PgmNr M5030: High-throughput multi-system phenotyping identifies pleiotropy and novel gene function.

Authors:
D. West 1 ; I. Morse 2 ; C. McKerlie 3 ; K. Lloyd 4 ; The DTCC-KOMP2 Consortium 1,2,3,4


Institutes
1) Children's Hospital of Oakland Research Institute, Oakland, CA; 2) Charles River Laboratories, Wilmington, MA; 3) The Center for Phenogenomics, Toronto, Canada; 4) University of California, Davis CA.


Abstract:

Our ongoing project’s goal is to discover and relate poorly-annotated genes to informative functional information and identify novel phenotypes associated with human disease as part of a high-throughput mouse production and phenotyping pipeline. Adult phenotyping data are now available for ~680 uniquely targeted mutant mouse strains produced and phenotyped by the DTCC Consortium, one of three members of the NIH KOMP2 project and a part of the International Mouse Phenotyping Consortium (IMPC). Mutants were produced from genes prioritized on the basis of interest expressed by the scientific community and selected for having little to no pre-existing functional annotation. Mice for this pipeline were generated by microinjection of targeted C57BL/6N stem cells, chimeric founders were backcrossed to C57BL/6NCrl mice, and target-confirmed heterozygous mutant mice were intercrossed to produce homozygotes for phenotyping. A total of 197 lines were either embryo lethal or partially subviable prior to weaning as homozygotes, and for a subset of these lines adult phenotyping was completed in heterozygous mice. The phenotyping pipeline uses 16 mice (8 females; F, 8 males; M) per strain and co-raised wild type controls. Mice enter the standardized IMPC comprehensive adult phenotyping pipeline (http://www.mousephenotype.org/impress) at ~9 weeks of age, with weekly tests ending in necropsy and blood collection at week 16. We found one or more phenotypic difference compared with controls in ~60% of the mutant lines (p< 0.0001; Mixed Model Analysis). Individual phenotypes (not including viability) were assigned to 16 broad categories. The most frequent phenodeviants were found in tests assessing metabolism (clinical chemistry, glucose tolerance test, and indirect calorimetery), behavior, the nervous system (auditory brain stem response and prepulse inhibition), and the musculoskeleton system (bone mineral content and bone morphology). Examples of pleiotropic, statistically validated, and characterized phenotypes from each of these categories will be presented. These findings provide significant novel functional annotation for many genes of interest to the scientific community. Data are readily available at: http://www.mousephenotype.org/; and mutants can be recovered from cryopreserved germplasm at https://www.komp.org/. Supported by NIH grants U42OD011175, U54HG006364. We thank the outstanding efforts of the DTCC Production and Phenotyping Teams.