High-throughput screening of mouse gene knockouts identifies established and novel skeletal phenotypes

Abstract

Screening gene function in vivo is a powerful approach to discover novel drug targets. We present high-throughput screening (HTS) data for 3 762 distinct global gene knockout (KO) mouse lines with viable adult homozygous mice generated using either gene-trap or homologous recombination technologies. Bone mass was determined from DEXA scans of male and female mice at 14 weeks of age and by microCT analyses of bones from male mice at 16 weeks of age. Wild-type (WT) cagemates/littermates were examined for each gene KO. Lethality was observed in an additional 850 KO lines. Since primary HTS are susceptible to false positive findings, additional cohorts of mice from KO lines with intriguing HTS bone data were examined. Aging, ovariectomy, histomorphometry and bone strength studies were performed and possible non-skeletal phenotypes were explored. Together, these screens identified multiple genes affecting bone mass: 23 previously reported genes (Calcr, Cebpb, Crtap, Dcstamp, Dkk1, Duoxa2, Enpp1, Fgf23, Kiss1/Kiss1r, Kl (Klotho), Lrp5, Mstn, Neo1, Npr2, Ostm1, Postn, Sfrp4, Slc30a5, Slc39a13, Sost, Sumf1, Src, Wnt10b), five novel genes extensively characterized (Cldn18, Fam20c, Lrrk1, Sgpl1, Wnt16), five novel genes with preliminary characterization (Agpat2, Rassf5, Slc10a7, Slc26a7, Slc30a10) and three novel undisclosed genes coding for potential osteoporosis drug targets.

Osteoporosis: Mouse screen reveals novel drug targets

A systematic analysis in mice has identified multiple genes affecting bone mass that could code for novel drug targets. Robert Brommage and his colleagues from Lexicon Pharmaceuticals in Texas, USA, scanned the bones f 3,776 mouse lines, each containing a global ‘knockout’ mutation that disables the function of a single gene. The researchers further examined bone architecture, strength and mineralization in any mice with signs of skeletal phenotypes. These analyses revealed dozens of genes that affected bone mass in some way, many of which were not previously known to impact skeletal health. Three of the newly identified genes were not disclosed. These genes code for enzymes or secreted proteins and, although their identity remains a secret, the authors suggest they could be potential therapeutic targets for treating osteoporosis.

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