonary neutrophilia. In this mouse strain, an R213G amino acid substitution leads to lower tissue binding affinity and elevated alveolar and plasma EC-SOD levels, though the redox-regulated mechanisms responsible for protection against S. aureus are not yet elucidated. Neutrophils are recruited to the areas of injury and inflammation, in part by activated platelets, which contain multiple redox-sensitive targets. Thus, we hypothesize that increased circulating EC-SOD due to the EC-SOD R213G variant protects against S. aureus pneumonia by reducing platelet activation and subsequent neutrophil recruitment to the lung. We demonstrate that, compared to WT mice with S. aureus pneumonia, platelet activation, formation of platelet-neutrophil aggregates (PNAs), and influx of neutrophils and PNAs into the lung are decreased in the infected R213G mice. Furthermore, pre-treatment with a MnTE-2-PyP SOD mimetic protects against S. aureus-induced platelet activation, pulmonary neutrophilia, and acute lung injury. Our data highlight the redox regulation of platelet activation as a driver of S. aureus-induced acute lung injury.

急性呼吸窘迫综合征(ARDS)是一种严重疾病,占重症监护病房入院人数的 10%,死亡率高达 31-45%,但药物治疗方法却很少。炎症失调和氧化应激是 ARDS 的标志性特征。我们之前研究发现,表达抗氧化酶细胞外超氧化物歧化酶(EC-SOD)自然发生多态性的转基因小鼠对金黄色葡萄球菌肺炎、急性肺损伤和肺中性粒细胞增多有保护作用。在这种小鼠品系中,R213G 氨基酸置换导致组织结合亲和力降低,肺泡和血浆中的 EC-SOD 水平升高,但氧化还原调控机制对金黄色葡萄球菌的保护作用尚未阐明。中性粒细胞被招募到损伤和炎症区域,部分原因是血小板被激活,而血小板含有多种氧化还原敏感靶点。因此,我们假设,EC-SOD R213G 变体导致的循环中 EC-SOD 增加可通过减少血小板活化及随后中性粒细胞被招募到肺部来预防金黄色葡萄球菌肺炎。我们证明,与患有金黄色葡萄球菌肺炎的 WT 小鼠相比,受感染的 R213G 小鼠的血小板活化、血小板-中性粒细胞聚集体(PNAs)的形成以及中性粒细胞和 PNAs 涌入肺部的情况均有所减少。此外,预处理 MnTE-2-PyP SOD 模拟物可防止金黄色葡萄球菌诱导的血小板活化、肺中性粒细胞增多和急性肺损伤。我们的数据强调了血小板活化的氧化还原调节是金黄色葡萄球菌诱导的急性肺损伤的驱动因素。
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引用次数: 0
The Bi-Steric Inhibitor RMC-5552 Reduces mTORC1 Signaling and Growth in Lymphangioleiomyomatosis. 双酯抑制剂 RMC-5552 可降低 mTORC1 信号转导并促进淋巴管瘤病的生长
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-12 DOI: 10.1165/rcmb.2024-0242OC
Jilly F Evans, Owen A Ledwell, Yan Tang, Ryan Rue, Alexander R Mukhitov, Rémi Diesler, Susan M Lin, Swaroop V Kanth, Maria C Basil, Edward Cantu, Elizabeth P Henske, Vera P Krymskaya

Mutations in the Tuberous Sclerosis Complex (TSC) genes result in the hyperactivation of the mechanistic/mammalian target of rapamycin 1 (mTORC1) growth pathway in mesenchymal pulmonary cells. Rapamycin (SirolimusTM), a naturally occurring macrolide, is the only therapeutic approved for women with lymphangioleiomyomatosis (LAM), a progressive, destructive lung disease caused by TSC gene mutations and mTORC1 hyperactivation. However, on cessation of the drug, lung function decline continues. We demonstrated here that pulmonary LAM cancer stem-like cells (SLS) most highly expressed the eukaryotic translation initiation factor 4E (eIF4E)-dependent translation initiation genes. We also showed that the eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) gene has the lowest expression in these cells, indicating that the 4E-BP1/eIF4E ratio in LAM SLS cells favors unrestrained eIF4E oncogenic mRNA translation. The bi-steric mTORC1-selective compound RMC-5552 prevented growth of LAM-associated fibroblasts (LAFs) and phosphorylation of proteins in the ribosomal protein S6K1/ribosomal protein S6 (S6K1/S6) and 4E-BP1/eIF4E translation mTORC1-driven pathways, whereas rapamycin only blocked the S6K/S6 axis. Rapamycin inhibition of LAF growth was rapidly reversed, but RMC-5552 inhibition was more durable. RMC-5552, through its potential to eradicate LAM cancer SLS cells, may have therapeutic benefit in LAM and other diseases with mTORC1 hyperactivity.

Tuberous Sclerosis Complex(TSC)基因突变会导致间质肺细胞中雷帕霉素 1(mTORC1)生长途径的机械/哺乳动物靶点过度激活。雷帕霉素(SirolimusTM)是一种天然大环内酯类药物,是唯一获准用于治疗淋巴管瘤(LAM)女性患者的药物,LAM 是一种由 TSC 基因突变和 mTORC1 过度激活引起的进行性破坏性肺部疾病。然而,停药后,肺功能会继续下降。我们在此证明,肺LAM癌干细胞(SLS)最高度表达真核翻译起始因子4E(eIF4E)依赖的翻译起始基因。我们还发现,真核起始因子 4E 结合蛋白 1(4E-BP1)基因在这些细胞中的表达量最低,这表明 LAM SLS 细胞中 4E-BP1/eIF4E 的比例有利于不受限制的 eIF4E 致癌 mRNA 翻译。双甾体 mTORC1 选择性化合物 RMC-5552 阻止了 LAM 相关成纤维细胞(LAFs)的生长以及核糖体蛋白 S6K1/核糖体蛋白 S6(S6K1/S6)和 4E-BP1/eIF4E 翻译 mTORC1 驱动通路中的蛋白磷酸化,而雷帕霉素只阻断了 S6K/S6 轴。雷帕霉素对 LAF 生长的抑制作用会迅速逆转,但 RMC-5552 的抑制作用更为持久。RMC-5552 具有根除 LAM 癌 SLS 细胞的潜力,可能对 LAM 和其他 mTORC1 活性亢进的疾病有治疗作用。
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引用次数: 0
Can ENaC "TIP" the Scales to Reduce Endothelial ROS and Vascular Leak During Pneumococcal Lung Injury? 在肺炎球菌肺损伤期间,ENaC 能否 "撬动 "天平以减少内皮 ROS 和血管渗漏?
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-08 DOI: 10.1165/rcmb.2024-0486ED
Alison W Ha, Eleftheria Letsiou, Steven M Dudek
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引用次数: 0
Circulating mtNFPs Are Associated with ARDS after CPB and Regulate Endothelial Barrier through FPR2. 循环中的 mtNFPs 与 CPB 后的 ARDS 有关,并通过 FPR2 调节内皮屏障。
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-08 DOI: 10.1165/rcmb.2024-0076OC
Peng Lu, Xiaopei Li, Jinqiang Wang, Xiangyu Li, Zihao Shen, Yuanpu Qi, Mingyu Chu, Xin Yao, Xiao Zhang, Yu Zheng, Faliang Zhan, Meijuan Song, Xiaowei Wang

Cardiopulmonary bypass (CPB) increases the risk of acute respiratory distress syndrome (ARDS) due to endothelial cell (EC) barrier dysfunction. However, the specific role of mitochondrial N-formyl peptides (mtNFPs) in ARDS following CPB remains unexplored. Here, we investigated the differential expression of circulating mtNFPs in patients after CPB, focusing on the novel role of FPR2 in ECs. Levels of circulating mtNFPs were assessed using enzyme-linked immunosorbent assay (ELISA). Several mtNFPs (ND4, ND5, ND6, and Cox1) were significantly upregulated in patients with ARDS at day 1 post-CPB compared to patients without ARDS. Higher levels of ND6 were correlated with worst PaO2/FiO2 (r=-0.2219 and P<0.0001) and cardiac Troponin T (r=2.107 and P<0.0001). Utilizing patient-derived serum and a rat lung ischemia reperfusion injury (LIRI) model, we observed a positive correlation between serum ND6 concentration and ARDS, which is also associated with EC barrier dysfunction. In vitro experiments, using trans-endothelial electric resistance (TEER) measurements and fluorescence microscopy with FITC-labeled VE-cadherin, demonstrated that ND6 disrupts the EC barrier through FPR2. Furthermore, FPR2 controls the release of ND6 out of mitochondria and cytoplasm under hypoxia reoxygenation (HR). Activated FPR2 leads to upregulation of nuclear transcription factor-kappa B (NF-κB) by inducing IκBα phosphorylation, promoting ICAM1 and VCAM1 expression, thereby compromising EC barrier integrity. Circulating pro-inflammatory and barrier-disruptive mtNFPs, particularly ND6, are associated with ARDS in patients undergoing CPB. The novel ND6-FPR2 axis regulates inflammation and EC permeability through the NF-κB pathway.

由于内皮细胞(EC)屏障功能障碍,心肺旁路(CPB)增加了急性呼吸窘迫综合征(ARDS)的风险。然而,线粒体 N-甲酰肽(mtNFPs)在 CPB 后 ARDS 中的特殊作用仍未得到研究。在此,我们研究了 CPB 后患者循环中 mtNFPs 的不同表达,重点关注 FPR2 在心肌中的新作用。我们使用酶联免疫吸附试验(ELISA)评估了循环中 mtNFPs 的水平。与无 ARDS 的患者相比,ARDS 患者在心肺复苏术后第 1 天的几种 mtNFPs(ND4、ND5、ND6 和 Cox1)明显上调。较高水平的 ND6 与最差的 PaO2/FiO2 相关(r=-0.2219 和 P
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引用次数: 0
Physiological Modeling of the Vascularized Human Lung Organoid. 血管化人体肺器官模型的生理建模
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-08 DOI: 10.1165/rcmb.2024-0413MA
Abdul S Qadir, Sukanta Das, Swathi Nedunchezian, Kaori Masuhara, Tushar J Desai, Jalees Rehman, Preetish Kadur Murthy, Yoshikazu Tsukasaki, Lijian Shao, Asrar B Malik

Studies using human lung organoids (hLO) have focused on differentiation of lung epithelial subtypes into distal alveolar unit. A major question has been whether introducing endothelial cells (EC) and resultant vascularization alter development of hLO. We describe herein a method for vessel infiltration of hLO in which we determined differences of these hLOs with standard avascular hLOs. hLO are generated by combining hiPSC-derived lung progenitor cells (LP) with EC at different LP:EC ratios. This results in vascularization of hLO and enables comparisons with hLO generated without EC. We observe red blood-filled vessels in hLOs generated post-implantation into the kidney capsule of NOD/SCID mice. Both human and mouse EC conjoin in the capsule to form chimeric vessels in hLOs. Vessel-infiltrating hLOs show robust generation of alveolar type II epithelial cells (ATII) and alveolar type I cells (ATI), although there was no difference in the observed 1:1 ATII/ATI cell ratio. Electron microscopy revealed better-developed surfactant production apparatus in ATII of vascularized hLOs compared to avascular hLOs. We observed prominent primitive airway sacs with alveolar epithelial cells lining lumen in vascularized vs. avascular hLOs. The vessel-infiltrating hLOs also mounted a robust inflammatory response characterized by mouse PMN influx after challenging host mice with lipopolysaccharide. Thus, interaction of EC with LP generated vascularized hLOs and drive ATII and ATI differentiation and hLOs also mount a robust inflammatory response upon LPS challenge of hLO-transplanted recipient mice. Our results show usefulness of generating hLOs in studying human lung development and mechanisms underlying inflammatory lung injury.

利用人体肺器官组织(hLO)进行的研究主要关注肺上皮亚型向远端肺泡单位的分化。一个主要问题是,引入内皮细胞(EC)和由此产生的血管化是否会改变 hLO 的发育。我们在本文中描述了一种对 hLO 进行血管浸润的方法,在这种方法中,我们确定了这些 hLO 与标准无血管 hLO 的差异。这导致了 hLO 的血管化,并能与不含 EC 的 hLO 进行比较。我们观察到植入 NOD/SCID 小鼠肾囊后生成的 hLO 中充满红色血管。人和小鼠的EC在囊内结合,在hLO中形成嵌合血管。血管浸润的 hLO 显示出肺泡 II 型上皮细胞(ATII)和肺泡 I 型细胞(ATI)的强劲生成,尽管观察到的 ATII/ATI 细胞比例为 1:1,但两者并无差异。电子显微镜显示,与无血管 hLO 相比,有血管 hLO 的 ATII 中表面活性物质生成器发育得更好。我们观察到,与无血管 hLO 相比,有血管 hLO 的原始气道囊突出,内腔有肺泡上皮细胞。在用脂多糖挑战宿主小鼠后,血管浸润的 hLO 还发起了以小鼠 PMN 大量涌入为特征的强烈炎症反应。因此,EC与LP的相互作用产生了血管化的hLO,并驱动了ATII和ATI的分化,当hLO移植的受体小鼠受到LPS挑战时,hLO也会产生强烈的炎症反应。我们的研究结果表明,生成 hLOs 有助于研究人类肺部发育和肺部炎症损伤的机制。
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引用次数: 0
Enhanced Gαq Signaling in TSC2-deficient Cells Is Required for Their Neoplastic Behavior. TSC2缺陷细胞的Gαq信号增强是其肿瘤行为的必要条件
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-08 DOI: 10.1165/rcmb.2024-0111OC
Aurélie Tréfier, Nihad Tousson-Abouelazm, Lama Yamani, Sajida Ibrahim, Kwang-Bo Joung, Adam Pietrobon, Julien Yockell-Lelievre, Terence E Hébert, Reese J Ladak, Tomoko Takano, Mark Nellist, Yoon Namkung, David Chatenet, William L Stanford, Stephane A Laporte, Arnold S Kristof

Inherited or sporadic loss of the TSC2 gene can lead to pulmonary lymphangioleiomyomatosis (LAM), a rare cystic lung disease caused by protease-secreting interstitial tumor nodules. The nodules arise by metastasis of cells that exhibit features of neural crest and smooth muscle lineage ('LAM cells'). Their aberrant growth is attributed to increased activity of 'mechanistic target of rapamycin complex 1' (mTORC1), an anabolic protein kinase that is normally suppressed by the TSC1-TSC2 protein complex. The mTORC1 inhibitor rapamycin slows the progression of LAM, but fails to eradicate disease, indicating a role for mTORC1-independent mechanisms in LAM pathogenesis. Our previous studies revealed G-protein coupled urotensin-II receptor (UT) signaling as a candidate mechanism, but how it promotes oncogenic signaling in TSC2-deficient cells remained unknown. Using a human pluripotent stem cell-derived in vitro model of LAM, we now show hyperactivation of UT, which was required for their enhanced migration and pro-neoplastic signaling in a rapamycin-insensitive mechanism that required heterotrimeric Gαq/11 (Gαq). Bioluminescence resonance energy transfer assays in HEK 293T cells lacking TSC2 demonstrated selective and enhanced activation of Gαq and its RhoA-associated effectors compared to wild-type control cells. By immunoprecipitation, recombinant UT was physically associated with Gαq and TSC2. The augmented Gαq signaling in TSC2-deleted cells was independent of mTOR activity, and associated with increased endosomal targeting of p63RhoGEF, a known RhoA-activating effector of Gαq. These studies identify potential mTORC1-independent pro-neoplastic mechanisms that can be targeted for prevention or eradication of pulmonary and extrapulmonary LAM tumors.

TSC2基因的遗传性或散发性缺失可导致肺淋巴管瘤病,这是一种罕见的囊性肺病,由分泌蛋白酶的间质肿瘤结节引起。这种结节是由具有神经嵴和平滑肌系特征的细胞("LAM 细胞")转移而来。它们的异常生长归因于 "雷帕霉素机理靶点复合体 1"(mTORC1)活性的增强,mTORC1 是一种合成代谢蛋白激酶,通常受到 TSC1-TSC2 蛋白复合体的抑制。mTORC1 抑制剂雷帕霉素能减缓 LAM 的进展,但无法根除疾病,这表明在 LAM 的发病机制中存在依赖于 mTORC1 的机制。我们之前的研究发现,G蛋白偶联尿促性素-II受体(UT)信号转导是一种候选机制,但它如何在TSC2缺陷细胞中促进致癌信号转导仍是未知数。现在,我们利用人多能干细胞衍生的LAM体外模型,显示了UT的过度激活,在雷帕霉素不敏感的机制中,UT是其增强迁移和促肿瘤信号转导所必需的,而这需要异三聚体Gαq/11(Gαq)。在缺乏 TSC2 的 HEK 293T 细胞中进行的生物荧光共振能量转移实验表明,与野生型对照细胞相比,Gαq 及其 RhoA 相关效应物的选择性激活增强。通过免疫沉淀,重组UT与Gαq和TSC2有物理关联。在 TSC2 缺失的细胞中,Gαq 信号的增强与 mTOR 活性无关,并且与 p63RhoGEF(一种已知的 Gαq 的 RhoA 激活效应因子)的内体靶向增加有关。这些研究发现了潜在的不依赖于 mTORC1 的促新陈代谢机制,可作为预防或根除肺部和肺外 LAM 肿瘤的靶点。
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引用次数: 0
Precision Cut Lung Slices: Emerging Tools for Preclinical and Translational Lung Research. An Official American Thoracic Society Workshop Report. 精确切割肺切片:临床前和转化肺研究的新兴工具。美国胸科学会官方研讨会报告。
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-05 DOI: 10.1165/rcmb.2024-0479ST
Mareike Lehmann, Ramaswamy Krishnan, Jennifer Sucre, Hrishikesh S Kulkarni, Ricardo H Pineda, Christopher Anderson, Nicholas E Banovich, Holger P Behrsing, Charlotte H Dean, Andrew Haak, Reinoud Gosens, Naftali Kaminski, Anna Zagorska, Cynthia Koziol-White, Jordan P Metcalf, Yong Ho Kim, Claudia Loebel, Enid Neptune, Alexandra Noel, Ganesh Raghu, Katherina Sewald, Ashish Sharma, Bela Suki, Anne Sperling, Amanda Tatler, Scott Turner, Ivan O Rosas, Pam van Ry, Timo Wille, Scott H Randell, Gloria Pryhuber, Mauricio Rojas, Jane Bourke, Melanie Königshoff

The urgent need for effective treatments for acute and chronic lung diseases underscores the significance of developing innovative preclinical human research tools. The 2023 ATS Workshop on Precision Cut Lung Slices (PCLS) brought together 35 experts to discuss and address the role of human tissue-derived PCLS as a unique tool for target and drug discovery and validation in pulmonary medicine. With increasing interest and usage, along with advancements in methods and technology, there is a growing need for consensus on PCLS methodology and readouts. The current document recommends standard reporting criteria and emphasizes the requirement for careful collection and integration of clinical metadata. We further discuss current clinically relevant readouts that can be applied to PCLS and highlight recent developments and future steps for implementing novel technologies for PCLS modeling and analysis. The collection and correlation of clinical metadata and multiomic analysis will further advent the integration of this preclinical platform into patient endotyping and the development of tailored therapies for lung disease patients.

急慢性肺部疾病迫切需要有效的治疗方法,这凸显了开发创新型临床前人体研究工具的重要性。2023 年美国肺科学学会(ATS)精密切肺切片(PCLS)研讨会汇聚了 35 位专家,共同讨论和探讨人体组织来源的 PCLS 作为一种独特工具在肺部医学的靶点和药物发现与验证中的作用。随着人们对 PCLS 的兴趣和使用日益增加,以及方法和技术的进步,人们越来越需要就 PCLS 方法和读数达成共识。本文件推荐了标准报告标准,并强调了仔细收集和整合临床元数据的要求。我们进一步讨论了目前可应用于 PCLS 的临床相关读数,并重点介绍了 PCLS 建模和分析新技术的最新进展和未来实施步骤。临床元数据的收集和关联以及多组学分析将进一步推动临床前平台与患者内分型的整合,并为肺病患者开发出量身定制的疗法。
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引用次数: 0
Novel Porcine Model Reveals Two Distinct LGR5 Cell Types During Lung Development and Homeostasis. 新型猪模型揭示肺发育和平衡过程中两种不同的 LGR5 细胞类型
IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-11-05 DOI: 10.1165/rcmb.2024-0040OC
Kathryn M Polkoff, Ross Lampe, Nithin K Gupta, Yanet Murphy, Jaewook Chung, Amber Carter, Jeremy M Simon, Katherine Gleason, Adele Moatti, Preetish K Murthy, Laura Edwards, Alon Greenbaum, Aleksandra Tata, Purushothama Rao Tata, Jorge A Piedrahita

Cells expressing LGR5 play a pivotal role in homeostasis, repair, and regeneration in multiple organs including skin and gastrointestinal tract, yet little is known about their role in the lung. Findings from mice, a widely used animal model, suggest that lung LGR5 expression differs from that of humans. In this work, using a new transgenic pig model, we identify two main populations of LGR5+ cells in the lung that are conserved in human, but not mouse lungs. Using RNA sequencing, 3D imaging and organoid models, we determine that in the fetal lung, epithelial LGR5 expression is transient in a subpopulation of SOX9+/ETV+/SFTPC+ progenitor lung tip cells. In contrast, epithelial LGR5 expression is absent from postnatal lung, but is reactivated in bronchioalveolar organoids derived from basal airway cells. We also describe a separate population of mesenchymal LGR5+ cells that surrounds developing and mature airways, lies adjacent to airway basal cells, and is closely associated with nerve fibers. Transcriptionally, mesenchymal LGR5+ cells include a subset of peribronchial fibroblasts (PBF) that express unique patterns of SHH, FGF, WNT and TGF-β signaling pathway genes. These results support distinct roles for LGR5+ cells in the lung and describe a physiologically relevant animal model for further studies on the function of these cells in repair and regeneration.

表达 LGR5 的细胞在皮肤和胃肠道等多个器官的稳态、修复和再生中发挥着关键作用,但人们对它们在肺部的作用却知之甚少。小鼠是一种广泛使用的动物模型,其研究结果表明肺部 LGR5 的表达与人类不同。在这项研究中,我们利用一种新的转基因猪模型,确定了肺部 LGR5+ 细胞的两个主要群体,它们在人类肺部是保守的,而在小鼠肺部则不是。通过使用 RNA 测序、三维成像和类器官模型,我们确定在胎儿肺中,上皮细胞 LGR5 的表达在 SOX9+/ETV+/SFTPC+ 原代肺尖细胞亚群中是短暂的。相反,出生后的肺中没有上皮细胞 LGR5 的表达,但在由基底气道细胞衍生的支气管肺泡器官组织中,上皮细胞 LGR5 的表达被重新激活。我们还描述了一个独立的间质 LGR5+ 细胞群,该细胞群围绕着发育和成熟的气道,毗邻气道基底细胞,并与神经纤维密切相关。从转录角度看,间质 LGR5+ 细胞包括支气管周围成纤维细胞(PBF)的一个亚群,它们表达 SHH、FGF、WNT 和 TGF-β 信号通路基因的独特模式。这些结果支持了 LGR5+ 细胞在肺中的独特作用,并描述了一个与生理相关的动物模型,以便进一步研究这些细胞在修复和再生中的功能。
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American Journal of Respiratory Cell and Molecular Biology
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