Lysosome-associated membrane glycoprotein 3 (LAMP3) is a type I transmembrane protein of the LAMP protein family with a cell-type-specific expression in alveolar type II cells in mice and hitherto unknown function. In type II pneumocytes, LAMP3 is localized in lamellar bodies, secretory organelles releasing pulmonary surfactant into the extracellular space to lower surface tension at the air/liquid interface. The physiological function of LAMP3, however, remains enigmatic. We generated Lamp3 knockout mice by CRISPR/Cas9. LAMP3 deficient mice are viable with an average life span and display regular lung function under basal conditions. The levels of a major hydrophobic protein component of pulmonary surfactant, SP-C, are strongly increased in the lung of Lamp3 knockout mice, and the lipid composition of the bronchoalveolar lavage shows mild but significant changes, resulting in alterations in surfactant functionality. In ovalbumin-induced experimental allergic asthma, the changes in lipid composition are aggravated, and LAMP3-deficient mice exert an increased airway resistance. Our data suggest a critical role of LAMP3 in the regulation of pulmonary surfactant homeostasis and normal lung function.
- MeSH
- bronchiální astma chemicky indukované genetika metabolismus patologie MeSH
- bronchoalveolární lavážní tekutina MeSH
- editace genu metody MeSH
- homeostáza genetika MeSH
- lipidomika MeSH
- modely nemocí na zvířatech MeSH
- myši knockoutované MeSH
- myši MeSH
- ovalbumin aplikace a dávkování MeSH
- plíce metabolismus patologie MeSH
- plicní alveoly metabolismus patologie MeSH
- plicní surfaktanty metabolismus MeSH
- pneumocyty metabolismus patologie MeSH
- protein - isoformy genetika metabolismus MeSH
- protein C asociovaný s plicním surfaktantem genetika metabolismus MeSH
- protein DC-LAMP nedostatek genetika MeSH
- regulace genové exprese MeSH
- respirační funkční testy MeSH
- rezistence dýchacích cest MeSH
- signální transdukce MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- ovalbumin MeSH
- plicní surfaktanty MeSH
- protein - isoformy MeSH
- protein C asociovaný s plicním surfaktantem MeSH
- protein DC-LAMP MeSH
- Sftpc protein, mouse MeSH Prohlížeč
Rationale: Several common and rare genetic variants have been associated with idiopathic pulmonary fibrosis, a progressive fibrotic condition that is localized to the lung. Objectives: To develop an integrated understanding of the rare and common variants located in multiple loci that have been reported to contribute to the risk of disease. Methods: We performed deep targeted resequencing (3.69 Mb of DNA) in cases (n = 3,624) and control subjects (n = 4,442) across genes and regions previously associated with disease. We tested for associations between disease and 1) individual common variants via logistic regression and 2) groups of rare variants via sequence kernel association tests. Measurements and Main Results: Statistically significant common variant association signals occurred in all 10 of the regions chosen based on genome-wide association studies. The strongest risk variant is the MUC5B promoter variant rs35705950, with an odds ratio of 5.45 (95% confidence interval, 4.91-6.06) for one copy of the risk allele and 18.68 (95% confidence interval, 13.34-26.17) for two copies of the risk allele (P = 9.60 × 10-295). In addition to identifying for the first time that rare variation in FAM13A is associated with disease, we confirmed the role of rare variation in the TERT and RTEL1 gene regions in the risk of IPF, and found that the FAM13A and TERT regions have independent common and rare variant signals. Conclusions: A limited number of common and rare variants contribute to the risk of idiopathic pulmonary fibrosis in each of the resequencing regions, and these genetic variants focus on biological mechanisms of host defense and cell senescence.
- Klíčová slova
- disease risk alleles, genetic variants, idiopathic pulmonary fibrosis, rare variants, targeted resequencing,
- MeSH
- ABC transportéry genetika MeSH
- celogenomová asociační studie MeSH
- DNA-helikasy genetika MeSH
- exoribonukleasy genetika MeSH
- genetická predispozice k nemoci MeSH
- genetická variace MeSH
- idiopatická plicní fibróza genetika MeSH
- interakce hostitele a patogenu genetika MeSH
- lidé MeSH
- logistické modely MeSH
- mucin 5B genetika MeSH
- promotorové oblasti (genetika) genetika MeSH
- protein A asociovaný s plicním surfaktantem genetika MeSH
- protein C asociovaný s plicním surfaktantem genetika MeSH
- proteiny aktivující GTPasu genetika MeSH
- proteiny vázající telomery genetika MeSH
- RNA genetika MeSH
- sekvenční analýza DNA MeSH
- stárnutí buněk genetika MeSH
- studie případů a kontrol MeSH
- telomerasa genetika MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- ABC transportéry MeSH
- ABCA3 protein, human MeSH Prohlížeč
- DNA-helikasy MeSH
- exoribonukleasy MeSH
- FAM13A protein, human MeSH Prohlížeč
- MUC5B protein, human MeSH Prohlížeč
- mucin 5B MeSH
- poly(A)-specific ribonuclease MeSH Prohlížeč
- protein A asociovaný s plicním surfaktantem MeSH
- protein C asociovaný s plicním surfaktantem MeSH
- proteiny aktivující GTPasu MeSH
- proteiny vázající telomery MeSH
- RNA MeSH
- RTEL1 protein, human MeSH Prohlížeč
- SFTPA2 protein, human MeSH Prohlížeč
- SFTPC protein, human MeSH Prohlížeč
- telomerasa MeSH
- telomerase RNA MeSH Prohlížeč
- TERT protein, human MeSH Prohlížeč
- TINF2 protein, human MeSH Prohlížeč
In this study, the effects of insulin and dexamethasone on the expression and mRNA transcription of 4 pulmonary surfactant-associated proteins [surfactant protein (SFTP)A, SFTPB, SFTPC and SFTPD] were examined. The commercially available cell lines, A549 and H441, were used as acceptable models of lung surfactant-producing cells. Subsequently, the effects of insulin on the expression of surfactant-associated proteins were examined in patients with lung adenocarcinoma during lung resection. Our results demonstrated the inhibitory effects of insulin on the transcription of the SFTPB, SFTPC and SFTPD genes in H441 cells and the SFTPB gene in A549 cells. Treatment with insulin significantly decreased the protein expression of SFTPA1 and SFTPA2 in the H441 cells and that of proSFTPB in the A549 cells. Dexamethasone promoted the transcription of the SFTPB, SFTPC and SFTPD genes in the A549 and H441 cells and reduced the transcription of the SFTPA1 and SFTPA2 genes in the H441 cells (SFTPA mRNA expression was not detected in A549 cells). Furthermore, we demonstrated that the mRNA levels of the selected genes were significantly lower in the cell lines compared to the lung tissue. A549 and H441 cells represent similar cell types. Yet, in our experiments, these cells reacted differently to insulin and/or dexamethasone treatment, and the mRNA levels of their main protein products, surfactant-associated proteins, were significantly lower than those in real tissue. Therefore, the results obtained in this study challenge the suitability of A549 and H441 cells as models of type II pneumocytes and Clara cells, respectively. However, we successfully demonstrate the possibility of studying the effects of insulin on pulmonary surfactant-associated genes and proteins in patients with lung adenocarcinoma.
- MeSH
- adenokarcinom plic MeSH
- adenokarcinom genetika MeSH
- dexamethason farmakologie MeSH
- inzulin farmakologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory plic genetika MeSH
- plíce účinky léků metabolismus MeSH
- protein A asociovaný s plicním surfaktantem genetika metabolismus MeSH
- protein B asociovaný s plicním surfaktantem genetika metabolismus MeSH
- protein C asociovaný s plicním surfaktantem genetika metabolismus MeSH
- protein D asociovaný s plicním surfaktantem genetika metabolismus MeSH
- proteiny asociované s plicním surfaktantem genetika metabolismus MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- regulace genové exprese účinky léků MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- dexamethason MeSH
- inzulin MeSH
- protein A asociovaný s plicním surfaktantem MeSH
- protein B asociovaný s plicním surfaktantem MeSH
- protein C asociovaný s plicním surfaktantem MeSH
- protein D asociovaný s plicním surfaktantem MeSH
- proteiny asociované s plicním surfaktantem MeSH
