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Genome-wide expression profiling of human lymphoblastoid cell lines implicates integrin beta-3 in the mode of action of antidepressants.

Oved K, Morag A, Pasmanik-Chor M, Rehavi M, Shomron N, Gurwitz D - Transl Psychiatry (2013)

Bottom Line: We compared genome-wide expression profiles of human lymphoblastoid cell lines from unrelated individuals following treatment with 1 μM paroxetine for 21 days with untreated control cells and examined which genes and microRNAs (miRNAs) showed the most profound and consistent expression changes.Using genome-wide miRNA arrays, we observed a corresponding decrease in the expression of two miRNAs, miR-221 and miR-222, both predicted to target ITGB3.Further genes whose expression was significantly modulated by chronic paroxetine are also implicated in neurogenesis.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel [2] Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel.

ABSTRACT
Selective serotonin reuptake inhibitors (SSRIs) are the first-line treatment for major depression. However, the link between inhibition of serotonin reuptake and remission from depression remains controversial: in spite of the rapid onset of serotonin reuptake inhibition, remission from depression takes several weeks, presumably reflecting synaptogenesis/neurogenesis and neuronal rewiring. We compared genome-wide expression profiles of human lymphoblastoid cell lines from unrelated individuals following treatment with 1 μM paroxetine for 21 days with untreated control cells and examined which genes and microRNAs (miRNAs) showed the most profound and consistent expression changes. ITGB3, coding for integrin beta-3, showed the most consistent altered expression (1.92-fold increase, P=7.5 × 10(-8)) following chronic paroxetine exposure. Using genome-wide miRNA arrays, we observed a corresponding decrease in the expression of two miRNAs, miR-221 and miR-222, both predicted to target ITGB3. ITGB3 is crucial for the activity of the serotonin transporter (SERT), the drug target of SSRIs. Moreover, it is presumably required for the neuronal guidance activity of CHL1, whose expression was formerly identified as a tentative SSRI response biomarker. Further genes whose expression was significantly modulated by chronic paroxetine are also implicated in neurogenesis. Surprisingly, the expression of SERT or serotonin receptors was not modified. Our findings implicate ITGB3 in the mode of action of SSRI antidepressants and provide a novel link between CHL1 and the SERT. Our observations suggest that SSRIs may relieve depression primarily by promoting neuronal synaptogenesis/neurogenesis rather than by modulating serotonin neurotransmission per se.

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Expression changes for MAL, HECW2, ITGB3 and KLHL24 following chronicparoxetine exposure. Data are shown for microarrays (a, b) and real-timePCR (c, d) experiments as averages for four lymphoblastoid cell lines(LCLs) (a, c) or for each individual cell line (b, d),respectively. See Table 1 and Materials and Methods forexperimental details. Note the close similarity for the altered gene expression in LCLsrepresenting four unrelated donors.
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fig1: Expression changes for MAL, HECW2, ITGB3 and KLHL24 following chronicparoxetine exposure. Data are shown for microarrays (a, b) and real-timePCR (c, d) experiments as averages for four lymphoblastoid cell lines(LCLs) (a, c) or for each individual cell line (b, d),respectively. See Table 1 and Materials and Methods forexperimental details. Note the close similarity for the altered gene expression in LCLsrepresenting four unrelated donors.

Mentions: Cultured human LCLs from four unrelated adult male donors (Ashkenazi Jewish ancestry)growing under optimal conditions (cell density of 3 to 8 × 105 cells perml) were exposed to 1 μM paroxetine for 21 days, whereas parallelcultures of the same LCLs received similar volumes of phosphate-buffered saline. FollowingRNA extraction, Affymetrix GeneChip Human Gene 1.0 ST arrays or GeneChip miRNA 2.0 arrayswere used for detecting the expression levels of genes and miRNAs, respectively, accordingto the manufacturer's protocol (see Methods). Data were analyzed by Partek GenomicsSuite and normalized as described in Materials and methods. Tables1 and 2 present genes and miRNAs, respectively,whose expression levels showed >1.5-fold (>1.4-fold for miRNAs) difference andP<0.001 in preparations from LCL cultures chronically exposed to paroxetinecompared with parallel controls. As shown, ITGB3 (coding for ITGB3; also known asplatelet glycoprotein IIIa and CD61) exhibited the most statistically significant changein expression levels following 21 days paroxetine exposure, (1.925-fold increasedexpression; P=7.50 × 10−8) for the four LCLs. Fourgenes (MAL, HECW2, ITGB3 and KLHL24) and two miRNAs (miR-221 andmiR-222) from Tables 1 and 2 wereselected for validation with real-time PCR in each of the four individual LCLs (Figures 1 and 2), confirming the effectsof paroxetine on their expression levels. The choice of genes and miRNAs was made as theyare known to be expressed in the brain tissues and implicated in neurogenesis. As shown inFigures 1 and 2, the alteredexpression levels of these selected genes and miRNAs following chronic paroxetine exposurewere closely similar in the LCLs of four unrelated donors.


Genome-wide expression profiling of human lymphoblastoid cell lines implicates integrin beta-3 in the mode of action of antidepressants.

Oved K, Morag A, Pasmanik-Chor M, Rehavi M, Shomron N, Gurwitz D - Transl Psychiatry (2013)

Expression changes for MAL, HECW2, ITGB3 and KLHL24 following chronicparoxetine exposure. Data are shown for microarrays (a, b) and real-timePCR (c, d) experiments as averages for four lymphoblastoid cell lines(LCLs) (a, c) or for each individual cell line (b, d),respectively. See Table 1 and Materials and Methods forexperimental details. Note the close similarity for the altered gene expression in LCLsrepresenting four unrelated donors.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3818017&req=5

fig1: Expression changes for MAL, HECW2, ITGB3 and KLHL24 following chronicparoxetine exposure. Data are shown for microarrays (a, b) and real-timePCR (c, d) experiments as averages for four lymphoblastoid cell lines(LCLs) (a, c) or for each individual cell line (b, d),respectively. See Table 1 and Materials and Methods forexperimental details. Note the close similarity for the altered gene expression in LCLsrepresenting four unrelated donors.
Mentions: Cultured human LCLs from four unrelated adult male donors (Ashkenazi Jewish ancestry)growing under optimal conditions (cell density of 3 to 8 × 105 cells perml) were exposed to 1 μM paroxetine for 21 days, whereas parallelcultures of the same LCLs received similar volumes of phosphate-buffered saline. FollowingRNA extraction, Affymetrix GeneChip Human Gene 1.0 ST arrays or GeneChip miRNA 2.0 arrayswere used for detecting the expression levels of genes and miRNAs, respectively, accordingto the manufacturer's protocol (see Methods). Data were analyzed by Partek GenomicsSuite and normalized as described in Materials and methods. Tables1 and 2 present genes and miRNAs, respectively,whose expression levels showed >1.5-fold (>1.4-fold for miRNAs) difference andP<0.001 in preparations from LCL cultures chronically exposed to paroxetinecompared with parallel controls. As shown, ITGB3 (coding for ITGB3; also known asplatelet glycoprotein IIIa and CD61) exhibited the most statistically significant changein expression levels following 21 days paroxetine exposure, (1.925-fold increasedexpression; P=7.50 × 10−8) for the four LCLs. Fourgenes (MAL, HECW2, ITGB3 and KLHL24) and two miRNAs (miR-221 andmiR-222) from Tables 1 and 2 wereselected for validation with real-time PCR in each of the four individual LCLs (Figures 1 and 2), confirming the effectsof paroxetine on their expression levels. The choice of genes and miRNAs was made as theyare known to be expressed in the brain tissues and implicated in neurogenesis. As shown inFigures 1 and 2, the alteredexpression levels of these selected genes and miRNAs following chronic paroxetine exposurewere closely similar in the LCLs of four unrelated donors.

Bottom Line: We compared genome-wide expression profiles of human lymphoblastoid cell lines from unrelated individuals following treatment with 1 μM paroxetine for 21 days with untreated control cells and examined which genes and microRNAs (miRNAs) showed the most profound and consistent expression changes.Using genome-wide miRNA arrays, we observed a corresponding decrease in the expression of two miRNAs, miR-221 and miR-222, both predicted to target ITGB3.Further genes whose expression was significantly modulated by chronic paroxetine are also implicated in neurogenesis.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel [2] Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel.

ABSTRACT
Selective serotonin reuptake inhibitors (SSRIs) are the first-line treatment for major depression. However, the link between inhibition of serotonin reuptake and remission from depression remains controversial: in spite of the rapid onset of serotonin reuptake inhibition, remission from depression takes several weeks, presumably reflecting synaptogenesis/neurogenesis and neuronal rewiring. We compared genome-wide expression profiles of human lymphoblastoid cell lines from unrelated individuals following treatment with 1 μM paroxetine for 21 days with untreated control cells and examined which genes and microRNAs (miRNAs) showed the most profound and consistent expression changes. ITGB3, coding for integrin beta-3, showed the most consistent altered expression (1.92-fold increase, P=7.5 × 10(-8)) following chronic paroxetine exposure. Using genome-wide miRNA arrays, we observed a corresponding decrease in the expression of two miRNAs, miR-221 and miR-222, both predicted to target ITGB3. ITGB3 is crucial for the activity of the serotonin transporter (SERT), the drug target of SSRIs. Moreover, it is presumably required for the neuronal guidance activity of CHL1, whose expression was formerly identified as a tentative SSRI response biomarker. Further genes whose expression was significantly modulated by chronic paroxetine are also implicated in neurogenesis. Surprisingly, the expression of SERT or serotonin receptors was not modified. Our findings implicate ITGB3 in the mode of action of SSRI antidepressants and provide a novel link between CHL1 and the SERT. Our observations suggest that SSRIs may relieve depression primarily by promoting neuronal synaptogenesis/neurogenesis rather than by modulating serotonin neurotransmission per se.

Show MeSH
Related in: MedlinePlus