biostudies-arrayexpressSingle Cell Omics Platform CBMRRattus norvegicusnf-core/rnaseq: 3.12.0https://www.ebi.ac.uk/biostudies/studies/E-MTAB-16695This study evaluates whether simultaneously activating five key metabolic hormone receptors can outperform current peptide therapies for obesity. Using diet‑induced obese rats, we test the combined administration of retatrutide, a GLP‑1, GIP, and glucagon triagonist, together with cagrilintide, an amylin and calcitonin receptor agonist, versus its monotherapies and evaluate the bulk transcriptomic profile of the hypothalamus and the dorsal vagal complex.biostudies-arrayexpressSample Collection - The termination was performed by decapitation for blood and tissue collection. Microdissections of the hypothalamus and dorsal vagal complex (DVC) was performed for RNA isolation and subsequent sequencing.Growth Protocol - Male Sprague Dawley rats were maintained on ad libitum high-fat, high-sucrose diet (HFHS, 59 kcal% fat, #E15772-347, ssniff spezialdiäten GmbH) from 10 weeks of age and for a minimum of 12 weeks. Rats had an average body weight of >700g before initiating pharmacological studies. The rats were double-housed (Department of Experimental Medicine Rigshospitalet Unit) in a humidity- (45-65%) and temperature-controlled environment (21-23 oC) with a 12-hour light/dark cycle (6:00 am – 6:00 pm).Nucleic Acid Extraction - For RNA extraction, all tissues were homogenized in TRIzol reagent (QIAzol Lysis Reagent, Qiagen, 79306) with a stainless-steel bead (Qiagen, 69989) using a TissueLyser II (Qiagen, 85300) set at 30 Hz for 3 min. Total RNA was isolated according to the manufacturer's protocol using QIAwave RNA Mini Kit (Qiagen, 74534) and RNA was quantified and RNA quality accessed using NanoDrop 2000 (Thermo Fischer, ND-2000). Total RNA was then converted to cDNA by mixing with FS buffer (Thermo Fisher Scientific, 18080-044), dithiothreitol (Thermo Fisher Scientific, 18080-044) and random primers (Sigma-Aldrich, 11034731001) followed by incubation at 70 oC for 3 min in a thermal cycler (Eppendorf Mastercycler Pro) for first strand synthesis. dNTPs (Thermo Fisher Scientific, R0192), RNase out (Thermo Fisher Scientific, 10777019) and Superscript III (Thermo Fisher Scientific, 18080-044) were subsequently added for cDNA synthesis using a thermal cycler (Eppendorf Mastercycler Pro) with the following steps: 5 min at 25 oC, 60 min at 50 oC and 15 min at 70 oC. cDNA was diluted 1:20 for all tissues and stored at−20 oC.Sequencing - 52-bp paired-end sequencing on a NovaSeq6000 sequencerSample Treatment - At an age of 23 weeks, DIO rats weighing 720g on average were divided into four experimental groups (n = 10): vehicle, 5 nmol kg-1 cagrilintide or retatrutide, and the loose equimolar combination of cagrilintide and retatrutide administered in a single dose. HFHS diet intake was measured and replenished daily for ad libitum feeding followed by measurements of body weight and s.c. o.d. compound administration for 16 days.Library Construction - Libraries were prepared using the Universal Plus mRNA-seq with NuQuant protocol (Tecan) as recommended by the manufacturer. Libraries were quantified with NuQuant using the CLARIOstar Plate Reader (BMG Labtech) and quality checked using a TapeStation instrument (Agilent Technologies)OrganizationMINSEQE ScoreAssays and DataProcessed DataMAGE-TAB FilesSequence Alignment - FASTQ-files were analyzed using the nf-core/rnaseq: 3.12.0 with the flags --with_umi --skip_umi_extract --umitools_umi_separator \":\" agains the rn7 genome and the ENSEMBL release 112 gene model.Data Transformation - Uploaded data is raw reads, each row is a gene and each column is a sample. No normalization has been performed.MetabolomicsUnknownTranscriptomicsGenomicsProteomicsIllumina NovaSeq 6000RNA-seq of coding RNARattus norvegicusSingle Cell Omics Platform CBMRShad HassanfalseCagrilintide and retatrutide combination therapy enhances weight loss and metabolic outcomes in obese ratsThis study evaluates whether simultaneously activating five key metabolic hormone receptors can outperform current peptide therapies for obesity. Using diet‑induced obese rats, we test the combined administration of retatrutide, a GLP‑1, GIP, and glucagon triagonist, together with cagrilintide, an amylin and calcitonin receptor agonist, versus its monotherapies and evaluate the bulk transcriptomic profile of the hypothalamus and the dorsal vagal complex.2026-07-20T00:00:00Z2026-07-20T11:23:55.172Z2026-02-27T09:16:00.1ZE-MTAB-16695ERP189703EFO_0002944EFO_0004170EFO_0003789EFO_0004917EFO_0005518EFO_0003816EFO_0003738EFO_0004184EFO_0003969