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INPP 4B overexpression and c‐ KIT downregulation in human achalasia
Author(s) -
Bonora E.,
Bianco F.,
Stanzani A.,
Giancola F.,
Astolfi A.,
Indio V.,
Evangelisti C.,
Martelli A. M.,
Boschetti E.,
Lugaresi M.,
Ioannou A.,
Torresan F.,
Stanghellini V.,
Clavenzani P.,
Seri M.,
Moonen A.,
Van Beek K.,
Wouters M.,
Boeckxstaens G. E.,
Zaninotto G.,
Mattioli S.,
De Giorgio R.
Publication year - 2018
Publication title -
neurogastroenterology and motility
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.489
H-Index - 105
eISSN - 1365-2982
pISSN - 1350-1925
DOI - 10.1111/nmo.13346
Subject(s) - achalasia , interstitial cell of cajal , immunohistochemistry , downregulation and upregulation , blot , protein kinase b , biology , ctgf , pathology , medicine , esophagus , gene , receptor , signal transduction , microbiology and biotechnology , genetics , growth factor
Background Achalasia is a rare motility disorder characterized by myenteric neuron and interstitial cells of Cajal ( ICC ) abnormalities leading to deranged/absent peristalsis and lack of relaxation of the lower esophageal sphincter. The mechanisms contributing to neuronal and ICC changes in achalasia are only partially understood. Our goal was to identify novel molecular features occurring in patients with primary achalasia. Methods Esophageal full‐thickness biopsies from 42 (22 females; age range: 16‐82 years) clinically, radiologically, and manometrically characterized patients with primary achalasia were examined and compared to those obtained from 10 subjects (controls) undergoing surgery for uncomplicated esophageal cancer (or upper stomach disorders). Tissue RNA extracted from biopsies of cases and controls was used for library preparation and sequencing. Data analysis was performed with the “edgeR” option of R‐Bioconductor. Data were validated by real‐time RT ‐ PCR , western blotting and immunohistochemistry. Key Results Quantitative transcriptome evaluation and cluster analysis revealed 111 differentially expressed genes, with a P ≤ 10 −3 . Nine genes with a P ≤ 10 −4 were further validated. CYR 61 , CTGF , c‐ KIT , DUSP 5 , EGR 1 were downregulated, whereas AKAP 6 and INPP 4B were upregulated in patients vs controls. Compared to controls, immunohistochemical analysis revealed a clear increase in INPP 4B, whereas c‐ KIT immunolabeling resulted downregulated. As INPP 4B regulates Akt pathway, we used western blot to show that phospho‐Akt was significantly reduced in achalasia patients vs controls. Conclusions & Inferences The identification of altered gene expression, including INPP 4B , a regulator of the Akt pathway, highlights novel signaling pathways involved in the neuronal and ICC changes underlying primary achalasia.