Mitral regurgitation (MR) is common with coronary artery disease (CAD), as altered myocardial substrate can impact valve performance. SPECT myocardial perfusion imaging (MPI) enables assessment of myocardial perfusion alterations. This study examined perfusion pattern in relation to MR. 2377 consecutive patients with known or suspected CAD underwent stress MPI and echocardiography (echo) within 1.6±2.3 days. MR was present on echo in 34% of patients, among whom 13% had advanced (≥moderate) MR. MR prevalence was higher among patients with abnormal MPI (44% vs. 29%, p<0.001), corresponding to increased global ischemia (p<0.001). Regional perfusion varied in left ventricular (LV) segments adjacent to each papillary muscle: Adjacent to the anterolateral papillary muscle, magnitude of baseline and stress-induced anterior/anterolateral perfusion abnormalities was greater among patients with MR (both p<0.001). Adjacent to the posteromedial papillary muscle, baseline inferior/inferolateral perfusion abnormalities were greater with MR (p<0.001), whereas stress inducibility was similar (p=0.39). In multivariate analysis, stress-induced anterior/anterolateral and rest inferior/inferolateral perfusion abnormalities were independently associated with MR (both p<0.05) even after controlling for perfusion in reference segments not adjacent to the papillary muscles. MR severity increased in relation to magnitude of perfusion abnormalities in each territory adjacent to the papillary muscles, as evidenced by greater prevalence of advanced MR among patients with ≥moderate anterior/anterolateral stress perfusion abnormalities (10.7% vs. 3.6%), with similar results when MR was stratified based on rest inferior/inferolateral perfusion (10.4% vs. 3.0%, both p<0.001). In conclusion, findings demonstrate that myocardial perfusion pattern in LV segments adjacent to the papillary muscles influences presence and severity of MR.