PMID- 24695667
OWN - NLM
STAT- MEDLINE
DCOM- 20150129
LR  - 20240322
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 9
IP  - 4
DP  - 2014
TI  - A novel humanized GLP-1 receptor model enables both affinity purification and 
      Cre-LoxP deletion of the receptor.
PG  - e93746
LID - 10.1371/journal.pone.0093746 [doi]
LID - e93746
AB  - Class B G protein-coupled receptors (GPCRs) are important regulators of endocrine 
      physiology, and peptide-based therapeutics targeting some of these receptors have 
      proven effective at treating disorders such as hypercalcemia, osteoporosis, and 
      type 2 diabetes mellitus (T2DM). As next generation efforts attempt to develop 
      novel non-peptide, orally available molecules for these GPCRs, new animal models 
      expressing human receptor orthologs may be required because small molecule 
      ligands make fewer receptor contacts, and thus, the impact of amino acid 
      differences across species may be substantially greater. The objective of this 
      report was to generate and characterize a new mouse model of the human 
      glucagon-like peptide-1 receptor (hGLP-1R), a class B GPCR for which established 
      peptide therapeutics exist for the treatment of T2DM. hGLP-1R knock-in mice 
      express the receptor from the murine Glp-1r locus. Glucose tolerance tests and 
      gastric emptying studies show hGLP-1R mice and their wild-type littermates 
      display similar physiological responses for glucose metabolism, insulin 
      secretion, and gastric transit, and treatment with the GLP-1R agonist, exendin-4, 
      elicits similar responses in both groups. Further, ex vivo assays show insulin 
      secretion from humanized islets is glucose-dependent and enhanced by GLP-1R 
      agonists. To enable additional utility, the targeting construct of the knock-in 
      line was engineered to contain both flanking LoxP sites and a C-terminal FLAG 
      epitope. Anti-FLAG affinity purification shows strong expression of hGLP-1R in 
      islets, lung, and stomach. We crossed the hGLP-1R line with Rosa26Cre mice and 
      generated global Glp-1r-/- animals. Immunohistochemistry of pancreas from 
      humanized and knock-out mice identified a human GLP-1R-specific antibody that 
      detects the GLP-1R in human pancreas as well as in the pancreas of hGLP-1r 
      knock-in mice. This new hGLP-1R model will allow tissue-specific deletion of the 
      GLP-1R, purification of potential GLP-1R partner proteins, and testing of novel 
      therapeutic agents targeting the hGLP-1R.
FAU - Jun, Lucy S
AU  - Jun LS
AD  - Endocrine Discovery, Lilly Research Laboratories, Eli Lilly and Co., 
      Indianapolis, Indiana, United States of America.
FAU - Showalter, Aaron D
AU  - Showalter AD
AD  - Endocrine Discovery, Lilly Research Laboratories, Eli Lilly and Co., 
      Indianapolis, Indiana, United States of America.
FAU - Ali, Nosher
AU  - Ali N
AD  - Endocrine Discovery, Lilly Research Laboratories, Eli Lilly and Co., 
      Indianapolis, Indiana, United States of America.
FAU - Dai, Feihan
AU  - Dai F
AD  - Department of Physiology, University of Toronto, Toronto, Ontario, Canada.
FAU - Ma, Wenzhen
AU  - Ma W
AD  - Endocrine Discovery, Lilly Research Laboratories, Eli Lilly and Co., 
      Indianapolis, Indiana, United States of America.
FAU - Coskun, Tamer
AU  - Coskun T
AD  - Endocrine Discovery, Lilly Research Laboratories, Eli Lilly and Co., 
      Indianapolis, Indiana, United States of America.
FAU - Ficorilli, James V
AU  - Ficorilli JV
AD  - Endocrine Discovery, Lilly Research Laboratories, Eli Lilly and Co., 
      Indianapolis, Indiana, United States of America.
FAU - Wheeler, Michael B
AU  - Wheeler MB
AD  - Department of Physiology, University of Toronto, Toronto, Ontario, Canada.
FAU - Michael, M Dodson
AU  - Michael MD
AD  - Endocrine Discovery, Lilly Research Laboratories, Eli Lilly and Co., 
      Indianapolis, Indiana, United States of America.
FAU - Sloop, Kyle W
AU  - Sloop KW
AD  - Endocrine Discovery, Lilly Research Laboratories, Eli Lilly and Co., 
      Indianapolis, Indiana, United States of America.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140402
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (GLP1R protein, human)
RN  - 0 (Glp1r protein, mouse)
RN  - 0 (Glucagon-Like Peptide-1 Receptor)
RN  - 0 (Insulin)
RN  - 0 (Receptors, Glucagon)
RN  - IY9XDZ35W2 (Glucose)
SB  - IM
EIN - PLoS One. 2015;10(7):e0132875. PMID: 26172935
MH  - Animals
MH  - Diabetes Mellitus, Type 2/*drug therapy/genetics/metabolism
MH  - Disease Models, Animal
MH  - Glucagon-Like Peptide-1 Receptor
MH  - Glucose/metabolism
MH  - Glucose Tolerance Test
MH  - Humans
MH  - Insulin/metabolism
MH  - Mice
MH  - Mice, Transgenic
MH  - Pancreas/metabolism
MH  - Receptors, Glucagon/genetics/*metabolism
PMC - PMC3973576
COIS- Competing Interests: We have read the journal's policy and have the following 
      conflicts: LSJ, ADS, NA, WM, TC, JVF, MDM, and KWS are paid employees of Eli 
      Lilly and Company and may own company stock or possess stock options. This does 
      not alter the authors' adherence to PLOS ONE policies on sharing data and 
      materials. FD and MBW have no conflicts of interest to disclose.
EDAT- 2014/04/04 06:00
MHDA- 2015/01/30 06:00
PMCR- 2014/04/02
CRDT- 2014/04/04 06:00
PHST- 2013/12/10 00:00 [received]
PHST- 2014/03/07 00:00 [accepted]
PHST- 2014/04/04 06:00 [entrez]
PHST- 2014/04/04 06:00 [pubmed]
PHST- 2015/01/30 06:00 [medline]
PHST- 2014/04/02 00:00 [pmc-release]
AID - PONE-D-13-51521 [pii]
AID - 10.1371/journal.pone.0093746 [doi]
PST - epublish
SO  - PLoS One. 2014 Apr 2;9(4):e93746. doi: 10.1371/journal.pone.0093746. eCollection 
      2014.