Journal: bioRxiv

Article Title: GLP1R agonists activate human POMC neurons

doi: 10.1101/2024.04.02.587825

Figure Lengend Snippet: A,B) Hypothalamic cells differentiated from human pluripotent stem cells (hPSCs) contain a significant number of POMC neurons as visualized by immunostaining for aMSH (A) and in live cultures derived from a POMC-GFP reporter cell line (B). Scale bars represent 25 and 12 µm in A and B, respectively. C) UMAP of single-cell RNAseq data from five genetically distinct hPSC lines differentiated into hypothalamic neurons, separated by cells annotated as being in POMC (dark blue) or non-POMC (light blue) clusters. D,E) Expression of GLP1R mRNA in hPSC-derived hypothalamic neurons (UMI ≥ 1), color-coded by expression level (D) and quantified for POMC or SST cell clusters, and median expression across all cell clusters (E). F) UMAP with 66 annotated clusters from mouse single-cell and single-nucleus RNAseq data, with the arrowhead indicating the Pomc-expressing cell cluster. G,H) Expression of Glp1r mRNA in mouse hypothalamic neurons (UMI ≥ 1), color-coded by expression level (G) or quantified across Pomc or Sst clusters, or the median across all cell clusters.

Article Snippet: To test for the transcriptional consequences of longer-term exposure to GLP1R agonists, HUES9-POMC-GFP hypothalamic cultures at 40 ± 2 days post-differentiation were cultured in N2B27 medium for 72 hours prior to treatment with vehicle (DMSO) or Semaglutide (2 µM, MedChem Express) in N2B27 medium for 20 hours.

Techniques: Immunostaining, Derivative Assay, Expressing

Journal: bioRxiv

Article Title: GLP1R agonists activate human POMC neurons

doi: 10.1101/2024.04.02.587825

Figure Lengend Snippet: A) Photomicrographs from a POMC-GFP reporter cell line showing (from left to right) neuronal morphology in brightfield, baseline fluorescence from the Cal-590 AM calcium sensitive dye, endogenous GFP fluorescence from the reporter, and a merge of these images. B-D) Representative traces from single neurons in response to administration of 200 nM GLP-1 for two minutes showing no response (B), decreased fluorescence (C), a prolonged increase in fluorescence (D), where the Y axis represents ΔF/F 0 . E) Representative trace showing how GLP1R responsive cells were classified by calculating whether values for ΔF/F 0 taken in a baseline period were significantly different from those taken after GLP1R agonist administration. F) Plot of the significance of response probability (Y axis) versus the magnitude of response (ΔF/F 0 ), where GFP+ POMC neurons are plotted in green, and a cutoff of P<0.01 was used to assign cells into inhibited (blue), activated (red), and non-responsive (gray) categories. Significance was calculated by multiple paired t-test from 18 pre-stimulus and 18 post-stimulus fluorescence intensity values. G) Representative trace showing how the magnitude of response to GLP1R was calculated by taking the area under the curve (AUC) in three-minute time intervals before and after its administration. H) Summary of the activated and inhibited cell responses for GFP- and POMC neurons. Non-responsive neurons are not shown.

Article Snippet: To test for the transcriptional consequences of longer-term exposure to GLP1R agonists, HUES9-POMC-GFP hypothalamic cultures at 40 ± 2 days post-differentiation were cultured in N2B27 medium for 72 hours prior to treatment with vehicle (DMSO) or Semaglutide (2 µM, MedChem Express) in N2B27 medium for 20 hours.

Techniques: Fluorescence

Journal: bioRxiv

Article Title: GLP1R agonists activate human POMC neurons

doi: 10.1101/2024.04.02.587825

Figure Lengend Snippet: A) Normalized expression levels (low to high = yellow to dark green) of candidate genes from scRNAseq data of hPSC-derived hypothalamic POMC neurons (top row), or non-POMC neurons (bottom row), where dot size indicates the fraction of cells in each population in which the candidate gene was detected (UMI ≥ 1). POMC , GLP1R , and CACNA1D are significantly enriched in POMC neurons. B) Table of voltage-gated calcium channel (VGCC) gene names, channel type, and the identity of drugs that selectively inhibit them. C) In a Semaglutide-activated cell, a cocktail of VGCC blockers abolishes agonist-induced fluorescence of the calcium indicator. D,E) Administration of the P/Q-type VGCC blocker ω-Agatoxin does not significantly decrease Semaglutide-induced calcium indicator fluorescence, as seen in a representative trace (D) and in a summary of all Semaglutide-activated cells (E). F,G) Panels as in D,E but with the T-type VGCC blocker TTA-P2. H,I) Panels as in D-G, but with the L-type VGCC blocker Benidipine, which abolished Semaglutide-induced calcium dye fluorescence, as observed with the drug cocktail in (C). J,K) Panels as in D-I, showing that the results from Benidipine (H,I) are replicated with Nifedipine, a second L-type calcium channel blocker. *p<0.05, **p<0.01, ***p<0.001 ****p<0.0001 by one-way ANOVA with repeated measures in E, G, I, K.

Article Snippet: To test for the transcriptional consequences of longer-term exposure to GLP1R agonists, HUES9-POMC-GFP hypothalamic cultures at 40 ± 2 days post-differentiation were cultured in N2B27 medium for 72 hours prior to treatment with vehicle (DMSO) or Semaglutide (2 µM, MedChem Express) in N2B27 medium for 20 hours.

Techniques: Expressing, Derivative Assay, Fluorescence

Journal: Indian Journal of Endocrinology and Metabolism

Article Title: Approved and Emerging Hormone-Based Anti-Obesity Medications: A Review Article

doi: 10.4103/ijem.ijem_442_23

Figure Lengend Snippet: GLP1R/GCGR in clinical development

Article Snippet: Cotadutide (MEDI0382) , AstraZeneca/Medimmune , GLP1R/GCGR dual agonist , Phase II (T2DM/Obesity) Phase II (T2DM) Phase II (T2DM) , • The daily injectable GLP1R/GCGR dual agonist, Cotadutide (MEDI0382), was being developed by AstraZeneca to treat T2DM, obesity, NAFLD, and NASH. Cotadutide exhibited a balanced agonist effect on the GLP1R and GCGR with a ratio of approximately 5:1 GLP1R to GCGR activity.[ ] .

Techniques: Activity Assay, Clinical Proteomics, Control, Drug discovery, Expressing, Gene Expression

Journal: Indian Journal of Endocrinology and Metabolism

Article Title: Approved and Emerging Hormone-Based Anti-Obesity Medications: A Review Article

doi: 10.4103/ijem.ijem_442_23

Figure Lengend Snippet: GLP1R/GIPR dual agonists in clinical development

Article Snippet: , , GLP1R/GIPR dual agonist , FDA/EMA approved (T2DM) Phase III (HF, SAS) Phase II (NASH) , • It is FDA/EMA approved for T2DM based on SURPASS trials.[ ] .

Techniques: Drug discovery