Background: The metabolic diseases have been increasingly reported as healthcare challenging diseases, increasing the health expenses and burdening the national health system. Identifying uniquely responsible mechanism is lured around insulin and vitamin D crosstalk. The present study aimed to loom the translation of the crosstalk between vitamin D level and insulin hyposensitivity linked genetic pathways. Methods: To do so, serum samples were collected from patients with diabetes, metabolic syndrome, and obesity versus control group with matched age, sex, and health status. Genes (NAT2, IRS1, and IGF1) were quantified by polymerase chain reaction (PCR) alongside their correlative analysis with measured glycemic parameters, including fasting blood sugar (FBS), glycated haemoglobin (HbA1c), and HOMA-IR value. Results:  Analysis of results demonstrated that vitamin D level is significantly (p<0.05) higher in control group [25±5.1] compared to diabetic [16.5±4.4], metabolic syndrome [18.6±3.4], and obesity [14.1±4.8] groups, conversely, these groups have significantly (p<0.05) higher HOMA, FBS, and HbA1c compared to control group. All patient groups have significantly (p<0.05) higher NAT2 and IRS1 compared to control, while IGF1 significantly (p<0.05) elevated in obese group. Conclusion: these finding provide a valuable insight into uncovering the mechanisms involved in vitamin D role linked to other plasma variables of proteins and genes including proteins (NAT2, IRS1, and IGF1) pertained to insulin desensitization.