Knockdown of Drosha in human alveolar type II cells alters expression of SP-A in culture

A pilot study

Patricia Silveyra, Zissis C. Chroneos, Susan L. Diangelo, Neal J. Thomas, Georgios Noutsios, Nikolaos Tsotakos, Judie A. Howrylak, Todd M. Umstead, Joanna Floros

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Human surfactant protein A (SP-A) plays an important role in surfactant metabolism and lung innate immunity. SP-A is synthesized and secreted by alveolar type II (ATII) cells, one of the two cell types of the distal lung epithelium (ATII and ATI). We have shown that miRNA interactions with sequence polymorphisms on the SP-A mRNA 3'UTRs mediate differential expression of SP-A1 and SP-A2 gene variants in vitro. In the present study, we describe a physiologically relevant model to study miRNA regulation of SP-A in human ATII. For these studies, we purified and cultured human ATII on an air-liquid interface matrix (A/L) or plastic wells without matrix (P). Gene expression analyses confirmed that cells cultured in A/L maintained the ATII phenotype for over 5 days, whereas P-cultured cells differentiated to ATI. When we transfected ATII with siRNAs to inhibit the expression of Drosha, a critical effector of miRNA maturation, the levels of SP-A mRNA and protein increased in a time dependent manner. We next characterized cultured ATII and ATI by studying expression of 1,066 human miRNAs using miRNA PCR arrays. We detected expression of >300 miRNAs with 24 miRNAs differentially expressed in ATII versus ATI, 12 of which predicted to bind SP-A 3'UTRs, indicating that these may be implicated in SP-A downregulation in ATI. Thus, miRNAs not only affect SP-A expression, but also may contribute to the maintenance of the ATII cell phenotype and/or the trans-differentiation of ATII to ATI cells, and may represent new molecular markers that distinguish ATII and ATI.

Original languageEnglish (US)
Pages (from-to)354-366
Number of pages13
JournalExperimental Lung Research
Volume40
Issue number7
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

Fingerprint

Pulmonary Surfactant-Associated Protein A
Alveolar Epithelial Cells
MicroRNAs
3' Untranslated Regions
Cultured Cells
Phenotype
Lung
Messenger RNA
Polymorphism
varespladib methyl
Innate Immunity
Metabolism
Gene expression
Surface-Active Agents
Plastics
Down-Regulation
Epithelium
Genes
Air
Cells

Keywords

  • Alveolar type II cells
  • Lung epithelium
  • MiRNAs
  • Surfactant protein A

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry

Cite this

Knockdown of Drosha in human alveolar type II cells alters expression of SP-A in culture : A pilot study. / Silveyra, Patricia; Chroneos, Zissis C.; Diangelo, Susan L.; Thomas, Neal J.; Noutsios, Georgios; Tsotakos, Nikolaos; Howrylak, Judie A.; Umstead, Todd M.; Floros, Joanna.

In: Experimental Lung Research, Vol. 40, No. 7, 01.01.2014, p. 354-366.

Research output: Contribution to journalArticle

Silveyra, P, Chroneos, ZC, Diangelo, SL, Thomas, NJ, Noutsios, G, Tsotakos, N, Howrylak, JA, Umstead, TM & Floros, J 2014, 'Knockdown of Drosha in human alveolar type II cells alters expression of SP-A in culture: A pilot study', Experimental Lung Research, vol. 40, no. 7, pp. 354-366. https://doi.org/10.3109/01902148.2014.929757
Silveyra, Patricia ; Chroneos, Zissis C. ; Diangelo, Susan L. ; Thomas, Neal J. ; Noutsios, Georgios ; Tsotakos, Nikolaos ; Howrylak, Judie A. ; Umstead, Todd M. ; Floros, Joanna. / Knockdown of Drosha in human alveolar type II cells alters expression of SP-A in culture : A pilot study. In: Experimental Lung Research. 2014 ; Vol. 40, No. 7. pp. 354-366.
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AU - Diangelo, Susan L.

AU - Thomas, Neal J.

AU - Noutsios, Georgios

AU - Tsotakos, Nikolaos

AU - Howrylak, Judie A.

AU - Umstead, Todd M.

AU - Floros, Joanna

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