Lesbian, gay, bisexual, transgender, two-spirit, and American Indian and Alaska Native community members share long histories of discrimination and poorer health status as compared to mainstream Americans. In particular, these groups experience bias-related victimization, a type of discrimination based on inherent traits such as race or ethnicity and sexual orientation. This cross-sectional study (N = 334) used a revised bias-related victimization measure and examined the relationship between self-reported bias-related victimization and generalized anxiety disorder, depression, and substance abuse among lesbian, gay, bisexual, transgender, and two-spirit American Indians and Alaska Natives. The results showed that 84.4% reported experiencing bias-related victimization. Those with the highest levels of bias-related victimization had 2.79 times (p = .009; 95% CI [1. 30, 6.02]) the risk of reporting symptoms of generalized anxiety disorder as compared to those with no bias-related victimization, controlling for income, education, sex, age, sexual orientation, and chronic disease. There was no significant relationship between bias-related victimization and major depression or substance dependence/abuse. Our results support a potential relationship between bias-related victimization and generalized anxiety disorder for lesbian, gay, bisexual, transgender, and two-spirit American Indians and Alaska Natives. Including diverse populations in research is essential to a better understanding of the impact on health outcomes. Inclusion of bias-related victimization questions in clinical treatment may help identify at-risk patients.