TET3 epigenetically controls feeding and stress response behaviors via AGRP neurons
Di Xie, … , Tamas L. Horvath, Yingqun Huang
Di Xie, … , Tamas L. Horvath, Yingqun Huang
Published October 3, 2022
Citation Information: J Clin Invest. 2022;132(19):e162365. https://doi.org/10.1172/JCI162365.
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Research Article Metabolism Neuroscience

TET3 epigenetically controls feeding and stress response behaviors via AGRP neurons

Abstract

The TET family of dioxygenases promote DNA demethylation by oxidizing 5-methylcytosine to 5-hydroxymethylcytosine (5hmC). Hypothalamic agouti-related peptide–expressing (AGRP-expressing) neurons play an essential role in driving feeding, while also modulating nonfeeding behaviors. Besides AGRP, these neurons produce neuropeptide Y (NPY) and the neurotransmitter GABA, which act in concert to stimulate food intake and decrease energy expenditure. Notably, AGRP, NPY, and GABA can also elicit anxiolytic effects. Here, we report that in adult mouse AGRP neurons, CRISPR-mediated genetic ablation of Tet3, not previously known to be involved in central control of appetite and metabolism, induced hyperphagia, obesity, and diabetes, in addition to a reduction of stress-like behaviors. TET3 deficiency activated AGRP neurons, simultaneously upregulated the expression of Agrp, Npy, and the vesicular GABA transporter Slc32a1, and impeded leptin signaling. In particular, we uncovered a dynamic association of TET3 with the Agrp promoter in response to leptin signaling, which induced 5hmC modification that was associated with a chromatin-modifying complex leading to transcription inhibition, and this regulation occurred in both the mouse models and human cells. Our results unmasked TET3 as a critical central regulator of appetite and energy metabolism and revealed its unexpected dual role in the control of feeding and other complex behaviors through AGRP neurons.

Authors

Di Xie, Bernardo Stutz, Feng Li, Fan Chen, Haining Lv, Matija Sestan-Pesa, Jonatas Catarino, Jianlei Gu, Hongyu Zhao, Christopher E. Stoddard, Gordon G. Carmichael, Marya Shanabrough, Hugh S. Taylor, Zhong-Wu Liu, Xiao-Bing Gao, Tamas L. Horvath, Yingqun Huang

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Figure 6

TET3 knockdown in AGRP neurons in female mice induces hyperphagia, obesity, and diabetes and reduces stress-like behaviors.

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TET3 knockdown in AGRP neurons in female mice induces hyperphagia, obesi...
(A) Cas9+ mice injected with AAV-sgTet3 or AAV bilaterally into the ARC at the age of 6 weeks became hyperphagic 2 weeks after injection. n = 8 animals per group. *P < 0.05 and **P < 0.01, by 2-tailed Student’s t test. (B) Representative images of mice 8 weeks after injection. (C) Post-injection body weight changes in mice. n = 8 animals per group. **P < 0.01, by 2-tailed Student’s t test. (D) Fat mass in mice 8 weeks after injection. n = 8 animals per group. ***P < 0.001, by 2-tailed Student’s t test. (E) Energy expenditure (EE) 2 weeks after injection. n = 8 animals per group. *P < 0.05 and **P < 0.01, by 2-tailed Student’s t test. (F) Blood insulin levels in ad libitum–fed mice 5 weeks after injection. n = 8 animals per group. ***P < 0.01, by 2-tailed Student’s t test. (G) Blood glucose levels in ad libitum–fed mice 6 weeks after injection. n = 8 animals per group. ***P < 0.01, by 2-tailed Student’s t test. (H) Blood leptin levels in ad libitum–fed mice 7 weeks after injection. n = 8 animals per group. **P < 0.01, by 2-tailed Student’s t test. (I) Glucose tolerance test (GTT) results for mice 8 weeks after injection. n = 8 animals per group. *P < 0.05, **P < 0.01, and ***P < 0.01, by 2-way ANOVA with Šidák’s post test. (J) Insulin tolerance test (ITT) results for mice 9 weeks after injection. n = 8 animals per group. *P < 0.05 and **P < 0.01, by 2-way ANOVA with Šidák’s post test. (K) Schematic diagram of experiments. Cas9+ mice were coinjected with AAV-sgTet3 and AAV-hM4Di bilaterally into the ARC on day 1 (D1), followed by implantation of an osmotic pump containing saline or C21 on day 4. Food intake measurements and ITTs were performed on day 5 and day 9, respectively. (L) Food intake data. “None” indicates age-matched Cas9+ mice without AAV injection or osmotic pump. n = 6 animals per group. **P < 0.01 and ***P < 0.001, by 1-way ANOVA with Tukey’s post test. (M) ITT data. n = 6 animals per group. *P < 0.05 and **P < 0.01, by 1-way ANOVA with Tukey’s post test. (N) Tail suspension test (TST) immobility scores for Cas9+ mice injected with AAV or AAV-sgTet3. n = 7 animals per group. ***P < 0.001, by 2-tailed Student’s t test. (O) Forced swim test (FST) immobility scores of Cas9+ mice injected with AAV or AAV-sgTet3. n = 7 animals per group. **P < 0.01, by 2-tailed Student’s t test. (P) Plasma corticosterone concentrations in Cas9+ mice injected with AAV or AAV-sgTet3. n = 7 animals per group. ***P < 0.001, by 2-tailed Student’s t test. All data represent the mean ± SEM.