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bioRxiv · 10.1101/2021.12.19.473196

Depletion of ALMS1 inhibits TGF-β/SMAD3/SNAI1 axis altering normal cell migration capacity and epithelial-mesenchyme transition.

Bea-Mascato, B.·Neira-Goyanes, E.·Iglesias-Rodriguez, A.·Valverde, D.
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Abstract

BackgroundALMS1 is a ubiquitous gene associated with Alstrom syndrome (ALMS). The main symptoms of ALMS affect multiple organs and tissues, generating at last, multi-organic fibrosis in the lungs, kidneys and liver. TGF-{beta} is one of the main pathways implicated in fibrosis, controlling the cell cycle, apoptosis, cell migration, cell adhesion and epithelial-mesenchymal transition (EMT). Nevertheless, the role of ALMS1 gene in fibrosis generation and other implicated processes such as cell migration or cell adhesion via the TGF-{beta} pathway has not been elucidated yet. MethodsInitially, we evaluated how depletion of ALMS1 affects different processes like apoptosis, cell cycle and mitochondrial activity in HeLa cells. Then, we performed proteomic profiling with TGF-{beta} stimuli in HeLa ALMS1 -/- cells and validated the results by examining different EMT biomarkers using qPCR. The expression of these EMT biomarkers were also studied in hTERT-BJ-5ta ALMS1 -/-. Finally, we also evaluated the SMAD3 and SMAD2 phosphorylation and cell migration capacity in both models. ResultsDepletion of ALMS1 generated apoptosis resistance to thapsigargin (THAP) and C2-Ceramide (C2-C), and G2/M cell cycle arrest in HeLa cells. For mitochondrial activity, results did not show significant differences between ALMS1 +/+ and ALMS1 -/-. Proteomic results showed inhibition of downstream pathways regulated by TGF-{beta}. The protein-coding genes (PCG) were associated with processes like focal adhesion or cell-substrate adherens junction in HeLa. SNAI1 showed an opposite pattern to what would be expected when activating the EMT in HeLa and BJ-5ta. Finally, in BJ-5ta model a reduced activation of SMAD3 but not SMAD2 were also observed. In HeLa models no alterations in the canonical TGF-{beta} pathway were observed but both cell lines showed a reduction in migration capacity. ConclusionALMS1 has a role in controlling the cell cycle and the apoptosis processes. Moreover, the depletion of ALMS1 affects the signal transduction through the TGF-{beta} and other processes like the cell migration and adhesion capacity.

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Bea-Mascato, B., Neira-Goyanes, E., Iglesias-Rodriguez, A., Valverde, D.. 2021-12-19. Depletion of ALMS1 inhibits TGF-β/SMAD3/SNAI1 axis altering normal cell migration capacity and epithelial-mesenchyme transition.. https://doi.org/10.1101/2021.12.19.473196

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