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Question: How frequently do beneficiaries lose Medicare Part D low-income subsidies (LIS), and is this associated with drug affordability and use? Findings: In this cohort study from 2007 to 2018, approximately one-fifth of those not deemed automatically eligible for LIS lost subsidies annually. Being younger than 65 years and a member of a racial and ethnic minority group was associated with increased temporary subsidy losses, out-of-pocket costs, and reduced prescription drug fills for chronic illnesses and overall. Meaning: Reducing barriers to maintaining Medicare Part D LIS could improve drug affordability and adherence, especially for disabled and racially and ethnically minoritized beneficiaries.