The current work investigates the impact of pre-deformation temperatures on the microstructure evolution and the subsequent cyclic stress-strain response of high-manganese steel showing twinning-induced plasticity (TWIP) at room temperature (RT). Deformation at low temperatures increases the hardening rate at low to medium degrees of deformation through concurrent martensitic transformation. In contrast, high temperatures promote dislocation slip. Thus, employing pre-treatments at temperatures below and above RT leads to the evolution of considerably different microstructures. Low-cycle fatigue experiments revealed distinct differences for the pre-treated TWIP steels.