Přejít k obsahu
Merck
  • Elevated de novo protein synthesis in FMRP-deficient human neurons and its correction by metformin treatment.

Elevated de novo protein synthesis in FMRP-deficient human neurons and its correction by metformin treatment.

Molecular autism (2020-05-29)
Kagistia Hana Utami, Nur Amirah Binte Mohammad Yusof, Jing Eugene Kwa, Ulla-Kaisa Peteri, Maija L Castrén, Mahmoud A Pouladi
ANOTACE

FXS is the most common genetic cause of intellectual (ID) and autism spectrum disorders (ASD). FXS is caused by loss of FMRP, an RNA-binding protein involved in the translational regulation of a large number of neuronal mRNAs. Absence of FMRP has been shown to lead to elevated protein synthesis and is thought to be a major cause of the synaptic plasticity and behavioural deficits in FXS. The increase in protein synthesis results in part from abnormal activation of key protein translation pathways downstream of ERK1/2 and mTOR signalling. Pharmacological and genetic interventions that attenuate hyperactivation of these pathways can normalize levels of protein synthesis and improve phenotypic outcomes in animal models of FXS. Several efforts are currently underway to trial this strategy in patients with FXS. To date, elevated global protein synthesis as a result of FMRP loss has not been validated in the context of human neurons. Here, using an isogenic human stem cell-based model, we show that de novo protein synthesis is elevated in FMRP-deficient neural cells. We further show that this increase is associated with elevated ERK1/2 and Akt signalling and can be rescued by metformin treatment. Finally, we examined the effect of normalizing protein synthesis on phenotypic abnormalities in FMRP-deficient neural cells. We find that treatment with metformin attenuates the increase in proliferation of FMRP-deficient neural progenitor cells but not the neuronal deficits in neurite outgrowth. The elevated level of protein synthesis and the normalization of neural progenitor proliferation by metformin treatment were validated in additional control and FXS patient-derived hiPSC lines. Overall, our results validate that loss of FMRP results in elevated de novo protein synthesis in human neurons and suggest that approaches targeting this abnormality are likely to be of partial therapeutic benefit in FXS.

MATERIÁLY
Číslo produktu
Značka
Popis produktu

Sigma-Aldrich
L-Ascorbic acid, powder, suitable for cell culture, γ-irradiated
Sigma-Aldrich
Anti-Microtubule-Associated Protein 2 (MAP2) Antibody, Chemicon®, from rabbit
Sigma-Aldrich
Anti-Ki-67 Antibody, clone Ki-S5, clone Ki-S5, Chemicon®, from mouse
Sigma-Aldrich
N6,2′-O-Dibutyryladenosine 3′,5′-cyclic monophosphate sodium salt, ≥97% (HPLC), powder
Sigma-Aldrich
Triton X-100, laboratory grade
Worthington Accessories, cap/necktube core for 25 L Dewar
Sigma-Aldrich
Anti-Tubulin Antibody, beta III isoform, CT, clone TU-20 (Similar to TUJ1), ascites fluid, clone TU-20 (Similar to TUJ1), Chemicon®
Sigma-Aldrich
Anti-Nestin Antibody, clone 10C2, clone 10C2, Chemicon®, from mouse
Sigma-Aldrich
Anti-Fragile X Mental Retardation Protein Antibody, clone 1C3, ascites fluid, clone 1C3, Chemicon®