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Safe, stable, and nurturing relationships and environments in early care and education (ECE) programs can buffer the negative consequences of trauma exposure and enhance children’s resilience and healing. However, research suggests that ECE staff typically have limited access to resources and supports for implementing trauma-informed care. This multiple case study investigated a quality improvement initiative that engaged six urban ECE programs over an 18-month period to implement a trauma-informed approach in their organizations. Results show that changes occurred at each level of the organizational system in participating ECE programs: individual, classroom, leadership and program levels, offering insights into the multi-level change that may be needed to effectively implement and sustain a culture of trauma-informed care in ECE programs.