Keratoconus (KC) is a chronic and degenerative condition marked by pathological weakening, thinning, and protrusion of the corneal tissue. Corneal biomechanical weakness constitutes a hallmark of early keratoconus (KC) stages, which has underlined the importance of the study of biomechanical response in the early clinical screening of KC. In this line, the family history of KC is consistently identified as the leading risk factor for KC development, and the first-order pediatric relatives of KC patients are considered the most vulnerable population for KC development. On the other hand, tear fluid has become an attractive and accessible source that could be useful in identifying the molecular targets of several ocular diseases, such as corneal ectasias and more specifically KC disease. So, based on the vulnerability of first-order pediatric KC relatives to develop the disease, as well as the potential of tear fluid as a biomarker source, the purpose of this study was to analyze quantitatively and qualitatively the tear proteome of young KC offspring using micro–liquid chromatography coupled to tandem mass spectrometry, trying to get closer to the molecular drivers underlying the alterations in corneal biomechanical properties at at-risk stages of KC and to provide new insights into the molecular mechanisms involved in the earlier stages of the disease.