Transgender women (TWs) face inequities in HIV and unique barriers to PrEP, an effective biomedical intervention to prevent HIV acquisition. To improve PrEP retention among TWs, we examined factors related to retention using a two-phase, sequential explanatory mixed-methods approach. In Phase I, we used data from a trial of 170 TWs who were provided oral PrEP to examine predictors of 24-week retention. In Phase II, we conducted 15 in-depth interviews with PrEP-experienced TWs and used thematic analysis to explain Phase I findings. In Phase I, more participants who were not retained at 24 weeks reported sex work engagement (18% versus 7%) and substantial/severe drug use (18% versus 8%). In Phase II, participants reported drug use as a barrier to PrEP, often in the context of sex work, and we identified two subcategories of sex work. TWs engaged in “non-survival sex work” had little difficulty staying on PrEP, while those engaged in “survival sex work” struggled to stay on PrEP. In Phase I, fewer participants not retained at 24 weeks reported gender-affirming hormone therapy (GAHT) use (56% versus 71%). In Phase II, participants prioritized medical gender affirmation services over PrEP but also described the bidirectional benefits of accessing GAHT and PrEP. TWs who engaged in “survival sex work” experience barriers to PrEP retention (e.g., unstable housing, drug use) and may require additional support to stay in PrEP care.