GIGYF1 disruption associates with autism and impaired IGF-1R signaling
Guodong Chen, … , Kun Xia, Hui Guo
Guodong Chen, … , Kun Xia, Hui Guo
Published August 2, 2022
Citation Information: J Clin Invest. 2022;132(19):e159806. https://doi.org/10.1172/JCI159806.
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Research Article Genetics Neuroscience

GIGYF1 disruption associates with autism and impaired IGF-1R signaling

Abstract

Autism spectrum disorder (ASD) represents a group of neurodevelopmental phenotypes with a strong genetic component. An excess of likely gene-disruptive (LGD) mutations in GIGYF1 was implicated in ASD. Here, we report that GIGYF1 is the second-most mutated gene among known ASD high–confidence risk genes. We investigated the inheritance of 46 GIGYF1 LGD variants, including the highly recurrent mutation c.333del:p.L111Rfs*234. Inherited GIGYF1 heterozygous LGD variants were 1.8 times more common than de novo mutations. Among individuals with ASD, cognitive impairments were less likely in those with GIGYF1 LGD variants relative to those with other high-confidence gene mutations. Using a Gigyf1 conditional KO mouse model, we showed that haploinsufficiency in the developing brain led to social impairments without significant cognitive impairments. In contrast, homozygous mice showed more severe social disability as well as cognitive impairments. Gigyf1 deficiency in mice led to a reduction in the number of upper-layer cortical neurons, accompanied by a decrease in proliferation and increase in differentiation of neural progenitor cells. We showed that GIGYF1 regulated the recycling of IGF-1R to the cell surface. KO of GIGYF1 led to a decreased level of IGF-1R on the cell surface, disrupting the IGF-1R/ERK signaling pathway. In summary, our findings show that GIGYF1 is a regulator of IGF-1R recycling. Haploinsufficiency of GIGYF1 was associated with autistic behavior, likely through interference with IGF-1R/ERK signaling pathway.

Authors

Guodong Chen, Bin Yu, Senwei Tan, Jieqiong Tan, Xiangbin Jia, Qiumeng Zhang, Xiaolei Zhang, Qian Jiang, Yue Hua, Yaoling Han, Shengjie Luo, Kendra Hoekzema, Raphael A. Bernier, Rachel K. Earl, Evangeline C. Kurtz-Nelson, Michaela J. Idleburg, Suneeta Madan-Khetarpal, Rebecca Clark, Jessica Sebastian, Alberto Fernandez-Jaen, Sara Alvarez, Staci D. King, Luiza L.P. Ramos, Mara Lucia S.F. Santos, Donna M. Martin, Dan Brooks, Joseph D. Symonds, Ioana Cutcutache, Qian Pan, Zhengmao Hu, Ling Yuan, Evan E. Eichler, Kun Xia, Hui Guo

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Figure 5

GIGYF1 KO disrupts IGF-1R/ERK pathway by regulation of IGF-1R recycling.

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GIGYF1 KO disrupts IGF-1R/ERK pathway by regulation of IGF-1R recycling...
(A) Coimmunoprecipitation assay for GIGYF1 and IGF-1R in HEK293T cells. (B) Double immunofluorescence of GIGYF1 and IGF-1R in HeLa cells. Scale bars: 10 μm. Inset scale bars: 5 μm. (C) Immunoblots of the whole cell lysate showing levels of pIGF-1R, IGF-1R, pERK1/2, ERK1/2, pAkt, and Akt at different duration of IGF-1 stimulation. Statistical data were analyzed using 2-way ANOVA. (D) Immunoblots of the whole-cell lysates showing levels of pERK1/2, ERK1/2 in HEK293T GIGYF1 KO cells expressing mock empty vector (pCAGGS-IRES-GFP) or HA-GIGYF1. The relative levels of pERK1/2 to ERK1/2 were quantified by densitometry and analyzed using 2-way ANOVA. (E) Immunoblot of pERK1/2 and ERK1/2 in the whole-cell lysates at 7 minutes of IGF-1 stimulation. Statistic data were analyzed using 1-way ANOVA. (F) Immunoblots of biotin-labelled IGF-1R, total IGF-1R and TMEM98. Total IGF-1R-α levels of unbiotinylated cells were determined. The protein levels of surface IGF-1R, surface IGF-1R/total IGF-1R, and total IGF-1R were quantified by densitometry from 3 biological replicates. Statistical data were analyzed using 2-tailed Student’s t tests. (G) Immunoblots of biotin-labeled IGF-1R and total IGF-1R at different conditions in a surface biotinylation recycling assay. GIGYF1 KO HEK293T cells and control HEK293T cells (lanes 2–8) were surface labeled with sulfo-NHS-S-S-biotin. Cells (lanes 3–8) were incubated to endocytosis. The remaining surface biotin was cleaved with glutathione cleavage buffer (lanes 2–8). Cells were incubated for a second time to recycle (lanes 4–5 and 7–8), then the surface biotin was stripped for a second time (lanes 3–4 and 6–7). Lanes 5 and 8 were incubated to recycle without a second cleavage. The stage of biotinylation recycling assay for each lysate is indicated with + and –. (H) Working model of GIGYF1 regulation of IGF-1R/ERK pathway. All data are represented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. See complete unedited blots in the supplemental material.