title: Study of CRTC2 pharmacology using antisense oligonuceotides.
abstract: The cAMP response element binding protein (CREB)-regulated transcriptional coactivator 2 (CRTC2) is a key component of the transcription complex regulating glucagon driven hepatic glucose production and previous evidence suggests that "inhibition" of CRTC2 improves glucose homeostasis in multiple rodent models of type 2 diabetes. Here we describe a process of identifying potential therapeutic antisense oligonucleotides (ASOs) directed against CRTC2. These ASOs were designed as locked nucleic acid (LNA) gapmers and a panel of approximately 400 sequences were first screened in vitro within both human and mouse liver cell lines. A group of active and selective compounds were then profiled in acute studies in mice to determine the level of CRTC2 mRNA reduction in liver as well as to obtain a preliminary indication of safety and tolerability. The compounds with the best activity and safety profiles were then evaluated in subchronic efficacy studies using the diet induced obese (DIO) mouse model of type 2 diabetes and primary human hepatocytes. Efficacy findings broadly confirmed the beneficial effect of reducing CRTC2 mRNA levels towards improving glucose control and other markers of metabolic function. Additionally, for the first time, translation to human cells has been established with demonstration of a reduction in glucagon-mediated glucose production in primary human hepatocytes and a potential clinical biomarker source identified to assess modulation of CRTC2 mRNA following ASO treatment. While the compounds identified herein did not demonstrate a therapeutic index sufficient for further development, this study should facilitate more efficient prosecution of compounds within an in vivo setting.
authors.affiliation: 1 Cardiovascular and Metabolic Disease Research Unit, Pfizer , Cambridge, Massachusetts.
authors.forename: Robert
authors.initials: R
authors.lastname: Dullea
authors.forename: Christopher
authors.initials: C
authors.lastname: Salatto
authors.forename: Simone
authors.initials: S
authors.lastname: Sciabola
authors.forename: Tracy
authors.initials: T
authors.lastname: Chen
authors.forename: Debra
authors.initials: D
authors.lastname: Dimattia
authors.forename: Harmeet
authors.initials: H
authors.lastname: Gandhok
authors.forename: John
authors.initials: J
authors.lastname: Kreeger
authors.forename: Yan
authors.initials: Y
authors.lastname: Weng
authors.forename: Tracey
authors.initials: T
authors.lastname: Clark
authors.forename: Chandra
authors.initials: C
authors.lastname: Vage
authors.forename: Robert
authors.initials: R
authors.lastname: Stanton
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D003924:Diabetes Mellitus, Type 2 / Q000209:etiology / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D059305:Diet, High-Fat / Q000009:adverse effects; D004041:Dietary Fats / Q000009:adverse effects; D005786:Gene Expression Regulation; D005934:Glucagon / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology; D057166:High-Throughput Screening Assays; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D016688:Mice, Inbred NOD; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000235:genetics* / Q000378:metabolism; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism
chemical_list: D001786:Blood Glucose; C504923:Crtc2 protein, mouse; D004041:Dietary Fats; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D014157:Transcription Factors; C477371:locked nucleic acid; D005934:Glucagon