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Objective: Although gender-specific evidence on Gambling Disorder (GD) is still limited, some studies reported specific differences, mainly in psychopathological profiles, gambling behavior patterns, and pathogenesis. In order to further examine the role of gender in GD, we conducted a multicenter investigation in a sample of Italian outpatients. Method: One hundred-four outpatients with a diagnosis of GD based on DSM-5 criteria were consecutively recruited at two clinics based in Milan. Socio-demographic and clinical variables were collected for the whole sample and analyzed for the effect of gender. The severity of illness was assessed using the Canadian Problem Gambling Index and the Gambling Attitudes and Beliefs (GABS). Results: Among females, a significantly higher mean age (52.23 ± 10.95 vs. 40.96 ± 15.76; p=0.005) and older age at illness onset emerged (43.5 ± 11.92 vs. 29.22 ± 14.26; p<0.001). Females showed a significantly higher rate of psychiatric comorbidities, lifetime suicide ideation, stressful events at GD onset, and positive family history for GD compared to males. A predictive effect of male gender was found for the GABS questionnaires by performing a linear regression model, with males showing a higher risk to reach higher scores compared to females (B= 11.833; t=2.177; p=0.034). Conclusions: Our study seems to confirm the hypotheses that gender in GD may influence psychopathological profiles, course, and comorbidity. GD in female gender is frequently a comorbid condition with other specific clinical characteristics compared to males. Identifying specific clinical factors by gender may prompt more focus on the public health of women in relation to gambling, while still recognizing that males are at-risk of earlier gambling problems. These findings should be considered in therapeutic perspectives. [ABSTRACT FROM AUTHOR]