Eine Punktmutation in der 5'-flankierenden Region vermindert die Transkription des Surfactant Protein B-Genes in vitro

Abstract Surfactant protein B (SFTPB) deficiency is a fatal disease affecting newborn infants. Surfactant is produced by alveolar type II cells which can be differentiated in vitro from patient specific induced pluripotent stem cell (iPSC)-derived lung organoids. Here we show the differentiation of patient specific iPSCs derived from a patient with SFTPB deficiency into lung organoids with mesenchymal and epithelial cell populations from both the proximal and distal portions of the human lung. We alter the deficiency by infecting the SFTPB deficient iPSCs with a lentivirus carrying the wild type SFTPB gene. After differentiating the mutant and corrected cells into lung organoids, we show expression of SFTPB mRNA during endodermal and organoid differentiation but the protein product only after organoid differentiation. We also show the presence of normal lamellar bodies and the secretion of surfactant into the cell culture medium in the organoids of lentiviral infected cells. These findings suggest that a lethal lung disease can be targeted and corrected in a human lung organoid model in vitro.

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Dissociation of surfactant protein B from canine surfactant large aggregates during formation of small surfactant aggregates by in vitro surface area cycling

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids ◽

10.1016/s1388-1981(99)00112-2 ◽

1999 ◽

Vol 1440 (1) ◽

pp. 49-58 ◽

Cited By ~ 12

Author(s):

Kevin Inchley ◽

Amanda Cockshutt ◽

Ruud Veldhuizen ◽

Fred Possmayer

Keyword(s):

Surface Area ◽

Surfactant Protein ◽

Surfactant Aggregates ◽

Surfactant Protein B ◽

Protein B

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A Combination of Short and Simple Surfactant Protein B and C Analogues as a New Synthetic Surfactant: In Vitro and Animal Experiments

Yonsei Medical Journal ◽

10.3349/ymj.2017.58.4.823 ◽

2017 ◽

Vol 58 (4) ◽

pp. 823 ◽

Cited By ~ 5

Author(s):

Yong-Sung Choi ◽

Sung-Hoon Chung ◽

Chong-Woo Bae

Keyword(s):

Animal Experiments ◽

Surfactant Protein ◽

Synthetic Surfactant ◽

Surfactant Protein B ◽

Protein B

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Membrane Activity of (Cys48Ser) Lung Surfactant Protein B Increases with Dimerisation

Biological Chemistry ◽

10.1515/bc.2001.116 ◽

2001 ◽

Vol 382 (6) ◽

pp. 933-939 ◽

Cited By ~ 12

Author(s):

Shahparak Zaltash ◽

William J. Griffiths ◽

David Beck ◽

Chen-Xia Duan ◽

Timothy E. Weaver ◽

...

Keyword(s):

Mass Spectrometry ◽

Transgenic Mice ◽

Ion Pairs ◽

Lung Surfactant ◽

Surfactant Protein ◽

Cd Spectroscopy ◽

Surfactant Protein B ◽

Lung Surfactant Protein ◽

Protein B

Abstract One of the possible functions of lung surfactant protein B (SPB), an hydrophobic membraneassociated saposinlike protein, is to reduce the alveolar surface tension by promoting insertion of phospholipids into the air/liquid interface of the lung. SPB is a covalent homodimer; Cys48 of two polypeptides form an intermolecular disulphide bond. In order to test whether dimerisation of SPB is important for surfactant function, transgenic mice which express (Cys48Ser) human SPB in a mouse SPB null background were generated. In previous studies (Cys48Ser)SPB showed a concentrationdependent in vitro activity, suggesting that it may form noncovalent dimers. Here (Cys48Ser)SPB isolated from bronchoalveolar lavage of transgenic mice was studied at different concentrations by circular dichroism (CD) spectroscopy, pulsating bubble surfactometry, mass spectrometry and reversedphase HPLC. The results indicate that (Cys48Ser)SPB, both in a phospholipid environment and in organic solvents, is largely monomeric and exhibits low activity at concentrations lower than 1 2 M, while at higher concentrations it forms noncovalent dimers, which are nearly functionally equivalent to native SPB in vitro. Furthermore, electrospray mass spectrometry showed that more dimers were found relative to the monomer when the polarity of the solvent was decreased, and when the concentration of SPB increased. (Cys48Ser)SPB also eluted earlier than native SPB in reversedphase HPLC. Taken together, these results indicate that a polar surface is buried upon dimerisation, thereby promoting formation of interchain ion pairs between Glu51 Arg52 and Glu51Arg52.

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