To investigate the effects of a clinical lytic defect on the structural response of human thoracolumbar functional spinal unit. A novel CT-compatible mechanical test system was used to image the deformation of a T12-L1 motion segment and measure the change in strain response under compressive loads ranging from 50 to 750 N. A lytic lesion (LM) with cortex involvement (33% by volume) was introduced to the upper vertebral body and the CT experiments were repeated. Finite element models, established from the CT volumes, were used to investigate the defect's effects on the structural response and the state of principal and shear stresses within the affected and adjacent vertebrae. The lytic lesion resulted in severe loss of the vertebral structural competence, resulting in significant, non-linear, and asymmetric increase in the experimentally measured strains and computed stresses within both vertebrae (p < 0.01). At the cortex, the tensile strains were significantly increased, while compressive strains significantly decreased, (p < 0.05). Both the vertebral bone and cortex regions adjacent to the defect showed significant increase in computed compressive, tensile, and shear stresses (p < 0.01). Changes in stress and strain distribution within the affected and adjacent vertebral bone and the experimentally observed bulging and buckling of the vertebral cortices suggested that initiation of catastrophic vertebral failure may occur under load magnitudes encountered in daily living. Although the effect of LM on the global deformation of the spine was well-predicted, our results show that FE predictions of local strain changes must be carefully assessed for clinical relevance. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1808-1819, 2016.