Orthologous Genetic Markers For Susceptibility To Human T2DM In A Cynomolgus Macaque Model

Type2diabetes mellitus （T2DM） is increasing in prevalence and has become a seriousthreat to human health worldwide. Appropriate animal models are needed to better understandthe pathogenesis of T2DM and evaluate potential therapeutic agents for its treatment.Homology of the genomes between humans and cynomolgus reached94%. What is more, thecynomolgus macaque （Macaca fascicularis） T2DM model suitably simulates the onset andprogression of human T2DM and is useful for the development of new drugs for theprevention and treatment of T2DM. Therefore, the cynomolgus macaque T2DM models arethe ideal T2DM animal models. However, the morbidity of T2DM in cynomolgus monkey isvery low, which is not able to meet research requirement. If genetic factors are considered,they might provide an improved cynomolgus T2DM animal model. The genetic backgroundof primates are highly similar to humans’, there are more common T2DM susceptibility locibetween humans and cynomolgus. In this study, cynomolgus individuals and cynomolgusDNA sequences corresponding to37human T2DM susceptibility loci were used forassociation studies to rapidly screen orthologous genetic markers in a primate disease model.The results are as follows:（1） Fifty-four human T2DM susceptibility SNPs with the highest levels of statisticallysignificant associations with T2DM （P value threshold of5.2108） were selected fromprevious reports. Sequence for37of the54human SNPs provided a single98%match to thecynomolgus macaque draft genome and was downloaded.Thirty-one of the37primer pairssuccessfully produced an amplicon of the appropriate size amplicons orthologus to humangenes containing SNPs rs4506565, rs10^-811661, rs7903146, rs1531343and rs7578326exhibited obvious sequence differences between A pool and B pool. For the fragmentcorresponding to rs1531343, four polymorphisms （SNP343A, SNP343B, SNP343C andSNP343D） were identified from the two DNA pools. In the fragments corresponding tors4506565, rs7578326, rs10^-811661and rs7903146, three （SNP565A, SNP565B, andSNP565C）, one （SNP8326A）, two （SNP661A, SNP661B） and three polymorphisms（SNP146A, SNP146B, and SNP146C） respectively, were identified.（2） Eighty-three cynomolgus macaques used in the present study were from at Landao Biotechnology Co. Ltd （Guangdong, China）, Gaoyao-Kang-da Laboratory Animal S&T Co.Ltd （Guangdong, China） and Jin-jie-kang Biotechnology Co. Ltd （Yunnan, China）. FPG andglycated hemoglobin （GHb） levels of all animals were monitored three times with One TouchUltraVue （Johnson&Johnson, USA） and ADAMSTMA1c HA8160（Arkray, Kyoto, Japan）,respectively. The83cynomolgus monkeys were genotyped for13SNPs by sequcening. Andwere involved association studies. Eight of these13cynomolgus SNPs in three of these fiveorthologs, SNP661A, SNP661B, SNP343A, SNP343B, SNP343C, SNP565A, SNP565B andSNP565C, exhibited a difference in allele frequencies between spontaneous cases andnon-progressors that was statistically significant. SNP661A, SNP661B, SNP343A, SNP343Band SNP343C also exhibited statistically significant associations with FPG levels （P <0.05）.No statistically significant association between any of the13SNPs and GHb were observed inthe spontaneous populations. Therefore, the SNP661A, SNP661B, SNP343A, SNP343B andSNP343C polymorphisms could be used as genetic markers for the production of acynomolgus T2DM animal model However, there were no significantly different allelefrequencies between the induced case and non-progressor groups, and no statisticallysignificant association has been found between any of the13SNPs and FPG or GHb in theinduced case and non-progresso groups either. Our study is the first to propose using humancandidate genes for susceptibility to human disease to screen orthologous genetic markers in aprimate disease model.