PMID- 34215333
OWN - NLM
STAT- MEDLINE
DCOM- 20210706
LR  - 20210707
IS  - 1756-0500 (Electronic)
IS  - 1756-0500 (Linking)
VI  - 14
IP  - 1
DP  - 2021 Jul 2
TI  - A modified protocol for successful miRNA profiling in human precision-cut lung 
      slices (PCLS).
PG  - 255
LID - 10.1186/s13104-021-05674-w [doi]
LID - 255
AB  - OBJECTIVE: Human precision cut lung slices (PCLS) are widely used as an ex vivo 
      model system for drug discovery and development of new therapies. PCLS reflect 
      the functional heterogeneity of lung tissue and possess relevant lung cell types. 
      We thus determined the use of PCLS in studying non-coding RNAs notably miRNAs, 
      which are important gene regulatory molecules. Since miRNAs play key role as 
      mediators of respiratory diseases, they can serve as valuable prognostic or 
      diagnostic biomarkers, and in therapeutic interventions, of lung diseases. A 
      technical limitation though is the vast amount of agarose in PCLS which impedes 
      (mi)RNA extraction by standard procedures. Here we modified our recently 
      published protocol for RNA isolation from PCLS to enable miRNA readouts. RESULTS: 
      The modified method relies on the separation of lysis and precipitation steps, 
      and a clean-up procedure with specific magnetic beads. We obtained successfully 
      quality miRNA amenable for downstream applications such as RTqPCR and whole 
      transcriptome miRNA analysis. Comparison of miRNA profiles in PCLS with published 
      data from human lung, identified all important miRNAs regulated in IPF, COPD, 
      asthma or lung cancer. Therefore, this shows suitability of the method for 
      analyzing miRNA targets and biomarkers in the valuable human PCLS model.
FAU - Niehof, Monika
AU  - Niehof M
AUID- ORCID: 0000-0001-5687-3052
AD  - Department of Preclinical Pharmacology and In Vitro Toxicology, Fraunhofer 
      Institute for Toxicology and Experimental Medicine, Biomedical Research in 
      Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German 
      Center for Lung Research (DZL), Nikolai-Fuchs-Str. 1, Hannover, 30625, Germany. 
      monika.niehof@item.fraunhofer.de.
FAU - Reamon-Buettner, Stella Marie
AU  - Reamon-Buettner SM
AD  - Department of Preclinical Pharmacology and In Vitro Toxicology, Fraunhofer 
      Institute for Toxicology and Experimental Medicine, Biomedical Research in 
      Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German 
      Center for Lung Research (DZL), Nikolai-Fuchs-Str. 1, Hannover, 30625, Germany.
FAU - Danov, Olga
AU  - Danov O
AD  - Department of Preclinical Pharmacology and In Vitro Toxicology, Fraunhofer 
      Institute for Toxicology and Experimental Medicine, Biomedical Research in 
      Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German 
      Center for Lung Research (DZL), Nikolai-Fuchs-Str. 1, Hannover, 30625, Germany.
FAU - Hansen, Tanja
AU  - Hansen T
AD  - Department of Preclinical Pharmacology and In Vitro Toxicology, Fraunhofer 
      Institute for Toxicology and Experimental Medicine, Biomedical Research in 
      Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German 
      Center for Lung Research (DZL), Nikolai-Fuchs-Str. 1, Hannover, 30625, Germany.
FAU - Sewald, Katherina
AU  - Sewald K
AD  - Department of Preclinical Pharmacology and In Vitro Toxicology, Fraunhofer 
      Institute for Toxicology and Experimental Medicine, Biomedical Research in 
      Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German 
      Center for Lung Research (DZL), Nikolai-Fuchs-Str. 1, Hannover, 30625, Germany.
LA  - eng
PT  - Journal Article
DEP - 20210702
PL  - England
TA  - BMC Res Notes
JT  - BMC research notes
JID - 101462768
RN  - 0 (MicroRNAs)
SB  - IM
MH  - Gene Expression Profiling
MH  - Humans
MH  - Lung
MH  - *Lung Diseases
MH  - *Lung Neoplasms
MH  - *MicroRNAs
PMC - PMC8252208
OTO - NOTNLM
OT  - Ex vivo
OT  - Lung material
OT  - Lung tissue
OT  - Microarray
OT  - Organotypic tissue
OT  - PCLS
OT  - RNA quality
OT  - RTqPCR
OT  - miRNA array
OT  - miRNA extraction
COIS- The authors declare that they have no competing interests.
EDAT- 2021/07/04 06:00
MHDA- 2021/07/07 06:00
PMCR- 2021/07/02
CRDT- 2021/07/03 05:27
PHST- 2021/03/22 00:00 [received]
PHST- 2021/06/24 00:00 [accepted]
PHST- 2021/07/03 05:27 [entrez]
PHST- 2021/07/04 06:00 [pubmed]
PHST- 2021/07/07 06:00 [medline]
PHST- 2021/07/02 00:00 [pmc-release]
AID - 10.1186/s13104-021-05674-w [pii]
AID - 5674 [pii]
AID - 10.1186/s13104-021-05674-w [doi]
PST - epublish
SO  - BMC Res Notes. 2021 Jul 2;14(1):255. doi: 10.1186/s13104-021-05674-w.