Background and Objectives: There is a paucity of longitudinal data documenting the temporal development of distress and burnout during medical school. The aim of this study was to examine trends and identify stressors associated with medical student distress over 4 years of medical education. Methods: Medical students from the class of 2016 at a Liaison Committee on Medical Education-accredited medical school completed surveys nine times from orientation through after the residency match. Surveys included demographic variables and measured distress domains using the Medical Student Well-Being Index. The authors used Microsoft Excel to calculate the proportion of students screening positive for individual distress domains at each of the nine acquisition periods for descriptive analysis. Results: Students completed 886 total surveys for an 85% response rate, which was relatively consistent across collection periods. Medical student distress and burnout increased from two (2%) to 12 (12%) respondents and from 19 (17%) to 37 (38%) respondents, respectively, from matriculation through after the residency match (P<0.01). Depersonalization increased from 15 (13%) to 34 (35%) respondents and emotional exhaustion increased from six (5%) to 22 (22%) respondents across 4 years of medical education (P<0.01). Emotional exhaustion peaked after medical school year 1, at 37 (45%), and year 3, at 45 (44%) respondents, with improvement after summer break and residency match. Conclusions: The results supported the literature demonstrating the development of burnout during medical school. Depersonalization increased early in the education process with minimal regression after development. Emotional exhaustion demonstrated a surprising increase after exposure to clinical clerkships. Further studies could support or refute the universality of these trends and evaluate prevention and intervention efforts targeting these key inflection points.