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 hom...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, Dkkl, Duoxa2, Enppl, Fgf23, Kissl/Kisslr, Kl （Klotho）, Lrp5, Mstn, Neol, Npr2, Ostml, Postn, Sfrp4, S1c30a5, Sic39a13, Sost, Sumf1, Src, Wnt10b）, five novel genes extensively characterized （Cldn18, Fam20c, Lrrkl, Sgpll, Wnt16）, five novel genes with preliminary characterization （Agpat2, RassfS, Slc10a7, Stc26a7, Slc30a10） and three novel undisclosed genes coding for potential osteoporosis drug targets.展开更多
TSC renal cystic disease is poorly understood and has no approved treatment.In a new principal cell-targeted murine model of Tsc cystic disease,the renal cystic epithelium is mostly composed of type A intercalated cel...TSC renal cystic disease is poorly understood and has no approved treatment.In a new principal cell-targeted murine model of Tsc cystic disease,the renal cystic epithelium is mostly composed of type A intercalated cells with an intact Tsc2 gene confirmed by sequencing,although these cells exhibit a Tsc-mutant disease phenotype.We used a newly derived targeted murine model in lineage tracing and extracellular vesicle(EV)characterization experiments and a cell culture model in EV characterization and cellular induction experiments to understand TSC cystogenesis.Using lineage tracing experiments,we found principal cells undergo clonal expansion but contribute very few cells to the cyst.We determined that cystic kidneys contain more interstitial EVs than noncystic kidneys,excrete fewer EVs in urine,and contain EVs in cyst fluid.Moreover,the loss of Tsc2 gene in EV-producing cells greatly changes the effect of EVs on renal tubular epithelium,such that the epithelium develops increased secretory and proliferative pathway activity.We demonstate that the mTORC1 pathway activity is independent form the EV production,and that the EV effects for a single cell line can vary significantly.TSC cystogenesis involves significant contribution from genetically intact cells conscripted to the mutant phenotype by mutant cell derived EVs.展开更多
文摘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, Dkkl, Duoxa2, Enppl, Fgf23, Kissl/Kisslr, Kl （Klotho）, Lrp5, Mstn, Neol, Npr2, Ostml, Postn, Sfrp4, S1c30a5, Sic39a13, Sost, Sumf1, Src, Wnt10b）, five novel genes extensively characterized （Cldn18, Fam20c, Lrrkl, Sgpll, Wnt16）, five novel genes with preliminary characterization （Agpat2, RassfS, Slc10a7, Stc26a7, Slc30a10） and three novel undisclosed genes coding for potential osteoporosis drug targets.
基金This work was supported by DoD(No.W81XWH-14-1-0343)(JJB)Federal Express Chair of Excellence(JJB),and Children’s Foundation Research Institute(JJB).
文摘TSC renal cystic disease is poorly understood and has no approved treatment.In a new principal cell-targeted murine model of Tsc cystic disease,the renal cystic epithelium is mostly composed of type A intercalated cells with an intact Tsc2 gene confirmed by sequencing,although these cells exhibit a Tsc-mutant disease phenotype.We used a newly derived targeted murine model in lineage tracing and extracellular vesicle(EV)characterization experiments and a cell culture model in EV characterization and cellular induction experiments to understand TSC cystogenesis.Using lineage tracing experiments,we found principal cells undergo clonal expansion but contribute very few cells to the cyst.We determined that cystic kidneys contain more interstitial EVs than noncystic kidneys,excrete fewer EVs in urine,and contain EVs in cyst fluid.Moreover,the loss of Tsc2 gene in EV-producing cells greatly changes the effect of EVs on renal tubular epithelium,such that the epithelium develops increased secretory and proliferative pathway activity.We demonstate that the mTORC1 pathway activity is independent form the EV production,and that the EV effects for a single cell line can vary significantly.TSC cystogenesis involves significant contribution from genetically intact cells conscripted to the mutant phenotype by mutant cell derived EVs.
