Congenital Zika Syndrome (CZS) was identified due to the increased incidence of congenital defects associated with Zika virus (ZIKV) infection, which include morphological, behavioral, neurological and ocular impairments. The eye has been described as a specialized CNS compartment being able to display symptoms of neurodegenerative diseases. Ocular lesions may result from defects that occurred during embryogenesis and are apparent in newborns exposed to ZIKV. However, late pregnancy infection or the absence of microcephaly cannot preclude the occurrence of ocular lesions and other CNS manifestations. Considering the need for surveillance of babies with possible ZIKV congenital exposure, we developed a method termed Cellular Imprinting Proteomics, CImP, for the identification and quantification of the ocular surface proteome of infants exposed to ZIKV during gestation.
The cohort was divided into CTRL (no infectious diseases), ZIKV (infants exposed to ZIKV, without microcephaly) and ZikvCZS (infants exposed to ZIKV, with microcephaly). The CImP is based on an improved impression cytology method and an optimized pipeline to extract and analyze the ocular proteome using mass spectrometry-based technology. A total of 2209 proteins were identified with modulation of neutrophil degranulation, cell death, ocular and neurodevelopment pathways in ZikvCZS compared to CTRL. Moreover, the molecular pattern of ocular surface cells retrieved from infants infected during the gestation but with no CZS was different from matched controls.
Molecular alterations in the ocular surface associated to ZIKV infection with and without CZS complications are reported for the first time. We predict that this method will be introduced successfully in the study of several neurological diseases with the aim to identify novel diagnostic and therapeutic biomarkers.